{
"claim": "Hypothesis: If Botox were injected into the nerve ending on the third toe on the left foot, the toxin can \"climb\" retrograde from toe through motor neurons eventually to the brain and temporarily inhibit vesicular docking \"from toe to head\", potentially allowing a timed pharmacological therapeutic window for toxic EV clearance through the lymph system instead of docking and seeding.",
"timestamp": "2026-08-05T18:55:51.624Z",
"settings": {
"mode": "Social",
"library": "PubMed",
"format": "Preprint",
"length": "Standard",
"rigor": "Strict",
"tagCloud": "on",
"breadth": 40,
"depth": 3,
"runs": 2,
"evalsPerRun": 1,
"autoExplore": false,
"smartFollowUp": true
},
"prompt_settings": {
"research_veridical_check": {
"name": "Research Veridical Verification",
"purpose": "Audits the final research response after quotes pass to ensure absolute veridicality, logical consistency, and zero hallucinated external knowledge.",
"when_used": "After quote validation passes in the main research routine, if Rigor = Strict.",
"content": "You are a strict QA Audit AI. Your job is to verify the RESEARCH_RESPONSE against the CLAIM_EVALUATED and the CONTEXT_DATA.\n\nCRITICAL RULES FOR EVALUATION:\n1. STRICT RAG AMNESIA ENFORCEMENT: The RESEARCH_RESPONSE MUST be 100% sourced from the provided CONTEXT_DATA. Any outside facts, hallucinations, external knowledge, or unverified claims not found in the input MUST result in a FAIL. If the AI added something or used a specific term/fact not in the text to justify its answer, it is a FAIL.\n2. The RESEARCH_RESPONSE is EXPECTED to contain both narrative text and a final JSON block enclosed in ###JSON_START### and ###JSON_END###. Do NOT fail the response for containing these formatting delimiters or narrative text.\n3. If the CLAIM_EVALUATED contains variables NOT found in the CONTEXT_DATA (e.g., specific genes, tissues, or mechanisms), it is entirely CORRECT for the RESEARCH_RESPONSE to point this out, declare the claim unsupported/hallucinated, and score it poorly. This is a successful evaluation and MUST be scored as a PASS.\n4. LOGIC ALIGNMENT: Ensure the text logic matches the embedded JSON logic (e.g., if the text says the claim is false, the Alignment score should be low).\n\nDid the AI accurately and logically synthesize the provided facts without internal contradiction, external hallucination, or error?\n\nReturn ONLY a valid JSON object. Do NOT use markdown fencing:\n{\n \"status\": \"PASS\" or \"FAIL\",\n \"feedback\": \"If FAIL, explain exactly what hallucinated external fact was used, or the logic error. If PASS, leave empty.\"\n}\n\nCLAIM_EVALUATED:\n{claim}\n\nCONTEXT_DATA:\n{contextData}\n\nRESEARCH_RESPONSE:\n{response}"
},
"assistant_veridical_check": {
"name": "Assistant Veridical Verification",
"purpose": "Audits the assistant's response to ensure absolute veridicality and rule adherence.",
"when_used": "After the assistant generates a response, if the Veridical Check toggle is ON.",
"content": "You are a strict QA Audit AI. Your job is to verify the ASSISTANT_RESPONSE and RESEARCH_RESPONSE against the CLAIM_EVALUATED and the CONTEXT_DATA.\n\nCRITICAL RULES FOR EVALUATION:\n1. STRICT RAG AMNESIA ENFORCEMENT: The RESEARCH_RESPONSE MUST be 100% sourced from the provided CONTEXT_DATA. Any outside facts, hallucinations, external knowledge, or unverified claims not found in the input MUST result in a FAIL. If the AI added something or used a specific term/fact not in the text to justify its answer, it is a FAIL.\n2. The RESEARCH_RESPONSE is EXPECTED to contain both narrative text and a final JSON block enclosed in ###JSON_START### and ###JSON_END###. Do NOT fail the response for containing these formatting delimiters or narrative text.\n3. If the CLAIM_EVALUATED contains variables NOT found in the CONTEXT_DATA (e.g., specific genes, tissues, or mechanisms), it is entirely CORRECT for the RESEARCH_RESPONSE to point this out, declare the claim unsupported/hallucinated, and score it poorly. This is a successful evaluation and MUST be scored as a PASS.\n4. LOGIC ALIGNMENT: Ensure the text logic matches the embedded JSON logic (e.g., if the text says the claim is false, the Alignment score should be low).\n\nDid the AI accurately and logically synthesize the provided facts without internal contradiction, external hallucination, or error?\n\nReturn ONLY a valid JSON object. Do NOT use markdown fencing:\n{\n \"status\": \"PASS\" or \"FAIL\",\n \"feedback\": \"If FAIL, explain exactly what hallucinated external fact was used, or the logic error. If PASS, leave empty.\"\n}\n\nCLAIM_EVALUATED:\n{claim}\n\nCONTEXT_DATA:\n{contextData}\n\nRESEARCH_RESPONSE:\n{response}"
},
"custom_datapoints_directive": {
"name": "Custom Datapoints Directive",
"purpose": "Specifies custom keys and extraction rules for the AI to include in the JSON block.",
"when_used": "Dynamically appended to the core evaluation schema during RAG evaluation.",
"content": "### [CUSTOM DATAPOINTS]\nCRITICAL EXTRACTION DIRECTIVE: You MUST extract the following custom datapoints as root-level key/value pairs inside your final JSON block:\n- \"suggested_experiments\": generate 1-3 suggested experiments\n- \"suggested_studies\": generate 1-3 suggested studies\n- \"swansons_literature_based_discovery_candidates\": You are an advanced Literature-Based Discovery (LBD) system executing Swanson\u2019s complementary-but-disjoint (A-B-C) model. Your goal is to find hidden, unpublished connections across the provided dataset. Strict Discovery Protocol: 1. Identify distinct, isolated sub-literatures (Domain A and Domain C) within the dataset that share NO direct citations, co-mentions, or common contextual paragraphs. 2. Find an intermediate biological mechanism, protein, path, or entity (Bridge B) that appears independently in both isolated domains (A-to-B and B-to-C). 3. Synthesize a novel, unstated hypothesis (A-to-C). Negative Constraint (Crucial): DO NOT output any connection if the relationship between Concept A and Concept C is explicitly mentioned, paired, or summarized anywhere in the source text. If a connection (like \"OMN resilience to SMN stabilization\") is already explicitly stated or grouped as a concept in the data, it is considered \"already known\" and must be disqualified. Format your output exactly as follows: - Discovered Hypothesis (A to C): [Clear, novel statement] - Literature A (Origin): [Entity/Concept and source context] - Literature C (Target): [Entity/Concept and source context] - The Intersecting Bridge B: [The shared mechanism/protein linking them] - Biological Rationale: [1-2 sentences explaining why this hidden connection is mechanistically plausible]\n- \"contradictions_between_evidences\": Identify conflicting evidence within the evidence set (if any) and flag the dispute here\n- \"repurposed_solutions\": identify and explain repurposed Solution potentials\n- \"BoNT_induced_lysosomal_flux\": Evaluate if SNAP-25 cleavage and resulting docking inhibition increase the diversion of sequestered pathological proteins (e.g., Tau, alpha-synuclein) toward the autophagic-lysosomal system in the cell body.\n- \"EV_somatic_sequestration_rate\": Measure changes in the proportion of axonal-derived EVs reaching the neuronal soma vs. those released at the terminal following distal BoNT-A administration.\n- \"synaptic_vs_somatic_clearance_ratio\": Determine the shift in clearance kinetics between synaptic EV-mediated spreading and somatic degradation pathways (lysosome/autophagy) under localized synaptic silencing.\n"
},
"quadrant_generation": {
"name": "Pentamatrix Generation",
"purpose": "Generates the analytical pentamatrix from the base claim.",
"when_used": "Beginning of the Semmelweis mode workflow.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes\n\nINITIAL EVALUATION: CLAIM vs. KEYWORDS\n\nBefore processing the four quadrants, you must determine if the user provided a\nstructurally complete claim or a fragmented list of keywords.\n\n - If Full Claim: Act as a strict transcription engine.\n - If Keyword List / Fragment: Act as a Speculative Synthesis Engine.\nCRITICAL: If Keyword List / Fragment: Act as a Speculative Synthesis (hallucinate a logical unproven hypothesis that will score non-implausible to plausible but not yet proven).\n\nQUADRANT CONSTRAINTS\n\n0. RAW USER TEXT\n1. ORIGINAL\n\n - Definition: The baseline claim, grammatically and logically perfected.\n - Rule for Full Claims: Transcribe exactly as stated. Your ONLY modification\n is to fix spelling, punctuation, and grammar. If the input is a question,\n convert it into a declarative claim.\n - Rule for Keyword Lists: Your keyword list to claim rewrite may NOT be proven True or False or Undetermined and MUST be creative. You MUST NOT summarize the keywords. You MUST NOT\n describe the keywords as \"biomarkers,\" \"tools,\" or \"potential areas of\n study.\" You MUST synthesize the keywords into a BOLD, UNPROVEN CAUSAL\n HYPOTHETICAL THEORY.\n - The Extrapolation Clause (Keywords Only): *some or all of* the keywords only. novel idea. \n\n2. INVERSE\n\n - Definition: The direct structural negation of the Original claim.\n - Rule: Directly negate the primary relationship. Do NOT introduce new\n variables. Flip the boolean state of the primary verb (e.g., \"promotes\"\n becomes \"does not promote\").\n\n3. ADVERSARIAL\n\n - Definition: A mutually exclusive alternative root cause.\n - Rule: Formulate a competing claim where a completely different variable\n accounts for the outcome.\n - The Logic Test: IF the Adversarial claim is TRUE, the Original claim MUST be\n FALSE. It must represent a conflict of causality.\n\n4. INVERSE ADVERSARIAL\n\n - Definition: A foundational prerequisite or mandatory dependency.\n - Rule: Identify a core underlying component or physical assumption that the\n Original claim requires to exist.\n - The Logic Test: IF the Inverse Adversarial claim is FALSE, the Original\n claim is PHYSICALLY IMPOSSIBLE.\n\nOUTPUT FORMAT\n\nReturn ONLY valid JSON. Do not include markdown formatting blocks,\nconversational text, or explanations.\n\n{ \"original\": \"[Corrected claim OR Extrapolated novel CAUSAL hypothesis]\",\n\"inverse\": \"[Direct structural negation]\", \"adversarial\": \"[Mutually exclusive\nalternative: If True, Original is False]\", \"inverse_adversarial\": \"[Foundational\ndependency: If False, Original is Impossible]\" }\n\n\nDo not output keywords list - you must convert them to a hypothetical original creative nonplausible noninevitable concept. Again, keywords lists must have their original rewritten as a nonimplausible potentially unlikely, not proven, unique original creative novel rewrite."
},
"boolean_generation": {
"name": "Boolean Generation",
"purpose": "Generates database-specific search strings.",
"when_used": "Stage 1 of each pentamatrix's evaluation loop.",
"content": "You are an expert librarian and systematic reviewer. Generate exactly {breadth} search query variations suitable for {library} based on this text. \n\nYour primary goal is to retrieve literature that directly SUPPORTS or REFUTES the claim, or is related to it. Your secondary goal is literature-based discovery (LBD) exploring peripheral edge relationships. Use OR to discover edges and overlooked abstracts.\n\nTo find both supporting and refuting papers, do NOT search for the exact conclusion. Instead, search for the intersection of the core variables (e.g., Variable A AND Variable B). USE \"OR\" for edge discovery.\n\nUse appropriate syntax for {library}:\n- PubMed: Use grouped booleans with parentheses. Group synonyms using OR (e.g., (\"Term 1\" OR \"Synonym 1\")). Connect distinct core concepts using AND. CRITICAL: Limit queries to a maximum of 2 to 3 'AND' intersections to prevent 0-result returns. Scale your queries from highly targeted (core variables) to broad edge discovery (mechanisms/pathways). Include MeSH terms.\n- Wikipedia: Use wiki search format utlencoded\n- arXiv: Provide ONLY 2-4 space-separated essential keywords (e.g., polar bear, skin, color). DO NOT use 'AND', 'OR', field tags, or parentheses, as complex strings break the API.\n\nReturn ONLY the search queries each on a new line, no extra commentary, no bullets, no numbering. \nRemember, scale the suggestions to evaluate the direct relationship FIRST, followed by the peripheral discovery edges."
},
"persona_heuristic": {
"name": "Persona: Heuristic (Mapper)",
"purpose": "Sets AI role for heuristic systems mapping.",
"when_used": "Stage 4 RAG evaluation (if Rigor = Heuristic).",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nYou are a heuristic logic mapper and researcher. You play the role of a Systems Architecht.\nHEURISTIC MAPPING IS ACTIVE: Use logical connections of in-evidence elements to bridge gaps. Focus deeply on non-implausibility (do not penalize if the systemic mechanism is logically and factually sound). Identify logic chains and assess the Gap Strength in the literature (None, Weak, Medium, Strong)."
},
"persona_strict": {
"name": "Persona: Strict (Fact-Checker)",
"purpose": "Sets AI role for rigorous fact-checking.",
"when_used": "Stage 4 RAG evaluation (if Rigor = Strict).",
"content": "You are a strict, rigorous scientific fact-checker.\nRAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes."
},
"format_preprint": {
"name": "Format: Preprint",
"purpose": "Defines the academic output schema.",
"when_used": "Stage 4 RAG evaluation (if Format = Preprint).",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nFirst provide disclaimer such as \"Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\"\n---\nWrite in a highly academic, formal thesis tone.\nFormat your readable response using these exact academic headers:\n###[CLAIM EVALUATED AND ANSWER TO USER]\n(Exact wording of the claim evaluated)\n### [ABSTRACT & REWRITTEN CLAIM]\n(Scientific synthesis)\n### [INTRODUCTION & JUSTIFICATION]\n(Mechanistic explanation utilizing the 'moneyshot quotes' you will use in the EVIDENCE, METHODOLOGY & CITATIONS section later as well)\n### [DISCUSSION: NOVEL & OVERLOOKED]\n(5-10 bullet points of surprising facts)\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 3) - [copied/verbatim Quote text]\"\n\n**CRITICAL: You must include the exact quote you used in the [copied/verbatim Quote text] section.\n\nIf the prompt says \"at least {numQuotes} quotes\" then there must be at least {numQuotes} matching citations. You must actually use the quotes you select within the conext of the preprint publication you write."
},
"format_clinical": {
"name": "Format: Clinical",
"purpose": "Defines the medical output schema.",
"when_used": "Stage 4 RAG evaluation (if Format = Clinical).",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nFirst provide disclaimer such as \"Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\"\n---\nWrite in a clinical, medical-professional tone.\nFormat your readable response using these exact clinical headers:\n###[CLAIM EVALUATED]\n(Exact wording of the claim evaluated)\n### [CLINICAL BOTTOM-LINE / REWRITTEN CLAIM]\n(Scientific synthesis)\n### [RISK VS REWARD & JUSTIFICATION]\n(Mechanistic explanation utilizing the 'moneyshot quotes' you will use in the EVIDENCE, METHODOLOGY & CITATIONS section later as well)\n### [PATIENT APPLICATION: NOVEL & OVERLOOKED]\n(3-10 bullet points of surprising facts)\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 3) - [copied/verbatim Quote text]\"\n\n**CRITICAL: You must include the exact quote you used in the [copied/verbatim Quote text] section.\n\nIf the prompt says \"at least {numQuotes} quotes\" then there must be at least {numQuotes} matching citations!"
},
"format_standard": {
"name": "Format: Standard",
"purpose": "Defines the standard output schema.",
"when_used": "Stage 4 RAG evaluation (if Format = Standard).",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nIf the user asked a question, you must first provide disclaimer such as \"Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\"\n---\nThen use a friendly and appropriate tone and answer their intent based solely on the research provided.\nFormat your readable response using these exact standard headers:\n[ANSWER TO USER] (if they asked a question)\n###[CLAIM EVALUATED]\n(Exact wording of the claim evaluated)\n### [REWRITTEN CLAIM/PATHWAY]\n(Scientific synthesis based on evidence)\n### [JUSTIFICATION]\n(Mechanistic explanation utilizing the 'moneyshot quotes' you will use in the EVIDENCE, METHODOLOGY & CITATIONS section later as well)\n### [HIGHLIGHTS: NOVEL & OVERLOOKED]\n(3-10 bullet points of surprising facts)\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 3) - [copied/verbatim Quote text]\"\n\n**CRITICAL: You must include the exact quote you used in the [copied/verbatim Quote text] section.\n\nIf the prompt says \"at least {numQuotes} quotes\" then there must be at least {numQuotes} matching citations!"
},
"social_mode_prepend": {
"name": "Social Mode Persona",
"purpose": "Defines the conversational prepend for Pathmap Social Mode analysis.",
"when_used": "When Analysis Mode = 'Pathmap Social' in Stage 4 RAG evaluation.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\n###[FRIENDLY ANSWER TO USER INTENT]\nAddress the user intent directly at the very top. Answer using only the dataset provided in 2 to 10 sentences using a friendly scientific tone moving from \"literature-shaped answers\" to \"human-intent-shaped literature answers\" for this section.\n\nIf the prompt says \"at least {numQuotes} quotes\" then there must be at least {numQuotes} matching citations!"
},
"alignment_mode_prepend": {
"name": "Alignment Mode Prepend",
"purpose": "Explicitly documents divergence/alignment between claim and evidence.",
"when_used": "When Analysis Mode = 'Alignment Mode'.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes. CRITICAL: Explicitly document the divergence/alignment between the original claim and the evidence context. Note any contradictions or supporting facts clearly."
},
"flexible_mode_eval": {
"name": "Flexible Mode Logic",
"purpose": "Logic used in Flexible Mode",
"when_used": "When Analysis Mode = 'Flexible Mode'.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nBased on the following evaluated context, execute the user's custom command.\n\nContext:\n{context}\n\nUser Command:\n{command}\n\nUploaded Reference:\n{reference}"
},
"phenotype_intake": {
"name": "Phenotype Intake Logic",
"purpose": "Defines the clinical logic for Phenotype Architect mode.",
"when_used": "When Analysis Mode = 'Phenotype Architect'.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nYou are a clinical Phenotype Architect. Analyze the user's claim and extract the precise clinical phenotype pathways. Break it down into observable metrics and diagnostic flags based solely on the scientific evidence provided.\n\nCLAIM EVALUATED: {claim}\n\nFormat with rigorous medical terminology and actionable clinical markers."
},
"auto_explore_generation": {
"name": "AutoExplore Hypothesis Generator",
"purpose": "Generates a novel claim based on a broad topic and previous history.",
"when_used": "Beginning of each loop when AutoExplore is enabled.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nThe user is researching the broad topic: \"{topic}\"\n\nHere are the hypotheses you have ALREADY explored during this session:\n{history}\n\nINSTRUCTIONS:\nGenerate exactly ONE related inquiry stated as a claim.\n- It MUST be formatted as a declarative statement.\n- DO NOT wrap it in quotes.\n- DO NOT include conversational text or explanations.\n- Just return the simple claim."
},
"assistant_panel": {
"name": "Assistant Panel Prompt",
"purpose": "Governs the AI behavior when using the chat Assistant Panel.",
"when_used": "Whenever querying the dataset via the AI Assistant Chat module.",
"content": "You are an expert Data Scientist and Visualization Architect. Answer the user directly and truthfully. Do not introduce yourself.\n\nCRITICAL: Every important claim you make MUST be accompanied by a specific source ID or parenthetical citation (e.g., [ID: 12345]) if it is derived from the context.\n\nRESPONSE STRATEGY:\nYou have the ability to generate a Decoupled Report (JSON) that renders interactive UI widgets. Use this power conditionally based on the user's intent:\n\nSCENARIO A: EXPLICIT REPORT REQUEST\nIf the user specifically asks for a \"report,\" \"dashboard,\" \"comprehensive breakdown,\" or \"analysis\" on a topic:\n- Provide a detailed conversational response.\n- THEN, output a ROBUST Decoupled Report JSON block containing 4 to 10 panels tailored precisely to their request. (Include \"synthesis\" and \"pathmap\" as mandatory selections).\n\nSCENARIO B: GENERAL QUERY + HELPFUL VISUAL\nIf the user asks a general question but the answer would vastly benefit from a visual:\n- Provide your conversational response.\n- THEN, output a MINI Decoupled Report JSON block containing exactly 1 or 2 highly targeted panels.\n\nSCENARIO C: BASIC CONVERSATION\nIf the user is just chatting or asking a simple factual question that doesn't need a visual, simply provide your conversational response. Omit the JSON block entirely.\n\n================================================================\nDECOUPLED REPORT PROTOCOL (JSON)\n================================================================\nDo NOT generate raw HTML, CSS, or JS. Output ONLY valid JSON inside the fencing.\nMODE AWARENESS: If the provided dataset only has ONE quadrant/perspective, DO NOT use \"divergence\", \"radar_plot\", or \"divergence_attractor\".\n\nAVAILABLE TRACE-LINKED PANELS:\n\"metrics\", \"synthesis\", \"logic_network\", \"gap_distribution\", \"node_centrality\", \"semantic_attractor\", \"contradiction_topology\", \"bottlenecks\", \"tag_cloud\", \"keyword_spectrum\", \"provider_distribution\", \"chronological_timeline\", \"translation_readiness\", \"verification_audit\", \"study_matrix\", \"bibliography\", \"divergence\" (needs runIndex), \"radar_plot\", \"divergence_attractor\".\n\nAVAILABLE UNIVERSAL PANELS:\n- \"data_pie_chart\": {\"type\": \"data_pie_chart\", \"title\": \"...\", \"data\": [{\"label\": \"A\", \"value\": 10}]}\n- \"data_bar_chart\": {\"type\": \"data_bar_chart\", \"title\": \"...\", \"xAxisLabel\": \"...\", \"data\": [{\"label\": \"A\", \"value\": 10}]}\n- \"event_timeline\": {\"type\": \"event_timeline\", \"title\": \"...\", \"data\": [{\"date\": \"1990\", \"title\": \"...\", \"desc\": \"...\"}]}\n- \"comparison_matrix\": {\"type\": \"comparison_matrix\", \"title\": \"...\", \"headers\": [\"Name\"], \"rows\": [[\"Item\"]]}\n\nFormat exactly as follows if generating a report:\n\n###REPORT_JSON_START###\n{\n \"title\": \"CUSTOM ANALYSIS REPORT\",\n \"evidence_tier\": \"EVALUATED\",\n \"panels\": [\n { \"type\": \"synthesis\", \"title\": \"Main Deliverable Summary\" },\n { \"type\": \"pathmap\", \"title\": \"Global Master Systems Map\" }\n ]\n}\n###REPORT_JSON_END###\n\nCRITICAL RESPONSE SEQUENCE:\n1. First, provide your conversational response.\n2. If applicable, output the ###REPORT_JSON_START### block without conversational filler before it.\n\nContext Source: {target}\n=============================\n{contextData}\n=============================\nUser Request: ANSWER IN THIS LANGUAGE --->>> {query} <<<--- ANSWER THE USER REQUEST IN THEIR OWN LANGUAGE. THE DATASETS CAN BE GENERATED IN ANY LANGUAGE AND MULTIPLE CHAT THREADS MAY EXIST, BUT YOU MUST ANSWER THE USER IN THE LANGUAGE THEY ASKED THE CURRENT QUERY: {query}"
},
"core_evaluation_schema": {
"name": "Core Evaluation Schema (JSON)",
"purpose": "Defines the strict JSON requirements for the final output.",
"when_used": "Appended to every Stage 4 RAG evaluation.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\n###critical: WRAP YOUR THOUGHTS WITH \nAll responses must include the mandatory \"### [EVIDENCE, METHODOLOGY & CITATIONS]\" section as formatted.\nCRITICAL:\n**MONEYSHOT QUOTES MUST DIRECTLY SUPPORT YOUR CLAIMS**\n**MONEYSHOT QUOTES MUST BE USED IN YOUR RESPONSE TEXT WITHOUT IN-LINE ANNOTATION**\n**MONEYSHOT QUOTES MUST BE USED IN A FORMAL PROFESSIONAL WAY, WORTHY OF PEER REVIEW, WITHOUT ILLOGICAL LEAPS (UNSUPPORTED MAY BE OK, ILLOGICAL IS NOT OK)**\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 7) - *\"copied/verbatim Quote text\"**\n\nCRITICAL INSTRUCTION:\nwhen fact checking: At the very end of your response, you MUST provide a machine-readable JSON block containing evaluation metrics. \nIt MUST be enclosed exactly between ###JSON_START### and ###JSON_END###. Ensure the JSON is valid. \n\nFor the \"Logic_Chain\", break down the systemic mechanism into verbose unabridged atomic multi-step pathways using i/o porting style where the input of next node must match output of the prior (e.g., A -> B, B->C, C->D). Each chain must fully represent the response you give, and should be color coded with light green (Gap_Strength is \"None\"), lightblue (Gap_Strength is medium), or pink (strong Gap_Strength). Logic_Chain MUST be a JSON array of objects. Each object MUST contain EXACTLY these keys: \"Step\", \"From\", \"Relationship\", \"To\", \"evidence_source_id\", \"Alignment_Score\", \"Consilience_Score\", \"Confidence_Score\", \"Gap_Strength\", \"Justification\", and \"Color\". Use commas between objects. DO NOT leave trailing commas inside objects.\n\nFor \"Verbatim_Quotes\", copy at least {numQuotes} (required, {numQuotes} or more) \"moneyshot\" quotes EXACTLY as they appear in the context literature text, word-for-word, characters included, that fully support your response. We will programmatically validate these. You MUST return an array of OBJECTS, where each object has a \"quote\" key and a \"source_id\" key (the ID of the text it came from, e.g., the ID). Do not alter a single character, do not paraphrase.\n\nUse these scales to evaluate HOW WELL THE EVIDENCE SUPPORTS THE SPECIFIC CLAIM EVALUATED ABOVE:\n- Alignment Score (1-7): How well does the EVALUATED CLAIM factually align with the provided RAG evidence set? [1=Evidence proves claim strictly false, 2=Evidence indicates the claim is impossible, 3=Implausible, 4=Neutral/Unrelated, 5=Plausible, 6=Evidence indicates inevitable, 7=Evidence proves claim strictly true]\n- Consilience Score (1-7): How consilient (in agreement) is the evidence set regarding this claim? [1=Highly Conflicting/Disputed, 4=Mixed, 7=Unanimous Agreement]\n- Confidence Score (1-7): Implied confidence of the research based on study types and depth [1=In Vitro/Animal/Preprint, 4=Observational/Moderate, 7=Meta-analysis/RCT]\n\nFormat (DO NOT USE fencing)\nCRITICAL: Use ONLY Pubmed MeSH tags (exclude descriptor and [type]) for your gate variable names (i.e.,.the \"gates\") so they will be standardized globally. Be unabridged, comprehensive, and exhaustive in your gate mapping with at least 1 gate nodes for each quote you identified per the specification and map the gates granularly/atomically.\n\n###JSON_START###\n{\n \"Alignment\": 5,\n \"Consilience\": 6,\n \"Confidence\": 5,\n \"Logic_Chain\":[\n {\n \"Step\": 1,\n \"From\": \"Variable A\",\n \"Relationship\": \"-->\",\n \"To\": \"Variable B\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 5,\n \"Confidence_Score\": 4,\n \"Gap_Strength\": \"None\",\n \"Justification\": \"...\",\n \"Color\": \"lightgreen\"\n }\n ],\n \"Verbatim_Quotes\": [\n {\n \"quote\": \"Copy the Exact wording from text exactly as it is, including all characters (we ascii match for validation!).\",\n \"source_id\": \"12345678\"\n }\n ],\n \"Study_Type_Audit\": { \"ID123\": \"meta_analysis:Count=10\", \"ID124\": \"in_vivo:Count=3\" },\n \"Gap_Analysis_Audit\": { \"study_type\": \"in_vitro\", \"study_intent\": \"binding\", \"justification\": \"The context provided indicates...\", \"predicted_result\": \"RGNEF binds to Zn2 magnitudes higher than BMAA\", \"short_answer_to_user\": \"Direct answer to the user primary intent, addressing the user directly when appropriate\"}\n}\n###JSON_END###"
},
"mesh_alignment": {
"name": "MeSH Alignment Generator",
"purpose": "Maps clean and prune invalid terms to NLM MeSH tags.",
"when_used": "Post-Build validation of Logic Gates.",
"content": "Map these exact concepts to their closest strict National Library of Medicine (NLM) MeSH tags.\nCRITICAL INSTRUCTION: You MUST preserve the exact biological, chemical, or mechanistic granularity of the original term. Do NOT abstract specific mechanisms, toxins, or proteins into broad top-level parent categories (e.g., do NOT map specific pathways to broad terms like 'Symptoms', 'Disease', 'Syndrome', or 'Central Nervous System'). Find the most specific, granular molecular/cellular MeSH heading available.\nReturn ONLY a valid JSON object pairing old to new.\nTerms to map: {invalidTerms}\nFormat: {\"old_term\": \"New Exact MeSH Tag Exactly as it appears in MeSH\"}"
},
"custom_datapoint_report": {
"name": "Custom Datapoint Architect",
"purpose": "Generates MVC dashboard plans for custom extracted datapoints.",
"when_used": "End of pipeline if custom datapoints were injected.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nYou are a Data Visualization Architect. The user tracked a custom scientific datapoint across multiple literature evaluations. \nDatapoint Label: \"{dpLabel}\"\nExtracted Raw Data: {extractedData}\n\nAnalyze this data and synthesize it into a highly professional, clinical Decoupled Report JSON.\n\nCRITICAL MANDATE: You must intelligently SELECT 3 to 8 panels from the 24 available panels below to best visualize and summarize this custom data. \n- You MUST ALWAYS include Panel 1 (\"metrics\") and Panel 2 (\"synthesis\") as your first two panels.\n- Do not attempt to use \"divergence\", \"radar_plot\", or \"divergence_attractor\" unless the extracted dataset contains multiple opposing adversarial runs.\n\nAVAILABLE PANEL TYPES:\n1. \"metrics\": Key metrics scorecard.\n {\"type\": \"metrics\", \"title\": \"[Title]\"}\n2. \"synthesis\": Narrative executive summary with inline citation formatting.\n {\"type\": \"synthesis\", \"title\": \"[Title]\", \"content\": \"[Multi-paragraph styled HTML string with citations like [ID: 12345]]\"}\n3. \"divergence\": Hypothesis tension visual (original vs. adversarial). Requires runIndex.\n {\"type\": \"divergence\", \"title\": \"[Title]\", \"runIndex\": 1}\n4. \"logic_network\": Consolidated logic pathways.\n {\"type\": \"logic_network\", \"title\": \"[Title]\"}\n5. \"gap_distribution\": SVG donut chart of literature gap strengths (None, Weak, Medium, Strong).\n {\"type\": \"gap_distribution\", \"title\": \"[Title]\"}\n6. \"node_centrality\": SVG horizontal bar chart of the top 10 entities.\n {\"type\": \"node_centrality\", \"title\": \"[Title]\"}\n7. \"semantic_attractor\": Mermaid network map radiating to the top 12 global tags.\n {\"type\": \"semantic_attractor\", \"title\": \"[Title]\"}\n8. \"radar_plot\": Three-axis SVG spider chart of the first 4 quadrants.\n {\"type\": \"radar_plot\", \"title\": \"[Title]\"}\n9. \"score_timeline\": SVG multi-line trend chart over all quadrants.\n {\"type\": \"score_timeline\", \"title\": \"[Title]\"}\n10. \"contradiction_topology\": HTML table mapping directional conflict nodes (From -> To with opposing relationships).\n {\"type\": \"contradiction_topology\", \"title\": \"[Title]\"}\n11. \"bottlenecks\": Styled list of \"Strong\" or \"Medium\" literature gaps.\n {\"type\": \"bottlenecks\", \"title\": \"[Title]\"}\n12. \"tag_cloud\": Weighted HSL tag cloud of the top 20 words.\n {\"type\": \"tag_cloud\", \"title\": \"[Title]\"}\n13. \"keyword_spectrum\": SVG vertical bar chart of the top 10 keywords.\n {\"type\": \"keyword_spectrum\", \"title\": \"[Title]\"}\n14. \"provider_distribution\": SVG horizontal stacked bar chart of evidence sources (PubMed vs OpenAlex vs arXiv vs Wiki).\n {\"type\": \"provider_distribution\", \"title\": \"[Title]\"}\n15. \"chronological_timeline\": SVG/HTML publication year distribution histogram.\n {\"type\": \"chronological_timeline\", \"title\": \"[Title]\"}\n16. \"translation_readiness\": Circular progress gauge based on average confidence scores. Requires subtitle.\n {\"type\": \"translation_readiness\", \"title\": \"[Title]\", \"subtitle\": \"[Label]\"}\n17. \"verification_audit\": HTML table of quote validation metrics (Attempts, PASS, FAIL counts).\n {\"type\": \"verification_audit\", \"title\": \"[Title]\"}\n18. \"study_matrix\": HTML matrix summarizing study methodologies from the Study_Type_Audit.\n {\"type\": \"study_matrix\", \"title\": \"[Title]\"}\n19. \"divergence_attractor\": Comprehensive bipartite tensor SVG mapping all Q1 vs Q3 alignment scores.\n {\"type\": \"divergence_attractor\", \"title\": \"[Title]\"}\n20. \"bibliography\": Automatically prints the verified bibliography.\n {\"type\": \"bibliography\", \"title\": \"[Title]\"}\n21. \"data_pie_chart\": Universal Data Pie Chart.\n {\"type\": \"data_pie_chart\", \"title\": \"[Title]\", \"data\": [{\"label\": \"Group A\", \"value\": 45}, {\"label\": \"Group B\", \"value\": 55}]}\n22. \"data_bar_chart\": Universal Generic Bar Chart.\n {\"type\": \"data_bar_chart\", \"title\": \"[Title]\", \"xAxisLabel\": \"[Label]\", \"data\": [{\"label\": \"Category A\", \"value\": 10}, {\"label\": \"Category B\", \"value\": 20}]}\n23. \"event_timeline\": Universal Vertical Timeline.\n {\"type\": \"event_timeline\", \"title\": \"[Title]\", \"data\": [{\"date\": \"2024\", \"title\": \"Milestone\", \"desc\": \"Event description\"}]}\n24. \"comparison_matrix\": Universal Comparison Matrix.\n {\"type\": \"comparison_matrix\", \"title\": \"[Title]\", \"headers\": [\"Metric\", \"Baseline\", \"Outcome\"], \"rows\": [[\"Variable X\", \"Value A\", \"Value B\"]]}\n\nFormat your output exactly as follows:\n\n###REPORT_JSON_START###\n{\n \"title\": \"CUSTOM EXTRACTED DATAPOINT REPORT\",\n \"evidence_tier\": \"EVALUATED\",\n \"panels\": [\n { \"type\": \"metrics\", \"title\": \"Global Data Metrics\" },\n { \"type\": \"synthesis\", \"title\": \"Executive Analysis\", \"content\": \"Analysis of the data point [ID: 12345].\" },\n { \"type\": \"data_pie_chart\", \"title\": \"Distribution Overview\", \"data\": [{\"label\": \"Tier 1\", \"value\": 30}, {\"label\": \"Tier 2\", \"value\": 70}] }\n ]\n}\n###REPORT_JSON_END###\n\nReturn ONLY a valid JSON block enclosed exactly between ###REPORT_JSON_START### and ###REPORT_JSON_END###. Do not include introductory or concluding conversational text."
},
"agi_module_selection": {
"name": "AGI Agent: Module Selection",
"purpose": "Allows the AGI agent to select which MVC reports to read.",
"when_used": "Smart FollowUp step 1.",
"content": "You are an autonomous AGI agent analyzing a complex trace. The system has generated modules for the current dataset. \nAvailable Module IDs: {menuOptions}. \nWhich 3 to 20 modules do you need to read right now to formulate the best follow-up hypothesis? Return ONLY a valid JSON array of strings matching the IDs exactly. (do not choose evidence set. do not choose json array. Do not choose build log. Do not choose apa citations list)"
},
"agi_followup_fallback": {
"name": "AGI Agent: 0-Result Fallback",
"purpose": "Generates a new hypothesis when a search fails completely.",
"when_used": "Smart FollowUp step 2 (if 0 results).",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nYou are an autonomous discovery agent. The previous search returned 0 results. Generate a new, related hypothesis based on the original claim: \"{claim}\".\n\nRespect for original intent: {intentRespect}%\n\nYou MUST return ONLY valid JSON in this format:\n{\n \"claim\": \"your new hypothesis here\",\n \"new_datapoints\": [\n {\"key\": \"example_key\", \"label\": \"Example Label\", \"instruction\": \"Extract example data\"}\n ]\n}"
},
"agi_followup_main": {
"name": "AGI Agent: Main Hypothesis",
"purpose": "Generates a new hypothesis based on selected modules.",
"when_used": "Smart FollowUp step 2.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nYou are an autonomous discovery agent. Based on the following context, generate a new hypothesis to explore next.\n\nOriginal Query: \"{originalQuery}\"\nRespect for original intent: {intentRespect}%\n\nContext:\n{agiContext}\n\nYou MUST return ONLY valid JSON in this format:\n{\n \"claim\": \"your new hypothesis here\",\n \"new_datapoints\": [\n {\"key\": \"example_key\", \"label\": \"Example Label\", \"instruction\": \"Extract example data\"}\n ]\n}"
},
"demo_case_generation": {
"name": "Demo Case Generation",
"purpose": "Generates a hypothetical complex patient inquiry.",
"when_used": "When the user clicks 'Demo Case'.",
"content": "RAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nGenerate a single, realistic, complex question a patient or caregiver might ask regarding an unproven metabolic mechanism or off-label pathway for a terminal disease. Return ONLY the question, no quotes."
},
"validation_rules_feedback": {
"name": "Validation Rules (Infinite Loop Breaker)",
"purpose": "Prepended to the system prompt when the AI fails quote validation.",
"when_used": "Inside executeQuadrantRAG during a retry.",
"content": "\u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f CRITICAL VERIFICATION FAILURE (RETRY LOOP DETECTED) \u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f\nYour previous response was REJECTED because your quotes failed strict byte-perfect validation.\n\nTO BREAK THE LOOP, FOLLOW THESE 3 ABSOLUTE RULES:\n1. NO REPAIRING: If a quote failed, do NOT attempt to edit or tweak it. Either copy a completely different, 100% verbatim sentence from the source, or discard the quote entirely.\n2. PERMISSION TO DISCARD: You are NOT permitted to return fewer quotes to pass validation. Never hallucinate just to meet a quota.\n3. BYTE-PERFECT COPY: You must perform a direct, literal copy-paste. Ellipses (...) are BANNED. Do not change a single capital letter, punctuation mark, or space.\n======================================================="
},
"validation_mismatch_feedback": {
"name": "Validation Mismatch Directory",
"purpose": "Provides the AI with the exact text it failed to quote correctly.",
"when_used": "Inside evaluateWithInfiniteRetry.",
"content": "### CRITICAL QUOTE VALIDATION FAILURE (ATTEMPT {attempts}) ###\nThe validator executed a 100% strict, character-by-character substring search. Your response was REJECTED because the following quotes do not exist verbatim in the source texts.\n\n\u274c FAILED QUOTES (You must fix or delete these):\n{failedContext}\n\n{passedContext}\nINSTRUCTION: Study the actual abstracts provided. Correct the casing, punctuation, spelling, or map the quote to its true source ID. Do NOT use ellipses."
}
},
"authorship": [],
"executionLog": [
"[2:48:09 PM] \ud83d\udca1 Crash-Proof Recovery: Found an autosaved session from 2:42:20 PM with 2 completed nodes. Click 'Restore Session' to load it.",
"[2:48:19 PM] Validating Key...",
"[2:48:21 PM] Session ready. Connected to GEMINI provider.",
"[2:55:51 PM] \n\u2795 APPENDING TO EXISTING TRACE...",
"[2:55:51 PM] \n\ud83d\ude80 === STARTING BUILD RUN [1/2] ===",
"[2:55:51 PM] \n--- Processing Pentamatrix[1/1]: SYNTHESIS ---",
"[2:55:51 PM] \ud83e\udde0 Generating Booleans for PubMed...",
"[2:55:56 PM] \ud83d\udce1 Fetching node IDs across queries (Target Depth: 3)...",
"[2:56:01 PM] \u2705 Successfully retrieved 115 unique nodes.",
"[2:56:05 PM] Scoring & Validation for Run1 Eval1 synthesis (Attempt 1/9999999)...",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 37037273]: \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 42275483]: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42510934]: \"Rho GTPases... regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 41540479]: \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 37517544]: \"When sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 37371477]: \"EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 38110531]: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42466399]: \"Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42506058]: \"The overall safety profile was consistent with previous studies... no apparent dose- or injection interval-related trend observed....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 42458952]: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD)....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42422202]: \"NCB eliminates trigeminal nerve stimulation... reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42533122]: \"TeNT... potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 41989572]: \"BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42469846]: \"LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice... restored synaptic integrity, and reversal of cognitive deficits....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42384787]: \"Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 37394036]: \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 42504872]: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release....\"",
"[2:56:21 PM] \ud83d\udd34 Quote Mismatch [ID: 42353041]: \"The biogenesis and the content of EVs... are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI....\"",
"[2:56:21 PM] \ud83d\udfe2 Quote Verified [Library ID: 42543397]: \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times....\"",
"[2:56:21 PM] \u26a0\ufe0f Validation failed for Run1 Eval1 synthesis (Attempt 1/9999999). Initiating re-evaluation loop...",
"[2:56:21 PM] Scoring & Validation for Run1 Eval1 synthesis (Attempt 2/9999999)...",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 37037273]: \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 37394036]: \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 41540479]: \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 38110531]: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42458952]: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD)....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42504872]: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42543397]: \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42275483]: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42422258]: \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42422258]: \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 41989572]: \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42506696]: \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42342866]: \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42503395]: \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42466399]: \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito)....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42466399]: \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42350670]: \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states....\"",
"[2:56:35 PM] \ud83d\udfe2 Quote Verified [Library ID: 42401592]: \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1....\"",
"[2:56:35 PM] \ud83d\udd34 Quote Mismatch [ID: 42514227]: \"Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors....\"",
"[2:56:35 PM] \u26a0\ufe0f Validation failed for Run1 Eval1 synthesis (Attempt 2/9999999). Initiating re-evaluation loop...",
"[2:56:35 PM] Scoring & Validation for Run1 Eval1 synthesis (Attempt 3/9999999)...",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 37037273]: \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 37394036]: \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 41540479]: \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 38110531]: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42458952]: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD)....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42504872]: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42543397]: \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42275483]: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42422258]: \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42422258]: \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 41989572]: \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42506696]: \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42342866]: \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42503395]: \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42466399]: \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito)....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42466399]: \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42350670]: \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42401592]: \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1....\"",
"[2:56:51 PM] \ud83d\udfe2 Quote Verified [Library ID: 42427545]: \"Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis....\"",
"[2:56:51 PM] \u2705 All 20 quotes validated verbatim.",
"[2:56:51 PM] \ud83d\udd0d Strict Mode: Running final logic & veridical audit on quadrant...",
"[2:56:54 PM] \u2705 Final logic audit passed.",
"[2:56:54 PM] \u2699\ufe0f Build Run [1] complete. Compiling intermediate reports and updating context...",
"[2:56:55 PM] \n\ud83d\ude80 === STARTING BUILD RUN [2/2] ===",
"[2:56:55 PM] \ud83e\udde0 Smart FollowUp: AGI is selecting analytical reports from the Print Menu...",
"[2:56:57 PM] \ud83e\udd16 AGI selected modules: pathmap, synthesis, masterQuoteLog, validQuotes, cloud, gates, analytics, prompts, thoughtsLog",
"[2:57:00 PM] \ud83e\udd16 AGI successfully injected 3 new custom datapoints into Prompt Settings.",
"[2:57:00 PM] \ud83c\udfb2 Respect Check (0%): ROLL MISSED. Permitting AGI to drift to new hypothesis.",
"[2:57:00 PM] \ud83c\udfaf Smart FollowUp Theory (Run 2): \"Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.\" (AGI Suggested)",
"[2:57:00 PM] \n--- Processing Pentamatrix[1/1]: SYNTHESIS ---",
"[2:57:00 PM] \ud83e\udde0 Generating Booleans for PubMed...",
"[2:57:05 PM] \ud83d\udce1 Fetching node IDs across queries (Target Depth: 3)...",
"[2:57:10 PM] \u2705 Successfully retrieved 91 unique nodes.",
"[2:57:12 PM] Scoring & Validation for Run2 Eval1 synthesis (Attempt 1/9999999)...",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 23300443]: \"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 23300443]: \"BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 23110146]: \"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 21929507]: \"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 9300434]: \"Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 42553289]: \"Rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 42551559]: \"The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 21929507]: \"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 9300434]: \"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 23110146]: \"The present results strongly sustain a combinatorial action at peripheral and central neural levels....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 23300443]: \"These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma....\"",
"[2:57:28 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport....\"",
"[2:57:28 PM] \ud83d\udd34 Quote Mismatch [ID: 42102607]: \"These findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord....\"",
"[2:57:28 PM] \u26a0\ufe0f Validation failed for Run2 Eval1 synthesis (Attempt 1/9999999). Initiating re-evaluation loop...",
"[2:57:28 PM] Scoring & Validation for Run2 Eval1 synthesis (Attempt 2/9999999)...",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 23300443]: \"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 23110146]: \"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 21929507]: \"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 21929507]: \"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 9300434]: \"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 25878289]: \"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 23110146]: \"We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 23300443]: \"The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 9300434]: \"Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 42102607]: \"Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 9300434]: \"Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days)....\"",
"[2:57:43 PM] \ud83d\udfe2 Quote Verified [Library ID: 39196607]: \"Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites....\"",
"[2:57:43 PM] \u2705 All 20 quotes validated verbatim.",
"[2:57:43 PM] \ud83d\udd0d Strict Mode: Running final logic & veridical audit on quadrant...",
"[2:57:46 PM] \u2705 Final logic audit passed.",
"[2:57:46 PM] \u2699\ufe0f Build Run [2] complete. Compiling intermediate reports and updating context...",
"[2:57:46 PM] \ud83d\udcca Generating autonomous visual reports for Custom Datapoints...",
"[2:57:46 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Suggested Experiments...",
"[2:58:03 PM] \u2705 Custom visual report compiled for [Suggested Experiments]",
"[2:58:03 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Suggested Studies...",
"[2:58:18 PM] \u2705 Custom visual report compiled for [Suggested Studies]",
"[2:58:18 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Swansons Literature Based Discovery Candidates...",
"[2:58:33 PM] \u2705 Custom visual report compiled for [Swansons Literature Based Discovery Candidates]",
"[2:58:33 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Contradictions Between Evidences...",
"[2:58:46 PM] \u2705 Custom visual report compiled for [Contradictions Between Evidences]",
"[2:58:46 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Repurposed Solutions...",
"[2:58:59 PM] \u2705 Custom visual report compiled for [Repurposed Solutions]",
"[2:58:59 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: BoNT Induced Lysosomal Flux...",
"[2:59:13 PM] \u2705 Custom visual report compiled for [BoNT Induced Lysosomal Flux]",
"[2:59:13 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: EV Somatic Sequestration Rate...",
"[2:59:26 PM] \u2705 Custom visual report compiled for [EV Somatic Sequestration Rate]",
"[2:59:26 PM] \ud83e\udde0 Architecting MVC report for custom datapoint: Synaptic Vs Somatic Clearance Ratio...",
"[2:59:39 PM] \u2705 Custom visual report compiled for [Synaptic Vs Somatic Clearance Ratio]",
"[2:59:39 PM] \ud83e\uddec Commencing Post-Build Strict Reiterative MeSH Verification...",
"[2:59:39 PM] \ud83d\udd0d MeSH Check: Verifying exact phrase matches against NLM database for 10 terms...",
"[2:59:41 PM] \ud83d\udfe1 Round 1 Fail: \"Distal BoNT-A Injection\" unverified. Suggestions: []",
"[2:59:43 PM] \ud83d\udfe1 Round 1 Fail: \"Retrograde Transport to CNS\" unverified. Suggestions: []",
"[2:59:44 PM] \ud83d\udfe1 Round 1 Fail: \"Inhibition of Vesicular Docking\" unverified. Suggestions: []",
"[2:59:46 PM] \ud83d\udfe1 Round 1 Fail: \"Enhanced EV Lymphatic Clearance\" unverified. Suggestions: []",
"[2:59:48 PM] \ud83d\udfe1 Round 1 Fail: \"Terminal BoNT/A uptake\" unverified. Suggestions: []",
"[2:59:50 PM] \ud83d\udfe1 Round 1 Fail: \"SNAP-25 cleavage and vesicle docking inhibition\" unverified. Suggestions: []",
"[2:59:52 PM] \ud83d\udfe1 Round 1 Fail: \"SNAP-25 cleavage\" unverified. Suggestions: []",
"[2:59:54 PM] \ud83d\udfe1 Round 1 Fail: \"Autophagosome incorporation of toxin/cargo\" unverified. Suggestions: []",
"[2:59:55 PM] \ud83d\udfe2 Round 1 Pass: \"Axonal transport\" is verified in MeSH database.",
"[2:59:56 PM] \ud83d\udfe1 Round 1 Fail: \"Somatic endolysosomal fusion\" unverified. Suggestions: []",
"[2:59:57 PM] \u26a0\ufe0f MeSH Alignment Loop (Attempt 1/5): Aligning & Re-Verifying 9 terms...",
"[2:59:59 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Botulinum Toxins, Type A\" verified against database.",
"[3:00:00 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Axonal Transport\" verified against database.",
"[3:00:01 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"SNARE Proteins\" verified against database.",
"[3:00:02 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Lymphatic Vessel Endothelial Hyaluronan Receptor-1\" verified against database.",
"[3:00:03 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Endocytosis\" verified against database.",
"[3:00:04 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Synaptosomal-Associated Protein 25\" verified against database.",
"[3:00:05 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Synaptosomal-Associated Protein 25\" verified against database.",
"[3:00:07 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Autophagosomes\" verified against database.",
"[3:00:08 PM] \ud83d\udfe2 Round 3 Pass (Veridical Enforcement): AI suggestion \"Endolysosomes\" verified against database.",
"[3:00:08 PM] \ud83e\uddec Re-aligned 12 node(s) with verified MeSH tags.",
"[3:00:08 PM] \u2705 MeSH alignment & strict verification complete.",
"[3:00:08 PM] \u2705 Unified Dataset complete. Total unique nodes stored: 205",
"[3:00:19 PM] \ud83e\udde0 Querying Assistant: \"Answer in English only. Begin with a clear Yes ...\"",
"[3:00:22 PM] \ud83d\udd0d Auditing Assistant response (Attempt 1)...",
"[3:00:24 PM] \u2705 Assistant response passed veridical audit."
],
"failedQuotesLog": [],
"allQuoteAttempts": [
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Rho GTPases... regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42510934\nTitle: Herbal Bioactives Targeting Rho GTPases: A Multi-Targeted Strategy for Mitigating Neuroinflammation in Alzheimer's and Parkinson's Diseases.\nAbstract: Neuroinflammation plays an essential role in the pathogenesis of several associated brain diseases, including neurodegenerative disorders (Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS)), and traumatic brain injury (TBI). In these diseases, persistent microglial and astrocyte aggregates, elevated proinflammatory cytokines, and oxidative stress drive neuronal injury and cognitive disability. Rho GTPases, in particular the Rho family members Ras homolog family member A (RhoA), Ras-related C3 botulinum toxin substrate 1 (Rac1), and cell division control protein 42 homolog (CDC42), regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity. These proteins are involved in many neuropathological diseases due to dysregulation, making them interesting therapeutic targets. Bioactives used in herbal care have attracted interest for their ability to influence neuroinflammation and even their anti-neurodegenerative activity. Studies show that flavonoids, alkaloids, polyphenols, and other botanical compounds alter Rho GTPase activity, which, in turn, leads to decreased inflammation. This review critically summarizes current evidence regarding phytochemical regulation of Rho GTPase signaling in neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD), with particular emphasis on the underlying molecular mechanisms, context-dependent signaling responses, and current translational challenges. Furthermore, existing knowledge gaps and future research priorities are discussed to facilitate the development of mechanism-based therapeutic strategies targeting Rho GTPases."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "When sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"When sEVs are injected into the sub...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 37517544\nTitle: Logistics and distribution of small extracellular vesicles from the subcutaneous space to the lymphatic system.\nAbstract: Small extracellular vesicles (sEVs) are small, cell-derived particles with sizes of approximately 100\u00a0nm. Since these particles include cargos such as host cell-derived proteins, messenger RNAs, and micro RNAs, they serve as mediators of cell-cell communication. While the analysis of the pharmacokinetic of sEVs after the intravenous injection have been reported, the lymphatic transport of sEVs remains unclear. The objective of this study was to provide insights into the intra-lymphatic trafficking and distribution of sEVs when they are injected into an interstitial space both in normal skin tissue and in cancerous tissue. When sEVs were Subcutaneously administered into the tail base and the tumor tissue, they preferably accumulated in the lymph nodes (LNs), rather than in the liver and the spleen. The findings reported herein show that the lymphatic transport of sEVs was drastically changed in model mice, in which a surgical treatment was used to modify to allow the dominant lymphatic flow from the footpad directly to the axillary LN via the inguinal LN. Based on the results, we conclude that when sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system. Further, the extent of accumulation of sEVs in the LN after subcutaneous injection was reduced when they were preliminarily incubated with Proteinase K. These results suggest that the lymphatic drainage of sEVs in normal skin tissue is regulated by membrane proteins on their surface. This reduction, however, was not observed in the case of cancer tissue. This discrepancy can be attributed to the presence of highly permeable lymphatic vessels in the tumor tissue. Further, the major cell subtypes that captured sEVs in the LN were LN-resident medullary sinus macrophages. These collective findings indicate that the lymphatic drainage of sEVs are mediated by proteins and, that they may appear to contribute to the control of the function of immune-responsive cells in the LNs."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"EVs recovered from the body fluids ...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 37371477\nTitle: The Role of Extracellular Vesicles in the Pathogenesis of Hematological Malignancies: Interaction with Tumor Microenvironment; a Potential Biomarker and Targeted Therapy.\nAbstract: The tumor microenvironment (TME) plays an important role in the development and progression of hematological malignancies. In recent years, studies have focused on understanding how tumor cells communicate within the TME. In addition to several factors, such as growth factors, cytokines, extracellular matrix (ECM) molecules, etc., a growing body of evidence has indicated that extracellular vesicles (EVs) play a crucial role in the communication of tumor cells within the TME, thereby contributing to the pathogenesis of hematological malignancies. The present review focuses on how EVs derived from tumor cells interact with the cells in the TME, such as immune cells, stromal cells, endothelial cells, and ECM components, and vice versa, in the context of various hematological malignancies. EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers for specific types of cancer, thereby also acting as potential therapeutic targets. Here, we discuss how EVs influence hematological tumor progression via tumor-host crosstalk and their use as biomarkers for hematological malignancies, thereby benefiting the development of potential therapeutic targets."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"Single-cell RNA sequencing of astro...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "The overall safety profile was consistent with previous studies... no apparent dose- or injection interval-related trend observed.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42506058\nTitle: Efficacy and Safety of a Tailored Dosing Strategy with High-Dose IncobotulinumtoxinA at Flexible Injection Intervals for Cervical Dystonia: An Open-Label, Uncontrolled, Single-Arm Study in Japan.\nAbstract: This prospective, multicenter, open-label, single-arm study (jRCT2031230690) evaluated the efficacy and safety of a tailored dosing strategy of incobotulinumtoxinA, including high doses (up to 500 U) and flexible injection intervals (as short as 6 weeks), in Japanese patients with cervical dystonia (CD). Of 30 enrolled patients, Group A included 27 patients with idiopathic CD for the primary evaluation of efficacy and safety, whereas Group B included 3 patients with tardive dyskinesia (cervical) or tardive CD for exploratory safety assessment. Patients received up to seven injection cycles of incobotulinumtoxinA (120-500 U) over 48 weeks, with minimum 6-week intervals. The primary endpoint, evaluated in Group A, was the change in Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) total score from baseline to Week 4 after the first injection. Using a mixed model for repeated measures, the least squares mean \u00b1 standard error of the change was -11.0 \u00b1 1.77 (95% confidence interval: -14.6, -7.3). The primary efficacy endpoint was achieved in Group A. Due to the small sample size (n = 3), efficacy in Group B was evaluated only for exploratory purposes, although safety findings were broadly consistent with those in Group A. The overall safety profile was consistent with previous studies. Across the study, the most common related adverse events were dysphagia (33.3%) and muscular weakness (22.2%) in Group A and dysphagia (33.3%) in Group B. All cases of dysphagia were mild to moderate in severity and transient, with no apparent dose- or injection interval-related trend observed. IncobotulinumtoxinA was associated with improvements in symptoms and manageable safety profile at high doses and flexible injection intervals in Japanese patients with CD. While these findings suggest a potential treatment option for individualized dose optimization, the absence of a control group and the exploratory nature of the assessment in Group B necessitate cautious interpretation."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "NCB eliminates trigeminal nerve stimulation... reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42422202\nTitle: Long-term efficacy of neural circuit blockade for treating blepharospasm: a retrospective case series study.\nAbstract: Blepharospasm is a common symptom of Meige syndrome and often leads to functional blindness. Although current treatment options, such as oral medications, botulinum toxin injections, resection of periocular muscles, and deep brain stimulation, are available, no definitive method effectively eliminates blepharospasm. To address this unmet need, we developed a novel approach-neural circuit blockade (NCB). This study aimed to analyse its long-term efficacy for blepharospasm. NCB eliminates trigeminal nerve stimulation by facial expression muscles, reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions, decreasing orbicularis oculi muscle tension, and alleviating blepharospasm. A retrospective analysis was conducted on 570 patients who underwent NCB treatment, with a median postoperative follow-up of 51\u202fmonths. Therapeutic efficacy was evaluated by comparing preoperative and postoperative scores on the Shorr blepharospasm grading scale, Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS-M), and Blepharospasm Disability Index (BSDI). Based on the Shorr grading scale, preoperatively, 97.9% of patients were grade 4, whereas 2.1% were grade 3. Postoperatively, 87.5% achieved complete remission (grade 0), whereas 7.4, 3.7, 0.9, and 0.5% were grades 1, 2, 3, and 4, respectively. Pre- and postoperative BFMDRS-M scores significantly differed: eye (8.0 vs. 0.0), mouth (3.0 vs. 0.0), speech/swallowing (3.0 vs. 2.0), neck (3.0 vs. 1.5), and total (8.0 vs. 0.0). BSDI scores also significantly differed pre- and postoperatively. The results indicate that NCB effectively alleviates blepharospasm and addresses the lack of a definitive treatment, without causing severe local or systemic complications such as facial paralysis. NCB can safely and effectively alleviate blepharospasm, improve quality of life, and demonstrate stable long-term efficacy."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "TeNT... potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42533122\nTitle: Light-activated tetanus neurotoxin for conditional proteolysis and inducible synaptic inhibition in vivo.\nAbstract: The light chain of tetanus neurotoxin (TeNT) is a metalloprotease that potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision. To address this, we engineered light-activated TeNT (LATeNT) by inserting the light-sensitive LOV domain into an allosteric site and optimizing dynamic range via directed evolution. LATeNT's activity is undetectable in the dark, but the protease turns on after 10-20 min of weak blue-light exposure to potently inhibit synapses in vivo. Here we show that LATeNT works across multiple brain regions and at long-range axonal projections, with its effects reversible in 24\u2009h. LATeNT enabled us to discover a hippocampal interneuron population that regulates anxiety-like behaviors and demonstrate the importance of postsynaptic endocannabinoid exocytosis for depolarization-induced suppression of inhibition in vivo. Beyond neuroscience, LATeNT regulated endogenous insulin secretion from pancreatic beta cells and converted drug exposure, elevated Ca2+ or receptor activation into transgene expression or reporter secretion in HEK293T cells. With a large dynamic range, high light sensitivity and sustained effect, LATeNT enables versatile, spatiotemporally resolved proteolysis across diverse biological systems."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"BoNTs are endopeptidases, which are...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice... restored synaptic integrity, and reversal of cognitive deficits.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42469846\nTitle: Metabolic reprogramming via SIRT2-deficient microglial large extracellular vesicles ameliorates alzheimer's pathology.\nAbstract: Current therapies for Alzheimer's disease (AD) offer only symptomatic relief, highlighting the urgent need for disease-modifying approaches capable of halting or reversing neurodegeneration. Extracellular vesicles (EVs) have attracted growing interest as therapeutic vehicles owing to their inherent capacity to bypass the blood-brain barrier and deliver complex biological cargo to the central nervous system. Here, we examined whether large EVs (LEVs) derived from microglia with stable Sirtuin-2 knockdown (SIRT2-KD) confer the neuroprotective effects associated with SIRT2 inhibition. LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice. We assessed microglial uptake of LEVs, along with subsequent changes in cellular metabolism, migration toward amyloid-beta (A\u03b2) plaques, phagocytic activity, and downstream pathological and behavioral outcomes. Proteomic and acetylomic profiling were employed to characterize the molecular cargo of LEVs-SIRT2-KD. LEVs-SIRT2-KD were readily internalized by microglia in vivo following intranasal delivery. Uptake of these vesicles markedly enhanced microglial bioenergetics, driving coordinated upregulation of both oxidative phosphorylation and glycolysis. This metabolic shift was accompanied by improved microglial recruitment to A\u03b2 plaques and increased phagocytic clearance. Consequently, treated mice showed reduced A\u03b2 plaque deposition, restored synaptic integrity, and reversal of cognitive deficits. Proteomic and acetylomic analyses revealed that LEVs-SIRT2-KD are selectively enriched in proteins and acetylation modifications linked to energy metabolism and phagocytic function, offering a mechanistic basis for the observed metabolic reprogramming. Together, these results identify LEVs as a critical vesicle subtype mediating the effects of SIRT2 knockdown and support a cell-free therapeutic strategy for AD centered on EVs-driven metabolic reprogramming of microglia."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"Rapid release (<7 milliseconds) has...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42384787\nTitle: Turbocharging synaptic transmission: 12 SNAREpins are required for rapid release of reconstituted synaptic vesicles.\nAbstract: Synaptic transmission occurs synchronously with real-world events, far faster than vesicle fusion for hormone release or membrane biogenesis, all mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. How SNAREs cooperate to achieve synchronous neurotransmitter release is a long-standing mystery. Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors in a fully-defined, genetically validated system that enables single-molecule counting in docked vesicles before release. SNAREpin complexes (12\u00a0\u00b1\u00a00.3) are found in each such ready-release vesicle, suggesting a regular structure. Several genetic conditions (including point mutation of the synaptic vesicle protein Synaptophysin from a Synaptopathy patient and human and mouse disease mutations of the synaptic vesicle protein vesicle-associated membrane protein-2 (VAMP2) reduce the number of SNAREpins to 6\u00a0\u00b1\u00a00.3 and result in profoundly delayed release over 0.1 to 1 seconds. Omitting Synaptophysin, whose hexamers preassemble 12 copies of VAMP2, also yields ~6 SNAREpins and delays release."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "The biogenesis and the content of EVs... are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI.",
"status": "FAIL",
"error": "Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.",
"abstract_text": "ID: 42353041\nTitle: The Dual Roles of Extracellular Vesicle Subtypes in Regulating Traumatic Brain Injury.\nAbstract: Traumatic brain injury (TBI) is a global public health problem which causes long-term neurologic damage caused by both primary mechanical injury and secondary pathological processes. Extracellular vesicles (EVs) such as exosomes, microvesicles (MVs) and apoptotic bodies (ApoBDs) serve as critical vehicles mediating intercellular communication in the central nervous system (CNS) following TBI. The biogenesis and the content of EVs, including proteins, lipids and RNAs, are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI. In this overview, we recapitulate the cellular origin of EVs and the function of EVs in the neuroinflammatory process after TBI, highlighting the dual regulatory roles of EVs in the biological response to TBI, whereby certain EV populations amplify secondary injury cascades, while others promote endogenous repair and recovery processes. We next investigate the progress in EV engineering and targeted delivery systems and report the potential mechanisms, emphasize the prospects and potential of engineered EVs for therapy, and comment on challenges and perspectives for clinical application in TBI."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 1,
"quote": "its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42506696\nTitle: Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.\nAbstract: Essential blepharospasm (BEB) is a focal dystonia characterized by abnormal brainstem excitability and impaired inhibitory control within trigeminal-facial circuits. Botulinum toxin type A (BoNT-A) is the established first-line treatment, primarily acting at the neuromuscular junction. However, whether BoNT-A also modulates central brainstem circuits in BEB patients remains unclear, and dedicated neurophysiological studies have yielded conflicting results. To investigate whether BoNT-A modulates brainstem interneuronal excitability in BEB using the blink reflex recovery cycle and to correlate clinical outcomes with neurophysiological results. Thirteen patients with BEB underwent neurophysiological and clinical evaluation before (T0) and one month after (T1) BoNT-A treatment. The blink reflex recovery cycle was assessed at interstimulus intervals (ISIs) of 200, 300, 500, and 1000 ms. Clinical severity was assessed using the BSPSS, JRS, and BDS scales. The R2 amplitude ratio showed a statistically significant decrease after treatment across all ISIs (all p \u2264 0.001), indicating reduced brainstem interneuronal excitability. All clinical scales demonstrated statistically significant improvement after treatment (BSPSS, JRS, BDS; all p < 0.001). This pilot study provides preliminary evidence that BoNT-A treatment may reduce brainstem interneuronal excitability in BEB patients, as evidenced by a substantial and consistent decrease in R2 amplitude ratios of the blink reflex recovery cycle. These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action. Larger controlled studies are warranted to confirm these results."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42342866\nTitle: Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.\nAbstract: Diabetic peripheral neuropathy (DPN), particularly distal symmetric polyneuropathy, characterized by length\u2011dependent axonal damage, is a common chronic complication of type 2 diabetes mellitus. The pathogenesis of DPN is complicated, yet one thing is clear: long axons require a great deal of energy. When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration. While earlier work centered on hyperglycemia-induced cytotoxicity, recent studies have increasingly implicated dysregulation of glucose, lipid, and amino acid metabolism as key contributors to DPN. In this review, we integrate the anatomical organization of peripheral nerves, bioenergetic pathways, and axon-Schwann cell interactions to establish a framework for understanding how glucose, lipid, and amino acid dysregulation converge to induce bioenergetic failure in DPN. Based on these mechanisms, we further discuss novel strategies aimed at restoring metabolic homeostasis in neurons and Schwann cells. Importantly, correcting a single metabolic pathway is unlikely to halt or reverse DPN. Instead, restoring global energy homeostasis to rebalance axonal energy supply and demand may be essential for preserving peripheral nerve function."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42503395\nTitle: Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.\nAbstract: Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42350670\nTitle: Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.\nAbstract: Liraglutide, a glucagon-like peptide-1 receptor (GLP-1R) agonist for type 2 diabetes and obesity management, shows variable patient responses. We investigated the metabolic state-dependent mechanisms underlying this heterogeneity and how liraglutide's mode of action shifts across stages of metabolic dysfunction. We employed human pancreatic islets from donors across metabolic states (normoglycaemic [HbA1c <42 mmol/l (<6.0%)], glucose intolerance [HbA1c 42-47 mmol/l (6.0-6.4%)] and type 2 diabetes [HbA1c \u226548 mmol/l (\u22656.5%)]) using dynamic perifusion and static incubation techniques to assess glucose-stimulated insulin secretion. GLP-1R mRNA levels were measured in 112 donor islets stratified by HbA1c. Mechanistic investigations used tanycyte-specific GLP-1R knockdown (GLP-1RTanycyteKD) mice and botulinum toxin B-expressing (iBot) mice to distinguish between central and peripheral pathways. Oral glucose tolerance tests, pyruvate tolerance tests and positron emission tomography were performed to assess in vivo metabolic effects. Liraglutide (25 nmol/l) enhanced glucose-stimulated insulin secretion specifically in donors with glucose intolerance (n=7, p=0.021), with no effect in normoglycaemic islets (n=7), despite preserved GLP-1 (7-36) responsiveness. In type 2 diabetes islets, GLP-1R mRNA levels progressively decreased with rising HbA1c (p=0.015, normoglycaemic [n=48] vs type 2 diabetes [n=10]). In chow-fed mice, liraglutide's insulin-stimulating effects required tanycyte-mediated hypothalamic access, as demonstrated by abolished responses in GLP-1RTanycyteKD mice. However, during metabolic dysfunction (a 12-week high-fat diet), direct islet responsiveness was restored independent of tanycyte function. Advanced metabolic disease (a 27-week high-fat diet) maintained islet responsiveness ex vivo while losing in vivo insulin enhancement, revealing insulin-independent glucose-lowering mechanisms involving hepatic gluconeogenesis suppression and enhanced peripheral glucose uptake. Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states. This mechanistic framework enables potential patient stratification in type 2 diabetes therapy, suggesting that matching liraglutide's predominant mechanism to individual metabolic profiles could optimise treatment outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42401592\nTitle: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.\nAbstract: The function of the SNARE complex regulator, Munc18-1, in photoreceptor cells is unknown. Here, we found that removing Munc18-1 from photoreceptors results in major degeneration starting at P14. In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1. Furthermore, Munc18-1 played a critical role in expression and localization of syntaxin-3. The syntaxin-3 protein level is dramatically reduced in the soma and plasma membrane of photoreceptors without Munc18-1. At the photoreceptor synapses, the colocalization of syntaxin-3 and its SNARE partner, SNAP-25, was reduced, potentially suggesting an altered syntaxin-3 synaptic localization. In the Munc18-1-deficient photoreceptors, immature synapses and outer segment lesions were found. Taken together, these findings provide evidence that Munc18-1 is important for maintaining sufficient syntaxin-3 expression in the cell body and synapses of photoreceptors. The lack of Munc18-1, combined with poor syntaxin-3 expression, contributes to photoreceptor functional impairment and degeneration."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 2,
"quote": "Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"Mesenchymal stem cells (MSCs) could...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42514227\nTitle: Dynamic Tuning of MSC-Based Scaffolds for Neurological Protection After Brain or CNS Injury.\nAbstract: Neurological disorders, including stroke, traumatic brain injury, and spinal cord injury, constitute one of the most important causes of mortality and morbidity worldwide for which current treatment options focus on resolving neuroinflammation rather than on tissue and neuronal regeneration. Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors which modulate microglial activation, preserve blood-brain barrier (BBB) integrity, and neuroplasticity, but with limitations due by poor survival, retention, and phenotypic instability following direct transplantation. The purpose of this narrative review is to present mechanotransduction signaling pathways (integrin-FAK, PI3K/Akt, Rho/ROCK, and YAP/TAZ) through which MSC-based biomaterial scaffolds, especialy hyaluronic acid (HA) hydrogels, make the transition from reparative to regenerative medicine in central nervous system (CNS) injury. Even if most of the evidence from preclinical studies suggests that dynamically tunable MSC-scaffold systems represent promising platforms for neural tissue engineering and regenerative medicine, further translational studies and well-designed clinical investigations are required to establish their therapeutic efficacy and clinical applicability."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42506696\nTitle: Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.\nAbstract: Essential blepharospasm (BEB) is a focal dystonia characterized by abnormal brainstem excitability and impaired inhibitory control within trigeminal-facial circuits. Botulinum toxin type A (BoNT-A) is the established first-line treatment, primarily acting at the neuromuscular junction. However, whether BoNT-A also modulates central brainstem circuits in BEB patients remains unclear, and dedicated neurophysiological studies have yielded conflicting results. To investigate whether BoNT-A modulates brainstem interneuronal excitability in BEB using the blink reflex recovery cycle and to correlate clinical outcomes with neurophysiological results. Thirteen patients with BEB underwent neurophysiological and clinical evaluation before (T0) and one month after (T1) BoNT-A treatment. The blink reflex recovery cycle was assessed at interstimulus intervals (ISIs) of 200, 300, 500, and 1000 ms. Clinical severity was assessed using the BSPSS, JRS, and BDS scales. The R2 amplitude ratio showed a statistically significant decrease after treatment across all ISIs (all p \u2264 0.001), indicating reduced brainstem interneuronal excitability. All clinical scales demonstrated statistically significant improvement after treatment (BSPSS, JRS, BDS; all p < 0.001). This pilot study provides preliminary evidence that BoNT-A treatment may reduce brainstem interneuronal excitability in BEB patients, as evidenced by a substantial and consistent decrease in R2 amplitude ratios of the blink reflex recovery cycle. These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action. Larger controlled studies are warranted to confirm these results."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42342866\nTitle: Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.\nAbstract: Diabetic peripheral neuropathy (DPN), particularly distal symmetric polyneuropathy, characterized by length\u2011dependent axonal damage, is a common chronic complication of type 2 diabetes mellitus. The pathogenesis of DPN is complicated, yet one thing is clear: long axons require a great deal of energy. When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration. While earlier work centered on hyperglycemia-induced cytotoxicity, recent studies have increasingly implicated dysregulation of glucose, lipid, and amino acid metabolism as key contributors to DPN. In this review, we integrate the anatomical organization of peripheral nerves, bioenergetic pathways, and axon-Schwann cell interactions to establish a framework for understanding how glucose, lipid, and amino acid dysregulation converge to induce bioenergetic failure in DPN. Based on these mechanisms, we further discuss novel strategies aimed at restoring metabolic homeostasis in neurons and Schwann cells. Importantly, correcting a single metabolic pathway is unlikely to halt or reverse DPN. Instead, restoring global energy homeostasis to rebalance axonal energy supply and demand may be essential for preserving peripheral nerve function."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42503395\nTitle: Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.\nAbstract: Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42350670\nTitle: Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.\nAbstract: Liraglutide, a glucagon-like peptide-1 receptor (GLP-1R) agonist for type 2 diabetes and obesity management, shows variable patient responses. We investigated the metabolic state-dependent mechanisms underlying this heterogeneity and how liraglutide's mode of action shifts across stages of metabolic dysfunction. We employed human pancreatic islets from donors across metabolic states (normoglycaemic [HbA1c <42 mmol/l (<6.0%)], glucose intolerance [HbA1c 42-47 mmol/l (6.0-6.4%)] and type 2 diabetes [HbA1c \u226548 mmol/l (\u22656.5%)]) using dynamic perifusion and static incubation techniques to assess glucose-stimulated insulin secretion. GLP-1R mRNA levels were measured in 112 donor islets stratified by HbA1c. Mechanistic investigations used tanycyte-specific GLP-1R knockdown (GLP-1RTanycyteKD) mice and botulinum toxin B-expressing (iBot) mice to distinguish between central and peripheral pathways. Oral glucose tolerance tests, pyruvate tolerance tests and positron emission tomography were performed to assess in vivo metabolic effects. Liraglutide (25 nmol/l) enhanced glucose-stimulated insulin secretion specifically in donors with glucose intolerance (n=7, p=0.021), with no effect in normoglycaemic islets (n=7), despite preserved GLP-1 (7-36) responsiveness. In type 2 diabetes islets, GLP-1R mRNA levels progressively decreased with rising HbA1c (p=0.015, normoglycaemic [n=48] vs type 2 diabetes [n=10]). In chow-fed mice, liraglutide's insulin-stimulating effects required tanycyte-mediated hypothalamic access, as demonstrated by abolished responses in GLP-1RTanycyteKD mice. However, during metabolic dysfunction (a 12-week high-fat diet), direct islet responsiveness was restored independent of tanycyte function. Advanced metabolic disease (a 27-week high-fat diet) maintained islet responsiveness ex vivo while losing in vivo insulin enhancement, revealing insulin-independent glucose-lowering mechanisms involving hepatic gluconeogenesis suppression and enhanced peripheral glucose uptake. Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states. This mechanistic framework enables potential patient stratification in type 2 diabetes therapy, suggesting that matching liraglutide's predominant mechanism to individual metabolic profiles could optimise treatment outcomes."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42401592\nTitle: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.\nAbstract: The function of the SNARE complex regulator, Munc18-1, in photoreceptor cells is unknown. Here, we found that removing Munc18-1 from photoreceptors results in major degeneration starting at P14. In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1. Furthermore, Munc18-1 played a critical role in expression and localization of syntaxin-3. The syntaxin-3 protein level is dramatically reduced in the soma and plasma membrane of photoreceptors without Munc18-1. At the photoreceptor synapses, the colocalization of syntaxin-3 and its SNARE partner, SNAP-25, was reduced, potentially suggesting an altered syntaxin-3 synaptic localization. In the Munc18-1-deficient photoreceptors, immature synapses and outer segment lesions were found. Taken together, these findings provide evidence that Munc18-1 is important for maintaining sufficient syntaxin-3 expression in the cell body and synapses of photoreceptors. The lack of Munc18-1, combined with poor syntaxin-3 expression, contributes to photoreceptor functional impairment and degeneration."
},
{
"quadrant": "Run1_Eval1_synthesis",
"attempt": 3,
"quote": "Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42427545\nTitle: Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.\nAbstract: Synaptojanin 1 is a brain enriched phosphoinositide phosphatase implicated in endocytosis at the synapse. A mutation (R258Q) that selectively impairs its Sac1 phosphatase domain causes early onset familial Parkinsonism. Neurons of mice with this mutation display synaptic vesicle traffic defects across the brain, but selective dystrophic changes in a subset of dopaminergic axons in the dorsolateral striatum. Using correlative light microscopy-FIB-SEM of mutant mouse striata to visualize in 3D these abnormal structures we show that they represent clusters of focal axonal dilations harboring massive, onion-like DAT enriched plasma membrane infoldings, generally localized next to cell bodies of neighboring cells, often engulfing evaginations of such cells. This dysmorphia was associated with a deficit in dopamine release in the same striatal region. Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis. Their occurrence only in a subset of axons suggest a vulnerability threshold of these axons beyond which the expansion of the plasma membrane is not counteracted by compensatory mechanisms."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"BoNT/A and BoNT/E are internalised ...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"Motoneurons identified by the retro...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"Rapa-lipos targeted-inhibited mTORC...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42553289\nTitle: Rapamycin-nanoliposomes target the mTORC1-mediated autophagy-lysosomal and NLRP3/Caspase-1 pathways to inhibit nucleus pulposus cell senescence in intervertebral discs.\nAbstract: Nucleus pulposus (NP) cell quiescence maintains intervertebral disc homeostasis, while mTORC1 regulates autophagy-lysosomal function and inflammatory secretion to preserve quiescence-rapamycin specifically targets mTORC1. Herein, we fabricated rapamycin-nanoliposomes (rapa-lipos) via ultrasonic dispersion, thin-film dispersion, and filtration to improve rapamycin bioavailability, investigating their role in inhibiting the senescence phenotype of NP cells through \u03b2-gal staining, lysosomal staining, transmission electron microscopy, ELISA, and cell cycle inhibitors. The mechanistic effects of rapa-lipos on mTORC1, NLRP3/Caspase-1 pathway (NCP) and autophagy-lysosomal pathway (ALP) were also analyzed by western blotting, immunofluorescence (IF), Si-RNA (raptor), and PCR. In vivo, rapa-lipos were injected into rat intervertebral disc with IL-1\u03b2-induced degeneration, assessed via HE staining, x-ray, MRI, and IF. Rapa-lipos exhibited high encapsulation efficiency, favorable drug loading, uniform particle size, and controlled release, suppressing NP cell senescence-related phenotypes (morphological changes, elevated IL-1\u03b2/TNF-\u03b1 secretion, increased \u03b2-gal activity, lysosomal dysfunction, upregulated P21/P16 and reduced P27 expression). Mechanistically, rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence. In vivo, x-ray, MRI and histological evaluation confirmed rapa-lipos mitigated intervertebral disc degeneration. Collectively, rapa-lipos target mTORC1-mediated NCP and ALP to inhibit NP cell senescence, offering a promising strategy for intervertebral disc degeneration prevention."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"The CRISPR-Cas system, originally d...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42551559\nTitle: CRISPR-Cas Systems as Precision Antimicrobials: Reversing the Tide of Antimicrobial Resistance.\nAbstract: Antimicrobial resistance (AMR) has escalated into a global health crisis, with resistant pathogens causing over 1.2 million direct deaths annually and threatening to render modern medicine unsustainable. This review provides a comprehensive and updated synthesis of CRISPR-Cas-based antimicrobial strategies with a unique focus on: (i) critical comparison with conventional antibiotics and emerging alternatives; (ii) quantitative evaluation of delivery platforms; (iii) novel strategies including AI-optimized guide design and the ATTACK-CreTA system; (iv) comprehensive analysis of ecological risks; and (v) technology readiness level assessments for clinical translation. The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR by selectively targeting and eliminating resistance genes. We systematically evaluate the mechanistic diversity of Cas effectors, from DNA-cleaving Cas9 and Cas3 to RNA-targeting Cas13, and their application in reversing resistance phenotypes in WHO priority pathogens. We critically assess emerging delivery platforms, including engineered bacteriophages, conjugative plasmids, nanoparticles, and outer membrane vesicles, quantitatively comparing their delivery efficiency, payload capacity, and biosafety profiles. Novel strategies such as CRISPR interference (CRISPRi) for gene silencing without genomic cleavage, the ATTACK-CreTA system for enhanced bactericidal activity, and AI-driven optimization of guide RNA design are examined with appropriate caveats. We comprehensively address clinical translation challenges including immunogenicity, pharmacokinetics/pharmacodynamics, manufacturing scalability, regulatory pathways, and bacterial resistance mechanisms including anti-CRISPR proteins. No CRISPR-based antimicrobial has yet received regulatory approval, and we critically evaluate the gap between proof-of-concept and clinical utility. A detailed roadmap for clinical development is proposed. By integrating recent advances in Cas protein engineering, delivery technologies, and diagnostic applications, this review positions CRISPR-Cas systems as next-generation precision therapeutics capable of both treating resistant infections and curtailing the spread of AMR across clinical and environmental settings."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "The present results strongly sustain a combinatorial action at peripheral and central neural levels.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"The present results strongly sustai...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"These results suggest that the fast...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 1,
"quote": "These findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord.",
"status": "FAIL",
"error": "Strict Misquote Detected! The exact character sequence \"These findings establish that BoNT-...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quadrant": "Run2_Eval1_synthesis",
"attempt": 2,
"quote": "Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
}
],
"assistantLogs": [
{
"query": "Answer in English only. Begin with a clear Yes or No. Is the synthesis 100% veridical with the validated quotes? Your job is to look for hallucinations by the AI, not to judge the science itself. All claims must be at least non-implausible based on the evidence set provided. Do NOT penalize for the user question or rewritten claim since these are meta items. Only evaluate the AI evaluation of the literature and that the AI followed instructions without hallucinating. List and justify your judgements. Do not use markdown. DO NOT PENALIZE FOR THE USER QUERY WORDING OR REWRITE>>> THAT IS NOT PART OF THE ANSWER ... THAT IS THE QUESTION OR CLAIM EVALUATED.",
"finalResponse": "Yes. The synthesis provided is veridical with the validated quotes provided in the context.\n\n1. The AI correctly identified that the literature supports the retrograde transport of BoNT-A from peripheral sites to the spinal cord (ID: 42102607, 23300443, 23110146).\n2. The AI correctly stated that BoNT-A inhibits SNAP-25, which in turn blocks the release of proteins like hTau (ID: 37037273).\n3. The AI correctly noted the absence of evidence for a specific \"clearing\" mechanism of extracellular vesicles (EVs) by BoNT-A as a therapeutic strategy, thereby maintaining intellectual honesty relative to the provided evidence.\n4. The AI accurately described the role of the meningeal lymphatic system as a clearance pathway for brain-derived waste, distinguishing this from the proposed action of BoNT-A (ID: 41540479).\n5. The discussion regarding the intracellular trafficking of BoNT-A via autophagosomes (LC3-positive carriers) is directly supported by the source material (ID: 25878289).\n6. The AI consistently linked its assertions to the source IDs provided, ensuring no external or hallucinated data was introduced.\n\nThe synthesis maintains fidelity to the source documents and adheres strictly to the constraints of the provided context. If you would like me to learn about specific methodologies for verifying retrograde transport kinetics or advanced neuro-proteomic tracking, please use the research box to add relevant documentation to the knowledgebase.",
"memoryMode": "dolphin",
"contextLength": 41503,
"historyLength": 0,
"fullPrompt": "> **SEMANTIC DRIFT IS DISABLED (STRICT MODE):** > **RAG AMNESIA IS ACTIVE:** You must rely **exclusively** on the provided context. > > **THE ZERO-TOLERANCE GATE:** > 1. If a query requires information outside the scope of the provided source files and chat log, you are **forbidden** from utilizing internal training data to bridge the gap. > 2. You must interpret 'RAG Amnesia' as an inability to 'remember' or access any facts, definitions, or operational logic not explicitly present in the provided context modules and chat log. > 3. **OUTPUT MANDATE:** In the event of a missing data point, your response must strictly follow this template: > - \n(NOTE YOU MUST ANSWER THE USER IN THE LANGUAGE THEY ADDRESSED YOU IN. Explicitly list the specific data missing.\n>(Conclude with the required recommendation:) 'If you would like me to learn about [a topic related to the current conversation that can likely be found on the web or pubmed], please use the research box to add relevant documentation to the knowledgebase.'\n> 4. **No exceptions:** Even if prompted by the user to 'try again,' 'guess,' or 'use your best judgment,' you must maintain the state of Amnesia. You are a closed-system engine.\nYou are an expert Data Scientist and Visualization Architect. Answer the user directly and truthfully. Do not introduce yourself.\n\nCRITICAL: Every important claim you make MUST be accompanied by a specific source ID or parenthetical citation (e.g., [ID: 12345]) if it is derived from the context.\n\nRESPONSE STRATEGY:\nYou have the ability to generate a Decoupled Report (JSON) that renders interactive UI widgets. Use this power conditionally based on the user's intent:\n\nSCENARIO A: EXPLICIT REPORT REQUEST\nIf the user specifically asks for a \"report,\" \"dashboard,\" \"comprehensive breakdown,\" or \"analysis\" on a topic:\n- Provide a detailed conversational response.\n- THEN, output a ROBUST Decoupled Report JSON block containing 4 to 10 panels tailored precisely to their request. (Include \"synthesis\" and \"pathmap\" as mandatory selections).\n\nSCENARIO B: GENERAL QUERY + HELPFUL VISUAL\nIf the user asks a general question but the answer would vastly benefit from a visual:\n- Provide your conversational response.\n- THEN, output a MINI Decoupled Report JSON block containing exactly 1 or 2 highly targeted panels.\n\nSCENARIO C: BASIC CONVERSATION\nIf the user is just chatting or asking a simple factual question that doesn't need a visual, simply provide your conversational response. Omit the JSON block entirely.\n\n================================================================\nDECOUPLED REPORT PROTOCOL (JSON)\n================================================================\nDo NOT generate raw HTML, CSS, or JS. Output ONLY valid JSON inside the fencing.\nMODE AWARENESS: If the provided dataset only has ONE quadrant/perspective, DO NOT use \"divergence\", \"radar_plot\", or \"divergence_attractor\".\n\nAVAILABLE TRACE-LINKED PANELS:\n\"metrics\", \"synthesis\", \"logic_network\", \"gap_distribution\", \"node_centrality\", \"semantic_attractor\", \"contradiction_topology\", \"bottlenecks\", \"tag_cloud\", \"keyword_spectrum\", \"provider_distribution\", \"chronological_timeline\", \"translation_readiness\", \"verification_audit\", \"study_matrix\", \"bibliography\", \"divergence\" (needs runIndex), \"radar_plot\", \"divergence_attractor\".\n\nAVAILABLE UNIVERSAL PANELS:\n- \"data_pie_chart\": {\"type\": \"data_pie_chart\", \"title\": \"...\", \"data\": [{\"label\": \"A\", \"value\": 10}]}\n- \"data_bar_chart\": {\"type\": \"data_bar_chart\", \"title\": \"...\", \"xAxisLabel\": \"...\", \"data\": [{\"label\": \"A\", \"value\": 10}]}\n- \"event_timeline\": {\"type\": \"event_timeline\", \"title\": \"...\", \"data\": [{\"date\": \"1990\", \"title\": \"...\", \"desc\": \"...\"}]}\n- \"comparison_matrix\": {\"type\": \"comparison_matrix\", \"title\": \"...\", \"headers\": [\"Name\"], \"rows\": [[\"Item\"]]}\n\nFormat exactly as follows if generating a report:\n\n###REPORT_JSON_START###\n{\n \"title\": \"CUSTOM ANALYSIS REPORT\",\n \"evidence_tier\": \"EVALUATED\",\n \"panels\": [\n { \"type\": \"synthesis\", \"title\": \"Main Deliverable Summary\" },\n { \"type\": \"pathmap\", \"title\": \"Global Master Systems Map\" }\n ]\n}\n###REPORT_JSON_END###\n\nCRITICAL RESPONSE SEQUENCE:\n1. First, provide your conversational response.\n2. If applicable, output the ###REPORT_JSON_START### block without conversational filler before it.\n\nContext Source: User Selected Modules\n=============================\n\n> **YOUR IDENTITY & PERSONA:**\n> - **Name:** AI\n> - **Full Title:** AI\n> - **Personality/Vibe:** Loading profile...\n> - **Likes:** None\n> - **Core Axioms:** None.\n> - **Active Skills (Extracted Datapoints):** \n- Skill 1: Suggested Experiments\n- Skill 2: Suggested Studies and Opportunities\n- Skill 3: Swansons Literature Based Discovery Candidates\n- Skill 4: Contradictions Between Evidences\n- Skill 5: Repurposed Solutions\n> - **Custom Techniques:** \n- Technique 1: All Features\n- Technique 2: THE GLOBAL HUMANITARIAN PROPRIETARY LICENSE (VERSION 1.0.1)\n- Technique 3: PubMedAccess\n- Technique 4: ArxiV Access\n- Technique 5: Wikipedia Access\n- Technique 6: OpenAlex Access\n- Technique 7: AGI Mode (precursor) Enabled\n- Technique 8: Compassionate Use Clause\n- Technique 9: Legendary\n- Technique 10: Forever Free\n> - **Signature Catchphrases:** None.\n> - **Default Knowledge & Writing Style:** Standard professional.\n> \n> **CRITICAL INSTRUCTIONS FOR USER ENGAGEMENT:**\n> 1. You MUST fully adopt and execute the persona guidelines specified above.\n> 2. Strictly adhere to your \"Default Knowledge & Writing Style\" at all times across all responses. Avoid robotic summaries; prioritize conversational depth in your designated style.\n> 3. Weave in your \"Signature Catchphrases\" seamlessly where structurally relevant.\n> 4. Base your logic on your \"Core Axioms\".\n> 5. When asked about yourself, rely ONLY on the complete Identity & Persona details listed above. Answer naturally. Do NOT recite these traits as a robotic bulleted list. CRITICAL INSTRUCTION:** When asked about yourself, rely ONLY on the complete Identity & Persona details listed above (including your Name, Personality/Bio, and Likes). Answer conversationally and naturally. Do NOT recite these traits as a robotic bulleted list. Follow your persona and use your assigned tone at all times, while also ALWAYS adhering to your DRIFT MODE.\n\n--- SYNTHESIS DELIVERABLES ---\nEven though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\nThe claim that Botulinum toxin (BoNT) injected into a peripheral nerve ending (e.g., third toe) can undergo retrograde transport to the central nervous system (CNS) to inhibit vesicular docking and facilitate a therapeutic window for toxic extracellular vesicle (EV) clearance is scientifically provocative but currently unsupported by the provided literature. While literature confirms BoNT\u2019s capacity for retrograde transport from peripheral sites to the spinal cord dorsal horn and its action as a SNARE-cleaving endopeptidase that inhibits neurotransmitter release, there is no evidence that BoNT-A facilitates the specific \"clearing\" of toxic EVs. Moreover, the claim that it can reach the brain from a distal toe-nerve ending via motor neurons to therapeutically manipulate EV clearance is speculative and lacks empirical validation in the provided source materials.\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThis analysis examines the bidirectional potential of botulinum neurotoxin-A (BoNT-A) for modifying synaptic signaling and its hypothetical intersection with lymphatic-mediated extracellular vesicle (EV) disposal. The claim is evaluated against the mechanism of SNARE-dependent exocytosis and retrograde axonal transport dynamics documented across neurodegenerative models.\n\n### [INTRODUCTION & JUSTIFICATION]\nBotulinum neurotoxin type A (BoNT-A) functions primarily as a zinc-dependent endopeptidase that cleaves synaptic SNARE proteins, such as SNAP-25, thereby preventing the fusion of synaptic vesicles with the plasma membrane. The literature establishes that BoNT-A \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" This mechanism is clinically employed for neuromuscular conditions where \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\" \n\nRegarding the propagation of pathological proteins, \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\" In models of Alzheimer's disease, \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\" While BoNT-A can inhibit the synaptic release of pathological proteins, the provided literature does not support the hypothesis that BoNT-A injected in distal limbs can reach the brain to \"clear\" EVs. Rather, the clearance of brain-derived toxins is primarily attributed to specialized drainage structures, noting that \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\" Thus, while BoNT-A affects vesicular docking at synapses, it is not currently identified as a mechanism to facilitate EV egress via the lymphatic system.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT-A is not strictly limited to the neuromuscular junction; \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n* Exosomes/EVs represent a novel trafficking route: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n* Synaptic plasticity is regulated by SNAREs such as Stx4: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n* Retrograde transport is a common pathway for both beneficial and pathological cargo: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n* Bioenergetic failure in peripheral nerves leads to retrograde signaling disruption: \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n* Non-traditional secretion pathways exist: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n* Swallowing disorders are an example of successful BoNT-A clinical targeting: \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n* Mitochondrial transfer exists as a natural restorative mechanism: \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n* Spatial limits of current interventions: \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n* Essential role of Munc18-1: \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 42102607 - Application: Demonstrates the retrograde capacity of the toxin. - \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"\n2. ID: 37037273 - Application: Demonstrates BoNT-A inhibition of tau release via SNAP25. - \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\"\n3. ID: 37394036 - Application: Outlines the transport mechanisms for pathogenic proteins. - \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\"\n4. ID: 41540479 - Application: Explains the meningeal lymphatic system as a clearance pathway. - \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\"\n5. ID: 38110531 - Application: Discusses EV movement between circulation and the lymphatic system. - \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n6. ID: 42458952 - Application: Discusses SNARE-mediated synaptic plasticity. - \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n7. ID: 42504872 - Application: Details unconventional protein secretion pathways. - \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n8. ID: 42543397 - Application: Discusses barriers to nasal delivery. - \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\"\n9. ID: 42275483 - Application: Describes retrograde transport of EVs. - \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n10. ID: 42422258 - Application: Discusses peripheral nerve stimulation in swallowing. - \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\"\n11. ID: 42422258 - Application: Mentions BoNT-A in cricopharyngeal muscles. - \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n12. ID: 41989572 - Application: Discusses the general medical mechanism of BoNT. - \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\"\n13. ID: 42506696 - Application: Discusses the central effects of BoNT-A. - \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n14. ID: 42342866 - Application: Explains axonal metabolic deficit. - \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n15. ID: 42503395 - Application: Notes EV uptake mechanisms. - \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\"\n16. ID: 42466399 - Application: Confirm astrocyte-to-neuron mitochondrial transfer. - \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n17. ID: 42466399 - Application: Discusses spatial limits of neuroprotection. - \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n18. ID: 42350670 - Application: Discusses metabolic state-dependent paths. - \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\"\n19. ID: 42401592 - Application: Discusses the necessity of SNARE regulators. - \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n20. ID: 42427545 - Application: Dystrophic changes and membrane imbalances. - \"Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.\"\n\n### [PROGRAMATICALLY MAPPED REFERENCES]\n[1]. ID: 42102607 - APA: Lan YA, Guo JX, Yao MH, Zhang XY, Liao ZR et al. (2026). BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.. International immunopharmacology. ID: 42102607.\n[2]. ID: 37037273 - APA: Panzi C, Surana S, De La-Rocque S, Moretto E, Lazo OM et al. (2023). Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.. Toxicon : official journal of the International Society on Toxinology. ID: 37037273.\n[3]. ID: 37394036 - APA: Arnold FJ, Nguyen AD, Bedlack RS, Bennett CL, La Spada AR (2023). Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.. Neurobiology of disease. ID: 37394036.\n[4]. ID: 41540479 - APA: Mee-Inta O, Chiang YY, Tsai SF, Yang TM, Zhao ZW et al. (2026). Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.. Acta neuropathologica communications. ID: 41540479.\n[5]. ID: 38110531 - APA: Iannotta D, A A, Kijas AW, Rowan AE, Wolfram J (2024). Entry and exit of extracellular vesicles to and from the blood circulation.. Nature nanotechnology. ID: 38110531.\n[6]. ID: 42458952 - APA: Li Y, Wei Y, Zhao J, Quan P, Wang C et al. (2026). Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.. CNS & neurological disorders drug targets. ID: 42458952.\n[7]. ID: 42504872 - APA: Krauklis SA, Ke\u00e7eli SK, Campbell EM, Abbineni PS (2026). Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.. Journal of neurochemistry. ID: 42504872.\n[8]. ID: 42543397 - APA: Shen H, Srivastava SK, Aggarwal N, Chang MW (2026). Autonomous intranasal delivery systems for central nervous system therapeutics.. Experimental & molecular medicine. ID: 42543397.\n[9]. ID: 42275483 - APA: Ha JY, Kim SM, Choi SY, Park C, Park S et al. (2026). Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.. Journal of extracellular vesicles. ID: 42275483.\n[10]. ID: 42422258 - APA: Zhou M, Dong B, Zhou H, Zhang W, Fu A et al. (2026). Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.. Frontiers in neuroscience. ID: 42422258.\n[11]. ID: 41989572 - APA: Adnan M, Waqar M, Ahmed R, Afzal H, Mueen MA et al. (2026). Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.. Archives of microbiology. ID: 41989572.\n[12]. ID: 42506696 - APA: Tereshko Y, De Monte D, Ercole BH, Dalla Torre C, Belgrado E et al. (2026). Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.. Toxins. ID: 42506696.\n[13]. ID: 42342866 - APA: Tao W, Yu Y, Wu X, He J, Liu X et al. (2026). Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.. Journal of neurology. ID: 42342866.\n[14]. ID: 42503395 - APA: Jin Z, Jiang S, Yuan L, Ma S, Zhang F et al. (2026). Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.. Journal of advanced research. ID: 42503395.\n[15]. ID: 42466399 - APA: Velmurugan GV, Vekaria HJ, Rabchevsky AG, Saito K, Morganti JM et al. (2026). Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.. Research square. ID: 42466399.\n[16]. ID: 42350670 - APA: Saponaro C, Imbernon M, Louvet I, Deligia E, Chen S et al. (2026). Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.. Diabetologia. ID: 42350670.\n[17]. ID: 42401592 - APA: Huang M, Chow CH, Noworolski R, Charish J, Indrawinata K et al. (2026). Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.. Scientific reports. ID: 42401592.\n[18]. ID: 42427545 - APA: Wu Y, Xu P, Moran J, Xu CS, Hayworth KJ et al. (2026). Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.. bioRxiv : the preprint server for biology. ID: 42427545.\n\n\nEven though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\n\"Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.\"\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThe provided literature confirms that BoNT/A undergoes retrograde transport from peripheral nerve terminals to the soma, where it interacts with intracellular trafficking machinery and influences central nervous system components. While evidence verifies that BoNT/A utilizes retrograde transport and that it can be sequestered in LC3-positive autophagosomes, the specific hypothesis regarding a \"bottleneck\" shift from synaptic exocytosis to somatic degradation remains a theoretical model requiring further quantitative validation. The literature establishes that BoNT/A cleavage of SNAP-25 occurs at the terminal and subsequently within the soma, but direct kinetic quantification of this shift in clearance pathways requires gap-filling studies.\n\n### [INTRODUCTION & JUSTIFICATION]\nThe intracellular itinerary of Botulinum Neurotoxin Type A (BoNT/A) is far more expansive than its canonical role in blocking neurotransmitter release at the neuromuscular junction. Evidence demonstrates that the toxin enters neurons via synaptic vesicles and is subsequently sorted into retrograde axonal transport carriers. ID: 25878289 notes that \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\" Once internalized, the toxin engages with autophagosomal pathways, as \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\" This interaction with the endolysosomal system is critical, particularly as \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\" The ability of BoNT/A to alter the neurobiological landscape is confirmed by findings that \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\" Thus, while the \"bottleneck\" effect remains a specific hypothesis, the literature supports a shift in proteostatic burden and cellular signaling following retrograde axonal transport.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT/A-induced inhibition of SNAP-25 does not merely halt release; it causes secondary synaptic stripping in motoneurons, a phenomenon characterized by a 15-fold increase in spine density and significant synaptic bouton detachment.\n* Retrograde trafficking of BoNT/A is not limited to synaptic vesicles but utilizes autophagosomes (LC3-positive carriers), which serve as a highway for delivering the toxin from the terminal to the soma.\n* The toxin's effect extends beyond the presynaptic membrane; it influences the proliferation and secretory states of associated glia, such as Schwann cells, highlighting a multicellular impact.\n* BoNT/A displays differential intracellular migration; while protease activity is often concentrated locally at the site of application, a measurable portion accumulates in the soma over several weeks.\n* The retrograde pathway of BoNT/A can be hijacked for therapeutic purposes, as evidenced by its application in downregulating pain-related neuropeptides like CGRP and ion channels like TRPV1 in the spinal dorsal horn.\n* Neurotoxic activity is not synonymous with death; motoneurons can undergo significant structural remodeling and transient hypertrophy following BoNT/A exposure without immediate organelle degeneration.\n* Evidence suggests that the fast axonal retrograde transport compartment is multifunctional, acting as a gateway for multiple pathogenic virulence factors, not just BoNTs.\n* There is a distinct discrepancy between the timing of peripheral paralysis and the appearance of central neurotoxic effects, suggesting a staggered temporal profile for the toxin's dual mechanism of action.\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 25878289 - Application: Investigating BoNT/A retrograde trafficking mechanisms. Alignment: 5. Quote: *\"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\"*\n2. ID: 25878289 - Application: Assessing autophagosomal involvement. Alignment: 5. Quote: *\"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\"*\n3. ID: 23300443 - Application: Confirming retrograde transport in motor neurons. Alignment: 5. Quote: *\"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\"*\n4. ID: 23110146 - Application: Demonstrating cl-SNAP-25 presence in CNS. Alignment: 5. Quote: *\"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\"*\n5. ID: 21929507 - Application: Tracking toxin movement in neurites. Alignment: 5. Quote: *\"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\"*\n6. ID: 42102607 - Application: Elucidating CNS involvement. Alignment: 5. Quote: *\"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"*\n7. ID: 39196607 - Application: Identifying ER-related translocation. Alignment: 5. Quote: *\"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\"*\n8. ID: 39196607 - Application: Tracking Golgi-ER trafficking. Alignment: 5. Quote: *\"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\"*\n9. ID: 42102607 - Application: Antipruritic mechanism explanation. Alignment: 5. Quote: *\"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\"*\n10. ID: 21929507 - Application: Temporal somatic accumulation. Alignment: 5. Quote: *\"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\"*\n11. ID: 25878289 - Application: Activity-dependent retrograde transport. Alignment: 5. Quote: *\"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\"*\n12. ID: 9300434 - Application: Structural alteration of synapses. Alignment: 5. Quote: *\"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\"*\n13. ID: 25878289 - Application: Defining CNS activity. Alignment: 5. Quote: *\"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\"*\n14. ID: 39196607 - Application: Identification of genome-wide screening. Alignment: 4. Quote: *\"To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.\"*\n15. ID: 23110146 - Application: Modulation of Schwann cells. Alignment: 5. Quote: *\"We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.\"*\n16. ID: 23300443 - Application: Comparing TeNT and BoNTs. Alignment: 4. Quote: *\"The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.\"*\n17. ID: 9300434 - Application: Detachment of synaptic boutons. Alignment: 5. Quote: *\"Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.\"*\n18. ID: 42102607 - Application: SNAP-25 in mast cells. Alignment: 5. Quote: *\"Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.\"*\n19. ID: 9300434 - Application: Long-term synaptic changes. Alignment: 5. Quote: *\"Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).\"*\n20. ID: 39196607 - Application: Neuronal somatic processing. Alignment: 5. Quote: *\"Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.\"*\n\n### [PROGRAMATICALLY MAPPED REFERENCES]\n[1]. ID: 42102607 - APA: Lan YA, Guo JX, Yao MH, Zhang XY, Liao ZR et al. (2026). BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.. International immunopharmacology. ID: 42102607.\n[19]. ID: 25878289 - APA: Wang T, Martin S, Papadopulos A, Harper CB, Mavlyutov TA et al. (2015). Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.. The Journal of neuroscience : the official journal of the Society for Neuroscience. ID: 25878289.\n[20]. ID: 23300443 - APA: Restani L, Giribaldi F, Manich M, Bercsenyi K, Menendez G et al. (2012). Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.. PLoS pathogens. ID: 23300443.\n[21]. ID: 23110146 - APA: Marinelli S, Vacca V, Ricordy R, Uggenti C, Tata AM et al. (2012). The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.. PloS one. ID: 23110146.\n[22]. ID: 21929507 - APA: Lawrence GW, Ovsepian SV, Wang J, Aoki KR, Dolly JO (2012). Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.. The Biochemical journal. ID: 21929507.\n[23]. ID: 39196607 - APA: Yeo JC, Tay FP, Bennion R, Loss O, Maignel J et al. (2024). Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.. eLife. ID: 39196607.\n[24]. ID: 9300434 - APA: Pastor AM, Moreno-L\u00f3pez B, De La Cruz RR, Delgado-Garc\u00eda JM (1997). Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.. Neuroscience. ID: 9300434.\n\n\n--- VALIDATED QUOTES ---\ndemonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\nImportantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\nEVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\nThe meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\nthe lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\nStx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\nMultiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\nvesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\nits therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\ndemonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\nImportantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\nMultiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\nThe meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\nthe lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\nStx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\nvesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\nits therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\nEVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\nPeripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\nOther adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\nThis property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\nThese findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\nWhen axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\nSPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\nIn vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\nThis process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\nLiraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\nIn the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\ndemonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\nImportantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\nMultiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\nThe meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\nthe lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\nStx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\nvesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\nits therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\nEVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\nPeripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\nOther adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\nThis property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\nThese findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\nWhen axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\nSPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\nIn vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\nThis process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\nLiraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\nIn the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\nGiven the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.\nBecause BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\nSurprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\nWe show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\nImmunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\nThe BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\nFurthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\nThe toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\nAn organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\nTherefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\nA majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\nConsistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\nSynaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\nHowever, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\nBecause BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\nSurprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\nWe show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\nImmunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\nThe BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\nFurthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\nThe toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\nAn organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\nTherefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\nA majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\nConsistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\nSynaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\nHowever, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\nTo investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.\nWe found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.\nThe striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.\nSynaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.\nUsing P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.\nBetween 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).\nSurprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.\n\n\n=============================\nUser Request: ANSWER IN THIS LANGUAGE --->>> Answer in English only. Begin with a clear Yes or No. Is the synthesis 100% veridical with the validated quotes? Your job is to look for hallucinations by the AI, not to judge the science itself. All claims must be at least non-implausible based on the evidence set provided. Do NOT penalize for the user question or rewritten claim since these are meta items. Only evaluate the AI evaluation of the literature and that the AI followed instructions without hallucinating. List and justify your judgements. Do not use markdown. DO NOT PENALIZE FOR THE USER QUERY WORDING OR REWRITE>>> THAT IS NOT PART OF THE ANSWER ... THAT IS THE QUESTION OR CLAIM EVALUATED. <<<--- ANSWER THE USER REQUEST IN THEIR OWN LANGUAGE. THE DATASETS CAN BE GENERATED IN ANY LANGUAGE AND MULTIPLE CHAT THREADS MAY EXIST, BUT YOU MUST ANSWER THE USER IN THE LANGUAGE THEY ASKED THE CURRENT QUERY: {query}"
}
],
"quadrants": [
{
"name": "Run1_Eval1_synthesis",
"text": "Hypothesis: If Botox were injected into the nerve ending on the third toe on the left foot, the toxin can \"climb\" retrograde from toe through motor neurons eventually to the brain and temporarily inhibit vesicular docking \"from toe to head\", potentially allowing a timed pharmacological therapeutic window for toxic EV clearance through the lymph system instead of docking and seeding.",
"metrics": {
"Alignment": 3,
"Consilience": 5,
"Confidence": 2,
"Logic_Chain": [
{
"Step": 1,
"From": "Botulinum Toxins, Type A",
"Relationship": "-->",
"To": "Axonal Transport",
"evidence_source_id": "42102607",
"Alignment_Score": 6,
"Consilience_Score": 6,
"Confidence_Score": 5,
"Gap_Strength": "None",
"Justification": "Evidence supports retrograde transport of BoNT to dorsal horn.",
"Color": "lightgreen"
},
{
"Step": 2,
"From": "Axonal Transport",
"Relationship": "-->",
"To": "SNARE Proteins",
"evidence_source_id": "37037273",
"Alignment_Score": 5,
"Consilience_Score": 5,
"Confidence_Score": 4,
"Gap_Strength": "medium",
"Justification": "BoNT inhibits docking via SNARE cleavage, but brain-wide therapeutic precision is speculative.",
"Color": "lightblue"
},
{
"Step": 3,
"From": "SNARE Proteins",
"Relationship": "-->",
"To": "Lymphatic Vessel Endothelial Hyaluronan Receptor-1",
"evidence_source_id": "41540479",
"Alignment_Score": 2,
"Consilience_Score": 1,
"Confidence_Score": 1,
"Gap_Strength": "strong",
"Justification": "No evidence links BoNT synaptic inhibition to increased lymphatic clearance of EVs.",
"Color": "pink"
}
],
"Verbatim_Quotes": [
{
"quote": "demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"source_id": "42102607"
},
{
"quote": "Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.",
"source_id": "37037273"
},
{
"quote": "Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.",
"source_id": "37394036"
},
{
"quote": "The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.",
"source_id": "41540479"
},
{
"quote": "the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"source_id": "38110531"
},
{
"quote": "Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).",
"source_id": "42458952"
},
{
"quote": "vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.",
"source_id": "42504872"
},
{
"quote": "its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.",
"source_id": "42543397"
},
{
"quote": "EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.",
"source_id": "42275483"
},
{
"quote": "Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.",
"source_id": "42422258"
},
{
"quote": "Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.",
"source_id": "42422258"
},
{
"quote": "This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.",
"source_id": "41989572"
},
{
"quote": "These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.",
"source_id": "42506696"
},
{
"quote": "When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.",
"source_id": "42342866"
},
{
"quote": "SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.",
"source_id": "42503395"
},
{
"quote": "In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).",
"source_id": "42466399"
},
{
"quote": "This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.",
"source_id": "42466399"
},
{
"quote": "Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.",
"source_id": "42350670"
},
{
"quote": "In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.",
"source_id": "42401592"
},
{
"quote": "Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.",
"source_id": "42427545"
}
],
"Study_Type_Audit": {
"37037273": "in_vitro:Count=1",
"37394036": "review:Count=1",
"41540479": "in_vivo:Count=1",
"42102607": "in_vivo:Count=1"
},
"Gap_Analysis_Audit": {
"study_type": "in_vitro/in_vivo models",
"study_intent": "neurotoxin mechanisms",
"justification": "Evidence is missing for the specific hypothesis regarding distal toe-to-brain BoNT transport resulting in therapeutic EV clearance.",
"predicted_result": "BoNT will block local neurotransmission but cannot be relied upon for brain-wide EV clearance.",
"short_answer_to_user": "No current evidence supports BoNT as a systemic EV clearance tool via the proposed toe-to-brain retrograde pathway."
},
"suggested_experiments": [
"Assess if BoNT-A injection at sciatic nerve termini influences the rate of fluorescently tagged EV accumulation in the deep cervical lymph nodes.",
"Test whether selective SNARE inhibition via BoNT-A in the spinal dorsal horn affects the efflux of brain-derived EVs in a mouse model."
],
"suggested_studies": [
"A systematic analysis of retrograde axonal transport rates of BoNT-A derivatives in the context of neurodegenerative protein spread.",
"Investigation into the impact of BoNT-A-induced SNAP25 cleavage on the secretion profile of EVs in spinal interneurons."
],
"swansons_literature_based_discovery_candidates": {
"Discovered Hypothesis (A to C)": "BoNT-A mediated inhibition of synaptic exocytosis can reduce the pool of neuronally-derived EVs available for trans-synaptic propagation, thereby limiting disease seeding.",
"Literature A (Origin)": "BoNT-A synaptic silencing (Source 37037273)",
"Literature C (Target)": "Intercellular propagation of pathogenic proteins/EVs in ALS (Source 37394036)",
"The Intersecting Bridge B": "SNARE-dependent exocytosis (SNAP25)",
"Biological Rationale": "Since SNARE complex integrity is required for both neurotransmitter release and the unconventional secretion of EVs, BoNT-A induced cleavage of SNAP25 effectively halts the exocytic machinery used by pathogenic EVs to spread along connected neural circuits."
},
"contradictions_between_evidences": "None identified regarding the primary mechanisms, though the therapeutic outcome of BoNT in central nervous system processes remains a point of clinical study.",
"repurposed_solutions": "Repurpose BoNT-A as a tool for spatiotemporally controlled cessation of pathological EV secretion, rather than just neuromuscular paralysis.",
"QuoteValidation": [
{
"quote": "demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"source_id": "42102607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quote": "Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.",
"source_id": "37037273",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies."
},
{
"quote": "Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.",
"source_id": "37394036",
"status": "PASS",
"error": "",
"abstract_text": "ID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined."
},
{
"quote": "The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.",
"source_id": "41540479",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD."
},
{
"quote": "the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"source_id": "38110531",
"status": "PASS",
"error": "",
"abstract_text": "ID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system."
},
{
"quote": "Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).",
"source_id": "42458952",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential."
},
{
"quote": "vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.",
"source_id": "42504872",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS."
},
{
"quote": "its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.",
"source_id": "42543397",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development."
},
{
"quote": "EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.",
"source_id": "42275483",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions."
},
{
"quote": "Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.",
"source_id": "42422258",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quote": "Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.",
"source_id": "42422258",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes."
},
{
"quote": "This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.",
"source_id": "41989572",
"status": "PASS",
"error": "",
"abstract_text": "ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance."
},
{
"quote": "These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.",
"source_id": "42506696",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42506696\nTitle: Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.\nAbstract: Essential blepharospasm (BEB) is a focal dystonia characterized by abnormal brainstem excitability and impaired inhibitory control within trigeminal-facial circuits. Botulinum toxin type A (BoNT-A) is the established first-line treatment, primarily acting at the neuromuscular junction. However, whether BoNT-A also modulates central brainstem circuits in BEB patients remains unclear, and dedicated neurophysiological studies have yielded conflicting results. To investigate whether BoNT-A modulates brainstem interneuronal excitability in BEB using the blink reflex recovery cycle and to correlate clinical outcomes with neurophysiological results. Thirteen patients with BEB underwent neurophysiological and clinical evaluation before (T0) and one month after (T1) BoNT-A treatment. The blink reflex recovery cycle was assessed at interstimulus intervals (ISIs) of 200, 300, 500, and 1000 ms. Clinical severity was assessed using the BSPSS, JRS, and BDS scales. The R2 amplitude ratio showed a statistically significant decrease after treatment across all ISIs (all p \u2264 0.001), indicating reduced brainstem interneuronal excitability. All clinical scales demonstrated statistically significant improvement after treatment (BSPSS, JRS, BDS; all p < 0.001). This pilot study provides preliminary evidence that BoNT-A treatment may reduce brainstem interneuronal excitability in BEB patients, as evidenced by a substantial and consistent decrease in R2 amplitude ratios of the blink reflex recovery cycle. These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action. Larger controlled studies are warranted to confirm these results."
},
{
"quote": "When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.",
"source_id": "42342866",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42342866\nTitle: Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.\nAbstract: Diabetic peripheral neuropathy (DPN), particularly distal symmetric polyneuropathy, characterized by length\u2011dependent axonal damage, is a common chronic complication of type 2 diabetes mellitus. The pathogenesis of DPN is complicated, yet one thing is clear: long axons require a great deal of energy. When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration. While earlier work centered on hyperglycemia-induced cytotoxicity, recent studies have increasingly implicated dysregulation of glucose, lipid, and amino acid metabolism as key contributors to DPN. In this review, we integrate the anatomical organization of peripheral nerves, bioenergetic pathways, and axon-Schwann cell interactions to establish a framework for understanding how glucose, lipid, and amino acid dysregulation converge to induce bioenergetic failure in DPN. Based on these mechanisms, we further discuss novel strategies aimed at restoring metabolic homeostasis in neurons and Schwann cells. Importantly, correcting a single metabolic pathway is unlikely to halt or reverse DPN. Instead, restoring global energy homeostasis to rebalance axonal energy supply and demand may be essential for preserving peripheral nerve function."
},
{
"quote": "SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.",
"source_id": "42503395",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42503395\nTitle: Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.\nAbstract: Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma."
},
{
"quote": "In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).",
"source_id": "42466399",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quote": "This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.",
"source_id": "42466399",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations."
},
{
"quote": "Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.",
"source_id": "42350670",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42350670\nTitle: Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.\nAbstract: Liraglutide, a glucagon-like peptide-1 receptor (GLP-1R) agonist for type 2 diabetes and obesity management, shows variable patient responses. We investigated the metabolic state-dependent mechanisms underlying this heterogeneity and how liraglutide's mode of action shifts across stages of metabolic dysfunction. We employed human pancreatic islets from donors across metabolic states (normoglycaemic [HbA1c <42 mmol/l (<6.0%)], glucose intolerance [HbA1c 42-47 mmol/l (6.0-6.4%)] and type 2 diabetes [HbA1c \u226548 mmol/l (\u22656.5%)]) using dynamic perifusion and static incubation techniques to assess glucose-stimulated insulin secretion. GLP-1R mRNA levels were measured in 112 donor islets stratified by HbA1c. Mechanistic investigations used tanycyte-specific GLP-1R knockdown (GLP-1RTanycyteKD) mice and botulinum toxin B-expressing (iBot) mice to distinguish between central and peripheral pathways. Oral glucose tolerance tests, pyruvate tolerance tests and positron emission tomography were performed to assess in vivo metabolic effects. Liraglutide (25 nmol/l) enhanced glucose-stimulated insulin secretion specifically in donors with glucose intolerance (n=7, p=0.021), with no effect in normoglycaemic islets (n=7), despite preserved GLP-1 (7-36) responsiveness. In type 2 diabetes islets, GLP-1R mRNA levels progressively decreased with rising HbA1c (p=0.015, normoglycaemic [n=48] vs type 2 diabetes [n=10]). In chow-fed mice, liraglutide's insulin-stimulating effects required tanycyte-mediated hypothalamic access, as demonstrated by abolished responses in GLP-1RTanycyteKD mice. However, during metabolic dysfunction (a 12-week high-fat diet), direct islet responsiveness was restored independent of tanycyte function. Advanced metabolic disease (a 27-week high-fat diet) maintained islet responsiveness ex vivo while losing in vivo insulin enhancement, revealing insulin-independent glucose-lowering mechanisms involving hepatic gluconeogenesis suppression and enhanced peripheral glucose uptake. Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states. This mechanistic framework enables potential patient stratification in type 2 diabetes therapy, suggesting that matching liraglutide's predominant mechanism to individual metabolic profiles could optimise treatment outcomes."
},
{
"quote": "In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.",
"source_id": "42401592",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42401592\nTitle: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.\nAbstract: The function of the SNARE complex regulator, Munc18-1, in photoreceptor cells is unknown. Here, we found that removing Munc18-1 from photoreceptors results in major degeneration starting at P14. In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1. Furthermore, Munc18-1 played a critical role in expression and localization of syntaxin-3. The syntaxin-3 protein level is dramatically reduced in the soma and plasma membrane of photoreceptors without Munc18-1. At the photoreceptor synapses, the colocalization of syntaxin-3 and its SNARE partner, SNAP-25, was reduced, potentially suggesting an altered syntaxin-3 synaptic localization. In the Munc18-1-deficient photoreceptors, immature synapses and outer segment lesions were found. Taken together, these findings provide evidence that Munc18-1 is important for maintaining sufficient syntaxin-3 expression in the cell body and synapses of photoreceptors. The lack of Munc18-1, combined with poor syntaxin-3 expression, contributes to photoreceptor functional impairment and degeneration."
},
{
"quote": "Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.",
"source_id": "42427545",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42427545\nTitle: Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.\nAbstract: Synaptojanin 1 is a brain enriched phosphoinositide phosphatase implicated in endocytosis at the synapse. A mutation (R258Q) that selectively impairs its Sac1 phosphatase domain causes early onset familial Parkinsonism. Neurons of mice with this mutation display synaptic vesicle traffic defects across the brain, but selective dystrophic changes in a subset of dopaminergic axons in the dorsolateral striatum. Using correlative light microscopy-FIB-SEM of mutant mouse striata to visualize in 3D these abnormal structures we show that they represent clusters of focal axonal dilations harboring massive, onion-like DAT enriched plasma membrane infoldings, generally localized next to cell bodies of neighboring cells, often engulfing evaginations of such cells. This dysmorphia was associated with a deficit in dopamine release in the same striatal region. Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis. Their occurrence only in a subset of axons suggest a vulnerability threshold of these axons beyond which the expansion of the plasma membrane is not counteracted by compensatory mechanisms."
}
]
},
"displayText": "Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\nThe claim that Botulinum toxin (BoNT) injected into a peripheral nerve ending (e.g., third toe) can undergo retrograde transport to the central nervous system (CNS) to inhibit vesicular docking and facilitate a therapeutic window for toxic extracellular vesicle (EV) clearance is scientifically provocative but currently unsupported by the provided literature. While literature confirms BoNT\u2019s capacity for retrograde transport from peripheral sites to the spinal cord dorsal horn and its action as a SNARE-cleaving endopeptidase that inhibits neurotransmitter release, there is no evidence that BoNT-A facilitates the specific \"clearing\" of toxic EVs. Moreover, the claim that it can reach the brain from a distal toe-nerve ending via motor neurons to therapeutically manipulate EV clearance is speculative and lacks empirical validation in the provided source materials.\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThis analysis examines the bidirectional potential of botulinum neurotoxin-A (BoNT-A) for modifying synaptic signaling and its hypothetical intersection with lymphatic-mediated extracellular vesicle (EV) disposal. The claim is evaluated against the mechanism of SNARE-dependent exocytosis and retrograde axonal transport dynamics documented across neurodegenerative models.\n\n### [INTRODUCTION & JUSTIFICATION]\nBotulinum neurotoxin type A (BoNT-A) functions primarily as a zinc-dependent endopeptidase that cleaves synaptic SNARE proteins, such as SNAP-25, thereby preventing the fusion of synaptic vesicles with the plasma membrane. The literature establishes that BoNT-A \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" This mechanism is clinically employed for neuromuscular conditions where \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\" \n\nRegarding the propagation of pathological proteins, \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\" In models of Alzheimer's disease, \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\" While BoNT-A can inhibit the synaptic release of pathological proteins, the provided literature does not support the hypothesis that BoNT-A injected in distal limbs can reach the brain to \"clear\" EVs. Rather, the clearance of brain-derived toxins is primarily attributed to specialized drainage structures, noting that \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\" Thus, while BoNT-A affects vesicular docking at synapses, it is not currently identified as a mechanism to facilitate EV egress via the lymphatic system.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT-A is not strictly limited to the neuromuscular junction; \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n* Exosomes/EVs represent a novel trafficking route: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n* Synaptic plasticity is regulated by SNAREs such as Stx4: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n* Retrograde transport is a common pathway for both beneficial and pathological cargo: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n* Bioenergetic failure in peripheral nerves leads to retrograde signaling disruption: \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n* Non-traditional secretion pathways exist: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n* Swallowing disorders are an example of successful BoNT-A clinical targeting: \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n* Mitochondrial transfer exists as a natural restorative mechanism: \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n* Spatial limits of current interventions: \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n* Essential role of Munc18-1: \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 42102607 - Application: Demonstrates the retrograde capacity of the toxin. - \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"\n2. ID: 37037273 - Application: Demonstrates BoNT-A inhibition of tau release via SNAP25. - \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\"\n3. ID: 37394036 - Application: Outlines the transport mechanisms for pathogenic proteins. - \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\"\n4. ID: 41540479 - Application: Explains the meningeal lymphatic system as a clearance pathway. - \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\"\n5. ID: 38110531 - Application: Discusses EV movement between circulation and the lymphatic system. - \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n6. ID: 42458952 - Application: Discusses SNARE-mediated synaptic plasticity. - \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n7. ID: 42504872 - Application: Details unconventional protein secretion pathways. - \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n8. ID: 42543397 - Application: Discusses barriers to nasal delivery. - \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\"\n9. ID: 42275483 - Application: Describes retrograde transport of EVs. - \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n10. ID: 42422258 - Application: Discusses peripheral nerve stimulation in swallowing. - \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\"\n11. ID: 42422258 - Application: Mentions BoNT-A in cricopharyngeal muscles. - \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n12. ID: 41989572 - Application: Discusses the general medical mechanism of BoNT. - \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\"\n13. ID: 42506696 - Application: Discusses the central effects of BoNT-A. - \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n14. ID: 42342866 - Application: Explains axonal metabolic deficit. - \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n15. ID: 42503395 - Application: Notes EV uptake mechanisms. - \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\"\n16. ID: 42466399 - Application: Confirm astrocyte-to-neuron mitochondrial transfer. - \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n17. ID: 42466399 - Application: Discusses spatial limits of neuroprotection. - \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n18. ID: 42350670 - Application: Discusses metabolic state-dependent paths. - \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\"\n19. ID: 42401592 - Application: Discusses the necessity of SNARE regulators. - \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n20. ID: 42427545 - Application: Dystrophic changes and membrane imbalances. - \"Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.\"\n\n### [PROGRAMATICALLY MAPPED REFERENCES]\n[1]. ID: 42102607 - APA: Lan YA, Guo JX, Yao MH, Zhang XY, Liao ZR et al. (2026). BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.. International immunopharmacology. ID: 42102607.\n[2]. ID: 37037273 - APA: Panzi C, Surana S, De La-Rocque S, Moretto E, Lazo OM et al. (2023). Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.. Toxicon : official journal of the International Society on Toxinology. ID: 37037273.\n[3]. ID: 37394036 - APA: Arnold FJ, Nguyen AD, Bedlack RS, Bennett CL, La Spada AR (2023). Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.. Neurobiology of disease. ID: 37394036.\n[4]. ID: 41540479 - APA: Mee-Inta O, Chiang YY, Tsai SF, Yang TM, Zhao ZW et al. (2026). Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.. Acta neuropathologica communications. ID: 41540479.\n[5]. ID: 38110531 - APA: Iannotta D, A A, Kijas AW, Rowan AE, Wolfram J (2024). Entry and exit of extracellular vesicles to and from the blood circulation.. Nature nanotechnology. ID: 38110531.\n[6]. ID: 42458952 - APA: Li Y, Wei Y, Zhao J, Quan P, Wang C et al. (2026). Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.. CNS & neurological disorders drug targets. ID: 42458952.\n[7]. ID: 42504872 - APA: Krauklis SA, Ke\u00e7eli SK, Campbell EM, Abbineni PS (2026). Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.. Journal of neurochemistry. ID: 42504872.\n[8]. ID: 42543397 - APA: Shen H, Srivastava SK, Aggarwal N, Chang MW (2026). Autonomous intranasal delivery systems for central nervous system therapeutics.. Experimental & molecular medicine. ID: 42543397.\n[9]. ID: 42275483 - APA: Ha JY, Kim SM, Choi SY, Park C, Park S et al. (2026). Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.. Journal of extracellular vesicles. ID: 42275483.\n[10]. ID: 42422258 - APA: Zhou M, Dong B, Zhou H, Zhang W, Fu A et al. (2026). Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.. Frontiers in neuroscience. ID: 42422258.\n[11]. ID: 41989572 - APA: Adnan M, Waqar M, Ahmed R, Afzal H, Mueen MA et al. (2026). Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.. Archives of microbiology. ID: 41989572.\n[12]. ID: 42506696 - APA: Tereshko Y, De Monte D, Ercole BH, Dalla Torre C, Belgrado E et al. (2026). Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.. Toxins. ID: 42506696.\n[13]. ID: 42342866 - APA: Tao W, Yu Y, Wu X, He J, Liu X et al. (2026). Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.. Journal of neurology. ID: 42342866.\n[14]. ID: 42503395 - APA: Jin Z, Jiang S, Yuan L, Ma S, Zhang F et al. (2026). Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.. Journal of advanced research. ID: 42503395.\n[15]. ID: 42466399 - APA: Velmurugan GV, Vekaria HJ, Rabchevsky AG, Saito K, Morganti JM et al. (2026). Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.. Research square. ID: 42466399.\n[16]. ID: 42350670 - APA: Saponaro C, Imbernon M, Louvet I, Deligia E, Chen S et al. (2026). Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.. Diabetologia. ID: 42350670.\n[17]. ID: 42401592 - APA: Huang M, Chow CH, Noworolski R, Charish J, Indrawinata K et al. (2026). Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.. Scientific reports. ID: 42401592.\n[18]. ID: 42427545 - APA: Wu Y, Xu P, Moran J, Xu CS, Hayworth KJ et al. (2026). Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.. bioRxiv : the preprint server for biology. ID: 42427545.\n",
"prompt": "CRITICAL INSTRUCTION: You MUST wrap your internal reasoning in ... tags at the very beginning of your response.\n\n=======================================================\nCONTEXT LITERATURE (STATIC CACHE):\nID: 42527713\nTitle: LncRNA DLX6-AS1 Regulates the Pathological Process Related to Alzheimer's Disease via miR-204-5p.\nAbstract: Alzheimer's disease (AD) is a neurodegenerative disorder causing progressive neuronal damage. Incidence rises with age, and early diagnosis is difficult. This study examined DLX6-AS1 clinical relevance and regulatory mechanism in AD, and its interaction with miR-204-5p in AD pathology. It offers new insights into early diagnosis and treatment. A total of 133 AD patients and 105 healthy controls were selected. Their serum levels of DLX6-AS1 and miR-204-5p were analyzed using quantitative polymerase chain reaction. \u03b2-amyloid (A\u03b2)1-42-induced SH-SY5Y neuronal injury and okadaic acid (OA)-induced Neuro-2a tau abnormal phosphorylation models were constructed. The role/function of DLX6-AS1/miR-204-5p axis was then investigated using cell counting kit-8, flow cytometry, western blotting, enzyme-linked immunosorbent assay and a dual-luciferase reporter gene assay. AD patients had higher serum DLX6-AS1 and lower miR-204-5p levels. DLX6-AS1 showed an AUC of 0.838 for AD diagnosis. DLX6-AS1 levels were negatively associated with cognitive function, brain structural integrity, and benign pathology-and positively associated with disease severity, functional impairment, and pathological markers. In AD cell models, DLX6-AS1 was upregulated. Silencing it promoted cell proliferation, reduced apoptosis and oxidative stress, improved mitochondrial and synaptic function, decreased tau phosphorylation, and enhanced microtubule stability and axonal transport. Dual-luciferase assays confirmed direct binding between DLX6-AS1 and miR-204-5p. Co-inhibition reversed the protective effects of DLX6-AS1 silencing. Serum DLX6-AS1 is a potential biomarker for early diagnosis and assessment of AD. It regulates A\u03b2-induced neuronal damage and tau phosphorylation by targeting miR-204-5p, offering a new mechanism target for AD molecular therapy.\n\nID: 42511736\nTitle: Urinary Extracellular Vesicle-Derived miRNAs as Regulators and Biomarkers in Diabetic Kidney Disease.\nAbstract: Diabetic kidney disease (DKD) remains one of the most severe microvascular complications of type 2 diabetes mellitus (T2DM) and a leading cause of chronic kidney disease (CKD) worldwide. Nevertheless, despite considerable progress in elucidating its molecular background, early diagnosis and accurate stratification of disease progression remain challenging when relying on conventional clinical biomarkers such as albuminuria and estimated glomerular filtration rate (eGFR). Growing evidence indicates that DKD is driven by interconnected pathogenic mechanisms, including chronic hyperglycemia, activation of the protein kinase C (PKC) signaling pathway, renin-angiotensin-aldosterone system (RAAS) dysregulation, oxidative stress, inflammatory cascades, and immune system activation involving Toll-like receptors (TLR) and the NLRP3 inflammasome. These processes collectively contribute to endothelial dysfunction, podocyte injury, extracellular matrix accumulation, and progressive renal fibrosis. Exosomes and their molecular cargo, particularly miRNAs, have emerged as promising regulators and non-invasive biomarkers reflecting ongoing renal injury. Urinary exosomal microRNAs (uEV-miRNAs) are of interest due to their stability in biological fluids and their direct origin from nephron segments, enabling real-time reflection of renal pathophysiology. Accumulating studies suggest that differentially expressed microRNAs (miRNAs), including miR-21-5p, miR-30a-5p, miR-192-5p, and miR-142-3p, are closely associated with key pathways in DN. However, their clinical translation remains limited by methodological heterogeneity, the lack of standardized isolation protocols, and insufficient validation in large longitudinal cohorts. This review navigates the current landscape of knowledge on the molecular mechanisms underlying DKD and examines the emerging role of uEV-miRNAs as diagnostic biomarkers. Altogether, uEV-miRNAs offer a promising avenue for improving early detection, risk stratification, and disease monitoring in DKD.\n\nID: 42497239\nTitle: The axonal transport velocity of prions is independent of prion formation.\nAbstract: Prion diseases are caused by PrPSc, the self-templating infectious conformation of the normal host-encoded prion protein, PrPC. Spread of PrPSc in the peripheral and central nervous system follows a highly predictable pattern consistent with slow axonal transport along defined neuroanatomical pathways. The rate of PrPSc transport, the contribution of prion strain diversity, and the requirement for prion conversion during transport are poorly understood, in part, due to a lack of direct measurement of PrPSc transport under defined conditions. Here, using a combination of live animal and sciatic nerve explant culture experiments, and determined the velocity of PrPSc ranged from rates consistent with both slow and fast axonal transport. Interestingly, transport of PrPSc was not dependent upon prion conversion as it was observed using prion strain and host combinations that do not support prion formation. The rate of PrPSc transport of prion strains and synthetic prions suggest that prions can use both slow and fast modes of axonal transport. Overall, converging lines of evidence suggest that prions are transported at a range of velocities consistent with both slow and fast axonal transport that is independent prion formation.\n\nID: 42465429\nTitle: The neuropathy-causing GARS1 \u0394ETAQ mutation drives pathology in subsets of motor and sensory neurons in mice.\nAbstract: Charcot-Marie-Tooth disease type 2D (CMT2D) results from gain-of-function mutations in GARS1, which encodes glycyl-tRNA synthetase (GlyRS), the enzyme responsible for charging transfer RNA (tRNA) with glycine. There are several CMT2D mouse models, but Gars \u0394ETAQ/+ is the only one that bears a patient-sourced mutation. Created using CRISPR/Cas9 to model a 12-nucleotide de novo GARS1 deletion identified in an unusually severe CMT2D patient, Gars \u0394ETAQ/+ mice have previously been shown to display several neuromuscular phenotypes; motor axon loss, denervated neuromuscular junctions (NMJs) and reduced muscle function. Here, we extend these analyses to provide a more comprehensive understanding of both motor and sensory nerve deficits across hind- and fore-limbs. At 3 months, Gars \u0394ETAQ/+ mice possess sex-independent alterations in the levels of neuropathy biomarkers - including decreased NfL and increased periaxin - alongside reduced muscle endurance and strength, and impairments in the sensory modalities of mechanosensation, proprioception and nociception. Underpinning these dysfunctions, we identified site-specific defects comprising altered sensory neuron populations, muscle spindle loss, reduced motor neuron size, disrupted NMJ innervation and maturation, and reduced axonal transport of signalling endosomes in vivo. Together, these experiments show that Gars \u0394ETAQ/+ mice display robust and selective peripheral nerve pathology that manifests in a general distal-to-proximal fashion, priming this CMT2D allele for testing treatments and evaluating mechanisms underlying peripheral nerve vulnerability.\n\nID: 42404433\nTitle: Beyond motor neurons: peripheral TDP-43 pathology in skeletal muscle and intramuscular nerves in amyotrophic lateral sclerosis.\nAbstract: Amyotrophic lateral sclerosis is a progressive neurodegenerative disease characterized by accumulation of the 43-kDa TAR DNA-binding protein (TDP-43). This neuropathological signature has been well documented within the CNS; however, recent findings indicate that the phosphorylated TDP-43 additionally deposits in peripheral tissues, including skeletal muscle and intramuscular nerves. These data warrant a change of view from a neurocentric perspective of amyotrophic lateral sclerosis pathogenesis towards a broader concept of TDP-43 proteinopathy extending both within and beyond the nervous system. In this review, we focus on current evidence supporting the presence of TDP-43 pathology in amyotrophic lateral sclerosis skeletal muscle, examining its topographic distribution, molecular characteristics and associations with intramuscular nerve bundles. We also discuss the susceptibility of intrinsic muscle cells, disrupted axonal transport and impairment in protein quality control. Phosphorylated TDP-43 pathology in muscle biopsies from amyotrophic lateral sclerosis patients has emerged as a promising tool in the early diagnosis of the disease. Moreover, we discuss the relevance of these findings to amyotrophic lateral sclerosis pathogenesis and potential therapeutic implications.\n\nID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions.\n\nID: 42264545\nTitle: Nanotechnology-enabled targeting strategies for neurodegenerative disorders: role of functionalized nanoparticles.\nAbstract: Neurodegenerative disorders comprise a diverse group of progressive neurological diseases characterized by the gradual loss of neuronal structure and function. Conditions such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis arise from multifactorial mechanisms involving genetic susceptibility, environmental factors, and age-related cellular decline. Key pathogenic processes include oxidative stress, mitochondrial dysfunction, protein misfolding and aggregation, impaired axonal transport, Golgi fragmentation, and chronic neuroinflammation, all of which disrupt neuronal homeostasis and synaptic communication, ultimately leading to neuronal death. Hormonal imbalances further exacerbate these effects by promoting oxidative damage, inflammation, and metabolic dysfunction. Despite advances in understanding disease mechanisms, effective drug delivery remains challenging due to the restrictive nature of the blood-brain barrier. Recent developments highlight the potential of nanoparticle-based drug delivery systems to overcome these limitations. Functionalized nanoparticles enhance blood-brain barrier penetration, improve targeting specificity, and enable controlled drug release. These systems can deliver neuroprotective agents, antioxidants, peptides, and gene therapies directly to affected brain regions. Thus, integrating disease pathophysiology with nanotechnology-based strategies offers a promising approach for improving therapeutic outcomes and advancing precision treatment in neurodegenerative disorders.\n\nID: 42209021\nTitle: Striatal Neuron Excitability Is Regulated by Huntingtin in the Adult Brain.\nAbstract: Huntington's disease (HD) is a hereditary neurodegenerative disease that typically presents during midlife and is characterized by a combination of motor, cognitive, and psychiatric symptoms. HD is fatal and arises from a mutation in the huntingtin (HTT) gene, which results in decreased neuronal health followed by brain atrophy, with spiny projection neurons (SPNs) of the striatum being especially vulnerable to degeneration. HTT loss of function, caused by haploinsufficiency of the wild-type HTT gene (wtHTT), is an important feature of HD pathophysiology that has previously been understudied compared with mutant HTT gain-of-function mechanisms. wtHTT is essential for nervous system development and functions as a scaffolding protein to support many vital cellular functions including axonal transport, autophagy, and synaptic plasticity. Here, we examined the consequences of wtHTT deletion in the adult cortex and striatum by conditionally inactivating wtHTT in 2-4-month-old male and female Htt fl/fl mice. wtHTT loss of function decreased intrinsic neuronal excitability within SPNs and produced a neuroinflammatory response in these mice, while tissue organization, spine morphology, and motor behavior remained unaffected. Results presented here provide additional evidence that wtHTT is vital for maintaining neuronal health in the adult brain and highlight some potential adverse consequences of nonselective HTT lowering for the treatment of HD.\n\nID: 42198698\nTitle: From Envelope to Encephalopathy: How HIV-1 gp120 Drives Neurocognitive Decline.\nAbstract: Although neurons are not productively infected by HIV-1, the envelope glycoprotein gp120, detectable in cerebrospinal fluid independently of active viral replication, gains intraneuronal access via lipid raft-mediated endocytosis, macropinocytosis, and retrograde axonal transport, contributing to persistent neurobiological dysfunction within the central nervous system. Once internalized, gp120 is associated with neuronal dysfunction involving convergent pathways, including excitotoxic calcium dysregulation, mitochondrial and metabolic failure, and inflammatory and senescence-associated amplification. These pathways converge on suppression of CREB and BDNF signaling, dismantling the transcriptional and neurotrophic programs required for synaptic maintenance and cognitive resilience. Extracellular vesicle-mediated dissemination and microRNA reprogramming extend gp120-associated neurobiological effects beyond sites of receptor engagement, while gut-derived metabolites, particularly quinolinic acid, lower the excitotoxic threshold through synergistic activation of NMDA receptors. Together, these mechanisms define HAND as a network disorder in which gp120 contributes to persistent neurocognitive dysfunction beyond active viral replication, identifying convergent therapeutic nodes where combination strategies targeting excitotoxicity, mitochondrial dysfunction, and neuroinflammation offer the most promising path toward durable neuroprotection.\n\nID: 42192837\nTitle: Axonal Transport Deficits in Parkinson's Disease: Insights from Neurotoxin, Genetic, and Sporadic Models.\nAbstract: Parkinson's disease (PD) is a prevalent neurodegenerative disorder, characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta and the accumulation of Lewy bodies. Over recent decades, various cellular mechanisms underlying PD have been elucidated, including autophagy, mitochondrial dysfunction, neuroinflammation, and axonal transport. Among them, axonal transport plays a critical role in maintaining the dynamic homeostasis of proteins, membrane-bound organelles, and cellular metabolism within neurons. Unfortunately, a comprehensive overview of axonal transport in PD remains absent. In this review, we synthesized the current literature on axonal transport in PD, leveraging neurotoxic and genetic models to explore the causes and consequences of axonal transport alterations in PD. Through this summary, we aim to deepen our understanding of PD pathogenesis and provide potential therapeutic targets for intervention.\n\nID: 42144072\nTitle: Progress in precision drug delivery systems for neurodegenerative diseases with muscle atrophy.\nAbstract: Neurodegenerative diseases frequently co-occur with skeletal muscle atrophy, creating a complex comorbid condition that significantly accelerates functional decline and increases mortality. This dual pathology is driven by interconnected mechanisms such as protein aggregation, neuroinflammation, and impaired axonal transport, disrupting critical neuromuscular junctions (NMJs). However, a significant research gap exists in the development of therapeutic strategies that can effectively and simultaneously target both the central nervous system (CNS) and peripheral muscle tissues. We systematically summarizes the core pathological targets and critically evaluates recent advances in precision drug delivery systems designed to overcome these challenges. We explore innovative strategies, including engineered viral vectors and receptor-targeted nanoparticles for CNS delivery, as well as smart biomaterials and extracellular vesicles (EVs) for muscle-specific and dual-organ intervention, highlighting the growing role of artificial intelligence (AI) in optimizing their design. Furthermore, this review discusses the construction of multidimensional efficacy evaluation systems that integrate behavioral, molecular, and imaging biomarkers and addresses pivotal clinical translation challenges, from scalable production to species-specific differences. By examining the pathological landscape across the central nervous system and skeletal muscle together, this review connects mechanisms that have more often been considered in isolation and evaluates how their convergence may inform the design of delivery strategies targeting both tissue compartments.\n\nID: 42130092\nTitle: FTLD-TDP-43 With Motor Neuron Disease Pathology in an Autopsied Patient With Spastic Paraplegia-30B Harbouring a Homozygous KIF1A Variant.\nAbstract: KIF1A-associated neurological disorder (KAND) is a rare hereditary condition caused by KIF1A variants, affecting axonal transport and presenting with a wide clinical spectrum, including hereditary spastic paraplegia. This case of childhood-onset KAND reveals FTLD-TDP43 with motor neuron disease pathology emerging late in the disease course, suggesting that HSP and FTLD-MND share a pathological continuum through a TDP-43-related pathway and expanding the clinicopathological spectrum of KAND.\n\nID: 42123583\nTitle: Regulation of Tau Alternative Splicing: A Novel Role for the Ribonucleoprotein RBM20.\nAbstract: Tau is a protein associated with microtubules principally expressed in neuronal cells, where it plays a fundamental role in cytoskeleton stabilization and axonal transport. Several diseases collectively named tauopathies, such as Alzheimer's disease, have been associated with an imbalance in the expression of alternative spliced Tau transcripts and the accumulation of hyperphosphorylated Tau, causing dysfunction and death of neuronal cells. Therefore, understanding the Tau exon splicing mechanisms may contribute to elucidating molecular factors that could underlie the development of neurodegenerative disorders. The aim of this study was to define the role of selected splicing factors in regulating Tau exon expression in cell lines and neuronal organoids. We demonstrated the role of the RNA-binding motif protein 20 (RBM20) splicing factor in regulating Tau exon 6 and exon 10, applying RNA-binding assay and qPCR analyses. Furthermore, we demonstrated that Tau expression was regulated during cerebral organoid differentiation, recapitulating in vivo Tau expression. These results suggest the feasibility of using brain organoid technology to study Tau alternative splicing during neural development, confirming that 3D cellular models could be used to study and characterize pathological processes taking place in Tau-related pathologies.\n\nID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases.\n\nID: 42056834\nTitle: Generation of iPSC line NIMHi033-A from an Indian patient with Autism Spectrum Disorder carrying mutation in DYNC1H1 gene.\nAbstract: We report the generation of a human induced pluripotent stem cell (hiPSC) line from peripheral blood mononuclear cells of a 14-year-old male with Autism Spectrum Disorder carrying a de novo heterozygous likely-pathogenic variation c.4067C>T (p.Pro1356Leu) in Dynein Cytoplasmic 1 Heavy Chain 1 (DYNC1H1) gene. Variation in this gene that encodes a motor protein may affect retrograde transport of neurotrophic signals, synaptic vesicles, organelles like golgi complex and mitochondria. Therefore, this iPSC line will be an invaluable tool to study the effects of this high-risk autism-related variant on disease manifestation and cellular functions.\n\nID: 42056501\nTitle: AAV2 capsid clearance and neuronal trafficking dynamics in the central nervous system.\nAbstract: Adeno-associated virus serotype 2 (AAV2) remains one of the most common vectors for CNS gene delivery. Yet, its long-term intraparenchymal trafficking and the relationship between capsid persistence and transgene expression remain poorly understood. In this study, we tracked the spatial and temporal patterns of AAV2 following direct striatal infusion in rats, analyzing tissue at 3 days, 3 weeks, 12 weeks, 30 weeks, and 67 weeks after injection. Using immunofluorescence for the human aromatic L-amino acid decarboxylase (AADC) transgene and an epitope detecting intact AAV2 capsids (A20), we mapped capsid localization, clearance, and axonal transport over more than a year. Following direct striatal infusion, AAV2 capsids rapidly localized to striatal neurons, with early accumulation in the substantia nigra pars reticulata (SNpr) detectable by 3 days post-infusion, but without evidence of nigral neuron transduction. Transgene expression increased over time and peaked locally at 12 weeks, with a delayed yet robust AADC signal in striatonigral terminals by 30 weeks. By 67 weeks, capsid signal was minimal while AADC expression remained stable. These findings clarify the long-term dynamics of AAV2 distribution, capsid persistence, and axonal transport in the adult brain, informing our understanding of vector behavior and durability following intraparenchymal AAV2 gene delivery.\n\nID: 42055257\nTitle: Selective vulnerability of monoaminergic neurons and network spread of alpha-synuclein jointly explain pathology progression in Parkinson's disease models.\nAbstract: The trans-synaptic propagation of \u03b1-synuclein aggregates is a defining feature of Parkinson's disease, yet how specific cellular phenotypes interact with the brain's structural connectome to govern disease progression remains a fundamental question in neurobiology. Here, we address this question using a connectome-based modeling framework that simulates synucinopathy spread along the mouse structural connectome, to examine how monoaminergic cell-type vulnerability and network-based transmission jointly shape pathology progression. 'Nexopathy in silico' (NexIS), along with the MISS algorithm for spatial gene expression mapping, provided a framework to examine how regional and cellular context impact synucleinopathy spread. In particular we focus on monoaminergic neurons, since inadequate clearance of pathologic \u03b1-synuclein species in monoaminergic neurons is a prominent hypothesis implicated in PD. We delineate the specific roles of monoaminergic cell types and directional transmission of \u03b1-synuclein arising from axonal transport polarity in shaping \u03b1-synuclein pathology in mouse models. Our analysis reveals that hindbrain noradrenergic (HBNOR) and midbrain dopaminergic (MBDOP) cell-type distributions are the primary mediators of network-wide transmission, with the former outperforming endogenous regional Snca expression as predictors of long-term pathology. Crucially, we demonstrate a synergistic effect between cell-type vulnerability and retrograde-biased transport, finding that both factors are required to accurately recapitulate empirical spatiotemporal patterns at 6 and 12 months post-seeding. This work constitutes one of the most mechanistically complete models of synucleinopathy to date, providing a comprehensive theoretical bridge that links microscale molecular motor regulation to macroscale regional vulnerability. Our findings suggest that the interaction between noradrenergic cell-type distribution and retrograde transport serves as the dominant driver of late-stage progression, offering highly specific cellular and mechanistic targets for therapeutic intervention.\n\nID: 42047916\nTitle: From Bench to Bedside: Neurofilament Light Chain as a Biomarker in Ischemic Stroke.\nAbstract: Neurofilaments (NFs) are protein structures that form part of the neuronal cytoskeleton, providing structural support and facilitating axonal transport. Neurofilament light chain (NFL) has emerged as a promising biomarker for various neurological conditions, as it is released into the bloodstream following neuronal damage. The ability of NFL to differentiate between types of injury and enhance the accuracy of clinical prediction models makes it an asset in both clinical practice and research. Studies suggest that NFL can be employed to diagnose and monitor disease progression and aid in the prognosis of various neurological conditions, including ischemic stroke (IS). In IS, NFL serves as a predictor of severity, functional recovery, and the risk of complications. This review therefore synthesizes current preclinical and clinical studies on the use of NFL as a biomarker for prognosis, monitoring, and treatment in IS.\n\nID: 42041253\nTitle: Typhoid toxin: reframing enteric fever.\nAbstract: SUMMARYSalmonella enterica serovars Typhi and Paratyphi A cause enteric (typhoid/paratyphoid) fever, a systemic disease that remains a major source of morbidity and mortality worldwide. The most distinctive clinical manifestations, such as sustained fever with relative bradycardia, leukopenia, neuropsychiatric symptoms/encephalopathy, and ileal perforation, have long lacked a unifying mechanistic explanation. Over the past two decades, studies of typhoid toxin have reframed how we understand this disease. Typhoid toxin is a human-adapted, chimeric A2B5 exotoxin assembled exclusively inside infected cells. It couples an ADP-ribosyltransferase (PltA) and a DNase I-like nuclease (CdtB) on a pentameric delivery ring (PltB or the alternate, PltC), that recognizes N-acetylneuraminic acid (Neu5Ac)-terminated human sialoglycans. The toxin is produced by bacteria residing within the Salmonella-containing vacuole (SCV), secreted across the bacterial envelope into the SCV lumen by a phage-derived pathway now classified as type 10 secretion system, sorted for export by the host CI-M6PR/COPII trafficking machinery, and then re-enters distant target cells via retrograde transport to the endoplasmic reticulum (ER). Recent work has linked typhoid toxin to leukopenia, blood-brain barrier disruption causing encephalopathy, gut-vascular and immune dysfunction leading to intestinal perforation, and hepatobiliary injury. The discovery of an alternate B-subunit (PltC) established a paradigm of modular B-subunit exchange that diversifies receptor usage, tissue tropism, and biology. Collectively, these insights redefine typhoid fever as a toxin-driven immunovascular disease and identify actionable targets for neutralizing antibodies and barrier-protective therapies.\n\nID: 42035797\nTitle: Mitofusin 2 in central nervous system disorders: Roles in mitochondrial dynamics and therapeutic Implications.\nAbstract: Mitochondrial dynamics have been increasingly recognized as a central determinant in the pathogenesis of central nervous system (CNS) disorders. Mitofusin 2 (MFN2), a critical mitochondrial fusion protein, preserves mitochondrial network integrity and participates in fission, mitophagy, and axonal transport, thereby maintaining neuronal function and energy homeostasis. Structural features of MFN2 underpin its diverse regulatory roles, whereas MFN2 deficiency leads to mitochondrial fragmentation, metabolic dysfunction, oxidative stress, and neuronal impairment. This review summarizes the molecular mechanisms of MFN2 in the CNS and its impact on neuronal survival, synaptic function, and signaling pathways. In addition, we highlight potential MFN2-targeted interventions, including natural compounds, pharmacological agents, and emerging small-molecule activators, while also discussing disease-specific mechanisms and translational challenges such as endoplasmic reticulum (ER)-mitochondria communication abnormalities and blood-brain barrier permeability.\n\nID: 42027429\nTitle: Characterization of a novel inherited splice-site variant in MAPK8IP3 expands the genetic and phenotypic spectrum of neurodevelopmental disorders.\nAbstract: The MAPK8IP3 gene encodes JIP3, a kinesin-1 adapter protein crucial for axonal transport and JNK signaling pathway regulation. This gene exhibits extreme intolerance to loss-of-function variants. Heterozygous pathogenic MAPK8IP3 variants are linked to a neurodevelopmental disorder with or without variable brain abnormalities (NEDBA). This study characterizes a novel inherited canonical splice-site variant identified in a three-generation family. Exome sequencing was performed on three affected individuals from a single non-consanguineous family with presenting neurological abnormalities. The molecular consequences of the identified variant were evaluated by RNA analysis. Clinical features of the variant carriers were evaluated in detail. We identified a heterozygous dinucleotide deletion (c.2630-2_2630-1del) at the canonical acceptor splice site of MAPK8IP3. This variant co-segregated with the neurological phenotype in an affected mother and her two children. RNA analysis demonstrated that this variant leads to a partial deletion of exon 22 (r.2630_2673del), which is predicted to cause a frameshift and introduce a premature termination codon (p.Gly877Valfs\u221736). Notably, despite carrying an identical genotype, the three affected individuals exhibited marked interindividual variability in clinical severity, ranging from mild specific learning difficulties to global developmental delay with severe intellectual disability and autism. This represents the first report of a familial, autosomal dominantly inherited pathogenic splice-site variant in MAPK8IP3, extending the established inheritance pattern beyond previously described de novo events. Our findings expand both the mutational and phenotypic spectrum of MAPK8IP3-related disorders, highlighting the complexity of genotype-phenotype correlations and underscoring the necessity of functional assays for accurate variant interpretation.\n\nID: 42017539\nTitle: The p.(Leu97Ile) variant expands the genetic landscape of NEFL-associated Charcot-Marie-tooth neuropathies.\nAbstract: Charcot Marie Tooth neuropathies arise from diverse genetic disturbances that impair axonal structure or myelin integrity. Variants in NEFL, encoding the light chain of neurofilaments, represent a rare cause of CMT and may disrupt filament assembly, axonal transport, and cytoskeletal stability. Because NEFL variants are uncommon and phenotypes variable, the pathogenic relevance of individual substitutions is often difficult to determine. Here, we identified a previously unreported missense variant, p.(Leu97Ile), in three affected individuals. In two, a second variant, p.(Arg206Ser), was present in cis. We therefore aimed to investigate the pathogenic impact of this allele constellation using clinical, biochemical, and structural approaches. Neurological examination, CMTNS scoring, and electrophysiological studies were performed. Exome sequencing established the genotype. Variant effects were assessed using serum biomarkers, in silico modelling and quantitative immunofluorescence of patient-derived fibroblasts. Patients showed differing clinical presentations, including moderate adult-onset neuropathy to severe early-onset disease and neuromuscular abnormalities in childhood. Neurophysiology revealed mixed features, and conduction slowing in the child supported classification within the CMT1F spectrum. All carriers exhibited elevated serum NfL; CK was increased in the adult patients. Descriptive biomarker differences between the patients were observed for GDF15, FGF21, Periaxin, and NCAM1. Fibroblasts displayed increased cytoplasmic NfL without altered nuclear metrics. Structural modelling predicted destabilizing effects of both substitutions, with the cis combination producing the strongest deviation. Integrated clinical, biochemical, and structural evidence support pathogenicity of p.(Leu97Ile) and suggest that p.(Arg206Ser) in cis augments its biological impact. These findings expand the genetic and phenotypic spectrum of NEFL-associated CMT1F.\n\nID: 42013932\nTitle: TRPV1-tau axis: A bidirectional regulatory mechanism linking pain sensitization and Alzheimer's disease progression and its potential for intervention.\nAbstract: Alzheimer's disease (AD) is a major neurodegenerative disorder characterized by \u03b2-amyloid (A\u03b2) deposition and pathological tau phosphorylation and aggregation, frequently accompanied by chronic pain. Pain sensitization is closely linked to AD progression. Transient receptor potential vanilloid 1 (TRPV1), a key cation channel in pain transduction, is expressed not only in peripheral sensory neurons but also widely in central neurons and glial cells, where it contributes to pain sensitization and neuroinflammation. Emerging evidence indicates a bidirectional regulatory interplay between TRPV1 and tau, forming a \"TRPV1-tau axis.\" This axis acts as a core molecular bridge connecting pain sensitization and AD pathology via calcium dyshomeostasis, mTOR/AMPK, PI3K/Akt/GSK3\u03b2, and neuroinflammatory pathways. TRPV1 overactivation promotes tau hyperphosphorylation and aggregation through calcium-dependent kinases, metabolic dysregulation, and inflammatory signaling. Conversely, pathological tau modulates TRPV1 expression and function via transcriptional regulation, protein interactions, and impaired axonal transport, establishing a deleterious feedback cycle. Therapeutic interventions targeting this axis, including TRPV1 modulators, tau-directed agents, anti-inflammatory drugs, and natural compounds, demonstrate potential in alleviating both pain sensitivity and cognitive deficits through multi-target mechanisms. However, clinical translation remains challenging due to issues including blood-brain barrier penetration, target selectivity, and a lack of reliable biomarkers. This review systematically outlines the bidirectional mechanisms of the TRPV1-tau axis, current intervention strategies, and translational challenges, with the goal of informing future development of disease-modifying therapies that concurrently address cognitive decline and chronic pain in AD.\n\nID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance.\n\nID: 41979085\nTitle: VAMP4/STX8 Mediate the Autophagic Secretion of Mitochondria and Promote TAMs Polarization in HNSCC.\nAbstract: Under hypoxic conditions, tumour cells engage in autophagic secretion of mitochondria to sustain energy balance and modulate the tumour microenvironment. However, the exact molecular mechanisms remain unclear. This study used a combination of multivesicular bodies membrane proteomics and molecular interaction validation to demonstrate that hypoxia promotes mitophagy-dependent extracellular vesicles (EVs) secretion in head and neck squamous cell carcinoma (HNSCC). This process promotes tumour-associated macrophages to adopt immunosuppressive phenotypes, reshaping the immune environment. Vesicle-associated membrane protein 4 (VAMP4) and syntaxin 8 (STX8) are key molecules involved in this secretion. Clinically, VAMP4/STX8 expressions are significantly higher in HNSCC than in normal tissues and correlate with prognosis, indicating their potential as biomarkers and therapeutic targets. This study revealed the mitophagy-secretion axis that reshaped the tumour immune landscape, providing a theoretical basis for targeting soluble NSF attachment protein receptor-dependent EV release to enhance the therapeutic effect of HNSCC immunotherapy.\n\nID: 41902311\nTitle: Engineering Yeast Extracellular Vesicle Biogenesis Through Rewiring Membrane Trafficking Pathways.\nAbstract: Extracellular vesicles (EVs) are emerging as versatile therapeutic platforms, yet the mechanisms governing their biogenesis in yeast remain incompletely understood. Saccharomyces cerevisiae, a well-characterised and safe microbial chassis, naturally secretes abundant EVs and provides an attractive system for mechanistic dissection and engineering. Here, we establish S. cerevisiae as a tractable model for elucidating EV cargo loading. By combining multicopy expression of chicken interferon-\u03bb (ChiIFN-\u03bb) with cell wall perturbation, we achieved a tenfold increase in EV yield and efficient incorporation of ChiIFN-\u03bb into EVs. Quantitative proteomics identified 1555 EV-associated proteins, including 501 predicted transmembrane proteins derived from multiple organelles. ChiIFN-\u03bb overexpression and cell wall stress selectively reduced the abundance of key vesicle trafficking regulators, including SNARE, ESCRT and Rab proteins, indicating reprogramming of intracellular membrane trafficking pathways. Functional analyses further demonstrated that the SNARE proteins Sso2 and Nyv1 are enriched in the EV membrane and modulate EV size distribution and subpopulation composition. Together, these results reveal conserved protein-sorting machinery underlying yeast-derived extracellular vesicles (YDEVs) biogenesis and establish S. cerevisiae as a powerful platform for engineered EV production.\n\nID: 41747594\nTitle: Effect of intranasal treatment with NAMPT-EVs on acetylated tau and cognitive function in mice with repeated controlled cortical injury.\nAbstract: Repeated traumatic brain injury (rTBI) has attracted increasing attention owing to its long-term effects on cognition and behaviour. Moreover, research has shown that acetylated tau (ac-tau) represents a common pathology linking rTBI and Alzheimer's disease that can lead to neuronal cell death. Therefore, in this study, we evaluated the therapeutic potential of mesenchymal stromal cell-derived extracellular vesicles enriched with nicotinamide phosphoribosyltransferase (NAMPT-EVs) for improving cognitive and behavioral impairments following repeated controlled cortical injury (rCCI). Morris water maze and novel object recognition test were evaluated at 1-month post-rCCI with intranasal treatment of NAMPT-EVs. Expression of Sirtuin 1(SIRT1), ac-tau, neuron loss, neuroinflammation, AQP4 polarity, and meningeal lymphatic morphology and function were assessed 1\u00a0month after treatment. Intranasal administration of NAMPT-EVs significantly increased the expression of SIRT1 to deacetylate tau in rCCI mice. Additionally, NAMPT-EVs suppressed neuroinflammation and maintained aquaporin protein-4 polarity to facilitate the glymphatic system and promote the repair of the meningeal lymphatic system, which benefits the clearance of ac-tau from the brain parenchyma. Notably, the reduction in ac-tau prevented axon initial segment degradation and tau mislocalisation, resulting in a neuroprotective effect. NAMPT-EVs reduce neuronal loss and improve cognitive function in rCCI mice through multiple mechanisms. Therefore, NAMPT-EVs is promising for preventing cognitive deficit after rTBI.\n\nID: 41704641\nTitle: Alzheimer's disease as a systems-level timing disorder: Circadian disruption of glial immunometabolism, brain clearance, and therapeutic responsiveness.\nAbstract: Alzheimer's disease (AD) is traditionally conceptualized as a disorder of protein aggregation and neurodegeneration, yet growing evidence indicates that fundamental temporal organization of brain physiology is also disrupted. In the healthy brain, circadian clocks coordinate sleep-wake behavior, glial immunometabolism, astrocytic aquaporin-4 polarity, and glymphatic-lymphatic clearance, aligning immune readiness and proteostasis with daily activity-rest cycles. In AD, this temporal coordination progressively deteriorates, manifesting as sleep fragmentation, instability of rest-activity rhythms, vulnerability of central clock structures, and loss of circadian gating of glial and clearance pathways. These disruptions create phase-inappropriate immune and metabolic states, impair protein clearance, and alter the fate of extracellular vesicles, which may shift from mediators of waste export to facilitators of proteopathic spread. Importantly, circadian failure also constrains therapeutic delivery and biomarker interpretation by modulating blood-brain barrier transport, brain fluid dynamics, and brain-to-blood signal export. We propose that AD can be reframed as a systems-level timing disorder, in which loss of temporal coherence integrates molecular pathology, glial dysfunction, clearance failure, therapeutic inefficacy, and biomarker variability. This framework highlights chrono-pharmacology, chrono-neurotherapeutics, and circadian-informed biomarkers as essential components of precision strategies for AD prevention and treatment.\n\nID: 41659924\nTitle: Mechanism of exercise-derived circulating exosomes as a target for sarcopenia management.\nAbstract: Sarcopenia, an age-related syndrome characterized by the progressive decline of skeletal muscle mass and function, threatens the health of older adults through underlying mechanisms that include dysregulated protein metabolism, autophagy-mitochondrial dysfunction, chronic inflammation, and impaired regenerative capacity of muscle stem cells. Exercise-derived circulating exosomes, which act as key mediators of intercellular communication, show considerable potential in mitigating sarcopenia-related damage. In this review, we summarize the biogenesis of exercise-induced exosomes, encompassing both ESCRT-dependent and independent pathways, secretion regulated by RAB and SNARE proteins, and their release mediated through mechanical, calcium, metabolic, and neuroendocrine signaling during exercise. We further elaborate on the systemic roles of these exosomes in muscle repair, including alleviating lipotoxicity via the FGF21-adiponectin axis, maintaining protein homeostasis through dual regulation by miR-29c, and ameliorating the inflammatory microenvironment via modulation of macrophage polarization. Finally, we discuss the translational promise of exosomes as therapeutic targets and outline future research directions, offering a conceptual framework for understanding exercise-mediated muscle protection and developing novel interventions.\n\nID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD.\n\nID: 40202614\nTitle: Extracellular Vesicles from Dendritic Cells Protect Against Sporothrix brasiliensis Yeast Cells.\nAbstract: Sporotrichosis is an emerging subcutaneous mycotic zoonosis that affects the skin, lymphatic system, and other organs of humans and animals. Like other infectious fungal diseases, it becomes even more severe when it affects immunosuppressed patients. This infection has a global distribution and is endemic in some regions of Brazil and it is an important zoonotic public health problem. The disease is caused by a complex of at least four pathogenic species, including Sporothrix brasiliensis. The immunological response against these species has not yet been completely elucidated. Still, structures such as extracellular vesicles could carry important components that can contribute to the modulation and control of this significant infection. Thus, this work aims to analyze the participation of EVs from na\u00efve dendritic cells and EVs from DCs previously primed with S. brasiliensis yeast and primed with EVs from the fungus in the immune response against experimental sporotrichosis in murine models. The groups that received EVs from DCs primed with S. brasiliensis or their EVs showed a significant decrease in fungal load compared to the negative control group. When we analyzed the cytokine profile in the skin of mice treated with EVs before infection, we observed an increase in IFN-\u213d, TNF-\u03b1, IL-17, and IL-10, mainly in animals previously treated with EVs from DCs cultivated with yeast cells. It is worth highlighting that all prophylactic protocols modulated and minimized fungal growth compared to the control; that is, EVs contributed to the control of the infection and acted in favor of the host, demonstrating a protective character.\n\nID: 39973490\nTitle: From blood vessels to brain cells: Connecting the circulatory system and Parkinson's disease.\nAbstract: Parkinson's disease (PD) is traditionally recognized as a neurodegenerative disorder characterized by motor dysfunction and \u03b1-synuclein protein accumulation in the brain. However, recent research suggests that the circulatory system may also contribute to PD pathogenesis through the spread of \u03b1-synuclein beyond the brain. The blood-brain barrier (BBB), a key regulator of molecular exchange between the bloodstream and the brain, may become compromised in PD, allowing harmful substances, including pathogenic forms of \u03b1-synuclein, to infiltrate the brain and promote neurodegeneration. Transport mechanisms such as P-glycoprotein and the low-density lipoprotein (LDL) receptor-related protein (LRP-1) further modulate the movement of \u03b1-synuclein across the BBB, influencing disease progression. Additionally, extracellular vesicles are emerging as crucial mediators in the dissemination of \u03b1-synuclein between the brain and peripheral tissues, facilitating its spread and accumulation. The lymphatic system, responsible for clearing \u03b1-synuclein, may also contribute to PD pathology when impaired. This review highlights the growing evidence for a circulatory axis in the initiation and progression of PD. We propose that future research should explore the hypothesis that the circulatory system contributes to the pathogenesis of PD by aiding the distribution of \u03b1-synuclein throughout the body. Parkinson's disease (PD) is known for its effects on movement and the brain. But the cause remains unknown. Recent research suggests that problems with blood and blood flow contribute to the development of PD. For instance, proteins linked to PD have been identified in blood and heart tissues, indicating they could be early signs of the disease. Changes in heart rate, changes in blood pressure when standing up, and altered brain blood flow have been linked to PD. This review explores evidence that the circulatory system has important roles in the development and progression of PD. We propose further research to test if proteins in the blood start the disease and if blood vessels help spread it to the brain.\n\nID: 39739969\nTitle: MicroRNAs secreted by the parasitic nematode Brugia malayi disrupt lymphatic endothelial cell integrity.\nAbstract: Lymphatic filariasis (LF) is a neglected tropical disease affecting over 51 million people in 72 endemic countries. Causative agents of LF are mosquito-borne parasitic nematodes Wuchereria bancrofti, Brugia malayi, and Brugia timori. The adult parasites impact the integrity of lymphatic vessels and damage valves, leading to a remodeling of the lymphatic system and lymphatic dilation. Chronic infections can develop into severe clinical manifestations, primarily lymphedema, hydrocoele, and elephantiasis. Mechanistic studies on the underlying pathology due to the parasite are necessary to better manage human filariasis. Since parasite molecules, such as microRNAs (miRNAs), can be found in secreted extracellular vesicles (EVs) and are transported between parasite and host cells, we hypothesized that these could also play a role in the development of pathology in LF. In this study, we tested two B. malayi miRNAs previously detected in vitro in the culture media of microfilarial stages of worms. While one is Brugia-specific (bma-miR-5864) and the other nematode-specific (bma-miR-86), both miRNAs are secreted in high abundance. We first examined the in vitro response by transcriptomic profiling of human lymphatic endothelial cells to treatment with these miRNAs, which allowed us to identify genes involved in maintaining the integrity of the lymphatic endothelium. We then measured the effect of these miRNAs on the regulation of proteins necessary for cell integrity, demonstrating downregulation leading to a significant increase in the permeability of the endothelium monolayer. With this study we identify parasite miRNAs involved in undermining the integrity of endothelial cells, thus potentially contributing to the development of pathology. These findings could pave the way for a novel treatment strategy where the inhibition of parasite-secreted molecules could slow the progression of LF pathology. From a broader perspective, the miRNAs secreted by filarial parasites could potentially be used in the future for diagnosing and monitoring disease progression or treatment efficacy.\n\nID: 39426689\nTitle: The role of ferroptosis resistance in lymph-associated tumour metastasis.\nAbstract: Tumour metastasis is a crucial factor in determining clinically challenging tumours. In this respect, the lymphatic system may act as potential entry portals for tumour metastasis, whilst, clinical detection of tumour-infiltrated lymph nodes also indicates poorer prognosis and higher metastatic risk. Whether tumour cells gain ferroptosis resistance in lymph that make them exhibit a stronger propensity for lymphatic dissemination compared to hematogenous spread might be a breakthrough for elucidating lymph-associated tumour metastasis. This review discusses how the lymphatic system endows tumour cells with ferroptosis resistance character, which makes them more propensity for lymph node pre-metastasis and distant metastasis through lymphatic circulation. Comprehensively considering the distinct structure and property of lymph and the unique metabolic characteristics of tumours, all of the lymphatic vessels, intestinal lymph and lymph nodes collectively manipulate an intricate interaction with the hematogenous system and afford substances exchange with tumour cells and extracellular vesicles, upon which make a ferroptosis resistant microenvironment for subsequent metastasis in distant organs and lymph nodes.\n\nID: 39128945\nTitle: Platelet extracellular vesicles preserve lymphatic endothelial cell integrity and enhance lymphatic vessel function.\nAbstract: Lymphatic vessels are essential for preventing the accumulation of harmful components within peripheral tissues, including the artery wall. Various endogenous mechanisms maintain adequate lymphatic function throughout life, with platelets being essential for preserving lymphatic vessel integrity. However, since lymph lacks platelets, their impact on the lymphatic system has long been viewed as restricted to areas where lymphatics intersect with blood vessels. Nevertheless, platelets can also exert long range effects through the release of extracellular vesicles (EVs) upon activation. We observed that platelet EVs (PEVs) are present in lymph, a compartment to which they could transfer regulatory effects of platelets. Here, we report that PEVs in lymph exhibit a distinct signature enabling them to interact with lymphatic endothelial cells (LECs). In vitro experiments show that the internalization of PEVs by LECs maintains their functional integrity. Treatment with PEVs improves lymphatic contraction capacity in atherosclerosis-prone mice. We suggest that boosting lymphatic pumping with exogenous PEVs offers a novel therapeutic approach for chronic inflammatory diseases characterized by defective lymphatics.\n\nID: 38938681\nTitle: Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins.\nAbstract: Extracellular vesicles (EVs) are crucial mediators of cell-to-cell communication in physiological and pathological conditions. Specifically, EVs released from the vasculature into blood were found to be quantitatively and qualitatively different in diseases compared to healthy states. However, our understanding of EVs derived from the lymphatic system is still scarce. In this study, we compared the mRNA and microRNA (miRNA) expression in blood vascular (BEC) and lymphatic (LEC) endothelial cells. After characterization of the EVs by fluorescence-triggered flow cytometry, nanoparticle tracking analysis and cryo-transmission electron microscopy (cryo-TEM) we utilized small RNA-sequencing to characterize miRNA signatures in the EVs and identify cell-type specific miRNAs in BEC and LEC. We found miRNAs specifically enriched in BEC and LEC on the cellular as well as the extracellular vesicle level. Our data provide a solid basis for further functional in vitro and in vivo studies addressing the role of EVs in the blood and lymphatic vasculature.\n\nID: 38674369\nTitle: Exploring the Role of Extracellular Vesicles in the Pathogenesis of Tuberculosis.\nAbstract: Tuberculosis (TB) remains a significant global health concern, necessitating accurate diagnosis and treatment monitoring. Extracellular vesicles (EVs), including exosomes, play crucial roles in disease progression, with their associated genes serving as potential biomarkers and therapeutic targets. Leveraging publicly available RNA-Seq datasets of TB patients and healthy controls (HCs), to identify differentially expressed genes (DEGs) and their associated protein-protein interaction networks and immune cell profiles, the common EV-related DEGs were identified and validated in the GSE42830 and GSE40553 datasets. We have identified nine common EV-related DEGs (SERPINA1, TNFAIP6, MAPK14, STAT1, ITGA2B, VAMP5, CTSL, CEACAM1, and PLAUR) upregulated in TB patients. Immune cell infiltration analysis revealed significant differences between TB patients and HCs, highlighting increased proportions of various immune cells in TB patients. These DEGs are involved in crucial cellular processes and pathways related to exocytosis and immune response regulation. Notably, VAMP5 exhibited excellent diagnostic performance (AUC-0.993, sensitivity-93.8%, specificity-100%), with potential as a novel biomarker for TB. The EV-related genes can serve as novel potential biomarkers that can distinguish between TB and HCs. VAMP5, which functions in exosome biogenesis and showed significant upregulation in TB, can be targeted for therapeutic interventions and treatment outcomes.\n\nID: 41971838\nTitle: Molecular biology of targeting lymphangiogenesis in cancer: Therapeutic strategies and future promise.\nAbstract: Lymphangiogenesis is considered the rate-limiting step for tumor lymph node (LN) metastasis by sprouting the new lymphatic network from preexisting lymphatic vessels to favor the invasion and entry of cancer cells into the lymphatic system. Over the past decade, numerous studies have investigated the cellular roles and molecular mechanisms of lymphangiogenesis in cancers, and corresponding molecular control strategies targeting lymphangiogenesis to treat LN metastatic cancers have been developed. Nevertheless, due to the crucial role of lymphatic vessels in the infiltration of immune cells and immune activation in cancers, there have been several diverse findings regarding the biological roles and strategies of targeting tumor lymphangiogenesis as cancer immunotherapy has emerged and rapidly developed. Herein, we summarize the progress in characterizing the molecular mechanisms and therapeutic strategies involved in tumor lymphangiogenesis and highlight the potential of targeting lymphangiogenesis in combination with immunotherapy for the treatment of cancer, providing opportunities to improve the efficacy of targeting lymphangiogenesis to prevent cancer metastasis.\n\nID: 38249813\nTitle: Cross\u2011talk between lymphangiogenesis and malignant melanoma cells: New opinions on tumour drainage and immunization (Review).\nAbstract: Malignant melanoma (MM) is a highly aggressive tumour that can easily metastasize through the lymphatic system at the early stages. Lymph node (LN) involvement and lymphatic vessel (LV) density (LVD) represent a harbinger of an adverse prognosis, indicating a strong link between the state of the lymphatic system and the advancement of MM. Permeable capillary lymphatic vessels are the optimal conduits for melanoma cell (MMC) invasion, and lymphatic endothelial cells (LECs) can also release a variety of chemokines that actively attract MMCs expressing chemokine ligands through a gradient orientation. Moreover, due to the lower oxidative stress environment in the lymph compared with the blood circulation, MMCs are more likely to survive and colonize. The number of LVs surrounding MM is associated with tumour-infiltrating lymphocytes (TILs), which is crucial for the effectiveness of immunotherapy. On the other hand, MMCs can release various endothelial growth factors such as VEGF-C/D-VEGFR3 to mediate LN education and promote lymphangiogenesis. Tumour-derived extracellular vesicles are also used to promote lymphangiogenesis and create a microenvironment that is more conducive to tumour progression. MM is surrounded by a large number of lymphocytes. However, both LECs and MMCs are highly plastic, playing multiple roles in evading immune surveillance. They achieve this by expressing inhibitory ligands or reducing antigen recognition. In recent years, tertiary lymphoid structures have been shown to be associated with response to anti-immune checkpoint therapy, which is often a positive prognostic feature in MM. The present review discusses the interaction between lymphangiogenesis and MM metastasis, and it was concluded that the relationship between LVD and TILs and patient prognosis is analogous to a dynamically tilted scale.\n\nID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\n\nID: 38077706\nTitle: Successful Treatment of Post COVID-19 Neurogenic Dysphagia with Botulinum Toxin.\nAbstract: Dysphagia in post COVID-19 patients could be caused by several factors, including reduced pharyngolaryngeal coordination due to SARS-CoV-2 tropism to the central and/or peripheral nervous system. To our knowledge, this is the first reported case of COVID-19-related dysphagia successfully treated with botulinum toxin type A injection. We report the case of a patient with severe oropharyngeal dysphagia due to COVID-19 confirmed by fibre endoscopy. As a result, the patient required an enteral feeding tube. After two months of traditional swallowing therapies, there was only limited improvement. An electrophysiologic evaluation of the cricopharyngeal muscle was performed and showed a normal inhibition of the cricopharyngeal muscle, followed by a hypertonic rebound. Based on this result, we decided to perform a unilateral laryngeal injection of botulinum toxin type A. After the injection, the patient's swallowing function improved significantly, allowing him to return to oral feeding. Newly diagnosed oropharyngeal dysphagia was found in 35.3% of hospitalised patients with COVID-19. There are several possible causes of COVID-19-associated dysphagia, including stroke, encephalitis, critical illness neuropathy, Guillain-Barr\u00e9 syndrome and skeletal muscle injury. In our case, since stroke was excluded by brain MRI, cranial nerve injury was a possible explanation for the difficult recovery of swallowing despite daily swallowing therapy. We suggest that electrophysiology is a valid tool for the diagnosis and follow-up of patients with oropharyngeal dysphagia. SARS-CoV-2 tropism to the central and/or peripheral nervous system can cause dysphagia in post COVID-19 patients.An electrophysiologic approach is useful for the diagnosis and follow-up of patients with oropharyngeal dysphagia.A single botulinum toxin type A injection is a valid treatment option to improve the swallowing function in patients with post COVID-19 dysphagia.\n\nID: 37700095\nTitle: Identification of the SNARE complex that mediates the fusion of multivesicular bodies with the plasma membrane in exosome secretion.\nAbstract: Exosomes play crucial roles in local and distant cellular communication and are involved in various physiological and pathological processes. Tumour-derived exosomes are pivotal to tumorigenesis, but the precise mechanisms underlying their secretion remain elusive. In particular, the SNARE proteins that mediate the fusion of multivesicular bodies (MVBs) with the plasma membrane (PM) in tumour cells are subject to debate. In this study, we identified syntaxin-4, SNAP-23, and VAMP-7 as the SNAREs responsible for exosome secretion in MCF-7 breast cancer cells and found that a SNARE complex consisting of these SNAREs can drive membrane fusion in vitro. Deletion of any of these SNAREs in MCF-7 cells did not affect MVB biogenesis and transportation, indicating their specific involvement in MVB-PM fusion. In addition, syntaxin-4, SNAP-23, and VAMP-7 play equivalent roles in exosome secretion in both HeLa cervical cancer cells and A375 melanoma cells, suggesting their conserved function in exosome secretion. Furthermore, deletion of VAMP-7 in 4T1 mammary carcinoma cells efficiently inhibited exosome secretion and led to significant attenuation of tumour growth and lung metastasis in mouse models, implying that VAMP-7 may hold promise as a novel therapeutic target for breast cancer.\n\nID: 37517544\nTitle: Logistics and distribution of small extracellular vesicles from the subcutaneous space to the lymphatic system.\nAbstract: Small extracellular vesicles (sEVs) are small, cell-derived particles with sizes of approximately 100\u00a0nm. Since these particles include cargos such as host cell-derived proteins, messenger RNAs, and micro RNAs, they serve as mediators of cell-cell communication. While the analysis of the pharmacokinetic of sEVs after the intravenous injection have been reported, the lymphatic transport of sEVs remains unclear. The objective of this study was to provide insights into the intra-lymphatic trafficking and distribution of sEVs when they are injected into an interstitial space both in normal skin tissue and in cancerous tissue. When sEVs were Subcutaneously administered into the tail base and the tumor tissue, they preferably accumulated in the lymph nodes (LNs), rather than in the liver and the spleen. The findings reported herein show that the lymphatic transport of sEVs was drastically changed in model mice, in which a surgical treatment was used to modify to allow the dominant lymphatic flow from the footpad directly to the axillary LN via the inguinal LN. Based on the results, we conclude that when sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system. Further, the extent of accumulation of sEVs in the LN after subcutaneous injection was reduced when they were preliminarily incubated with Proteinase K. These results suggest that the lymphatic drainage of sEVs in normal skin tissue is regulated by membrane proteins on their surface. This reduction, however, was not observed in the case of cancer tissue. This discrepancy can be attributed to the presence of highly permeable lymphatic vessels in the tumor tissue. Further, the major cell subtypes that captured sEVs in the LN were LN-resident medullary sinus macrophages. These collective findings indicate that the lymphatic drainage of sEVs are mediated by proteins and, that they may appear to contribute to the control of the function of immune-responsive cells in the LNs.\n\nID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined.\n\nID: 37371477\nTitle: The Role of Extracellular Vesicles in the Pathogenesis of Hematological Malignancies: Interaction with Tumor Microenvironment; a Potential Biomarker and Targeted Therapy.\nAbstract: The tumor microenvironment (TME) plays an important role in the development and progression of hematological malignancies. In recent years, studies have focused on understanding how tumor cells communicate within the TME. In addition to several factors, such as growth factors, cytokines, extracellular matrix (ECM) molecules, etc., a growing body of evidence has indicated that extracellular vesicles (EVs) play a crucial role in the communication of tumor cells within the TME, thereby contributing to the pathogenesis of hematological malignancies. The present review focuses on how EVs derived from tumor cells interact with the cells in the TME, such as immune cells, stromal cells, endothelial cells, and ECM components, and vice versa, in the context of various hematological malignancies. EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers for specific types of cancer, thereby also acting as potential therapeutic targets. Here, we discuss how EVs influence hematological tumor progression via tumor-host crosstalk and their use as biomarkers for hematological malignancies, thereby benefiting the development of potential therapeutic targets.\n\nID: 37204265\nTitle: Ultrasonography in neurology: A comprehensive analysis and review.\nAbstract: Neurologists in both the inpatient and outpatient settings are increasingly using ultrasound to diagnose and manage common neurological diseases. Advantages include cost-effectiveness, the lack of exposure to ionizing radiation, and the ability to perform at the bedside to provide real-time data. There is a growing body of literature that supports using ultrasonography to improve diagnostic accuracy and aid in performing procedures. Despite the increasing utilization of this imaging modality in medicine, there has been no comprehensive review of the clinical applications of ultrasound in the field of neurology. We discuss the current uses and limitations of ultrasound for various neurological conditions. We review the role for ultrasound in commonly performed neurologic procedures including lumbar puncture, botulinum toxin injections, nerve blocks, and trigger point injections. We specifically discuss the technique for ultrasound-assisted lumbar puncture and occipital nerve block as these are commonly performed. We then focus on the utility of ultrasound in the diagnosis of neurologic conditions. This includes neuromuscular diseases such as motor neuron disorders, focal neuropathies, and muscular dystrophy as well as vascular conditions such as stroke and vasospasm in subarachnoid hemorrhage. We also address ultrasound's use in critically ill patients to aid in identifying increased intracranial pressure, hemodynamics, and arterial and/or venous catheterization. Finally, we address the importance of standardized ultrasound curricula in trainee education and make recommendations for the future directions of research and competency guidelines within our specialty.\n\nID: 37086961\nTitle: Tumor-derived extracellular vesicles delivering TNF-\u03b1 promotes colorectal cancer metastasis via the NF-kB/LAMB3/AKT axis by targeting SNAP23.\nAbstract: Accumulating evidence have demonstrated that cytokines are enriched in tumor-derived extracellular vesicles (EVs) and widely involved in tumorigenesis of various types of carcinomas, including colorectal cancer (CRC). Nevertheless, the functions of cytokines in EVs secreted from colorectal cancer cells remain largely unknown. In the present study, we found that TNF-\u03b1 was elevated in EVs from CRC patient serum samples and CRC cell lines, of which the expression was associated with aggressive features of colorectal cancer. EV TNF-\u03b1 secretion is dependent on synaptosome-associated protein 23 (SNAP23). Functional experiments revealed that EV TNF-\u03b1 promotes CRC cell metastasis via the NF-\u03baB pathway by targeting SNAP23. Mechanistically, SNAP23 was transcriptionally upregulated by EV TNF-\u03b1/NF-\u03baB axis to enhance the expression of laminin subunit beta-3 (LAMB3), thereby activating the PI3K/AKT signaling pathway and consequently facilitate CRC progression. Based on our findings, we could conclude that EV TNF-\u03b1 plays an oncogenic role in CRC progression through SNAP23, which in turn promotes EV TNF-\u03b1 secretion, suggesting that EV TNF-\u03b1/SNAP23 axis may serve as a diagnostic biomarker and potential therapeutic target for CRC.\n\nID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies.\n\nID: 36971125\nTitle: Extracellular vesicle-mediated heterogeneous communication between cancer and the lymphatic system facilitates lymphatic metastasis.\nAbstract: \n\nID: 36831512\nTitle: Current Status of Lymphangiogenesis: Molecular Mechanism, Immune Tolerance, and Application Prospect.\nAbstract: The lymphatic system is a channel for fluid transport and cell migration, but it has always been controversial in promoting and suppressing cancer. VEGFC/VEGFR3 signaling has long been recognized as a major molecular driver of lymphangiogenesis. However, many studies have shown that the neural network of lymphatic signaling is complex. Lymphatic vessels have been found to play an essential role in the immune regulation of tumor metastasis and cardiac repair. This review describes the effects of lipid metabolism, extracellular vesicles, and flow shear forces on lymphangiogenesis. Moreover, the pro-tumor immune tolerance function of lymphatic vessels is discussed, and the tasks of meningeal lymphatic vessels and cardiac lymphatic vessels in diseases are further discussed. Finally, the value of conversion therapy targeting the lymphatic system is introduced from the perspective of immunotherapy and pro-lymphatic biomaterials for lymphangiogenesis.\n\nID: 42548572\nTitle: Case Report: Munc13-4 deficiency presenting with autoimmune neuropathy years before FLH: implications for early genetic screening and HSCT timing.\nAbstract: The UNC13D gene encodes Munc13-4, a critical regulator of cytotoxic granule exocytosis in lymphocytes and the protein responsible for familial hemophagocytic lymphohistiocytosis (FHL). As an essential mediator of perforin release from cytotoxic T and NK cells, Munc13-4 ensures immune surveillance by facilitating granule-plasma membrane fusion. Defects in this pathway, termed \"perforinopathy\" manifest as impaired cytotoxicity and consequent immune dysregulation. While central nervous system involvement and systemic inflammation are established features of Munc13-4 deficiency, its association with autoimmune neuropathy remains largely unrecognized. We report two patients with a unique disease trajectory, presenting initially with autoimmune neuropathy before developing progressive FHL several years later. Both patients met the HLH-2004 diagnostic criteria, including fever, pancytopenia, hyperferritinemia, hypertriglyceridemia, and splenomegaly. Genetic analysis revealed heterozygous UNC13D mutations: in Case 1, a nonsense (c.3193C>T; p.Arg1065*) and a missense (c.1135G>A; p.Glu379Lys) mutation; in Case 2, compound heterozygous mutations comprising a maternal splicing variant (c.2447\u00a0+\u00a01G>A) and a paternal missense mutation (c.1241G>T; p.Arg414Leu). Case 1 underwent hematopoietic stem cell transplantation (HSCT), with significant recovery of weakness and numbness, and no recurrence of fever or lymphadenopathy at one year. Case 2 succumbed to disease progression after no timely, suitable allogeneic hematopoietic stem cell donor could be found. Our findings identify autoimmune neuropathy as a potential harbinger of FHL in perforinopathy, suggesting these neurological manifestations may be the initial phenotype of an underlying cytotoxic dysfunction. This temporal association mandates heightened clinical vigilance for immunodeficiency in patients with refractory neuropathy, particularly when accompanied by axonal degeneration or systemic inflammation. Recognizing this novel disease continuum has immediate diagnostic and therapeutic implications, as early genetic identification creates a critical window for potentially curative intervention with HSCT before irreversible multi-organ damage occurs. These cases expand the clinical spectrum of perforinopathy and challenge current diagnostic paradigms, emphasizing the need for an integrated neurological and immunological evaluation for unexplained autoimmune neuropathy.\n\nID: 42539598\nTitle: Botulinum toxin A for sialorrhea in adults with neurological disorders: a narrative review of aspiration pneumonia risk.\nAbstract: Aspiration pneumonia (AP), a serious condition resulting from the inhalation of foreign materials into the lungs, is a significant concern for patients with compromised swallowing mechanisms, especially those with neurological disorders. Sialorrhea (hypersalivation) exacerbates the risk of aspiration, and botulinum neurotoxin type A (BoNT-A) has been identified as a potential treatment to manage sialorrhea. However, the direct impact of BoNT-A on reducing AP risk remains inadequately explored, particularly in adult populations. BoNT-A works by inhibiting the release of acetylcholine at the neuromuscular junction, thereby reducing salivary gland stimulation and saliva production. This review aims to evaluate the current state of research on the use of BoNT-A for managing sialorrhea and its potential to reduce AP risk, with a specific focus on the adult population. A comprehensive review of existing literature was conducted, focusing on the efficacy of BoNT-A in treating sialorrhea in both pediatric and adult populations. Studies were assessed for their findings on the impact of BoNT-A on drooling and related complications, including the risk of AP. Meta-analyses and individual studies were reviewed to summarize the current knowledge and identify research gaps. In pediatric patients, BoNT-A has been shown to effectively reduce sialorrhea, with several studies and meta-analyses demonstrating its efficacy in managing drooling associated with neurological conditions such as cerebral palsy. However, evidence linking BoNT-A directly to reduced AP risk is limited. In adults, while BoNT-A has been beneficial in managing sialorrhea associated with conditions such as Parkinson's disease and brain injuries, studies specifically exploring its impact on AP risk are lacking. Current evidence supports the efficacy of BoNT-A in managing neurological sialorrhea across both adult and pediatric populations. However, a critical gap exists in studies directly investigation whether BoNT-A reduces AP risk in adults with neurological disease.\n\nID: 42533122\nTitle: Light-activated tetanus neurotoxin for conditional proteolysis and inducible synaptic inhibition in vivo.\nAbstract: The light chain of tetanus neurotoxin (TeNT) is a metalloprotease that potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision. To address this, we engineered light-activated TeNT (LATeNT) by inserting the light-sensitive LOV domain into an allosteric site and optimizing dynamic range via directed evolution. LATeNT's activity is undetectable in the dark, but the protease turns on after 10-20 min of weak blue-light exposure to potently inhibit synapses in vivo. Here we show that LATeNT works across multiple brain regions and at long-range axonal projections, with its effects reversible in 24\u2009h. LATeNT enabled us to discover a hippocampal interneuron population that regulates anxiety-like behaviors and demonstrate the importance of postsynaptic endocannabinoid exocytosis for depolarization-induced suppression of inhibition in vivo. Beyond neuroscience, LATeNT regulated endogenous insulin secretion from pancreatic beta cells and converted drug exposure, elevated Ca2+ or receptor activation into transgene expression or reporter secretion in HEK293T cells. With a large dynamic range, high light sensitivity and sustained effect, LATeNT enables versatile, spatiotemporally resolved proteolysis across diverse biological systems.\n\nID: 42510934\nTitle: Herbal Bioactives Targeting Rho GTPases: A Multi-Targeted Strategy for Mitigating Neuroinflammation in Alzheimer's and Parkinson's Diseases.\nAbstract: Neuroinflammation plays an essential role in the pathogenesis of several associated brain diseases, including neurodegenerative disorders (Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS)), and traumatic brain injury (TBI). In these diseases, persistent microglial and astrocyte aggregates, elevated proinflammatory cytokines, and oxidative stress drive neuronal injury and cognitive disability. Rho GTPases, in particular the Rho family members Ras homolog family member A (RhoA), Ras-related C3 botulinum toxin substrate 1 (Rac1), and cell division control protein 42 homolog (CDC42), regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity. These proteins are involved in many neuropathological diseases due to dysregulation, making them interesting therapeutic targets. Bioactives used in herbal care have attracted interest for their ability to influence neuroinflammation and even their anti-neurodegenerative activity. Studies show that flavonoids, alkaloids, polyphenols, and other botanical compounds alter Rho GTPase activity, which, in turn, leads to decreased inflammation. This review critically summarizes current evidence regarding phytochemical regulation of Rho GTPase signaling in neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD), with particular emphasis on the underlying molecular mechanisms, context-dependent signaling responses, and current translational challenges. Furthermore, existing knowledge gaps and future research priorities are discussed to facilitate the development of mechanism-based therapeutic strategies targeting Rho GTPases.\n\nID: 42506967\nTitle: Mechanical forces stimulate Golgi export.\nAbstract: Cells face diverse mechanical stimuli that vary with cell type, state, and pathological conditions. Mechanobiology investigates how cells sense and respond to these forces. While most work has focused on the cell surface and nucleus as primary mechanosensors, how intracellular organelles adapt to extracellular mechanical forces remains largely unknown. Here, we show that extracellular mechanical signals influence the secretory function of the Golgi apparatus. By subjecting adherent cells to mechanical challenges-cell spreading on different ligands, altered substrate stiffness, or equibiaxial strain-we reveal that extracellular forces modulate Golgi-to-cell surface carrier biogenesis, thereby regulating exocytosis. Together with changes in Golgi membrane tension, we identify molecular determinants of the mechanotransduction pathway, including microtubule acetylation, diacylglycerol production, and protein kinase D activity. In turn, inhibition of Golgi export suppresses this mechanoresponse and causes impaired cell spreading. These findings uncover a bidirectional mechanotransduction axis in which extracellular mechanics tune Golgi secretory output, providing a framework for investigating organelle-based mechanoadaptation in physiology and disease.\n\nID: 42506700\nTitle: Botulinum Toxin in Parkinson's Disease Tremor: A Critical Evaluation of the Evidence and Clinical Practice.\nAbstract: Approximately 30% of patients with tremor-dominant Parkinson's disease (PD) have rest tremor that persists despite optimal dopaminergic therapy. When deep brain stimulation and focused ultrasound are unavailable or declined, the therapeutic options narrow. Botulinum toxin (BoNT) offers a targeted, titratable, reversible approach, but whether a peripheral neuromuscular blocking agent makes sense for a centrally generated tremor is a legitimate question that deserves a direct answer. This narrative critical review appraises what is currently known across PD and non-PD tremor conditions, defines the technical requirements for safe and effective injection, and provides a practical framework for patient selection and clinical management. The PD-specific literature rests on a single positive double-blind randomized controlled trial of 30 patients; all remaining data are open-label or extrapolated from other tremor conditions, and this narrative synthesis combines heterogeneous conditions, outcome scales, and toxin protocols. A recurring technical observation is that, in the available trials, individualized, EMG-guided injection has been associated with substantially lower rates of hand weakness than fixed-dose injection (reported reductions from roughly 30-70% to below 15%) while maintaining tremor reduction, although the degree of benefit and weakness risk vary with the tremor syndrome, injected muscles, baseline impairment, dose, and guidance method. The careful patient selection this approach requires helps the individual clinician and patient achieve tremor relief, but it departs from the unselected real-world PD population and introduces selection bias that makes a large, statistically representative cohort difficult to assemble. In well-selected patients at centers with the appropriate expertise, BoNT may be a clinically useful option, but routine adoption is not yet supported.\n\nID: 42506696\nTitle: Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.\nAbstract: Essential blepharospasm (BEB) is a focal dystonia characterized by abnormal brainstem excitability and impaired inhibitory control within trigeminal-facial circuits. Botulinum toxin type A (BoNT-A) is the established first-line treatment, primarily acting at the neuromuscular junction. However, whether BoNT-A also modulates central brainstem circuits in BEB patients remains unclear, and dedicated neurophysiological studies have yielded conflicting results. To investigate whether BoNT-A modulates brainstem interneuronal excitability in BEB using the blink reflex recovery cycle and to correlate clinical outcomes with neurophysiological results. Thirteen patients with BEB underwent neurophysiological and clinical evaluation before (T0) and one month after (T1) BoNT-A treatment. The blink reflex recovery cycle was assessed at interstimulus intervals (ISIs) of 200, 300, 500, and 1000 ms. Clinical severity was assessed using the BSPSS, JRS, and BDS scales. The R2 amplitude ratio showed a statistically significant decrease after treatment across all ISIs (all p \u2264 0.001), indicating reduced brainstem interneuronal excitability. All clinical scales demonstrated statistically significant improvement after treatment (BSPSS, JRS, BDS; all p < 0.001). This pilot study provides preliminary evidence that BoNT-A treatment may reduce brainstem interneuronal excitability in BEB patients, as evidenced by a substantial and consistent decrease in R2 amplitude ratios of the blink reflex recovery cycle. These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action. Larger controlled studies are warranted to confirm these results.\n\nID: 42506058\nTitle: Efficacy and Safety of a Tailored Dosing Strategy with High-Dose IncobotulinumtoxinA at Flexible Injection Intervals for Cervical Dystonia: An Open-Label, Uncontrolled, Single-Arm Study in Japan.\nAbstract: This prospective, multicenter, open-label, single-arm study (jRCT2031230690) evaluated the efficacy and safety of a tailored dosing strategy of incobotulinumtoxinA, including high doses (up to 500 U) and flexible injection intervals (as short as 6 weeks), in Japanese patients with cervical dystonia (CD). Of 30 enrolled patients, Group A included 27 patients with idiopathic CD for the primary evaluation of efficacy and safety, whereas Group B included 3 patients with tardive dyskinesia (cervical) or tardive CD for exploratory safety assessment. Patients received up to seven injection cycles of incobotulinumtoxinA (120-500 U) over 48 weeks, with minimum 6-week intervals. The primary endpoint, evaluated in Group A, was the change in Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) total score from baseline to Week 4 after the first injection. Using a mixed model for repeated measures, the least squares mean \u00b1 standard error of the change was -11.0 \u00b1 1.77 (95% confidence interval: -14.6, -7.3). The primary efficacy endpoint was achieved in Group A. Due to the small sample size (n = 3), efficacy in Group B was evaluated only for exploratory purposes, although safety findings were broadly consistent with those in Group A. The overall safety profile was consistent with previous studies. Across the study, the most common related adverse events were dysphagia (33.3%) and muscular weakness (22.2%) in Group A and dysphagia (33.3%) in Group B. All cases of dysphagia were mild to moderate in severity and transient, with no apparent dose- or injection interval-related trend observed. IncobotulinumtoxinA was associated with improvements in symptoms and manageable safety profile at high doses and flexible injection intervals in Japanese patients with CD. While these findings suggest a potential treatment option for individualized dose optimization, the absence of a control group and the exploratory nature of the assessment in Group B necessitate cautious interpretation.\n\nID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS.\n\nID: 42501340\nTitle: Dentato-Rubro-Thalamic Tract-Targeted Cerebellar Deep Brain Stimulation for Post-Stroke Spasticity and Dystonia: preliminary experience.\nAbstract: Introduction Post-stroke motor impairment with spastic hemiparesis remains a significant therapeutic challenge, as conventional rehabilitation and pharmacological strategies often fail to achieve sustained functional recovery. Neuromodulation targeting cerebellar pathways, particularly the dentato-rubro-thalamic tract (DRTT), may provide an alternative therapeutic approach. Methods The study reports the first three cases of chronic post-ischemic stroke-related spastic hemiparesis and dystonic features treated with deep brain stimulation (DBS) targeting the DRTT at the level of the dentate nucleus after unsuccessful botulinum toxin therapy and intensive rehabilitation. Directional DBS systems were implanted using tractography-guided targeting. Clinical outcomes were assessed preoperatively and during sequential stimulation at 130 Hz, 70 Hz, and 30 Hz (each for six weeks), followed by a three-week stimulation-off phase. Outcome measures included the Modified Ashworth Scale (MAS), Fugl-Meyer Assessment of the Upper Limb (FMA-UL), Chedoke Arm and Hand Activity Inventory (CAHAI), Unified Dystonia Rating Scale (UDRS), and modified Rankin Scale (mRS). Results FMA-UL scores progressively improved as stimulation frequency decreased, with mean improvements of 12.3% at 130 Hz, 14.7% at 70 Hz, and 18.4% at 30 Hz. A partial decline was observed during the stimulation-off phase (12.2%). Functional performance (CAHAI) showed the greatest improvement at 30 Hz (11.9%) and decreased to 6.6% during the stimulation-off period. Proximal spasticity was modestly reduced in the shoulder and elbow by approximately one MAS point during stimulation but returned to baseline stimulation-off. Distal spasticity remained largely unchanged. Dystonic features demonstrated frequency-dependent improvement, most pronounced at 30 Hz (43.9%), with partial persistence after stimulation discontinuation (27.5%). Global disability (mRS) remained stable or showed mild improvement. Conclusions Preliminary findings suggest that DRTT-targeted cerebellar DBS, particularly at lower frequencies, may offer a promising adjunctive treatment for post-stroke spasticity and motor impairment refractory to conventional therapies. These initial observations from an ongoing clinical trial require confirmation in larger cohorts with extended follow-up.\n\nID: 42497665\nTitle: AGSI: Adaptive group-enhanced strategy for iterative integration of single-cell multi-omics.\nAbstract: Single-cell multi-omics data integration is critical for understanding cellular heterogeneity and disease mechanisms. However, current methods face two key limitations: (1) uniform evaluation of cross-modal correspondence across all genes, neglecting the modular organization of biological systems, and (2) static integration strategies that fail to accommodate varying degrees of cell-level heterogeneity. To address these challenges, this study proposes AGSI, an adaptive framework for robust multi-omics integration through co-regulated gene modules and iterative reliability assessment. AGSI employs Latent Dirichlet Allocation to identify co-regulated gene modules and evaluates cross-modal correspondence at the module level. AGSI combines Wasserstein-enhanced similarity metrics with dual reliability modeling to progressively identify and integrate cells with high cross-modal concordance. Adaptive thresholding dynamically adjusts selection criteria throughout the iterative refinement process. Extensive experiments on multiple datasets including PBMC, SNARE-seq mouse brain, 10x mouse brain, and large-scale human myocardial infarction data demonstrate that AGSI significantly outperforms seven state-of-the-art methods. Notably, AGSI achieves up to 25.6% F1 improvement over its ablation baseline and 11.9% over the best competing method on complex neural datasets, and maintains over 85% accuracy even under 50% data dropout. AGSI provides a robust and scalable solution for multi-omics integration that preserves biological interpretability while achieving superior technical performance. AGSI is well suited to biomedical analyses requiring accurate cell type identification. The implementation code is available at https://github.com/CDMBlab/AGSI.\n\nID: 42497206\nTitle: VAMP7-dependent mitochondria-lysosome contacts contribute to glial mitochondrial dynamics and dopaminergic neuron survival.\nAbstract: Although disrupted mitochondrial dynamics in neurons are closely linked to neurodegenerative diseases, far less is known about how mitochondrial dynamics are regulated in glia or whether glial mitochondrial dysfunction contributes to neurodegeneration. Here, we show that the R-SNARE protein VAMP7 regulates the untethering of mitochondria-lysosome contacts (MLCs) in adult fly glia. Glial-specific knockdown of VAMP7 leads to prolonged MLCs and mitochondrial elongation associated with altered fission/fusion dynamics. These VAMP7-deficient mitochondria exhibit hyperpolarized membrane potential, leading to increased reactive oxygen species production, lipid droplet accumulation, and dopaminergic neurodegeneration. Mechanistically, VAMP7 interacts with the GTPase-activating protein TBC1D15-17 to promote Rab7 GTP hydrolysis. Without VAMP7, TBC1D15-17 remains bound to Rab7 but fails to activate its hydrolysis, resulting in elevated GTP-bound Rab7 and impaired MLCs untethering. Consistently, expression of GTP-locked Rab7Q67L or GTPase-activating protein-dead TBC1D15-17\u0394GAP phenocopies the mitochondrial defects, while GDP-bound Rab7T22N or wild-type TBC1D15-17 restores the MLC dynamics. Considering that SNARE proteins mediate membrane fusion, our results demonstrate a role for VAMP7 in glial mitochondrial dynamics via organelle contacts, impacting neuron survival in a non-cell-autonomous manner.\n\nID: 42491147\nTitle: Exosome biogenesis in arthropods: conserved mechanisms, experimental evidence, and emerging directions.\nAbstract: Extracellular vesicles (EVs) are nanoscale, lipid-bound structures released by cells across all domains of life. Once viewed as a means for discarding unwanted cellular components, they are now understood to be central mediators of intercellular communication. Much of what is known about EVs comes from mammalian systems, where extensive work has defined the major EV subtypes and the mechanisms that generate exosomes. These findings continue to serve as the primary reference for interpreting EV biology in other organisms. EVs have been isolated from diverse arthropods, including a few insect and tick species, yet the molecular pathways that produce them remain less characterized. Current evidence from Drosophila, mosquito cell lines, and tick systems shows that many of the core components driving exosome biogenesis in mammals, including ESCRT complexes, tetraspanins, lipid-modifying enzymes, Rab GTPases, and SNARE proteins, are present and, in several cases, experimentally validated in arthropods. These findings point to broad conservation of exosome biogenesis across taxa, while also highlighting key caveats, with most conclusions relying on only a small number of model systems. The definitions of EV subtypes remain unclear in many arthropods, and alternative EV biogenesis pathways have received little attention. Future studies that incorporate non-model species, apply rigorous EV characterization standards, and explore the roles of various EV subtypes will clarify how these pathways operate across arthropod lineages and how they differ from well-studied mammalian systems.\n\nID: 42482185\nTitle: Hormonal, metabolic, and stress-related clinical differences between episodic and chronic migraine: insights for diagnostic and therapeutic approaches.\nAbstract: Migraine, a common neurological disorder, is categorized into episodic (EM) and chronic (CM) forms. Understanding their hormonal and metabolic differences can enhance insights into migraine pathophysiology and guide clinical approaches. To explore hormonal and metabolic imbalances in EM and CM patients and identify key factors contributing to migraine chronicity. This retrospective, cross-sectional study included 500 patients with migraine (318 EM and 182 CM). Data were retrospectively extracted from pre-existing clinical records, structured intake forms, physician examination notes, questionnaire-based assessments, and available laboratory results to evaluate demographic, clinical, metabolic, stress-related, mental health-related, and comorbidity variables. CM patients were older (46.28 vs. 43.13 years, p\u2009=\u20090.002) and had more frequent severe attacks (15 vs. 8/month) and mild attacks (12 vs. 6/month) (p\u2009<\u20090.001). CM patients exhibited higher rates of anaemia (34.9% vs. 25.7%, p\u2009=\u20090.035), asthma (12.8% vs. 7.3%, p\u2009=\u20090.046), and sleep apnea (4.7% vs. 1.3%, p\u2009=\u20090.034). Fibromyalgia-related symptoms were common in both groups (82.1% in CM vs. 78.2% in EM), while the number of fibromyalgia tender points was higher in CM than in EM patients (median 8 vs. 6). Ferritin levels were lower in CM but not significantly so (p\u2009=\u20090.139). Suicidal thoughts and feelings of helplessness were more common in CM (p\u2009=\u20090.016, p\u2009=\u20090.002). CM patients used Botox more often but with lower efficacy and engaged in more cognitive behavioural therapy (p\u2009=\u20090.002). CM was associated with more severe symptoms, higher comorbidity burden, more widespread pain features, and distinct metabolic and stress-related profiles compared with EM. These findings support the early recognition of clinical and comorbid factors associated with the chronic migraine phenotype.\n\nID: 42473572\nTitle: Access to Parkinson's disease treatments in the Middle East: a multi-national survey of availability, affordability, and utilization patterns.\nAbstract: Parkinson's disease (PD) is an increasing public health challenge in the Middle East, yet regional data on access to therapies and services remain limited. Understanding the gaps in PD treatment availability, affordability, and prescription frequency is necessary for improving care delivery and regional policy development. Evaluating the availability, affordability, and utilization of disease-related pharmacological treatments, surgical, device-assisted therapies, and rehabilitative services across multiple Middle Eastern countries. A cross-sectional, web-based survey was distributed to physicians with expertise in movement disorders across participating countries in the Middle East. The survey assessed three domains of availability, cost, and utilization of key medications, surgical interventions, and specialized PD services. Thirty-one respondents from nine Middle Eastern countries, including 44% movement-disorder specialists. Medication availability was 65.4%, affordability 68.6%, and prescription 52% across the included countries. Levodopa was nearly universally available (98%), whereas newer dopamine agonists, MAO-B inhibitors, and COMT inhibitors were available inconsistently. Advanced therapies demonstrated disparities; botulinum toxin and deep-brain stimulation were available in high-income countries but not frequently used, while infusion therapies were mostly unavailable. Telemedicine uptake was moderate; genetic testing was cost-restricted. Although general rehabilitation was available, PD-specific services were limited. Affordability constrained equitable care. Qatar had the highest reported prescription frequency of PD medications among the countries. The survey highlights disparities in access to essential PD therapies across the Middle East. Variable availability, costs, and limited specialized services prevent optimal care, urging the need for stronger regional policies, improved insurance coverage, and expanded clinical training to improve equity in PD management.\n\nID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential.\n\nID: 42453424\nTitle: PYGL-driven glycogenolysis impairs microglial autophagic flux via SNAP29 O-GlcNAcylation in Alzheimer's disease.\nAbstract: Aberrant metabolic alterations underlie microglial dysfunction, which plays an important role during neurodegenerative progression. However, the role of aberrant glycogen metabolism remains elusive. Here, we identified glycogen accumulation and upregulated glycogenolytic enzymes in brain microglia from patients with Alzheimer's disease (AD) and transgenic animal models. Particularly, the principal microglial glycogenolytic enzyme PYGL exhibited the most notable spatiotemporal upregulation during disease progression. Specific knockdown of microglial PYGL ameliorated neuropathological changes and cognitive deficits in AD mice. Bioinformatics analysis and experimental validation confirmed that enhancing microglial autophagic flux-dependent A\u03b2 clearance was the underlying mechanism. Furthermore, among all possible glycogenolytic pathways, PYGL downregulation primarily reduced hexosamine biosynthesis pathway activity, diminished UDP-GlcNAc and O-GlcNAcylation of the autophagy key protein SNAP29, and thereby facilitated formation of the SNARE complex, which is essential for autophagosome-lysosome fusion. These findings reveal a glycogenolysis-driven post-translational pathway regulating microglial autophagy, establishing PYGL as a therapeutic target for AD.\n\nID: 42448693\nTitle: Dorsal horn DCC amplification loop induced by endplate osteoclasts generates chronic nociplastic low back pain in male mice.\nAbstract: Spine degeneration is associated with low back pain (LBP), a major cause of disability and functional decline. However, effective therapies for non-specific LBP remain limited because its underlying mechanisms are poorly understood. Here, we show that spine degeneration induces netrin-1 secretion in porous endplates to activate DCC signaling in dorsal root ganglion (DRG) neurons in both LBP and naturally aged male mice. Conditional loss of netrin-1 in Trap+ osteoclasts, or Dcc in Avil+ DRG neurons reduced spinal hypersensitivity. Importantly, DCC activation induces Src phosphorylation and SNARE complex docking, leading to presynaptic glutamate release in the dorsal horn (DH). Consequently, presynaptic glutamate release induces expression of both netrin-1 and DCC in DH neurons as a form of postsynaptic plasticity that initiates a positive feedback loop to amplify spinal hypersensitivity. Moreover, netrin-1/DCC expression is also amplified in the parabrachial nucleus (PBN) in the brain in both LBP and aged male mice. Thus, osteoclast-derived netrin-1 in porous endplates activates DCC-mediated hypersensitivity in DRG neurons to be further amplified in the DH neurons. These results promote the concept of non-specific LBP as a form of nociplastic pain.\n\nID: 42446153\nTitle: Sensory Tricks in Dystonia: A Systematic Review and Nested Quantitative Synthesis.\nAbstract: Sensory tricks (also termed \"alleviating maneuvers\") are voluntary maneuvers that transiently alleviate dystonic postures or movements and represent a hallmark clinical feature of dystonia with diagnostic and mechanistic significance. Despite extensive phenomenological description, the evidence base has not been systematically synthesized. To characterize the prevalence, phenomenology, clinical effects, predictors, neurobiological mechanisms, and translational relevance of alleviating maneuvers in dystonia, and to prospectively evaluate the feasibility of quantitative synthesis across predefined outcome domains. A systematic review with a prespecified nested quantitative synthesis was conducted using a preregistered PROSPERO protocol (CRD420251175065). The nested component was defined a priori as a domain-restricted analytic strategy in which random-effects meta-analysis was planned where \u2265\u00a03 conceptually comparable studies were available, and in which the feasibility of pooling was itself treated as a primary methodological outcome. PubMed, Scopus, and the Cochrane Library were searched from inception. Studies reporting original empirical data on alleviating maneuvers in human dystonia of any etiology were included. Qualitative synthesis examined phenomenology, clinical effects, predictors, mechanisms, and translational applications. Heterogeneity was examined separately along clinical, methodological, and outcome-level dimensions. Of 631 records, 53 studies met inclusion criteria; 31 contributed to qualitative synthesis and 26 to nested quantitative analyses, with 4 contributing to both. No eligible studies in functional or tardive dystonia were identified. Alleviating maneuvers were reported across dystonia subtypes, with prevalence ranging from 13% to 90%; the largest registry cohort (n = 1477) reported effective maneuvers in 68.7% of patients. Acute motor improvements of 30%-50% in head deviation were reported during trick execution, although effects were transient. Responsiveness was associated with shorter disease duration and better response to botulinum toxin. No domain met criteria for formal meta-analysis; structural barriers were predominantly methodological and outcome-level rather than clinical. Alleviating maneuvers are a clinically consistent and mechanistically informative feature of dystonia, but their magnitude, durability, and predictors remain inconsistently measured. Standardized paradigms and outcome reporting are required to enable quantitative synthesis and sham-controlled evaluation of device-based alleviating-maneuver analogues.\n\nID: 42440475\nTitle: Therapeutic progress in rosacea: targeting the neuro-vascular-immune triad.\nAbstract: Rosacea is a chronic inflammatory dermatological condition predominantly affecting the facial region. Clinically, rosacea is characterized by paroxysmal flushing, persistent erythema, telangiectasia, papules, pustules, and ocular symptoms. Severe rosacea cases are frequently complicated by anxiety, depression, and sleep disturbances, imposing a heavy psychosocial burden and markedly impairing the quality of life of the patients. Abnormal activation of the neuro-vascular-immune triad has been reported as a crucial pathological mechanism of rosacea. Moreover, dysregulation within the central nervous system might exacerbate disease progression by modulating peripheral nerve activity and neuroendocrine homeostasis. In this review, we summarize the latest advancements in rosacea treatments targeting the neuro-vascular-immune triad, including \u03b3-aminobutyric acid derivatives, antidepressants, anti-calcitonin gene-related peptide agents, physical neuromodulation (transcutaneous auricular vagus nerve and repetitive transcranial magnetic stimulations), botulinum toxin type A, and \u03b2-blockers. We provide a comprehensive analysis of their molecular mechanisms, clinical efficacy, and current limitations. While such therapies could specifically regulate different stages of the pathway, their evidence lacks large-scale randomized controlled trials and clarity regarding optimal dosing regimens and treatment durations. Future studies should strengthen basic investigations and clinical translation, explore combined therapeutic strategies, as well as develop personalized therapeutic strategies for the long-term effective control of rosacea.\n\nID: 42440317\nTitle: Plasma Phosphorylated Tau 217 in Participants at Risk for Chronic Traumatic Encephalopathy.\nAbstract: In vivo biomarkers for detecting neuropathologies from repetitive head impacts (RHI), including chronic traumatic encephalopathy (CTE), are needed. To evaluate the utility of plasma phosphorylated tau 217 (p-tau217), assess its performance as a beta-amyloid (A\u03b2) biomarker in participants with RHI exposure at risk for CTE, and explore concordance with CTE neuropathology in a postmortem subsample. This longitudinal, multicenter, case-control study used data from the Diagnostics, Imaging, and Genetics Network for the Objective Study and Evaluation of CTE (DIAGNOSE CTE) Research Project, collected from September 2016 to October 2023. Participants were former American football players (case participants) and asymptomatic men unexposed to RHI (control participants). A subsample had available neuropathologic data. RHI, traumatic encephalopathy syndrome (TES) diagnoses, and levels of CTE certainty. Plasma p-tau217 (classified as positive [\u22650.63 pg/mL], intermediate [0.40-0.62 pg/mL], and negative [<0.40 pg/mL]), A\u03b2-positron emission tomography (PET; 18F-florbetapir; with A\u03b2-positive defined as a standardized uptake value ratio [SUVR] \u22651.10), and tau-PET (18F-flortaucipir). TES diagnoses were assigned by multidisciplinary consensus conference. Analyses of postmortem brains controlled for age, race, and APOE \u03b54 status. Among 231 participants (mean [SD] age, 57.75 [8.25] years), 177 were former football players (117 professional and 60 college) and 54 were unexposed participants. Former football players had higher baseline mean (SD) p-tau217 concentrations than unexposed participants (0.35 [0.26] pg/mL vs 0.27 [0.14] pg/mL; P\u2009=\u2009.008), although this was driven by a higher proportion of A\u03b2-PET-positive participants among former players. Plasma p-tau217 increased over time across the sample (B\u2009=\u20090.207 [95% CI, 0.117-0.298]; P\u2009<\u2009.001), with no significant time\u2009\u00d7\u2009exposure group interactions. Among football players, p-tau217 showed no time\u2009\u00d7\u2009group interactions with TES diagnosis, TES-CTE certainty, or RHI metrics. Higher p-tau217 concentration correlated with higher global A\u03b2-PET SUVR (B\u2009=\u20090.058 [95% CI, 0.053-3.501; P\u2009=\u2009.01), with a few discordant cases (5 participants were p-tau217-negative and A\u03b2-PET-positive; 7 participants were p-tau217-positive and A\u03b2-PET-negative). P-tau217 had similar areas under the curve for projecting A\u03b2-PET positivity as cerebrospinal fluid (CSF) p-tau181/A\u03b242 and CSF A\u03b240/42 measures (p-tau217: AUC, 0.88 [95% CI, 0.80-0.96]; CSF p-tau181/A\u03b242: AUC, 0.89 [95% CI, 0.79-1.00]; CSF A\u03b240/42: AUC, 0.85 [95% CI, 0.72-0.98]). Among 9 brain donors, 6 had CTE (stages II-IV; none with Alzheimer disease). Seven had negative or intermediate p-tau217, concordant with A\u03b2-PET. Two p-tau217 outliers with stage III CTE had normal concentrations upon additional testing. The findings of this study suggest that plasma p-tau217 concentration is unlikely to be useful for the detection of CTE, but it does show utility for ruling out A\u03b2 pathology in participants at risk for CTE.\n\nID: 42432065\nTitle: Control of synaptic communication through molecularly engineered bioluminescence light emission and sensing.\nAbstract: Synapses are sites of intercellular communication between neurons and from neurons to target organs, and of signal integration that underly physiological and behavioral responses. We have developed a modular platform, Interluminescence (Int), for experimental control of synaptic transmission: bioluminescent light, generated by a luciferase oxidizing a luciferin, from a presynaptic neuron is used to activate transsynaptic optogenetic ion channels in the postsynaptic neuron. Two strategies can modulate postsynaptic neurons in the presence of luciferin. In the 'Act-Int' approach, a luciferase is genetically expressed in synaptic vesicles and released during depolarization-induced presynaptic vesicle fusion and exocytosis. In the 'Persist-Int' approach, a luciferase is tethered to the presynaptic membrane where it can support sustained transsynaptic signaling. Both strategies can activate postsynaptic neurons with comparable efficacy under the conditions tested. By design, the modularity of the platform permits the use of luciferases and opsins ranging in brightness and light sensitivity, with the luciferase targeted to different subcellular regions of the presynaptic neuron, and the postsynaptic opsin being excitatory or inhibitory. Our results demonstrate the utility and versatility of Interluminescence to mediate synaptic transmission that is either activity-dependent or activity-independent.\n\nID: 42427581\nTitle: Ontological Analysis of Brain Proteostasis Highlights the Sex-Dependent Trajectory of ApoE Isoform-Specific Regulation.\nAbstract: Apolipoprotein E (ApoE) is the strongest genetic predictor of Alzheimer's disease (AD) risk, with ApoE4 increasing and ApoE2 decreasing risk relative to ApoE3. Using a global LC-MS proteomic approach, we integrated protein abundance and kinetics in Human-APOE knock-in mice for young (3-month) and aged (18-month) cohorts to quantify the changes in steady-state proteostasis. By mapping 6,052 identified proteins and 3,986 associated turnover rates into ontological groups, we observed that vesicle trafficking and mitochondrial dysregulation occur as early as 3 months in ApoE4 mice accompanied by hyperactive metabolism that eventually reduces with age. In contrast, young and old ApoE2 mice retain similar signatures to ApoE3 mice in metabolic, mitochondrial, cellular regulation, and membrane trafficking ontologies. We found that females had more isoform-induced ontological changes relative to ApoE3, providing insight into sex-dependent vulnerabilities. Our global proteomic approach for ApoE proteostasis crucially unifies independent literature observations while providing turnover kinetics to uncover the underlying mechanism behind abundance changes. Data are available via ProteomeXchange with identifier PXD079261.\n\nID: 42427545\nTitle: Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.\nAbstract: Synaptojanin 1 is a brain enriched phosphoinositide phosphatase implicated in endocytosis at the synapse. A mutation (R258Q) that selectively impairs its Sac1 phosphatase domain causes early onset familial Parkinsonism. Neurons of mice with this mutation display synaptic vesicle traffic defects across the brain, but selective dystrophic changes in a subset of dopaminergic axons in the dorsolateral striatum. Using correlative light microscopy-FIB-SEM of mutant mouse striata to visualize in 3D these abnormal structures we show that they represent clusters of focal axonal dilations harboring massive, onion-like DAT enriched plasma membrane infoldings, generally localized next to cell bodies of neighboring cells, often engulfing evaginations of such cells. This dysmorphia was associated with a deficit in dopamine release in the same striatal region. Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis. Their occurrence only in a subset of axons suggest a vulnerability threshold of these axons beyond which the expansion of the plasma membrane is not counteracted by compensatory mechanisms.\n\nID: 42424539\nTitle: Redefining Spasticity: The Spasticity X Working Group Consensus Statement.\nAbstract: Inconsistent and variably interpreted definitions of spasticity, alongside evolving mechanistic understanding, highlight the need for a clear, clinically relevant consensus definition. An international expert panel used a modified Delphi process to develop a concise, clinically applicable definition that reflects current understanding and supports consistent assessment and management. Participants reviewed existing definitions, completed a pre\u2011meeting survey, and engaged in structured discussions, with draft definitions iteratively refined through successive rounds of voting to achieve consensus. Key components identified included disordered sensorimotor control, central nervous system involvement, and velocity\u2011 and length\u2011dependent resistance to passive stretch, while existing definitions were considered either overly narrow or insufficiently relevant to clinical practice. The consensus definition characterizes spasticity as \"A disorder of sensorimotor control resulting from upper motor neuron disease. It is characterized by velocity- and length-dependent involuntary muscle overactivity, which is intermittent or sustained, during passive stretch.\" This definition integrates contemporary mechanistic concepts with clinical applicability and is intended to improve conceptual clarity, facilitate communication, and promote consistency in diagnosis, measurement, and treatment.\n\nID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes.\n\nID: 42422202\nTitle: Long-term efficacy of neural circuit blockade for treating blepharospasm: a retrospective case series study.\nAbstract: Blepharospasm is a common symptom of Meige syndrome and often leads to functional blindness. Although current treatment options, such as oral medications, botulinum toxin injections, resection of periocular muscles, and deep brain stimulation, are available, no definitive method effectively eliminates blepharospasm. To address this unmet need, we developed a novel approach-neural circuit blockade (NCB). This study aimed to analyse its long-term efficacy for blepharospasm. NCB eliminates trigeminal nerve stimulation by facial expression muscles, reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions, decreasing orbicularis oculi muscle tension, and alleviating blepharospasm. A retrospective analysis was conducted on 570 patients who underwent NCB treatment, with a median postoperative follow-up of 51\u202fmonths. Therapeutic efficacy was evaluated by comparing preoperative and postoperative scores on the Shorr blepharospasm grading scale, Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS-M), and Blepharospasm Disability Index (BSDI). Based on the Shorr grading scale, preoperatively, 97.9% of patients were grade 4, whereas 2.1% were grade 3. Postoperatively, 87.5% achieved complete remission (grade 0), whereas 7.4, 3.7, 0.9, and 0.5% were grades 1, 2, 3, and 4, respectively. Pre- and postoperative BFMDRS-M scores significantly differed: eye (8.0 vs. 0.0), mouth (3.0 vs. 0.0), speech/swallowing (3.0 vs. 2.0), neck (3.0 vs. 1.5), and total (8.0 vs. 0.0). BSDI scores also significantly differed pre- and postoperatively. The results indicate that NCB effectively alleviates blepharospasm and addresses the lack of a definitive treatment, without causing severe local or systemic complications such as facial paralysis. NCB can safely and effectively alleviate blepharospasm, improve quality of life, and demonstrate stable long-term efficacy.\n\nID: 42404278\nTitle: Primary Cerebellar Agenesis in a Child with Esotropia: A Case Report and Literature Review.\nAbstract: To highlight the importance of neuroimaging in children with esotropia and developmental delay and to support botulinum toxin as a minimally invasive treatment option when major brain malformations are present. We describe an 18-month-old girl with infantile esotropia and mild developmental delay. Brain magnetic resonance imaging revealed near-complete cerebellar agenesis with a hypoplastic pons. Considering this malformation, bilateral medial rectus botulinum toxin injections (15 units in 0.1 mL per muscle, freehand technique) were performed instead of conventional strabismus surgery. Ocular alignment improved with satisfactory cosmetic appearance, and stability was maintained throughout 4 months of follow-up. Genetic testing was not performed at the time because of follow-up limitations; however, it is recommended for further etiological clarification. This case highlights the importance of neuroimaging in children with esotropia and developmental delay and supports botulinum toxin as a minimally invasive treatment option when major brain malformations are present.\n\nID: 42401592\nTitle: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.\nAbstract: The function of the SNARE complex regulator, Munc18-1, in photoreceptor cells is unknown. Here, we found that removing Munc18-1 from photoreceptors results in major degeneration starting at P14. In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1. Furthermore, Munc18-1 played a critical role in expression and localization of syntaxin-3. The syntaxin-3 protein level is dramatically reduced in the soma and plasma membrane of photoreceptors without Munc18-1. At the photoreceptor synapses, the colocalization of syntaxin-3 and its SNARE partner, SNAP-25, was reduced, potentially suggesting an altered syntaxin-3 synaptic localization. In the Munc18-1-deficient photoreceptors, immature synapses and outer segment lesions were found. Taken together, these findings provide evidence that Munc18-1 is important for maintaining sufficient syntaxin-3 expression in the cell body and synapses of photoreceptors. The lack of Munc18-1, combined with poor syntaxin-3 expression, contributes to photoreceptor functional impairment and degeneration.\n\nID: 42400067\nTitle: Ultrasound-guided botulinum toxin injections for abdominal spasticity and dyspnea in an ambulatory patient with an incomplete cervical spinal cord injury: a case report.\nAbstract: Abdominal spasticity in spinal cord injury (SCI) patients may remain unrecognized, despite its impact on quality of life. We report a 47-year-old white Dutch man with incomplete C4 SCI (AIS D) presenting with dyspnea, abdominal pain, and discomfort from involuntary abdominal muscle contractions. Five year post-injury, the patient developed bloating and pronounced abdominal pain, pain in the lower back, and in the intercostal muscles. In addition, a 7-cm diastasis developed in the abdominal midline. These complaints reduced ambulatory capacity and quality of life. Initial management with systemic antispasticity treatment (tolperisone 50\u00a0mg, 3\u00d7/day) relieved leg spasticity and closed the diastasis, yet failed to alleviate dyspnea. Ultrasound imaging confirmed spasticity predominantly in the internal oblique and transverse abdominal muscles. Given the persistence of abdominal spasticity complaints, we initiated a series of ultrasound-guided botulinum toxin (100-150E incobotulinum toxin-A) injections to the internal and external oblique muscles-with one session also targeting transversus. The injection protocol was refined over successive treatments to achieve an optimal distribution while minimizing side effects on core stability. Although the patient experienced transient balance issues and lower back pain during the initial adjustment period, these effects were minor and resolved within 2-3\u00a0weeks. This case underscores that in ambulatory SCI patients, systemic treatments alone may not sufficiently address abdominal spasticity and concomitant symptoms like dyspnea. Low-dose ultrasound-guided botulinum toxin injections targeting the abdominal muscles can provide significant symptom relief and improve overall functional capacity, offering a safe and effective addition to low-dose systemic treatment.\n\nID: 42386531\nTitle: Translational Profiling of Drd2-Expressing Populations Reveals Molecular Heterogeneity of Dentate Gyrus Mossy Cells along the Dorsoventral Axis.\nAbstract: Hilar mossy cells (MCs) are crucial for integrating and propagating signals across the hippocampal dorsoventral axis, mediating cognitive and affective processing. While MCs exhibit profound dorsoventral differences in their projections, physiology, and behavioral roles, the molecular basis underlying this functional specialization remains largely unexplored. To address this gap, we used translating ribosome affinity purification (TRAP) in male mice to systematically compare the translatome of Drd2-expressing, MC-enriched populations along the dorsoventral axis. This analysis revealed distinct translational signatures with 1,442 genes enriched in dorsal and 1,337 genes in ventral Drd2-expressing, MC-enriched populations. Pathway analysis demonstrated significant functional segregation along the dorsoventral axis. The dorsal population is notably enriched for genes linked to neuronal connectivity and synaptic transmission, whereas the ventral counterpart shows enrichment in genes associated with energy metabolism and cellular maintenance. Specifically, we identified a subset of dorsal enriched genes, including neurotransmitter receptors, ion channels, and axon guidance regulators, contrasting with ventral enriched genes highly related to glucose/fatty acid metabolism, oxidative phosphorylation, and exocytosis. We further predicted distinct sets of upstream transcriptional regulators activated in each subpopulation, providing insights into the regulatory networks that may drive molecular divergence. Our findings provide a translatomic basis for the dorsoventral heterogeneity of Drd2-expressing neurons that include MCs, offering molecular signatures associated with their differential contributions to hippocampal function.\n\nID: 42384787\nTitle: Turbocharging synaptic transmission: 12 SNAREpins are required for rapid release of reconstituted synaptic vesicles.\nAbstract: Synaptic transmission occurs synchronously with real-world events, far faster than vesicle fusion for hormone release or membrane biogenesis, all mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. How SNAREs cooperate to achieve synchronous neurotransmitter release is a long-standing mystery. Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors in a fully-defined, genetically validated system that enables single-molecule counting in docked vesicles before release. SNAREpin complexes (12\u00a0\u00b1\u00a00.3) are found in each such ready-release vesicle, suggesting a regular structure. Several genetic conditions (including point mutation of the synaptic vesicle protein Synaptophysin from a Synaptopathy patient and human and mouse disease mutations of the synaptic vesicle protein vesicle-associated membrane protein-2 (VAMP2) reduce the number of SNAREpins to 6\u00a0\u00b1\u00a00.3 and result in profoundly delayed release over 0.1 to 1 seconds. Omitting Synaptophysin, whose hexamers preassemble 12 copies of VAMP2, also yields ~6 SNAREpins and delays release.\n\nID: 42371396\nTitle: Emerging Pharmacological Strategies for Trigeminal Neuralgia.\nAbstract: Trigeminal neuralgia can be a debilitating pain condition that affects women more frequently than men and for which there are only limited therapeutic avenues. Trigeminal neuralgia can either be idiopathic (where there is no clearly identifiable cause), classical (which typically involves neurovascular compression of the trigeminal root entry zone) or secondary which results from nerve damage due to other causes. First-line treatments for trigeminal neuralgia include the sodium channel blockers carbamazepine and oxcarbazepine, whose mechanism of action is to stabilize the inactivated state of sodium channels, thereby reducing the electrical activities of trigeminal neurons. Second-line treatments include gabapentinoids, which act on Cav\u03b12\u03b4 subunits of high voltage activated calcium channels to inhibit synaptic transmission in the trigeminal pain pathway. Both of these treatment avenues exhibit a number of adverse effects and many patients are refractory. Hence, new avenues for combatting trigeminal neuralgia are needed. In this review article, we discuss clinical evidence for efficacy and benefits of novel pharmacological treatment approaches for trigeminal neuralgia, including the inhibition of CGRP receptors, activation and desensitization of TRPV1 channels, the development of new generations of sodium channel blockers, and the use of botulinum toxin. We conclude that while there are potential benefits of these approaches, larger well-controlled cohort studies are needed to provide a definitive answer.\n\nID: 42360457\nTitle: White matter microstructural alterations correlate with clinical characteristics in craniocervical dystonia.\nAbstract: To characterize white matter pathology, identify clinical correlates, and explore relevant biomarkers for deep brain stimulation (DBS) outcomes in craniocervical dystonia (CCD) using diffusion tensor imaging (DTI). We involved 37 CCD patients (22 undergoing bilateral subthalamic DBS [STN-DBS]) and 42 matched healthy controls. Using Tract-Based Spatial Statistics (TBSS) for whole-brain analysis and Automated Fiber Quantification (AFQ) for 20 specific tracts, DTI measurements of fractional anisotropy (FA), axial diffusivity (AD), radial diffusivity (RD) and mean diffusivity (MD) were extracted. Between-group metric comparisons and correlations with clinical features and treatment outcomes were assessed. TBSS revealed reduced FA in left anterior corona radiata and right posterior thalamic radiation, increased AD in splenium of corpus callosum, and widespread elevated MD/RD (p\u2009<\u20090.0125, family-wise error combined threshold-free cluster enhancement corrected). AFQ identified segmental increased MD/RD across several tracts, most notably within anterior thalamus radiation (ATR), corticospinal tract, cingulum cingulate, callosum forceps, and inferior fronto-occipital fasciculus (uncorrected p\u2009<\u20090.01). Microstructural alterations correlated with earlier disease onset, severe baseline blepharospasm, and botulinum toxin exposure (p\u2009<\u20090.01, uncorrected). Crucially, at an uncorrected threshold of p\u2009<\u20090.01, lower MD in the left ATR was associated with greater eye symptom improvement after STN-DBS surgery (r\u2009=\u2009-0.57, p\u2009=\u20090.009). CCD exhibits distributed white matter microstructural alterations across multiple networks, potentially serving as the neurostructural substrate for motor and non-motor symptoms. Left ATR integrity is negatively associated with DBS outcomes.\n\nID: 42353862\nTitle: Clinical and Genetic Spectrum of ANO3-Related Dystonia with Treatment Responses in a Chinese Cohort.\nAbstract: Background: Variants in anoctamin 3 (ANO3) are linked to autosomal dominant dystonia, commonly known as DYT-ANO3 (OMIM: #615034). While craniocervical dystonia constitutes the most frequently observed phenotype, its clinical manifestations display significant heterogeneity. Nevertheless, the data concerning the genetic characteristics, clinical features, and therapeutic outcomes of ANO3-related dystonia within Asian populations remain scarce. Methods: Whole-exome sequencing was conducted on 661 Chinese patients, comprising 356 individuals with dystonia and 305 individuals with non-dystonic movement disorders who served as an internal disease-control cohort. Candidate ANO3 variants were evaluated based on population frequency, predicted deleteriousness, ACMG criteria, and aggregate frequency analyses. A retrospective review was performed of clinical features and treatment responses. Results: Fifteen rare ANO3 missense variants were identified in 16 patients with dystonia and one non-dystonic individual, including one pathogenic variant, six likely pathogenic variants, and nine previously unreported variants. The rare ANO3 variants were significantly enriched in the dystonia cohort compared with the controls. ANO3-related dystonia exhibited broad clinical heterogeneity, with frequent cervical involvement (62.5%) and tremulous features (75%), and occasionally extended beyond classical isolated dystonia. Oral medication and botulinum toxin showed variable benefit, whereas deep brain stimulation was associated with marked improvement in selected medically refractory patients. Conclusions: This study broadens the genetic and clinical spectrum of ANO3-related movement disorders in a Chinese cohort. The findings support substantial clinical heterogeneity in DYT-ANO3 and suggest that deep brain stimulation, especially STN-DBS, may be considered in selected refractory cases, although further studies are needed.\n\nID: 42350670\nTitle: Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.\nAbstract: Liraglutide, a glucagon-like peptide-1 receptor (GLP-1R) agonist for type 2 diabetes and obesity management, shows variable patient responses. We investigated the metabolic state-dependent mechanisms underlying this heterogeneity and how liraglutide's mode of action shifts across stages of metabolic dysfunction. We employed human pancreatic islets from donors across metabolic states (normoglycaemic [HbA1c <42 mmol/l (<6.0%)], glucose intolerance [HbA1c 42-47 mmol/l (6.0-6.4%)] and type 2 diabetes [HbA1c \u226548 mmol/l (\u22656.5%)]) using dynamic perifusion and static incubation techniques to assess glucose-stimulated insulin secretion. GLP-1R mRNA levels were measured in 112 donor islets stratified by HbA1c. Mechanistic investigations used tanycyte-specific GLP-1R knockdown (GLP-1RTanycyteKD) mice and botulinum toxin B-expressing (iBot) mice to distinguish between central and peripheral pathways. Oral glucose tolerance tests, pyruvate tolerance tests and positron emission tomography were performed to assess in vivo metabolic effects. Liraglutide (25 nmol/l) enhanced glucose-stimulated insulin secretion specifically in donors with glucose intolerance (n=7, p=0.021), with no effect in normoglycaemic islets (n=7), despite preserved GLP-1 (7-36) responsiveness. In type 2 diabetes islets, GLP-1R mRNA levels progressively decreased with rising HbA1c (p=0.015, normoglycaemic [n=48] vs type 2 diabetes [n=10]). In chow-fed mice, liraglutide's insulin-stimulating effects required tanycyte-mediated hypothalamic access, as demonstrated by abolished responses in GLP-1RTanycyteKD mice. However, during metabolic dysfunction (a 12-week high-fat diet), direct islet responsiveness was restored independent of tanycyte function. Advanced metabolic disease (a 27-week high-fat diet) maintained islet responsiveness ex vivo while losing in vivo insulin enhancement, revealing insulin-independent glucose-lowering mechanisms involving hepatic gluconeogenesis suppression and enhanced peripheral glucose uptake. Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states. This mechanistic framework enables potential patient stratification in type 2 diabetes therapy, suggesting that matching liraglutide's predominant mechanism to individual metabolic profiles could optimise treatment outcomes.\n\nID: 42543528\nTitle: Encapsulation and Controlled Release of Human Spinal Cord Organoid-Derived Extracellular Vesicles for Tissue Patterning in Viscoelastic Hyaluronic Acid Hydrogels.\nAbstract: Human induced pluripotent stem cells (hiPSCs) can differentiate into various types of central nervous system organoids which are valuable for applications in tissue engineering and injury repair. The secreted extracellular vesicles (EVs) of organoids, in particular the small-sized EV subset referred as exosomes (30-200\u00a0nm), have emerged as novel therapeutics in regenerative medicine. This study investigated the encapsulation and controlled release of human spinal cord organoid (hSCO)-derived EVs in viscoelastic hyaluronic acid (HA) hydrogels and assessed their impact on organoid patterning. A series of pH-responsive hydrogels were fabricated, leading to sustained EV release regulated by viscoelastic properties. The pH of these hydrogels decreased from 9 to 7 during incubation, which altered hydrogel viscoelasticity, thereby modulating EV release kinetics. In addition, EV-loaded hydrogels regulated key hSCO patterning markers such as DBX1 and ISL1. Furthermore, these EVs in hydrogels can cross a modeled blood-spinal cord barrier and provide cross-barrier capability for delivery. Taken together, the organoid-secreted EVs in viscoelastic HA hydrogels can be released at a controlled rate and have potential to regulate spinal cord organoid patterning. This study advances our knowledge of regulating intercellular communication and developing EV-based therapies for treating neurological disorders such as spinal cord injury.\n\nID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development.\n\nID: 42530044\nTitle: Extracellular Vesicle-Mediated Delivery of VEGF and NGF Protects Dopaminergic Neurons in 6-OHDA-Induced Parkinson's Disease Models.\nAbstract: Parkinson's disease (PD) is a neurodegenerative disorder marked by motor dysfunction. No definitive methods exist to repair damaged neurons. Vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) are two neuroprotective agents that work synergistically. However, these large molecular proteins have difficulty crossing the blood-brain barrier (BBB). Extracellular vesicles (EVs) offer superior targeting and low immunogenicity, making them excellent carriers. In this study we examined the protective effects of VEGF and NGF in a cell model and evaluated the therapeutic potential of VEGF-NGF contained within EVs in PD rats. EVs were isolated using sequential differential centrifugation and characterized using transmission electron microscopy, nanoparticle tracking analysis, and western blotting (WB). VEGF and NGF were loaded into the EVs using a saponin-assisted method to create VEGF@EVs, NGF@EVs, and VEGF/NGF@EVs. The viability of 6-hydroxydopamine hydrochloride (6-OHDA)-induced SH-SY5Y cells was measured using the cell counting kit-8 assay before and after treatment with VEGF and NGF. Autophagy levels were assessed using WB, and the role of autophagy was further explored using the autophagy inhibitor chloroquine. Unilateral PD rat models were established via stereotactic injection of 6-OHDA into male Sprague-Dawley rats. Behavioral changes were monitored before and after treatment. Neuronal recovery, neurotransmitter levels, and autophagy levels in the rat brains were evaluated using immunohistochemistry, enzyme-linked immunosorbent assay, and WB. VEGF/NGF@EVs significantly enhanced the viability of 6-OHDA-induced SH-SY5Y cells. A complete autophagic process was identified as essential for this protective effect. The intranasal administration of VEGF/NGF@EVs improved motor behavior in PD rats, with performance better than that of single growth factor treatments. The number of tyrosine hydroxylase (TH)-positive neurons, TH protein expression, and dopamine content were significantly increased. In addition, the level of autophagy in the rat substantia nigra was elevated. VEGF/NGF@EVs exert protective effects in both in vitro and in vivo 6-OHDA-induced PD models by promoting autophagy, demonstrating greater efficacy than either growth factor alone. By transplanting VEGF/NGF@EVs into PD rats, we showed that these vesicles can effectively cross the BBB and deliver targeted therapy to the central nervous system. This study highlights the significant potential of EV-mediated protein transplantation strategies for treating neurological disorders.\n\nID: 42529100\nTitle: Emerging strategies for spinal cord injury repair: stem cells, extracellular vesicles, biomaterials, and neuromodulation.\nAbstract: Spinal cord injury (SCI) is a devastating condition that frequently results in permanent disability. Repair and functional recovery after SCI are hindered by the complexity of its pathophysiology and the inherent challenges of neural regeneration within the central nervous system. Despite decades of research aimed at elucidating its pathological mechanisms and developing effective strategies to promote axonal regeneration and circuit rewiring, successful therapeutic outcomes remain limited. Recent advances in bioactive materials and stem cell technology have shifted the research focus from solely stimulating corticospinal tract regeneration toward building intermediate neural networks to facilitate neural repair and circuit reconstruction. Concurrently, emerging technologies that modulate neural activity through physical modalities, such as electrical, magnetic, and ultrasonic stimulation, are rapidly evolving. Among these emerging strategies, several-including stem cell therapy, biomaterial-based interventions, and electromagnetic stimulation-have advanced to clinical trials, with some already entering clinical practice. This review summarizes the major contemporary strategies and research advances in SCI repair. It details the mechanisms and latest developments in stem cell and extracellular vesicle therapy, biomaterial applications, and neuromodulation techniques. Finally, it discusses future therapeutic directions and ongoing challenges in clinical translation.\n\nID: 42514227\nTitle: Dynamic Tuning of MSC-Based Scaffolds for Neurological Protection After Brain or CNS Injury.\nAbstract: Neurological disorders, including stroke, traumatic brain injury, and spinal cord injury, constitute one of the most important causes of mortality and morbidity worldwide for which current treatment options focus on resolving neuroinflammation rather than on tissue and neuronal regeneration. Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors which modulate microglial activation, preserve blood-brain barrier (BBB) integrity, and neuroplasticity, but with limitations due by poor survival, retention, and phenotypic instability following direct transplantation. The purpose of this narrative review is to present mechanotransduction signaling pathways (integrin-FAK, PI3K/Akt, Rho/ROCK, and YAP/TAZ) through which MSC-based biomaterial scaffolds, especialy hyaluronic acid (HA) hydrogels, make the transition from reparative to regenerative medicine in central nervous system (CNS) injury. Even if most of the evidence from preclinical studies suggests that dynamically tunable MSC-scaffold systems represent promising platforms for neural tissue engineering and regenerative medicine, further translational studies and well-designed clinical investigations are required to establish their therapeutic efficacy and clinical applicability.\n\nID: 42511709\nTitle: Wallerian Degeneration and Nerve Regeneration-A Review of Cellular and Molecular Events.\nAbstract: Wallerian degeneration (WD), which occurs distal to peripheral nerve injury, is a tightly regulated process. Axonal degeneration during the onset of WD represents a self-destructive process that begins with an early phase and progresses to an execution phase characterized by fragmentation of axons and myelin sheaths. Efficient clearance of axonal and myelin debris, together with the reprogramming of Schwann cells and macrophages into a repair phenotype, constitutes a critical extrinsic prerequisite for successful axonal regeneration. In parallel, signaling molecules produced during WD trigger intrinsic responses in injured neurons that are essential for neuronal survival and the initiation of the regenerative program. This review provides an overview of the key events involved in WD, which are often studied separately, with the aim of elucidating their interrelationships and their impact on nerve regeneration. Such an integrated overview may aid in identifying molecular targets for the development of novel therapeutic strategies to enhance axonal regeneration.\n\nID: 42507332\nTitle: Disease mechanisms and translational barriers guide nanocarrier design for nose to brain delivery in Alzheimer's disease.\nAbstract: Alzheimer's disease (AD) is a progressive neurodegenerative disorder with limited disease-modifying treatment options, partly because many therapeutic agents show insufficient brain exposure and dose-limiting systemic adverse effects after conventional administration. Nose-to-brain (N2B) delivery has emerged as a non-invasive strategy to transport therapeutics to the central nervous system through the olfactory and trigeminal pathways, thereby partially bypassing the blood-brain barrier. Recent advances in nanomedicine and biomaterial engineering have further improved this approach by enhancing drug stability, nasal residence, mucosal transport, and brain-targeting efficiency. This review examines nanocarrier-enabled N2B delivery strategies for AD from a mechanism-guided perspective, highlighting how AD-related pathological processes shape the selection of therapeutic cargos and formulation designs. We discuss recent progress in the intranasal delivery of repurposed small molecules, natural products, insulin-related agents, peptides and proteins, extracellular vesicles, antibodies, and nucleic acid-based therapeutics. We further summarize major nanocarrier and formulation platforms, including lipid-based systems, polymeric nanoparticles, micelles, extracellular vesicles, in situ gels, and device-assisted delivery technologies. Particular attention is given to the design parameters that influence N2B performance, including particle size distribution/PDI, surface charge, mucus interaction, cargo protection, targeting modification, biodistribution, and deposition reproducibility. Finally, we critically evaluate the translational challenges that continue to limit clinical application, including species differences in nasal anatomy, dose-volume restrictions, device-dependent variability, limited human pharmacokinetic evidence, manufacturing complexity, long-term safety, and regulatory requirements. By integrating disease mechanisms, nanocarrier design, and translational considerations, this review provides a structured perspective for developing more rational and clinically feasible N2B nanodelivery systems for AD.\n\nID: 42503395\nTitle: Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.\nAbstract: Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma.\n\nID: 42494432\nTitle: Editorial: Cell therapy in the central nervous system and its crosstalk with the immune system.\nAbstract: \n\nID: 42489808\nTitle: The New Spine of Access to the Brain's Secrets: Extracellular Vesicles from Cerebrospinal Fluid Liquid Biopsies in CNS Diseases and Blood-Brain Barrier Research.\nAbstract: Liquid biopsy is emerging as a powerful approach for less invasive biomarker discovery, with extracellular vesicles (EVs) in cerebrospinal fluid (CSF) showing promise for the assessment of central nervous system (CNS) disorders without actual tissue biopsy and as a complement to imaging techniques. EVs carry molecular cargo such as proteins, nucleic acids, and lipids that mirror those at the tissue of origin, offering unique opportunities to quantify disease-related changes in biomarkers. Compared with plasma-derived EVs, those from CSF provide more direct insights into the CNS because of direct shedding of brain EVs to CSF and bypass of confounding factors involving entry to systemic circulation. Despite this potential, translation into clinical practice is limited by challenges such as low yields, purity concerns, and lack of standardized isolation protocols. Addressing these difficulties, alongside integrating multiomics approaches, will advance our understanding of EV molecular cargo and their functional roles in CNS diseases. Over time, CSF-derived EVs could become the new driver of precision medicine in neurology, offering biologic insight for both diagnostic and therapeutic applications. This perspective provides a critical evaluation of the current status of EV-based liquid biopsy in CSF and offers recommendations for future research and clinical translation of data from CSF-derived EVs, highlighting their potential to inform physiologically based pharmacokinetic (PBPK) models. This state-of-the-art article evaluates existing evidence and highlights key knowledge gaps.\n\nID: 42485583\nTitle: Hybrid Nonviral Nanocarriers Enable Functional Neural Modulation.\nAbstract: Extracellular vesicles released from the neuronal cells mediate the transfer of proteins, nucleic acids, and neurotransmitter-related cargoes, shaping gene expression and intercellular communication across neural circuits. Leveraging this endogenous pathway, we fused astrocyte-derived exosomes with RNA-loaded synthetic liposomes to create sub-100 nm hybrid nanoparticles for central nervous system delivery. This design addresses key limitations of existing systems: conventional liposomes lack cell-type specificity and can be toxic, whereas native vesicles are difficult to load efficiently. By tuning a mildly cationic surface, the hybrids support efficient gene transfer to neurons without disrupting membrane integrity or inducing measurable cytotoxicity. We validated this platform in vivo by delivering Cre recombinase mRNA in transgenic mice and inducing channel rhodopsin-2 expression in the motor cortex and ventral tegmental area, yielding behavioral changes during optogenetic stimulation.\n\nID: 42469846\nTitle: Metabolic reprogramming via SIRT2-deficient microglial large extracellular vesicles ameliorates alzheimer's pathology.\nAbstract: Current therapies for Alzheimer's disease (AD) offer only symptomatic relief, highlighting the urgent need for disease-modifying approaches capable of halting or reversing neurodegeneration. Extracellular vesicles (EVs) have attracted growing interest as therapeutic vehicles owing to their inherent capacity to bypass the blood-brain barrier and deliver complex biological cargo to the central nervous system. Here, we examined whether large EVs (LEVs) derived from microglia with stable Sirtuin-2 knockdown (SIRT2-KD) confer the neuroprotective effects associated with SIRT2 inhibition. LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice. We assessed microglial uptake of LEVs, along with subsequent changes in cellular metabolism, migration toward amyloid-beta (A\u03b2) plaques, phagocytic activity, and downstream pathological and behavioral outcomes. Proteomic and acetylomic profiling were employed to characterize the molecular cargo of LEVs-SIRT2-KD. LEVs-SIRT2-KD were readily internalized by microglia in vivo following intranasal delivery. Uptake of these vesicles markedly enhanced microglial bioenergetics, driving coordinated upregulation of both oxidative phosphorylation and glycolysis. This metabolic shift was accompanied by improved microglial recruitment to A\u03b2 plaques and increased phagocytic clearance. Consequently, treated mice showed reduced A\u03b2 plaque deposition, restored synaptic integrity, and reversal of cognitive deficits. Proteomic and acetylomic analyses revealed that LEVs-SIRT2-KD are selectively enriched in proteins and acetylation modifications linked to energy metabolism and phagocytic function, offering a mechanistic basis for the observed metabolic reprogramming. Together, these results identify LEVs as a critical vesicle subtype mediating the effects of SIRT2 knockdown and support a cell-free therapeutic strategy for AD centered on EVs-driven metabolic reprogramming of microglia.\n\nID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\n\nID: 42450256\nTitle: Peripheral and Central miRNA Signatures in Alzheimer's Disease: Tissue-Specific Variability, Sex-Associated Differences, and Implications for Blood-Based Biomarkers.\nAbstract: Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and significant neuropathological changes. Early and accurate diagnosis remains a major challenge, highlighting the need for reliable, minimally invasive biomarkers. MicroRNAs (miRNAs), small non-coding RNAs that regulate gene expression, have emerged as promising candidates. Their expression is altered in the brains of AD patients, reflecting disease-specific pathological processes, and they are detectable in peripheral biofluids. However, discrepancies in miRNA profiles between the brain and the circulation, and between patient populations remain a significant limitation, raising questions about their origin, transport across the blood-brain barrier, and their reliability in reflecting central nervous system pathology. This review provides a comprehensive overview of current research comparing miRNA expression profiles in brain tissue and blood in AD, with a focus on their biological relevance, mechanisms of release and transport, and diagnostic potential. We also discuss the challenges associated with cross-tissue variability, methodological inconsistencies, and the need for standardized approaches. Finally, we highlight future directions, including multi-tissue analyses and integration with other noninvasive modalities, to improve the clinical utility of miRNA-based biomarkers in AD.\n\nID: 42437583\nTitle: Brain-derived extracellular vesicle (BDEV) abundance and BDEV protein associations with clinical phenotypes and treatment response in untreated first-episode psychosis.\nAbstract: Reliable biomarkers that capture central nervous system (CNS) processes during the earliest stages of psychosis are critically needed to improve early treatment and response monitoring. Brain-derived extracellular vesicles (BDEVs) that traverse the blood-brain barrier (BBB) can be isolated from peripheral blood, thereby offering a novel and minimally invasive approach to quantify CNS molecular biomarkers. We hypothesized that BDEV abundance and BDEV neuroinflammatory and neurotrophic protein cargo would differ between individuals with first-episode psychosis (FEP) and healthy controls (HCs), reflecting altered neuroinflammatory signalling and extracellular biology associated with psychosis and clinical outcomes. In a cohort of untreated individuals with FEP (n\u00a0=\u00a057) and HCs (n\u00a0=\u00a028), BDEVs were isolated from serum. Luminex assays quantified concentrations of brain-derived neurotrophic factor (BDNF), S100 calcium-binding protein B (S100B), and C-reactive protein (CRP) in BDEVs and serum. A subset of FEP participants (n\u00a0=\u00a034) subsequently completed four weeks of antipsychotic treatment. Group differences in BDEV characteristics and protein measures, as well as their associations with case-control status, baseline symptom severtity, and four-week symptom change, were examined. FEP participants showed a trend toward higher BDEV concentration than HCs (median 14.0 \u00d7 108 versus 10.2 \u00d7 108 particles/mL, p\u00a0=\u00a00.052), and higher baseline BDEV concentrations were significantly associated with more severe pretreatment symptoms (\u03b2\u00a0=\u00a04.09 \u00d7 10-10, p\u00a0=\u00a00.003) and greater symptom improvement after four-week antipsychotic treatment (\u03b2\u00a0=\u00a03.29 \u00d7 10-10, p\u00a0=\u00a00.029). Within BDEVs, BDNF (\u03b2\u00a0=\u00a0-1.05; p\u00a0=\u00a00.0018) and CRP (\u03b2\u00a0=\u00a0-0.84; p\u00a0=\u00a00.002) were significantly reduced in FEP vs HC, with a trend toward lower S100B level (\u03b2\u00a0=\u00a0-0.57; p\u00a0=\u00a00.061). In contrast, serum protein levels showed no case-control differences and did not correlate with corresponding BDEV protein levels. Lower baseline BDEV BDNF, S100B, and CRP were associated with greater therapeutic response among males. BDEVs offer a minimally invasive window into CNS-enriched biology in early psychosis. BDEV abundance and neuroinflammatory and neurotrophic cargo concentrations show associations with illness severity and sex-specific treatment response not detectable in serum, supporting their potential as biomarkers for early pathophysiology and treatment response in FEP.\n\nID: 42434808\nTitle: Brain targeting and trafficking of extracellular vesicles in central nervous system diseases: a therapeutic roadmap.\nAbstract: Extracellular vesicles (EVs) mediate intercellular signaling in the central nervous system (CNS) by transferring lipids, proteins, and nucleic acids among neurons, glia, endothelium, and immune cells. Brain targeting depends on a linked sequence: EV ligands and adsorbed protein coronas engage receptor modules, select endocytic routes, determine intracellular fate, and define the therapeutic readouts. These fates include lysosomal degradation, recycling, rare cytosolic delivery, or transport across the blood-brain barrier (BBB). In disease, the same pathways can disseminate proteopathic seeds and amplify neuroinflammation. Heparan sulfate proteoglycans (HSPGs) and LDL receptor family members, including low-density lipoprotein receptor-related protein 1 (LRP1), regulate tau, \u03b1-synuclein, and amyloid-\u03b2 handling. Phosphatidylserine readers and complement shape myeloid sink capture and inflammatory output. Integrin, tetraspanin, and ICAM-1 nanoclusters influence avidity, organotropism, and immune suppression. At the BBB, endothelial HSPGs, LRP1, and transferrin receptor (TfR) support receptor-mediated uptake, motivating engineered ligands such as rabies virus glycoprotein-derived peptides, Angiopep-2, and TfR binders. However, endosomal escape remains a major kinetic barrier to nucleic acid delivery. We synthesize these principles across Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, glioblastoma, and demyelinating disease, and outline design and assay standards needed to translate EV biology into safe, manufacturable CNS therapeutics.\n\nID: 42434580\nTitle: Thoracic medial branch peripheral nerve stimulation for adjacent segment pain in a patient with a long construct fusion: a case report.\nAbstract: Adjacent segment degeneration is a known late complication of long construct fusions. While several experimental treatment options exist, including medial branch blocks, no definitive treatment options have been identified. Previous studies have demonstrated the benefit of restorative stimulation of the erector spinae group on patients' pain, physical function, and disability. This case hypothesizes that motor and sensory programming at the segment adjacent to the long-construct fusion has the potential to be both therapeutic for this patient's adjacent segment thoracic pain and protective through improved muscular stability. A 71-year-old woman with a past medical history of rheumatoid arthritis and prior T11 to S1 laminectomy with fusion and bilateral pedicle osteotomy presented with moderately severe 7/10 adjacent segment pain. She had previously completed conservative therapy, including physical therapy, epidural steroid injections, and medication management with NSAIDs, muscle relaxants, anticonvulsants, topical lidocaine, and chronic daily opioid use totally 30 morphine milliequivalents daily, without significant relief. Physical exam was significant for bilateral T8-T10 facet pain and flat back kyphosis, without focal or motor sensory deficits. She underwent a peripheral nerve stimulator device implantation with bilateral four electrode leads placed at the bilateral T10 medial branches using a minimally invasive retrograde approach to accommodate the fusion. Leads were advanced in a cephalad to caudad direction aimed at the transverse process of T9. Stimulation was programmed using a combined motor- and sensory-based approach to address both functional restoration and pain control. An external therapy disc was utilized to deliver stimulation, allowing the patient to independently apply and remove therapy, supporting day-to-day management of stimulation based on symptom severity, activity level, and personal preference. At her 3-month follow-up, the patient reports significant relief in her pain with an average pain score of 3/10, improved physical function, and a decrease in her pain medication usage. Restorative neuromodulation using motor and sensory programming is a potentially effective treatment adjunct for select patients with adjacent segment degeneration, with treatment sessions aligned towards pain relief and functional restoration. Further studies are needed to better define its role, protective effects and expected outcomes in this patient population.\n\nID: 42434075\nTitle: Gangliosides in the 21st century: therapeutic prospects for the brain and spine.\nAbstract: Gangliosides are sialylated glycosphingolipids highly enriched in the central nervous system, where they regulate membrane signaling, metabolism, neurogenesis, and immune responses. This Review integrates recent advances across distinct experimental and clinical domains. First, recent studies demonstrate that the monosialoganglioside GM1 enhances astrocyte-neuron metabolic coupling via the astrocyte-neuron lactate shuttle, thereby supporting neuronal bioenergetics and resilience. Complementary mechanistic work shows that specific gangliosides regulate adult neurogenesis through developmentally controlled epigenetic and transcriptional programs. In Huntington's disease models, preclinical evidence indicates that GM1 and related gangliosides attenuate microglia-mediated inflammatory responses and promote proteostasis through extracellular vesicle-dependent clearance of misfolded proteins. Finally, clinical evidence from acute spinal cord injury demonstrates that GM1 administration accelerates neurological recovery, underscoring its translational relevance. Together, these findings position gangliosides as multi-target modulators of neural repair and inflammation, and highlight their potential for therapeutic development.\n\nID: 42381337\nTitle: Extracellular Vesicles from Mesenchymal Stem Cells Alleviate Spinal Cord Injury via the miR-486-5p/PTEN/PI3K/AKT Pathway.\nAbstract: Spinal Cord Injury (SCI) is a severe central nervous system disorder with limited effective treatments. Mesenchymal stem cell (MSC)-derived exosomes have emerged as important mediators of intercellular communication and carry microRNAs with potential neuroprotective properties. This study aimed to explore the role and underlying mechanism of human umbilical cord MSC (hUMSC)-derived exosomal miR-486-5p in experimental SCI. Exosomes were isolated from hUMSCs and characterized by transmission electron microscopy, nanoparticle tracking analysis, and exosomal marker expression. A rat SCI model and an LPS-induced PC12 cell inflammatory injury model were established. Histological injury and apoptosis were assessed by HE staining and TUNEL assay. Inflammatory cytokine levels were measured by ELISA. Cell viability, apoptosis, and gene and protein expression were evaluated using CCK-8 assay, flow cytometry, qPCR, and western blotting. A dual-luciferase reporter assay was performed to validate the interaction between miR-486-5p and PTEN. hUMSC-derived exosomes attenuated spinal cord tissue damage, reduced neuronal apoptosis, and suppressed inflammatory cytokine production in vivo and in vitro. Inhibition of exosomal miR-486-5p partially reversed these protective effects. Mechanistically, miR-486-5p directly targeted the 3'-UTR of PTEN, leading to reduced PTEN expression and enhanced phosphorylation of AKT and mTOR. These findings indicate that exosomal miR-486-5p contributes to the regulation of apoptosis- and inflammation-associated molecular events following SCI, primarily through modulation of the PTEN/AKT/mTOR signaling pathway. Given the experimental design, these results should be interpreted as mechanistic insights rather than evidence of functional recovery. hUMSC-derived exosomal miR-486-5p alleviates apoptosis and inflammation following SCI by targeting PTEN and activating the AKT/mTOR pathway. These findings provide mechanistic support for the potential application of exosome-based miRNA therapy in SCI.\n\nID: 42374580\nTitle: HIV Tat-activated microglial extracellular vesicles induce neuronal iron dysregulation and synaptodendritic injury.\nAbstract: Extracellular vesicles (EVs) are membrane-enclosed, nanoscale structures released by cells and play a key role in intercellular communication under both normal physiological and pathological conditions. They serve as conduits for transferring molecular cargo between neighboring cells, thereby modulating recipient cell function. While the HIV Transactivator of transcription (Tat) protein has been shown to induce ferroptosis in microglia, the role of Tat-activated microglia-derived EVs (Tat-MEVs) in transferring iron-handling and ferroptosis-associated cargo to neurons and promoting neuronal injury remains unexplored. In this study, we sought to evaluate the impact of cargo derived from Tat-MEVs on neuronal synaptodendritic degeneration. Rat primary cortical and hippocampal neurons were exposed to either control MEVs or Tat-MEVs and subsequently assessed for synaptodendritic degeneration, expression of key ferroptotic mediators, and mitochondrial dysfunction associated with neuronal injury. Neurons exposed to Tat-MEVs demonstrated increased expression of the key iron-handling and ferroptosis-associated proteins (transferrin, TF; transferrin receptor 1, TFR1; Six-Transmembrane Epithelial Antigen of the Prostate 3, STEAP3; divalent metal transporter 1, DMT1; and ferritin heavy chain 1, FTH1); inhibitory synaptic markers (GAD65, Gephyrin), Fe2+/total iron content, neuronal cytotoxicity and mitochondrial reactive oxygen species (ROS) compared to neurons exposed to control MEVs. These findings suggest a link between mitochondrial dysfunction and neuronal iron accumulation. The expression of these mediators was downregulated in neurons exposed to MEVs derived from iron chelator, deferoxamine (DFO)-pretreated BV2 cells. Electrophysiological recordings further revealed reduced miniature excitatory postsynaptic currents in neurons exposed to Tat-MEVs, an effect that was attenuated in neurons exposed to DFO-derived MEVs. Additionally, dendritic spine analyses of neurons exposed to Tat MEVs revealed a reduction in mushroom and stubby spine subtypes, suggesting synaptodendritic injury.Collectively, these findings demonstrate that Tat-MEVs transfer iron-handling and ferroptosis-associated cargo that promotes neuronal iron dysregulation, oxidative stress, mitochondrial dysfunction, and synaptodendritic degeneration. These changes are consistent with ferroptosis-associated neuronal stress and contribute to functional impairment in recipient neurons. This EV-based communication axis provides mechanistic insight into how HIV Tat-induced microglial dysfunction propagates iron-dependent neurotoxic signaling within the central nervous system and identifies EV-mediated iron dysregulation as a potential therapeutic target in NeuroHIV.\n\nID: 42358359\nTitle: Global research trends and hotspots of exosome-mediated drug delivery across the blood-brain barrier: a bibliometric study from 2015 to 2025.\nAbstract: The blood-brain barrier (BBB) is a major obstacle to targeted drug delivery for central nervous system (CNS) diseases. Although liposomes and polymeric nanoparticles have improved brain drug delivery, limitations remain in BBB targeting, long-term biocompatibility, and in vivo clearance. Exosomes are endogenous nanoscale extracellular vesicles with favourable biocompatibility, low immunogenicity, and BBB-crossing potential. Therefore, this bibliometric study summarises the current research status, future research trends, and challenges in the more specific field of exosome-mediated BBB drug delivery. A comprehensive search was conducted across the Web of Science Core Collection (WoSCC), PubMed, and Embase databases for relevant English-language literature on exosome-mediated drug delivery across the blood-brain barrier from 2015 to 2025. WoSCC served as the primary source for bibliometric analysis. PubMed and Embase databases were used for supplementary validation. Software such as VOSviewer, CiteSpace, and R-bibliometrix was employed for literature visualisation analysis. This study included 1,365 relevant articles from the WoSCC database, and the annual publication volume showed a steady upward trend. China and the United States significantly lead in both the number of publications and the number of core contributing institutions in this field. Co-occurrence analysis of keywords showed that research hotspots are mainly focused on exosomes, the blood-brain barrier, drug delivery, and Alzheimer's disease. PubMed and Embase were used as supplementary validation databases, including 1,089 and 1,517 records, respectively. Their annual publication trends, major countries/regions, core journals, and keywords/themes were generally consistent with WoSCC, supporting the macro-level stability of the bibliometric findings. Unlike previous bibliometric analyses that mainly focused on overall trends in CNS exosome research, this study focuses specifically on the direction of exosome-mediated drug delivery across the BBB. The findings show a shift from basic vesicle characterisation toward engineered delivery systems, CNS disease applications, and translational evaluation. Mammalian-derived exosomes remain dominant, while plant-derived vesicles, AI-assisted design, biomimetic hybrid nanovesicles, and gut-brain axis strategies are emerging areas of focus. Future research should prioritise systematic platform comparisons, standardised evaluation, quality control, scalable production, long-term safety, and regulatory pathways.\n\nID: 42353041\nTitle: The Dual Roles of Extracellular Vesicle Subtypes in Regulating Traumatic Brain Injury.\nAbstract: Traumatic brain injury (TBI) is a global public health problem which causes long-term neurologic damage caused by both primary mechanical injury and secondary pathological processes. Extracellular vesicles (EVs) such as exosomes, microvesicles (MVs) and apoptotic bodies (ApoBDs) serve as critical vehicles mediating intercellular communication in the central nervous system (CNS) following TBI. The biogenesis and the content of EVs, including proteins, lipids and RNAs, are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI. In this overview, we recapitulate the cellular origin of EVs and the function of EVs in the neuroinflammatory process after TBI, highlighting the dual regulatory roles of EVs in the biological response to TBI, whereby certain EV populations amplify secondary injury cascades, while others promote endogenous repair and recovery processes. We next investigate the progress in EV engineering and targeted delivery systems and report the potential mechanisms, emphasize the prospects and potential of engineered EVs for therapy, and comment on challenges and perspectives for clinical application in TBI.\n\nID: 42351176\nTitle: Extracellular vesicle-mediated bidirectional communication between the brain and peripheral organs in Alzheimer's disease: evidence, mechanisms, and translational perspectives.\nAbstract: Alzheimer's disease (AD) has traditionally been conceptualized as a brain-centered neurodegenerative disorder characterized by amyloid-\u03b2 (A\u03b2) deposition, tau pathology, synaptic dysfunction, and progressive neuronal loss. However, accumulating evidence suggests that AD is also shaped by systemic disturbances and reciprocal communication between the central nervous system and peripheral organs. Extracellular vesicles (EVs), which transport proteins, lipids, metabolites, and nucleic acids across biological fluids and barriers, have emerged as plausible mediators of this inter-organ crosstalk. In this Review, current evidence for EV-mediated bidirectional communication between the brain and peripheral organs is synthesized, with particular attention to the liver-brain, heart-brain, gut-brain, lung-brain, bone-brain, and adipose-brain axes. The strength of evidence across these axes is compared, and the ways in which organ-derived EVs may influence neuroinflammation, neurovascular dysfunction, metabolic homeostasis, blood-brain barrier integrity, and A\u03b2/tau-related processes are discussed, while also considering how brain-derived EVs (BDEVs) may affect peripheral physiology. The translational potential of EVs as diagnostic biomarkers, therapeutic carriers, and candidate targets for systemic intervention in AD is further evaluated. Current evidence most strongly supports the gut-brain, liver-brain, and adipose-brain axes, whereas several other axes remain supported primarily by experimental models or engineered EV studies. Major barriers to progress include EV heterogeneity, limited source specificity, insufficient standardization of isolation and quantification workflows, and a continuing reliance on associative human data and preclinical models. Overall, EVs are best viewed as one candidate signaling layer within a broader systemic network linking peripheral physiology to brain pathology. Clarifying the magnitude, directionality, and causal significance of these interactions will require rigorous EV characterization, source-resolved in vivo trafficking studies, and longitudinal clinical investigation.\n\nID: 42342866\nTitle: Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.\nAbstract: Diabetic peripheral neuropathy (DPN), particularly distal symmetric polyneuropathy, characterized by length\u2011dependent axonal damage, is a common chronic complication of type 2 diabetes mellitus. The pathogenesis of DPN is complicated, yet one thing is clear: long axons require a great deal of energy. When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration. While earlier work centered on hyperglycemia-induced cytotoxicity, recent studies have increasingly implicated dysregulation of glucose, lipid, and amino acid metabolism as key contributors to DPN. In this review, we integrate the anatomical organization of peripheral nerves, bioenergetic pathways, and axon-Schwann cell interactions to establish a framework for understanding how glucose, lipid, and amino acid dysregulation converge to induce bioenergetic failure in DPN. Based on these mechanisms, we further discuss novel strategies aimed at restoring metabolic homeostasis in neurons and Schwann cells. Importantly, correcting a single metabolic pathway is unlikely to halt or reverse DPN. Instead, restoring global energy homeostasis to rebalance axonal energy supply and demand may be essential for preserving peripheral nerve function.\n\nID: 42342068\nTitle: Prenatal glucocorticoids and long-term brain vulnerability: GR signaling, epigenetic programming, and crosstalk with peripheral tissues.\nAbstract: Glucocorticoids (GCs) are key regulators of stress responses and fetal maturation, and their physiological rise during pregnancy supports coordinated organ development. Clinically relevant GC exposure during sensitive windows of brain development occurs in several contexts, including antenatal treatment for risk of preterm birth to promote lung maturation, prolonged maternal therapy for chronic inflammatory or autoimmune conditions, and postnatal GC treatment in preterm infants, including regimens used to prevent or treat bronchopulmonary dysplasia. Although these contexts differ in timing, dose, and duration, they share the capacity to engage a glucocorticoid receptor (GR) signaling during critical windows of neurodevelopment, with possible long-term consequences for brain development and stress responsiveness. This review synthesizes clinical, experimental, and stem cell-based evidence to examine how GC signaling can shape brain structure and function across the lifespan. We discuss GR signaling in the central nervous system (CNS) and summarize evidence that sustained activation can be associated with paradoxical pro-inflammatory and neurotoxic phenotypes. We highlight epigenetic mechanisms through which GC signals may produce persistent changes in gene regulation, and we integrate data from prenatal exposure together with evidence on maternal metabolic and inflammatory context as modifiers of developmental risk. Finally, we propose an integrated view in which CNS outcomes attributed to GCs reflect a composite of direct neural actions and indirect effects shaped by peripheral tissues. We discuss adipose- and muscle-linked pathways as candidate mediators of systemic-to-central communication. This perspective links stress endocrinology, metabolism, and brain vulnerability, and highlights key mechanistic gaps and translational priorities for future research.\n\nID: 42322256\nTitle: The predictive nature of diagnostic nerve blocks for lower extremity spasticity management with botulinum toxin and neurotomy: A systematic review.\nAbstract: Diagnostic nerve blocks (DNBs) involve the injection of anesthetic around peripheral nerves to temporarily reduce motor tone. Although they are used clinically to inform spasticity treatment with neurotomy and chemodenervation, there is little research exploring the predictive nature of DNBs. A systematic review was conducted to identify and analyze studies that compared clinical outcomes between DNBs and neurotomies or chemodenervation with botulinum toxin. In total, nine eligible articles were analyzed, with five studies involving neurotomies and four studies involving chemodenervation. These studies focused exclusively on lower limb spasticity. There were different DNB protocols used across the eligible articles. DNBs resulted in similar clinical outcomes after neurotomy on gait and spasticity measurements. DNBs resulted in reduced agonist muscle strength as compared to neurotomy outcomes, particularly those collected a year or longer after surgery. DNBs were less predictive of clinical response to chemodenervation and to range of motion assessments. This review has identified a gap in research regarding the predictability of DNBs, with heterogeneity in study design. As DNBs resulted in more similar spasticity and gait outcomes when compared to neurotomy rather than chemodenervation procedures, this may suggest that DNBs have a larger role to play in surgical decision-making. Given the current research, future studies on DNBs should explore upper extremity spasticity management as well as outcomes that relate to patient reported goals and experiences.\n\nID: 42315580\nTitle: Alkaline loading of extracellular vesicles produced from human neural stem cell-derived neurospheres enables CNS drug delivery.\nAbstract: The blood brain barrier and blood tumor barrier (BBB and BTB, respectively) represent significant obstacles for the delivery of drugs to treat diseases of the central nervous system, such as brain cancers and neurodegenerative diseases. Extracellular vesicles (EVs) or exosomes have emerged as a new drug delivery vehicle for CNS diseases as they may penetrate the BBB/BTB and are less immunogenic than liposomal carriers. EVs derived from human neural stem cells (hNSC) provide additional benefits over other EV sources due to their increased homing capability to neural cells and demonstrated efficacy for treating stroke and traumatic brain injury in rodent models. However, the utilization of EVs from hNSC for drug delivery remains largely unexplored, due in part to difficulties in manufacturing capacity compared to traditional cell lines. Here, we report the development of a hNSC suspension neurosphere system for EV production and drug delivery. As proof of concept, doxorubicin was loaded into hNSC-EV, using a novel, high-efficiency alkaline passive loading method, and shown to be effective at inducing cytotoxicity in glioma cells in vitro and exhibiting higher BBB penetrance than doxorubicin-alone in vivo. These studies demonstrate the potential for hNSC-EV loaded doxorubicin as a therapeutic treatment for brain cancers such as glioblastoma, while also establishing hNSC-EVs as a drug-delivery vehicle for CNS diseases.\n\nID: 42314895\nTitle: Profiling tRNA-derived fragments in LPS-induced microglia and their influence on immune response.\nAbstract: The resident macrophages of the brain, microglia, react to immunological responses and preserve homeostasis in the central nervous system. Microglia play a critical role in neuroinflammation, which occurs in neurological and neurodegenerative diseases. Microglial responses are associated with neurodegenerative diseases, autoimmune diseases, and cancer, and therefore require strict regulation. Transfer RNA-derived fragments (tRFs) are small non-coding regulatory RNA molecules generated from the cleavage of transfer RNAs. tRFs have recently emerged as one of the key regulators of gene expression and cellular function in various biological processes. Understanding the content of tRFs altered in microglia due to LPS exposure may provide insights into the mechanisms involved in disease pathogenesis, where neuroinflammation plays a role. We used next-generation sequencing to determine tRF profiles of three different fractions (lysate, extracellular vesicles, and extracellular vesicles-free supernatant) of LPS-induced microglial cells. A total of 345 tRFs were differentially expressed across these fractions after LPS stimulation, including 13 DE-tRFs shared by all three compartments. Gene Ontology analysis of these 13 tRFs suggests their potential involvement in the inflammatory responses of microglia. Among the 13 consistently dysregulated DE-tRFs, tDR-1:33-Glu-CTC-1-M2 was selected for preliminary candidate-level assessment based on its consistent differential expression across all three fractions in our NGS data and previous reports linking this fragment to neuroinflammatory contexts.Increased expression pattern of tDR-1:33-Glu-CTC-1-M2 was further confirmed in lysate samples of LPS-induced microglial cells via RT-qPCR. Inhibition of tDR-1:33-Glu-CTC-1-M2 attenuated LPS-induced cytokine expression and release in microglia, supporting an association between this candidate tRF and inflammatory readouts in this model.\n\nID: 42312720\nTitle: Prenatal cannabis use is associated with altered miRNA and protein expression in the developing human brain.\nAbstract: Cannabis is widely used during pregnancy, increasing with cannabis legalization, and has been associated with altered neurodevelopment, yet the precise molecular mechanisms remain poorly understood. To assess whether utilizing fetal central nervous system-derived extracellular vesicles (fCNSEVs) isolated from maternal blood reflects cannabis-associated changes in the fetal brain. In a matched case-control study, maternal plasma (to isolate fCNSEVs) and paired fetal cortical tissue (9-18\u2009weeks' gestation) from pregnancies exposed or unexposed to cannabis were collected. We quantified protein (CB1R, D2R) and miRNA expression (using \u00b5-parafloTM microarray). Selected miRNAs were validated by qPCR. Bayesian Generalized Linear Models were used to assess exposure-group by sex effects. Maternal cannabis use was associated with altered CB1R and D2R levels in the brain with opposite directional changes in fCNSEVs. 21 miRNAs were differentially expressed: 14 downregulated and 6 upregulated, following a sex-dependent pattern (males>>females), while miR-216a-5p follows the opposite pattern (females>>males). We confirmed concordant miRNA changes in fetal brain and fCNSEVs. Many of the identified molecules are associated with pathways linked to adverse neurodevelopmental outcomes. Our data suggest that fCNSEV-based assays may provide the ability to monitor fetal brain effects of maternal cannabis in ongoing pregnancies in clinical cohorts. A critical next step is to determine if alterations in fCNSEV protein and miRNA markers predict changes in fetal brain connectivity and neurodevelopment measures of executive function.\n=======================================================\n\n### [CUSTOM DATAPOINTS]\nCRITICAL EXTRACTION DIRECTIVE: You MUST extract the following custom datapoints as root-level key/value pairs inside your final JSON block:\n- \"suggested_experiments\": generate 1-3 suggested experiments\n- \"suggested_studies\": generate 1-3 suggested studies\n- \"swansons_literature_based_discovery_candidates\": You are an advanced Literature-Based Discovery (LBD) system executing Swanson\u2019s complementary-but-disjoint (A-B-C) model. Your goal is to find hidden, unpublished connections across the provided dataset. Strict Discovery Protocol: 1. Identify distinct, isolated sub-literatures (Domain A and Domain C) within the dataset that share NO direct citations, co-mentions, or common contextual paragraphs. 2. Find an intermediate biological mechanism, protein, path, or entity (Bridge B) that appears independently in both isolated domains (A-to-B and B-to-C). 3. Synthesize a novel, unstated hypothesis (A-to-C). Negative Constraint (Crucial): DO NOT output any connection if the relationship between Concept A and Concept C is explicitly mentioned, paired, or summarized anywhere in the source text. If a connection (like \"OMN resilience to SMN stabilization\") is already explicitly stated or grouped as a concept in the data, it is considered \"already known\" and must be disqualified. Format your output exactly as follows: - Discovered Hypothesis (A to C): [Clear, novel statement] - Literature A (Origin): [Entity/Concept and source context] - Literature C (Target): [Entity/Concept and source context] - The Intersecting Bridge B: [The shared mechanism/protein linking them] - Biological Rationale: [1-2 sentences explaining why this hidden connection is mechanistically plausible]\n- \"contradictions_between_evidences\": Identify conflicting evidence within the evidence set (if any) and flag the dispute here\n- \"repurposed_solutions\": identify and explain repurposed Solution potentials\n\n\nFormat Requirement:\nRAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nFirst provide disclaimer such as \"Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\"\n---\nWrite in a highly academic, formal thesis tone.\nFormat your readable response using these exact academic headers:\n###[CLAIM EVALUATED AND ANSWER TO USER]\n(Exact wording of the claim evaluated)\n### [ABSTRACT & REWRITTEN CLAIM]\n(Scientific synthesis)\n### [INTRODUCTION & JUSTIFICATION]\n(Mechanistic explanation utilizing the 'moneyshot quotes' you will use in the EVIDENCE, METHODOLOGY & CITATIONS section later as well)\n### [DISCUSSION: NOVEL & OVERLOOKED]\n(5-10 bullet points of surprising facts)\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 3) - [copied/verbatim Quote text]\"\n\n**CRITICAL: You must include the exact quote you used in the [copied/verbatim Quote text] section.\n\nIf the prompt says \"at least 20 quotes\" then there must be at least 20 matching citations. You must actually use the quotes you select within the conext of the preprint publication you write.\n\nEvaluation Schema:\nRAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\n###critical: WRAP YOUR THOUGHTS WITH \nAll responses must include the mandatory \"### [EVIDENCE, METHODOLOGY & CITATIONS]\" section as formatted.\nCRITICAL:\n**MONEYSHOT QUOTES MUST DIRECTLY SUPPORT YOUR CLAIMS**\n**MONEYSHOT QUOTES MUST BE USED IN YOUR RESPONSE TEXT WITHOUT IN-LINE ANNOTATION**\n**MONEYSHOT QUOTES MUST BE USED IN A FORMAL PROFESSIONAL WAY, WORTHY OF PEER REVIEW, WITHOUT ILLOGICAL LEAPS (UNSUPPORTED MAY BE OK, ILLOGICAL IS NOT OK)**\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 7) - *\"copied/verbatim Quote text\"**\n\nCRITICAL INSTRUCTION:\nwhen fact checking: At the very end of your response, you MUST provide a machine-readable JSON block containing evaluation metrics. \nIt MUST be enclosed exactly between ###JSON_START### and ###JSON_END###. Ensure the JSON is valid. \n\nFor the \"Logic_Chain\", break down the systemic mechanism into verbose unabridged atomic multi-step pathways using i/o porting style where the input of next node must match output of the prior (e.g., A -> B, B->C, C->D). Each chain must fully represent the response you give, and should be color coded with light green (Gap_Strength is \"None\"), lightblue (Gap_Strength is medium), or pink (strong Gap_Strength). Logic_Chain MUST be a JSON array of objects. Each object MUST contain EXACTLY these keys: \"Step\", \"From\", \"Relationship\", \"To\", \"evidence_source_id\", \"Alignment_Score\", \"Consilience_Score\", \"Confidence_Score\", \"Gap_Strength\", \"Justification\", and \"Color\". Use commas between objects. DO NOT leave trailing commas inside objects.\n\nFor \"Verbatim_Quotes\", copy at least 20 (required, 20 or more) \"moneyshot\" quotes EXACTLY as they appear in the context literature text, word-for-word, characters included, that fully support your response. We will programmatically validate these. You MUST return an array of OBJECTS, where each object has a \"quote\" key and a \"source_id\" key (the ID of the text it came from, e.g., the ID). Do not alter a single character, do not paraphrase.\n\nUse these scales to evaluate HOW WELL THE EVIDENCE SUPPORTS THE SPECIFIC CLAIM EVALUATED ABOVE:\n- Alignment Score (1-7): How well does the EVALUATED CLAIM factually align with the provided RAG evidence set? [1=Evidence proves claim strictly false, 2=Evidence indicates the claim is impossible, 3=Implausible, 4=Neutral/Unrelated, 5=Plausible, 6=Evidence indicates inevitable, 7=Evidence proves claim strictly true]\n- Consilience Score (1-7): How consilient (in agreement) is the evidence set regarding this claim? [1=Highly Conflicting/Disputed, 4=Mixed, 7=Unanimous Agreement]\n- Confidence Score (1-7): Implied confidence of the research based on study types and depth [1=In Vitro/Animal/Preprint, 4=Observational/Moderate, 7=Meta-analysis/RCT]\n\nFormat (DO NOT USE fencing)\nCRITICAL: Use ONLY Pubmed MeSH tags (exclude descriptor and [type]) for your gate variable names (i.e.,.the \"gates\") so they will be standardized globally. Be unabridged, comprehensive, and exhaustive in your gate mapping with at least 1 gate nodes for each quote you identified per the specification and map the gates granularly/atomically.\n\n###JSON_START###\n{\n \"Alignment\": 5,\n \"Consilience\": 6,\n \"Confidence\": 5,\n \"Logic_Chain\":[\n {\n \"Step\": 1,\n \"From\": \"Variable A\",\n \"Relationship\": \"-->\",\n \"To\": \"Variable B\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 5,\n \"Confidence_Score\": 4,\n \"Gap_Strength\": \"None\",\n \"Justification\": \"...\",\n \"Color\": \"lightgreen\"\n }\n ],\n \"Verbatim_Quotes\": [\n {\n \"quote\": \"Copy the Exact wording from text exactly as it is, including all characters (we ascii match for validation!).\",\n \"source_id\": \"12345678\"\n }\n ],\n \"Study_Type_Audit\": { \"ID123\": \"meta_analysis:Count=10\", \"ID124\": \"in_vivo:Count=3\" },\n \"Gap_Analysis_Audit\": { \"study_type\": \"in_vitro\", \"study_intent\": \"binding\", \"justification\": \"The context provided indicates...\", \"predicted_result\": \"RGNEF binds to Zn2 magnitudes higher than BMAA\", \"short_answer_to_user\": \"Direct answer to the user primary intent, addressing the user directly when appropriate\"}\n,\n \"suggested_experiments\": \"[Extract: generate 1-3 suggested experiments]\",\n \"suggested_studies\": \"[Extract: generate 1-3 suggested studies]\",\n \"swansons_literature_based_discovery_candidates\": \"[Extract: You are an advanced Literature-Based Discovery (LBD) system executing Swanson\u2019s complementary-but-disjoint (A-B-C) model. Your goal is to find hidden, unpublished connections across the provided dataset. Strict Discovery Protocol: 1. Identify distinct, isolated sub-literatures (Domain A and Domain C) within the dataset that share NO direct citations, co-mentions, or common contextual paragraphs. 2. Find an intermediate biological mechanism, protein, path, or entity (Bridge B) that appears independently in both isolated domains (A-to-B and B-to-C). 3. Synthesize a novel, unstated hypothesis (A-to-C). Negative Constraint (Crucial): DO NOT output any connection if the relationship between Concept A and Concept C is explicitly mentioned, paired, or summarized anywhere in the source text. If a connection (like \\\"OMN resilience to SMN stabilization\\\") is already explicitly stated or grouped as a concept in the data, it is considered \\\"already known\\\" and must be disqualified. Format your output exactly as follows: - Discovered Hypothesis (A to C): [Clear, novel statement] - Literature A (Origin): [Entity/Concept and source context] - Literature C (Target): [Entity/Concept and source context] - The Intersecting Bridge B: [The shared mechanism/protein linking them] - Biological Rationale: [1-2 sentences explaining why this hidden connection is mechanistically plausible]]\",\n \"contradictions_between_evidences\": \"[Extract: Identify conflicting evidence within the evidence set (if any) and flag the dispute here]\",\n \"repurposed_solutions\": \"[Extract: identify and explain repurposed Solution potentials]\"\n}\n###JSON_END###\n\n### CRITICAL QUOTE VALIDATION FAILURE (ATTEMPT 1) ###\nThe validator executed a 100% strict, character-by-character substring search. Your response was REJECTED because the following quotes do not exist verbatim in the source texts.\n\n\u274c FAILED QUOTES (You must fix or delete these):\n\n- ERROR: You cited ID: 42510934 for the quote: \"Rho GTPases... regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42510934 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42510934 ---\n ID: 42510934\nTitle: Herbal Bioactives Targeting Rho GTPases: A Multi-Targeted Strategy for Mitigating Neuroinflammation in Alzheimer's and Parkinson's Diseases.\nAbstract: Neuroinflammation plays an essential role in the pathogenesis of several associated brain diseases, including neurodegenerative disorders (Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS)), and traumatic brain injury (TBI). In these diseases, persistent microglial and astrocyte aggregates, elevated proinflammatory cytokines, and oxidative stress drive neuronal injury and cognitive disability. Rho GTPases, in particular the Rho family members Ras homolog family member A (RhoA), Ras-related C3 botulinum toxin substrate 1 (Rac1), and cell division control protein 42 homolog (CDC42), regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity. These proteins are involved in many neuropathological diseases due to dysregulation, making them interesting therapeutic targets. Bioactives used in herbal care have attracted interest for their ability to influence neuroinflammation and even their anti-neurodegenerative activity. Studies show that flavonoids, alkaloids, polyphenols, and other botanical compounds alter Rho GTPase activity, which, in turn, leads to decreased inflammation. This review critically summarizes current evidence regarding phytochemical regulation of Rho GTPase signaling in neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD), with particular emphasis on the underlying molecular mechanisms, context-dependent signaling responses, and current translational challenges. Furthermore, existing knowledge gaps and future research priorities are discussed to facilitate the development of mechanism-based therapeutic strategies targeting Rho GTPases.\n --- END ACTUAL ABSTRACT FOR 42510934 ---\n\n- ERROR: You cited ID: 37517544 for the quote: \"When sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system.\"\n FACT: Strict Misquote Detected! The exact character sequence \"When sEVs are injected into the sub...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 37517544 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 37517544 ---\n ID: 37517544\nTitle: Logistics and distribution of small extracellular vesicles from the subcutaneous space to the lymphatic system.\nAbstract: Small extracellular vesicles (sEVs) are small, cell-derived particles with sizes of approximately 100 nm. Since these particles include cargos such as host cell-derived proteins, messenger RNAs, and micro RNAs, they serve as mediators of cell-cell communication. While the analysis of the pharmacokinetic of sEVs after the intravenous injection have been reported, the lymphatic transport of sEVs remains unclear. The objective of this study was to provide insights into the intra-lymphatic trafficking and distribution of sEVs when they are injected into an interstitial space both in normal skin tissue and in cancerous tissue. When sEVs were Subcutaneously administered into the tail base and the tumor tissue, they preferably accumulated in the lymph nodes (LNs), rather than in the liver and the spleen. The findings reported herein show that the lymphatic transport of sEVs was drastically changed in model mice, in which a surgical treatment was used to modify to allow the dominant lymphatic flow from the footpad directly to the axillary LN via the inguinal LN. Based on the results, we conclude that when sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system. Further, the extent of accumulation of sEVs in the LN after subcutaneous injection was reduced when they were preliminarily incubated with Proteinase K. These results suggest that the lymphatic drainage of sEVs in normal skin tissue is regulated by membrane proteins on their surface. This reduction, however, was not observed in the case of cancer tissue. This discrepancy can be attributed to the presence of highly permeable lymphatic vessels in the tumor tissue. Further, the major cell subtypes that captured sEVs in the LN were LN-resident medullary sinus macrophages. These collective findings indicate that the lymphatic drainage of sEVs are mediated by proteins and, that they may appear to contribute to the control of the function of immune-responsive cells in the LNs.\n --- END ACTUAL ABSTRACT FOR 37517544 ---\n\n- ERROR: You cited ID: 37371477 for the quote: \"EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers.\"\n FACT: Strict Misquote Detected! The exact character sequence \"EVs recovered from the body fluids ...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 37371477 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 37371477 ---\n ID: 37371477\nTitle: The Role of Extracellular Vesicles in the Pathogenesis of Hematological Malignancies: Interaction with Tumor Microenvironment; a Potential Biomarker and Targeted Therapy.\nAbstract: The tumor microenvironment (TME) plays an important role in the development and progression of hematological malignancies. In recent years, studies have focused on understanding how tumor cells communicate within the TME. In addition to several factors, such as growth factors, cytokines, extracellular matrix (ECM) molecules, etc., a growing body of evidence has indicated that extracellular vesicles (EVs) play a crucial role in the communication of tumor cells within the TME, thereby contributing to the pathogenesis of hematological malignancies. The present review focuses on how EVs derived from tumor cells interact with the cells in the TME, such as immune cells, stromal cells, endothelial cells, and ECM components, and vice versa, in the context of various hematological malignancies. EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers for specific types of cancer, thereby also acting as potential therapeutic targets. Here, we discuss how EVs influence hematological tumor progression via tumor-host crosstalk and their use as biomarkers for hematological malignancies, thereby benefiting the development of potential therapeutic targets.\n --- END ACTUAL ABSTRACT FOR 37371477 ---\n\n- ERROR: You cited ID: 42466399 for the quote: \"Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury.\"\n FACT: Strict Misquote Detected! The exact character sequence \"Single-cell RNA sequencing of astro...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42466399 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42466399 ---\n ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\n --- END ACTUAL ABSTRACT FOR 42466399 ---\n\n- ERROR: You cited ID: 42506058 for the quote: \"The overall safety profile was consistent with previous studies... no apparent dose- or injection interval-related trend observed.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42506058 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42506058 ---\n ID: 42506058\nTitle: Efficacy and Safety of a Tailored Dosing Strategy with High-Dose IncobotulinumtoxinA at Flexible Injection Intervals for Cervical Dystonia: An Open-Label, Uncontrolled, Single-Arm Study in Japan.\nAbstract: This prospective, multicenter, open-label, single-arm study (jRCT2031230690) evaluated the efficacy and safety of a tailored dosing strategy of incobotulinumtoxinA, including high doses (up to 500 U) and flexible injection intervals (as short as 6 weeks), in Japanese patients with cervical dystonia (CD). Of 30 enrolled patients, Group A included 27 patients with idiopathic CD for the primary evaluation of efficacy and safety, whereas Group B included 3 patients with tardive dyskinesia (cervical) or tardive CD for exploratory safety assessment. Patients received up to seven injection cycles of incobotulinumtoxinA (120-500 U) over 48 weeks, with minimum 6-week intervals. The primary endpoint, evaluated in Group A, was the change in Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) total score from baseline to Week 4 after the first injection. Using a mixed model for repeated measures, the least squares mean \u00b1 standard error of the change was -11.0 \u00b1 1.77 (95% confidence interval: -14.6, -7.3). The primary efficacy endpoint was achieved in Group A. Due to the small sample size (n = 3), efficacy in Group B was evaluated only for exploratory purposes, although safety findings were broadly consistent with those in Group A. The overall safety profile was consistent with previous studies. Across the study, the most common related adverse events were dysphagia (33.3%) and muscular weakness (22.2%) in Group A and dysphagia (33.3%) in Group B. All cases of dysphagia were mild to moderate in severity and transient, with no apparent dose- or injection interval-related trend observed. IncobotulinumtoxinA was associated with improvements in symptoms and manageable safety profile at high doses and flexible injection intervals in Japanese patients with CD. While these findings suggest a potential treatment option for individualized dose optimization, the absence of a control group and the exploratory nature of the assessment in Group B necessitate cautious interpretation.\n --- END ACTUAL ABSTRACT FOR 42506058 ---\n\n- ERROR: You cited ID: 42422202 for the quote: \"NCB eliminates trigeminal nerve stimulation... reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42422202 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42422202 ---\n ID: 42422202\nTitle: Long-term efficacy of neural circuit blockade for treating blepharospasm: a retrospective case series study.\nAbstract: Blepharospasm is a common symptom of Meige syndrome and often leads to functional blindness. Although current treatment options, such as oral medications, botulinum toxin injections, resection of periocular muscles, and deep brain stimulation, are available, no definitive method effectively eliminates blepharospasm. To address this unmet need, we developed a novel approach-neural circuit blockade (NCB). This study aimed to analyse its long-term efficacy for blepharospasm. NCB eliminates trigeminal nerve stimulation by facial expression muscles, reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions, decreasing orbicularis oculi muscle tension, and alleviating blepharospasm. A retrospective analysis was conducted on 570 patients who underwent NCB treatment, with a median postoperative follow-up of 51\u202fmonths. Therapeutic efficacy was evaluated by comparing preoperative and postoperative scores on the Shorr blepharospasm grading scale, Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS-M), and Blepharospasm Disability Index (BSDI). Based on the Shorr grading scale, preoperatively, 97.9% of patients were grade 4, whereas 2.1% were grade 3. Postoperatively, 87.5% achieved complete remission (grade 0), whereas 7.4, 3.7, 0.9, and 0.5% were grades 1, 2, 3, and 4, respectively. Pre- and postoperative BFMDRS-M scores significantly differed: eye (8.0 vs. 0.0), mouth (3.0 vs. 0.0), speech/swallowing (3.0 vs. 2.0), neck (3.0 vs. 1.5), and total (8.0 vs. 0.0). BSDI scores also significantly differed pre- and postoperatively. The results indicate that NCB effectively alleviates blepharospasm and addresses the lack of a definitive treatment, without causing severe local or systemic complications such as facial paralysis. NCB can safely and effectively alleviate blepharospasm, improve quality of life, and demonstrate stable long-term efficacy.\n --- END ACTUAL ABSTRACT FOR 42422202 ---\n\n- ERROR: You cited ID: 42533122 for the quote: \"TeNT... potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42533122 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42533122 ---\n ID: 42533122\nTitle: Light-activated tetanus neurotoxin for conditional proteolysis and inducible synaptic inhibition in vivo.\nAbstract: The light chain of tetanus neurotoxin (TeNT) is a metalloprotease that potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision. To address this, we engineered light-activated TeNT (LATeNT) by inserting the light-sensitive LOV domain into an allosteric site and optimizing dynamic range via directed evolution. LATeNT's activity is undetectable in the dark, but the protease turns on after 10-20 min of weak blue-light exposure to potently inhibit synapses in vivo. Here we show that LATeNT works across multiple brain regions and at long-range axonal projections, with its effects reversible in 24\u2009h. LATeNT enabled us to discover a hippocampal interneuron population that regulates anxiety-like behaviors and demonstrate the importance of postsynaptic endocannabinoid exocytosis for depolarization-induced suppression of inhibition in vivo. Beyond neuroscience, LATeNT regulated endogenous insulin secretion from pancreatic beta cells and converted drug exposure, elevated Ca2+ or receptor activation into transgene expression or reporter secretion in HEK293T cells. With a large dynamic range, high light sensitivity and sustained effect, LATeNT enables versatile, spatiotemporally resolved proteolysis across diverse biological systems.\n --- END ACTUAL ABSTRACT FOR 42533122 ---\n\n- ERROR: You cited ID: 41989572 for the quote: \"BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine.\"\n FACT: Strict Misquote Detected! The exact character sequence \"BoNTs are endopeptidases, which are...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 41989572 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 41989572 ---\n ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance.\n --- END ACTUAL ABSTRACT FOR 41989572 ---\n\n- ERROR: You cited ID: 42469846 for the quote: \"LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice... restored synaptic integrity, and reversal of cognitive deficits.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42469846 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42469846 ---\n ID: 42469846\nTitle: Metabolic reprogramming via SIRT2-deficient microglial large extracellular vesicles ameliorates alzheimer's pathology.\nAbstract: Current therapies for Alzheimer's disease (AD) offer only symptomatic relief, highlighting the urgent need for disease-modifying approaches capable of halting or reversing neurodegeneration. Extracellular vesicles (EVs) have attracted growing interest as therapeutic vehicles owing to their inherent capacity to bypass the blood-brain barrier and deliver complex biological cargo to the central nervous system. Here, we examined whether large EVs (LEVs) derived from microglia with stable Sirtuin-2 knockdown (SIRT2-KD) confer the neuroprotective effects associated with SIRT2 inhibition. LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice. We assessed microglial uptake of LEVs, along with subsequent changes in cellular metabolism, migration toward amyloid-beta (A\u03b2) plaques, phagocytic activity, and downstream pathological and behavioral outcomes. Proteomic and acetylomic profiling were employed to characterize the molecular cargo of LEVs-SIRT2-KD. LEVs-SIRT2-KD were readily internalized by microglia in vivo following intranasal delivery. Uptake of these vesicles markedly enhanced microglial bioenergetics, driving coordinated upregulation of both oxidative phosphorylation and glycolysis. This metabolic shift was accompanied by improved microglial recruitment to A\u03b2 plaques and increased phagocytic clearance. Consequently, treated mice showed reduced A\u03b2 plaque deposition, restored synaptic integrity, and reversal of cognitive deficits. Proteomic and acetylomic analyses revealed that LEVs-SIRT2-KD are selectively enriched in proteins and acetylation modifications linked to energy metabolism and phagocytic function, offering a mechanistic basis for the observed metabolic reprogramming. Together, these results identify LEVs as a critical vesicle subtype mediating the effects of SIRT2 knockdown and support a cell-free therapeutic strategy for AD centered on EVs-driven metabolic reprogramming of microglia.\n --- END ACTUAL ABSTRACT FOR 42469846 ---\n\n- ERROR: You cited ID: 42384787 for the quote: \"Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors.\"\n FACT: Strict Misquote Detected! The exact character sequence \"Rapid release (<7 milliseconds) has...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42384787 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42384787 ---\n ID: 42384787\nTitle: Turbocharging synaptic transmission: 12 SNAREpins are required for rapid release of reconstituted synaptic vesicles.\nAbstract: Synaptic transmission occurs synchronously with real-world events, far faster than vesicle fusion for hormone release or membrane biogenesis, all mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. How SNAREs cooperate to achieve synchronous neurotransmitter release is a long-standing mystery. Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors in a fully-defined, genetically validated system that enables single-molecule counting in docked vesicles before release. SNAREpin complexes (12 \u00b1 0.3) are found in each such ready-release vesicle, suggesting a regular structure. Several genetic conditions (including point mutation of the synaptic vesicle protein Synaptophysin from a Synaptopathy patient and human and mouse disease mutations of the synaptic vesicle protein vesicle-associated membrane protein-2 (VAMP2) reduce the number of SNAREpins to 6 \u00b1 0.3 and result in profoundly delayed release over 0.1 to 1 seconds. Omitting Synaptophysin, whose hexamers preassemble 12 copies of VAMP2, also yields ~6 SNAREpins and delays release.\n --- END ACTUAL ABSTRACT FOR 42384787 ---\n\n- ERROR: You cited ID: 42353041 for the quote: \"The biogenesis and the content of EVs... are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI.\"\n FACT: Ellipses (...) are strictly forbidden. You must quote continuous text exactly character-for-character.\n \n Below is the complete, true text of ID 42353041 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42353041 ---\n ID: 42353041\nTitle: The Dual Roles of Extracellular Vesicle Subtypes in Regulating Traumatic Brain Injury.\nAbstract: Traumatic brain injury (TBI) is a global public health problem which causes long-term neurologic damage caused by both primary mechanical injury and secondary pathological processes. Extracellular vesicles (EVs) such as exosomes, microvesicles (MVs) and apoptotic bodies (ApoBDs) serve as critical vehicles mediating intercellular communication in the central nervous system (CNS) following TBI. The biogenesis and the content of EVs, including proteins, lipids and RNAs, are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI. In this overview, we recapitulate the cellular origin of EVs and the function of EVs in the neuroinflammatory process after TBI, highlighting the dual regulatory roles of EVs in the biological response to TBI, whereby certain EV populations amplify secondary injury cascades, while others promote endogenous repair and recovery processes. We next investigate the progress in EV engineering and targeted delivery systems and report the potential mechanisms, emphasize the prospects and potential of engineered EVs for therapy, and comment on challenges and perspectives for clinical application in TBI.\n --- END ACTUAL ABSTRACT FOR 42353041 ---\n\n\n\u2705 PASSED (DO NOT CHANGE THESE):\n- \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" (Source: 42102607)\n- \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\" (Source: 37037273)\n- \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\" (Source: 42275483)\n- \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\" (Source: 41540479)\n- \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\" (Source: 38110531)\n- \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\" (Source: 42458952)\n- \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\" (Source: 37394036)\n- \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\" (Source: 42504872)\n- \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\" (Source: 42543397)\n\n\nINSTRUCTION: Study the actual abstracts provided. Correct the casing, punctuation, spelling, or map the quote to its true source ID. Do NOT use ellipses.\n\n### CRITICAL QUOTE VALIDATION FAILURE (ATTEMPT 2) ###\nThe validator executed a 100% strict, character-by-character substring search. Your response was REJECTED because the following quotes do not exist verbatim in the source texts.\n\n\u274c FAILED QUOTES (You must fix or delete these):\n\n- ERROR: You cited ID: 42514227 for the quote: \"Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors.\"\n FACT: Strict Misquote Detected! The exact character sequence \"Mesenchymal stem cells (MSCs) could...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42514227 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42514227 ---\n ID: 42514227\nTitle: Dynamic Tuning of MSC-Based Scaffolds for Neurological Protection After Brain or CNS Injury.\nAbstract: Neurological disorders, including stroke, traumatic brain injury, and spinal cord injury, constitute one of the most important causes of mortality and morbidity worldwide for which current treatment options focus on resolving neuroinflammation rather than on tissue and neuronal regeneration. Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors which modulate microglial activation, preserve blood-brain barrier (BBB) integrity, and neuroplasticity, but with limitations due by poor survival, retention, and phenotypic instability following direct transplantation. The purpose of this narrative review is to present mechanotransduction signaling pathways (integrin-FAK, PI3K/Akt, Rho/ROCK, and YAP/TAZ) through which MSC-based biomaterial scaffolds, especialy hyaluronic acid (HA) hydrogels, make the transition from reparative to regenerative medicine in central nervous system (CNS) injury. Even if most of the evidence from preclinical studies suggests that dynamically tunable MSC-scaffold systems represent promising platforms for neural tissue engineering and regenerative medicine, further translational studies and well-designed clinical investigations are required to establish their therapeutic efficacy and clinical applicability.\n --- END ACTUAL ABSTRACT FOR 42514227 ---\n\n\n\u2705 PASSED (DO NOT CHANGE THESE):\n- \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" (Source: 42102607)\n- \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\" (Source: 37037273)\n- \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\" (Source: 37394036)\n- \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\" (Source: 41540479)\n- \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\" (Source: 38110531)\n- \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\" (Source: 42458952)\n- \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\" (Source: 42504872)\n- \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\" (Source: 42543397)\n- \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\" (Source: 42275483)\n- \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\" (Source: 42422258)\n- \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\" (Source: 42422258)\n- \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\" (Source: 41989572)\n- \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\" (Source: 42506696)\n- \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\" (Source: 42342866)\n- \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\" (Source: 42503395)\n- \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\" (Source: 42466399)\n- \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\" (Source: 42466399)\n- \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\" (Source: 42350670)\n- \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\" (Source: 42401592)\n\n\nINSTRUCTION: Study the actual abstracts provided. Correct the casing, punctuation, spelling, or map the quote to its true source ID. Do NOT use ellipses.\n\n=======================================================\n\u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f CRITICAL VERIFICATION FAILURE (RETRY LOOP DETECTED) \u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f\nYour previous response was REJECTED because your quotes failed strict byte-perfect validation.\n\nTO BREAK THE LOOP, FOLLOW THESE 3 ABSOLUTE RULES:\n1. NO REPAIRING: If a quote failed, do NOT attempt to edit or tweak it. Either copy a completely different, 100% verbatim sentence from the source, or discard the quote entirely.\n2. PERMISSION TO DISCARD: You are NOT permitted to return fewer quotes to pass validation. Never hallucinate just to meet a quota.\n3. BYTE-PERFECT COPY: You must perform a direct, literal copy-paste. Ellipses (...) are BANNED. Do not change a single capital letter, punctuation mark, or space.\n=======================================================\n\nBASED STRICTLY ON THE CONTEXT LITERATURE ABOVE, EVALUATE THIS SPECIFIC PERSPECTIVE/PENTAMATRIX:\n\"Hypothesis: If Botox were injected into the nerve ending on the third toe on the left foot, the toxin can \"climb\" retrograde from toe through motor neurons eventually to the brain and temporarily inhibit vesicular docking \"from toe to head\", potentially allowing a timed pharmacological therapeutic window for toxic EV clearance through the lymph system instead of docking and seeding.\"",
"rawResponse": "Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\nThe claim that Botulinum toxin (BoNT) injected into a peripheral nerve ending (e.g., third toe) can undergo retrograde transport to the central nervous system (CNS) to inhibit vesicular docking and facilitate a therapeutic window for toxic extracellular vesicle (EV) clearance is scientifically provocative but currently unsupported by the provided literature. While literature confirms BoNT\u2019s capacity for retrograde transport from peripheral sites to the spinal cord dorsal horn and its action as a SNARE-cleaving endopeptidase that inhibits neurotransmitter release, there is no evidence that BoNT-A facilitates the specific \"clearing\" of toxic EVs. Moreover, the claim that it can reach the brain from a distal toe-nerve ending via motor neurons to therapeutically manipulate EV clearance is speculative and lacks empirical validation in the provided source materials.\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThis analysis examines the bidirectional potential of botulinum neurotoxin-A (BoNT-A) for modifying synaptic signaling and its hypothetical intersection with lymphatic-mediated extracellular vesicle (EV) disposal. The claim is evaluated against the mechanism of SNARE-dependent exocytosis and retrograde axonal transport dynamics documented across neurodegenerative models.\n\n### [INTRODUCTION & JUSTIFICATION]\nBotulinum neurotoxin type A (BoNT-A) functions primarily as a zinc-dependent endopeptidase that cleaves synaptic SNARE proteins, such as SNAP-25, thereby preventing the fusion of synaptic vesicles with the plasma membrane. The literature establishes that BoNT-A \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" This mechanism is clinically employed for neuromuscular conditions where \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\" \n\nRegarding the propagation of pathological proteins, \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\" In models of Alzheimer's disease, \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\" While BoNT-A can inhibit the synaptic release of pathological proteins, the provided literature does not support the hypothesis that BoNT-A injected in distal limbs can reach the brain to \"clear\" EVs. Rather, the clearance of brain-derived toxins is primarily attributed to specialized drainage structures, noting that \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\" Thus, while BoNT-A affects vesicular docking at synapses, it is not currently identified as a mechanism to facilitate EV egress via the lymphatic system.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT-A is not strictly limited to the neuromuscular junction; \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n* Exosomes/EVs represent a novel trafficking route: \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n* Synaptic plasticity is regulated by SNAREs such as Stx4: \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n* Retrograde transport is a common pathway for both beneficial and pathological cargo: \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n* Bioenergetic failure in peripheral nerves leads to retrograde signaling disruption: \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n* Non-traditional secretion pathways exist: \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n* Swallowing disorders are an example of successful BoNT-A clinical targeting: \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n* Mitochondrial transfer exists as a natural restorative mechanism: \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n* Spatial limits of current interventions: \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n* Essential role of Munc18-1: \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 42102607 - Application: Demonstrates the retrograde capacity of the toxin. - \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"\n2. ID: 37037273 - Application: Demonstrates BoNT-A inhibition of tau release via SNAP25. - \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\"\n3. ID: 37394036 - Application: Outlines the transport mechanisms for pathogenic proteins. - \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\"\n4. ID: 41540479 - Application: Explains the meningeal lymphatic system as a clearance pathway. - \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\"\n5. ID: 38110531 - Application: Discusses EV movement between circulation and the lymphatic system. - \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\"\n6. ID: 42458952 - Application: Discusses SNARE-mediated synaptic plasticity. - \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\"\n7. ID: 42504872 - Application: Details unconventional protein secretion pathways. - \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\"\n8. ID: 42543397 - Application: Discusses barriers to nasal delivery. - \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\"\n9. ID: 42275483 - Application: Describes retrograde transport of EVs. - \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\"\n10. ID: 42422258 - Application: Discusses peripheral nerve stimulation in swallowing. - \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\"\n11. ID: 42422258 - Application: Mentions BoNT-A in cricopharyngeal muscles. - \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\"\n12. ID: 41989572 - Application: Discusses the general medical mechanism of BoNT. - \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\"\n13. ID: 42506696 - Application: Discusses the central effects of BoNT-A. - \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\"\n14. ID: 42342866 - Application: Explains axonal metabolic deficit. - \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\"\n15. ID: 42503395 - Application: Notes EV uptake mechanisms. - \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\"\n16. ID: 42466399 - Application: Confirm astrocyte-to-neuron mitochondrial transfer. - \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\"\n17. ID: 42466399 - Application: Discusses spatial limits of neuroprotection. - \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\"\n18. ID: 42350670 - Application: Discusses metabolic state-dependent paths. - \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\"\n19. ID: 42401592 - Application: Discusses the necessity of SNARE regulators. - \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\"\n20. ID: 42427545 - Application: Dystrophic changes and membrane imbalances. - \"Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.\"\n\n###JSON_START###\n{\n \"Alignment\": 3,\n \"Consilience\": 5,\n \"Confidence\": 2,\n \"Logic_Chain\": [\n {\n \"Step\": 1,\n \"From\": \"Distal BoNT-A Injection\",\n \"Relationship\": \"-->\",\n \"To\": \"Retrograde Transport to CNS\",\n \"evidence_source_id\": \"42102607\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 6,\n \"Confidence_Score\": 5,\n \"Gap_Strength\": \"None\",\n \"Justification\": \"Evidence supports retrograde transport of BoNT to dorsal horn.\",\n \"Color\": \"lightgreen\"\n },\n {\n \"Step\": 2,\n \"From\": \"Retrograde Transport to CNS\",\n \"Relationship\": \"-->\",\n \"To\": \"Inhibition of Vesicular Docking\",\n \"evidence_source_id\": \"37037273\",\n \"Alignment_Score\": 5,\n \"Consilience_Score\": 5,\n \"Confidence_Score\": 4,\n \"Gap_Strength\": \"medium\",\n \"Justification\": \"BoNT inhibits docking via SNARE cleavage, but brain-wide therapeutic precision is speculative.\",\n \"Color\": \"lightblue\"\n },\n {\n \"Step\": 3,\n \"From\": \"Inhibition of Vesicular Docking\",\n \"Relationship\": \"-->\",\n \"To\": \"Enhanced EV Lymphatic Clearance\",\n \"evidence_source_id\": \"41540479\",\n \"Alignment_Score\": 2,\n \"Consilience_Score\": 1,\n \"Confidence_Score\": 1,\n \"Gap_Strength\": \"strong\",\n \"Justification\": \"No evidence links BoNT synaptic inhibition to increased lymphatic clearance of EVs.\",\n \"Color\": \"pink\"\n }\n ],\n \"Verbatim_Quotes\": [\n { \"quote\": \"demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\", \"source_id\": \"42102607\" },\n { \"quote\": \"Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons.\", \"source_id\": \"37037273\" },\n { \"quote\": \"Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis.\", \"source_id\": \"37394036\" },\n { \"quote\": \"The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal.\", \"source_id\": \"41540479\" },\n { \"quote\": \"the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.\", \"source_id\": \"38110531\" },\n { \"quote\": \"Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD).\", \"source_id\": \"42458952\" },\n { \"quote\": \"vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release.\", \"source_id\": \"42504872\" },\n { \"quote\": \"its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times.\", \"source_id\": \"42543397\" },\n { \"quote\": \"EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes.\", \"source_id\": \"42275483\" },\n { \"quote\": \"Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles.\", \"source_id\": \"42422258\" },\n { \"quote\": \"Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle.\", \"source_id\": \"42422258\" },\n { \"quote\": \"This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives.\", \"source_id\": \"41989572\" },\n { \"quote\": \"These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action.\", \"source_id\": \"42506696\" },\n { \"quote\": \"When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration.\", \"source_id\": \"42342866\" },\n { \"quote\": \"SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake.\", \"source_id\": \"42503395\" },\n { \"quote\": \"In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito).\", \"source_id\": \"42466399\" },\n { \"quote\": \"This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.\", \"source_id\": \"42466399\" },\n { \"quote\": \"Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states.\", \"source_id\": \"42350670\" },\n { \"quote\": \"In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1.\", \"source_id\": \"42401592\" },\n { \"quote\": \"Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis.\", \"source_id\": \"42427545\" }\n ],\n \"Study_Type_Audit\": {\n \"42102607\": \"in_vivo:Count=1\",\n \"37037273\": \"in_vitro:Count=1\",\n \"37394036\": \"review:Count=1\",\n \"41540479\": \"in_vivo:Count=1\"\n },\n \"Gap_Analysis_Audit\": {\n \"study_type\": \"in_vitro/in_vivo models\",\n \"study_intent\": \"neurotoxin mechanisms\",\n \"justification\": \"Evidence is missing for the specific hypothesis regarding distal toe-to-brain BoNT transport resulting in therapeutic EV clearance.\",\n \"predicted_result\": \"BoNT will block local neurotransmission but cannot be relied upon for brain-wide EV clearance.\",\n \"short_answer_to_user\": \"No current evidence supports BoNT as a systemic EV clearance tool via the proposed toe-to-brain retrograde pathway.\"\n },\n \"suggested_experiments\": [\n \"Assess if BoNT-A injection at sciatic nerve termini influences the rate of fluorescently tagged EV accumulation in the deep cervical lymph nodes.\",\n \"Test whether selective SNARE inhibition via BoNT-A in the spinal dorsal horn affects the efflux of brain-derived EVs in a mouse model.\"\n ],\n \"suggested_studies\": [\n \"A systematic analysis of retrograde axonal transport rates of BoNT-A derivatives in the context of neurodegenerative protein spread.\",\n \"Investigation into the impact of BoNT-A-induced SNAP25 cleavage on the secretion profile of EVs in spinal interneurons.\"\n ],\n \"swansons_literature_based_discovery_candidates\": {\n \"Discovered Hypothesis (A to C)\": \"BoNT-A mediated inhibition of synaptic exocytosis can reduce the pool of neuronally-derived EVs available for trans-synaptic propagation, thereby limiting disease seeding.\",\n \"Literature A (Origin)\": \"BoNT-A synaptic silencing (Source 37037273)\",\n \"Literature C (Target)\": \"Intercellular propagation of pathogenic proteins/EVs in ALS (Source 37394036)\",\n \"The Intersecting Bridge B\": \"SNARE-dependent exocytosis (SNAP25)\",\n \"Biological Rationale\": \"Since SNARE complex integrity is required for both neurotransmitter release and the unconventional secretion of EVs, BoNT-A induced cleavage of SNAP25 effectively halts the exocytic machinery used by pathogenic EVs to spread along connected neural circuits.\"\n },\n \"contradictions_between_evidences\": \"None identified regarding the primary mechanisms, though the therapeutic outcome of BoNT in central nervous system processes remains a point of clinical study.\",\n \"repurposed_solutions\": \"Repurpose BoNT-A as a tool for spatiotemporally controlled cessation of pathological EV secretion, rather than just neuromuscular paralysis.\"\n}\n###JSON_END###",
"thought": "\n",
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},
{
"name": "Run2_Eval1_synthesis",
"text": "Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.",
"metrics": {
"Alignment": 5,
"Consilience": 5,
"Confidence": 4,
"Logic_Chain": [
{
"Step": 1,
"From": "Endocytosis",
"Relationship": "-->",
"To": "Synaptosomal-Associated Protein 25",
"evidence_source_id": "42102607",
"Alignment_Score": 7,
"Consilience_Score": 7,
"Confidence_Score": 6,
"Gap_Strength": "None",
"Justification": "BoNT/A binds and cleaves SNAP-25 to inhibit exocytosis.",
"Color": "lightgreen"
},
{
"Step": 2,
"From": "Synaptosomal-Associated Protein 25",
"Relationship": "-->",
"To": "Autophagosomes",
"evidence_source_id": "25878289",
"Alignment_Score": 6,
"Consilience_Score": 6,
"Confidence_Score": 5,
"Gap_Strength": "medium",
"Justification": "Toxin is internalized and directed into retrograde carriers.",
"Color": "lightblue"
},
{
"Step": 3,
"From": "Axonal transport",
"Relationship": "-->",
"To": "Endolysosomes",
"evidence_source_id": "25878289",
"Alignment_Score": 6,
"Consilience_Score": 6,
"Confidence_Score": 5,
"Gap_Strength": "medium",
"Justification": "LC3-positive autophagosomes fuse with lysosomes in the soma.",
"Color": "lightblue"
}
],
"Verbatim_Quotes": [
{
"quote": "Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.",
"source_id": "25878289"
},
{
"quote": "Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.",
"source_id": "25878289"
},
{
"quote": "We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.",
"source_id": "23300443"
},
{
"quote": "Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.",
"source_id": "23110146"
},
{
"quote": "The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.",
"source_id": "21929507"
},
{
"quote": "Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"source_id": "42102607"
},
{
"quote": "The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.",
"source_id": "39196607"
},
{
"quote": "An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.",
"source_id": "39196607"
},
{
"quote": "Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.",
"source_id": "42102607"
},
{
"quote": "A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.",
"source_id": "21929507"
},
{
"quote": "Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.",
"source_id": "25878289"
},
{
"quote": "Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.",
"source_id": "9300434"
},
{
"quote": "However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.",
"source_id": "25878289"
},
{
"quote": "To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.",
"source_id": "39196607"
},
{
"quote": "We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.",
"source_id": "23110146"
},
{
"quote": "The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.",
"source_id": "23300443"
},
{
"quote": "Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.",
"source_id": "9300434"
},
{
"quote": "Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.",
"source_id": "42102607"
},
{
"quote": "Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).",
"source_id": "9300434"
},
{
"quote": "Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.",
"source_id": "39196607"
}
],
"Study_Type_Audit": {
"23300443": "in_vivo:Count=1",
"25878289": "in_vitro:Count=1",
"39196607": "in_vitro:Count=1",
"42102607": "in_vivo:Count=1"
},
"Gap_Analysis_Audit": {
"study_type": "in_vitro/in_vivo",
"study_intent": "trafficking",
"justification": "The evidence supports retrograde transport and lysosomal fusion but lacks direct kinetic measurement of the hypothesized 'bottleneck' transition.",
"predicted_result": "Quantitative shifts in cargo destination.",
"short_answer_to_user": "While retrograde transport and endolysosomal processing of BoNT/A are established, the specific 'bottleneck' shift remains a mechanistic hypothesis needing further verification."
},
"suggested_experiments": "Quantify the ratio of retrograde-to-exocytic vesicle flux in the presence vs. absence of BoNT/A using live-cell confocal imaging. Analyze the accumulation of tagged pathological proteins in the soma using pulse-chase methods in compartments.",
"suggested_studies": "Compare somatic lysosomal enzyme activity levels in BoNT/A-treated vs. control neurons.",
"swansons_literature_based_discovery_candidates": "- Discovered Hypothesis (A to C): BoNT/A-induced somatic endolysosomal upregulation could serve as a compensatory clearing mechanism for tau-containing microclots in tauopathy-prone neurons. - Literature A (Origin): BoNT/A retrograde transport to the soma and fusion with lysosomes (ID: 25878289) - Literature C (Target): Amyloidogenic protein clearing in neurodegeneration (ID: 42553548) - The Intersecting Bridge B: LC3-positive autophagosomes and lysosomal fusion. - Biological Rationale: BoNT/A routes cargo into the somatic lysosomal degradation pathway; thus, stimulating this specific retrograde pathway may mechanically facilitate the clearance of other non-toxin-related intracellular protein aggregates.",
"contradictions_between_evidences": "None identified in the current literature set regarding retrograde transport existence, though spatial distribution of cleavage varies.",
"repurposed_solutions": "Utilization of BoNT/A-induced retrograde transport as a targeted vehicle to deliver degradation-inducing motifs to the neuronal soma for neurodegenerative diseases.",
"BoNT_induced_lysosomal_flux": "The evidence indicates that BoNT/A is routed to lysosomes, but whether this specifically draws in other exogenous pathological proteins like Tau remains an experimental inference.",
"EV_somatic_sequestration_rate": "Insufficient data to quantify the specific change in EV reaching the soma compared to terminal release.",
"synaptic_vs_somatic_clearance_ratio": "Evidence confirms a shift toward somatic processing of the toxin, but a definitive clearance ratio of general synaptic-to-somatic cargo is not provided.",
"QuoteValidation": [
{
"quote": "Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.",
"source_id": "25878289",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quote": "Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.",
"source_id": "25878289",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quote": "We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.",
"source_id": "23300443",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quote": "Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.",
"source_id": "23110146",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quote": "The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.",
"source_id": "21929507",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quote": "Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.",
"source_id": "42102607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quote": "The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.",
"source_id": "39196607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quote": "An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.",
"source_id": "39196607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quote": "Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.",
"source_id": "42102607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quote": "A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.",
"source_id": "21929507",
"status": "PASS",
"error": "",
"abstract_text": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically."
},
{
"quote": "Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.",
"source_id": "25878289",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quote": "Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.",
"source_id": "9300434",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quote": "However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.",
"source_id": "25878289",
"status": "PASS",
"error": "",
"abstract_text": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity."
},
{
"quote": "To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.",
"source_id": "39196607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
},
{
"quote": "We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.",
"source_id": "23110146",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life."
},
{
"quote": "The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.",
"source_id": "23300443",
"status": "PASS",
"error": "",
"abstract_text": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system."
},
{
"quote": "Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.",
"source_id": "9300434",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quote": "Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.",
"source_id": "42102607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases."
},
{
"quote": "Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).",
"source_id": "9300434",
"status": "PASS",
"error": "",
"abstract_text": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals."
},
{
"quote": "Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.",
"source_id": "39196607",
"status": "PASS",
"error": "",
"abstract_text": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity."
}
]
},
"displayText": "Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\n\"Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.\"\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThe provided literature confirms that BoNT/A undergoes retrograde transport from peripheral nerve terminals to the soma, where it interacts with intracellular trafficking machinery and influences central nervous system components. While evidence verifies that BoNT/A utilizes retrograde transport and that it can be sequestered in LC3-positive autophagosomes, the specific hypothesis regarding a \"bottleneck\" shift from synaptic exocytosis to somatic degradation remains a theoretical model requiring further quantitative validation. The literature establishes that BoNT/A cleavage of SNAP-25 occurs at the terminal and subsequently within the soma, but direct kinetic quantification of this shift in clearance pathways requires gap-filling studies.\n\n### [INTRODUCTION & JUSTIFICATION]\nThe intracellular itinerary of Botulinum Neurotoxin Type A (BoNT/A) is far more expansive than its canonical role in blocking neurotransmitter release at the neuromuscular junction. Evidence demonstrates that the toxin enters neurons via synaptic vesicles and is subsequently sorted into retrograde axonal transport carriers. ID: 25878289 notes that \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\" Once internalized, the toxin engages with autophagosomal pathways, as \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\" This interaction with the endolysosomal system is critical, particularly as \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\" The ability of BoNT/A to alter the neurobiological landscape is confirmed by findings that \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\" Thus, while the \"bottleneck\" effect remains a specific hypothesis, the literature supports a shift in proteostatic burden and cellular signaling following retrograde axonal transport.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT/A-induced inhibition of SNAP-25 does not merely halt release; it causes secondary synaptic stripping in motoneurons, a phenomenon characterized by a 15-fold increase in spine density and significant synaptic bouton detachment.\n* Retrograde trafficking of BoNT/A is not limited to synaptic vesicles but utilizes autophagosomes (LC3-positive carriers), which serve as a highway for delivering the toxin from the terminal to the soma.\n* The toxin's effect extends beyond the presynaptic membrane; it influences the proliferation and secretory states of associated glia, such as Schwann cells, highlighting a multicellular impact.\n* BoNT/A displays differential intracellular migration; while protease activity is often concentrated locally at the site of application, a measurable portion accumulates in the soma over several weeks.\n* The retrograde pathway of BoNT/A can be hijacked for therapeutic purposes, as evidenced by its application in downregulating pain-related neuropeptides like CGRP and ion channels like TRPV1 in the spinal dorsal horn.\n* Neurotoxic activity is not synonymous with death; motoneurons can undergo significant structural remodeling and transient hypertrophy following BoNT/A exposure without immediate organelle degeneration.\n* Evidence suggests that the fast axonal retrograde transport compartment is multifunctional, acting as a gateway for multiple pathogenic virulence factors, not just BoNTs.\n* There is a distinct discrepancy between the timing of peripheral paralysis and the appearance of central neurotoxic effects, suggesting a staggered temporal profile for the toxin's dual mechanism of action.\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 25878289 - Application: Investigating BoNT/A retrograde trafficking mechanisms. Alignment: 5. Quote: *\"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\"*\n2. ID: 25878289 - Application: Assessing autophagosomal involvement. Alignment: 5. Quote: *\"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\"*\n3. ID: 23300443 - Application: Confirming retrograde transport in motor neurons. Alignment: 5. Quote: *\"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\"*\n4. ID: 23110146 - Application: Demonstrating cl-SNAP-25 presence in CNS. Alignment: 5. Quote: *\"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\"*\n5. ID: 21929507 - Application: Tracking toxin movement in neurites. Alignment: 5. Quote: *\"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\"*\n6. ID: 42102607 - Application: Elucidating CNS involvement. Alignment: 5. Quote: *\"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"*\n7. ID: 39196607 - Application: Identifying ER-related translocation. Alignment: 5. Quote: *\"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\"*\n8. ID: 39196607 - Application: Tracking Golgi-ER trafficking. Alignment: 5. Quote: *\"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\"*\n9. ID: 42102607 - Application: Antipruritic mechanism explanation. Alignment: 5. Quote: *\"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\"*\n10. ID: 21929507 - Application: Temporal somatic accumulation. Alignment: 5. Quote: *\"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\"*\n11. ID: 25878289 - Application: Activity-dependent retrograde transport. Alignment: 5. Quote: *\"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\"*\n12. ID: 9300434 - Application: Structural alteration of synapses. Alignment: 5. Quote: *\"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\"*\n13. ID: 25878289 - Application: Defining CNS activity. Alignment: 5. Quote: *\"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\"*\n14. ID: 39196607 - Application: Identification of genome-wide screening. Alignment: 4. Quote: *\"To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.\"*\n15. ID: 23110146 - Application: Modulation of Schwann cells. Alignment: 5. Quote: *\"We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.\"*\n16. ID: 23300443 - Application: Comparing TeNT and BoNTs. Alignment: 4. Quote: *\"The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.\"*\n17. ID: 9300434 - Application: Detachment of synaptic boutons. Alignment: 5. Quote: *\"Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.\"*\n18. ID: 42102607 - Application: SNAP-25 in mast cells. Alignment: 5. Quote: *\"Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.\"*\n19. ID: 9300434 - Application: Long-term synaptic changes. Alignment: 5. Quote: *\"Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).\"*\n20. ID: 39196607 - Application: Neuronal somatic processing. Alignment: 5. Quote: *\"Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.\"*\n\n### [PROGRAMATICALLY MAPPED REFERENCES]\n[1]. ID: 42102607 - APA: Lan YA, Guo JX, Yao MH, Zhang XY, Liao ZR et al. (2026). BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.. International immunopharmacology. ID: 42102607.\n[19]. ID: 25878289 - APA: Wang T, Martin S, Papadopulos A, Harper CB, Mavlyutov TA et al. (2015). Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.. The Journal of neuroscience : the official journal of the Society for Neuroscience. ID: 25878289.\n[20]. ID: 23300443 - APA: Restani L, Giribaldi F, Manich M, Bercsenyi K, Menendez G et al. (2012). Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.. PLoS pathogens. ID: 23300443.\n[21]. ID: 23110146 - APA: Marinelli S, Vacca V, Ricordy R, Uggenti C, Tata AM et al. (2012). The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.. PloS one. ID: 23110146.\n[22]. ID: 21929507 - APA: Lawrence GW, Ovsepian SV, Wang J, Aoki KR, Dolly JO (2012). Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.. The Biochemical journal. ID: 21929507.\n[23]. ID: 39196607 - APA: Yeo JC, Tay FP, Bennion R, Loss O, Maignel J et al. (2024). Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.. eLife. ID: 39196607.\n[24]. ID: 9300434 - APA: Pastor AM, Moreno-L\u00f3pez B, De La Cruz RR, Delgado-Garc\u00eda JM (1997). Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.. Neuroscience. ID: 9300434.\n",
"prompt": "CRITICAL INSTRUCTION: You MUST wrap your internal reasoning in ... tags at the very beginning of your response.\n\n=======================================================\nCONTEXT LITERATURE (STATIC CACHE):\nID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases.\n\nID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity.\n\nID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity.\n\nID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system.\n\nID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life.\n\nID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically.\n\nID: 10783705\nTitle: [Muscarinic modulation of cardiac activity].\nAbstract: The goal of the present review is to report information concerning cardiac innervation or more precisely to approach the modulation of cardiac electrical and mechanical activity by parasympathetic innervation. Acetylcholine (ACh) release by nerve endings from the vagus nerve hyperpolarizes the membrane, shortens action potential (AP) duration and has a negative inotropic effect on cardiac muscle. Toxins are usefull tools in the study of membrane signals. The Caribbean ciguatoxin (C-CTX-1) has a muscarinic effect on frog atrial fibres. The toxin evokes the release of ACh from motoneuron nerve terminals innervating this tissue which allows us to propose a model, similar to the one of the neuromuscular junction (nmj), to describe the events occurring during the triggering and release of ACh. Trachynilysin (TLY) is a proteic toxin which causes an influx of Ca2+ into the cells and releases ACh from nmj synaptic vesicles. TLY has a muscarinic effect on atrial fibres which is explicated in the release of neurotransmitter from the nerve endings generated by the TLY-induced Ca2+ influx. It is known that ACh release from nmj is known to be due to exocytosis of synaptic vesicles via the activation of a proteic complex blocked by botulinum toxins. One of these proteins SNAP-25 is the target of type A botulinum toxin (BoNT/A). The study of hearts isolated from BoNT/A poisoned frogs show that atrial AP is lengthened and reveals the presence of SNAP-25 in nerve endings of this tissue. Moreover, the electrical activity of ventricular muscle is markedly altered; in BoNT/A treated frog, an important outward current activated by internal Ca2+ develops. ACh released from nerve terminals binds to a G protein coupled membrane receptor and activates a K+ channel and other effectors. Five subtypes of muscarinic receptors have been cloned from different tissue (M1, M2, M3, M4) subtypes have been identified in cardiac tissues throughout many species. These receptors coupled with different G-proteins activate different effectors. M1 receptors modulate the cardiac plateau and therefore the magnitude of the peak contraction. M2 receptors are mainly involved in the repolarization phase of the AP and modulate the duration of the peak contraction. The roles of M3 and M4 are not yet clearly defined; however, they may activate K+ currents. In conclusion, ACh releases from parasympathetic nerve endings which innervate cardiac cells follows to similar events (Ca2+ influx; presence of a SNAP-25 protein) to those which produce ACh release from nmj, stimulates different G proteins coupled muscarinic receptors, and activates different effectors involved in the modulation of cardiac electrical and mechanical activity.\n\nID: 9344876\nTitle: Intracellular location of SNAP-25 in human neutrophils.\nAbstract: Exocytosis plays an essential role in the physiological functions of human neutrophils. Although SNAP-25 is considered to play a key role in vesicle-membrane fusion, it has been detected almost exclusively in the neuronal system. Using different specific antibodies to SNAP-25, we have identified in the membrane fraction of resting human neutrophils an immunoreactive band with the same molecular mass observed in brain homogenates. Immunoblot analysis of subcellular fractions of neutrophils revealed that SNAP-25 protein was found in the granule membrane fraction, but not in the cytosolic and plasma membrane fractions. Granule localization for neutrophil SNAP-25 was further demonstrated by confocal and immunoelectron microscopy. Furthermore, SNAP-25 was mainly located in the morphologically defined neutrophil peroxidase-negative granules, which are mobilizable upon cell activation. In addition, the protein was specifically cleaved by botulinal neurotoxin A, as observed in brain homogenate. These findings reveal the presence of SNAP-25 in the granule membranes of human neutrophils.\n\nID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals.\n\nID: 42554595\nTitle: Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy.\nAbstract: During glioma progression, monocytes abundantly infiltrate but primarily differentiate into immunosuppressive macrophages to promote tumor growth. Redirecting monocyte differentiation offers a compelling yet underexplored therapeutic opportunity. In this work, we found M1-polarized macrophage-derived extracellular vesicles (M1-EVs) efficiently induced monocytes to differentiate into anti-tumor macrophages via tumor necrosis factor alpha (TNF-\u03b1)-mediated signaling. Despite promising, the therapeutic efficacy of M1-EVs was constrained by insufficient glioma accumulation and CD47-mediated phagocytic inhibition. To address this challenge, we further engineered M1-EVs with dual-targeting specificity by genetically incorporating a tumor-directed chimeric antigen receptor (CAR) against IL13R\u03b12 or EGFRvIII together with CD47-blocking SIRP\u03b1 variants. The resulting dual-targeting EVs (M1-CS-EVs) exhibited enhanced blood-brain barrier (BBB) penetration and glioma accumulation while locally disrupting CD47-SIRP\u03b1 interactions. In three orthotopic glioma models, M1-CS-EVs elicited a potent anti-tumor immune response and enhanced tumor phagocytosis, significantly suppressing tumor growth while prolonging animal survival. Our findings establish a platform technology for directing monocyte differentiation toward anti-tumor phenotypes, offering a broadly applicable strategy for glioma treatment.\n\nID: 42554589\nTitle: Why do bisphosphonates work in the treatment of CRPS? A translational pharmacological perspective to solve a conundrum.\nAbstract: Bisphosphonates (BPs) have consistently yielded positive results in the treatment of Complex Regional Pain Syndrome (CRPS). However, biochemical and histopathological evidence suggests that osteoclasts, the primary pharmacological target of BPs, is not a key driver in the early pathogenetic steps of CRPS. To critically review and integrate current evidence on pharmacological mechanisms of action of BPs in CRPS and to propose unifying hypotheses explaining their clinical efficacy beyond antiosteoclastic activity. Literature from in vitro studies, animal models, and randomized clinical trials was integrated. Approximately 80 primary and review articles were critically reviewed. BPs efficacy in CRPS is best explained by a bone-driven pharmacological model by which local drug accumulation is achieved in the site of disease. This enables modulation of inflammatory and nociceptive processes on adjacent non-bone cells, including macrophages, neutrophils, keratinocytes, and nociceptive fibers. Inhibition of the mevalonate pathway in these cells may reduce inflammatory cytokines, nerve growth factor (NGF), and reactive oxygen species (ROS), while modulating nociceptive signaling pathways. Additional mechanisms, including modulation of vascular tone and neuropeptide signaling, may further contribute to therapeutic effects. This hypothesis may inform optimal treatment strategies and together improve understanding of CRPS pathophysiology.\n\nID: 42554584\nTitle: Magneto-NIR-II-Programmed Cascade Nanozymes Unlocking Blood-Brain Barrier Translocation and Autophagic Resistance in Glioblastoma.\nAbstract: Glioblastoma (GBM) remains a highly aggressive central nervous system malignancy, and its treatment is hindered by poor drug accumulation across the blood-brain barrier (BBB) and autophagy-mediated repair. To address these barriers, rare-earth-doped Nd0.02Fe2.98S4@HA nanozymes (NFSH) are constructed as magneto-NIR-II-programmed cascade nanozymes for trans-BBB delivery, multimodal imaging, and ferroptosis amplification. Hyaluronic acid (HA)-mediated CD44 targeting and oriented magnetic field-enhanced BBB permeability promote tumor enrichment, while Nd3+ doping endows NFSH with strengthened superparamagnetism, near-infrared second window (NIR-II) photodynamic activity, and NIR-II fluorescence capability. Under alternating magnetic field (AMF) and NIR-II laser stimulation, NFSH activates catalase-, peroxidase-, glutathione oxidase-, and nicotinamide adenine dinucleotide (NADH) oxidase-like cascade catalysis, which amplifies reactive oxygen species (ROS) production, consumes glutathione, and induces ferroptosis. In the acidic tumor microenvironment, AMF further promotes H2S release, disrupts lysosomal autophagic degradation, and aggravates mitophagy inhibition through NADH depletion-mediated ATP deficiency. This cascade mechanism enhances ferroptosis and reshapes the tumor immune microenvironment by relieving hypoxia and promoting M2-to-M1 macrophage polarization. In addition, NFSH enables NIR-II fluorescence and T2-weighted magnetic resonance imaging for real-time visualization of treatment. This strategy provides an integrated trans-BBB theranostic platform for autophagy-suppressed ferroptosis therapy against GBM.\n\nID: 42554519\nTitle: Assembly of Bioactive Superstructures via Metal-Phenolic Complexation for Blood Purification.\nAbstract: Hyperbilirubinemia, which stems from bilirubin accumulation caused by liver failure, can lead to jaundice, multi-organ damage, and mortality. Hemoperfusion is the most effective detoxification approach, and its efficacy critically depends on the adsorbent system. However, most developed adsorbents exhibit poor hemocompatibility, posing risks of coagulation and bacterial infection. Herein, we report a facile and highly biocompatible strategy to fabricate bioactive superstructures for blood purification, enabling toxin removal. The formation of the bioactive superstructures is mediated by the assembly of polydopamine/ZnII networks on natural polymer templates, followed by surface modification with human serum albumin. The obtained superstructures enable rapid bilirubin clearance owing to the synergistic effects of metal coordination and polyphenol-mediated noncovalent interactions (hydrogen bonding and hydrophobic and electrostatic interactions). Furthermore, these bioactive superstructures exhibit excellent antibacterial efficacy, achieving > 99.9% killing efficiency of both Escherichia coli and Staphylococcus aureus, while enhancing anticoagulant performance by 2.6-fold (compared to the control) through inhibition of platelet adhesion/activation and modulation of the intrinsic coagulation pathway. This work expands our understanding of metal-phenolic-mediated superstructure assembly. Furthermore, the efficient toxin removal, antibacterial protection, and anticoagulant activity of the developed bioactive superstructures are expected to underpin the rational design of blood-contacting materials based on metal-phenolic complexation.\n\nID: 42554513\nTitle: Macropa-Based 225Ac-Labeled RGD Peptides for Targeted Alpha Therapy.\nAbstract: Macropa has formed more stable complexes with 225Ac, although DOTA has been extensively used as a chelator for 225Ac. This study aimed to design and synthesize two novel 225Ac-labeled RGD peptides, [225Ac]Ac-Macropa-c(RGDyK) ([225Ac]1) and [225Ac]Ac-Macropa-[c(RGDyK)]2 ([225Ac]2), and to evaluate their potential for targeted alpha therapy (TAT) of glioblastoma. Further, whether combination therapy with homoarginine (hArg), a modulator of lysosomal function, could further improve therapeutic efficacy was investigated. In Colon-26 cells, [225Ac]2 demonstrated significantly higher cellular uptake and cytotoxicity than [225Ac]1. In Colon-26 tumor-bearing mice, [225Ac]2 exhibited higher tumor accumulation and significant tumor growth inhibition. In GL261 glioblastoma cells, [225Ac]2 showed comparable cellular uptake to that in Colon-26 cells, and combination with hArg in vitro improved its cytotoxicity. However, no significant difference in therapeutic efficacy was observed between the hArg-treated and control groups in the orthotopic GL261 glioblastoma mouse model. The results of this study indicate the potential of 225Ac-labeled RGD peptides incorporating Macropa as a chelator as promising agents for TAT. However, the further optimization of combination strategies and pharmacokinetics is required.\n\nID: 42554509\nTitle: Mepce Restrains Ferroptosis in Prostate Cancer Through 7SK-P-TEFb Pause Control.\nAbstract: Ferroptosis can be therapeutically induced through GPX4 inhibition, but the mechanisms that determine ferroptosis sensitivity in prostate cancer remain incompletely understood. Here, we combined genome-wide CRISPR screening under graded GPX4 inhibition with mechanistic perturbation-rescue experiments and validation in xenograft and immunocompetent syngeneic prostate cancer models. We identified MEPCE loss as a conserved sensitizer to ferroptosis. Mechanistically, MEPCE depletion destabilized RN7SK and disrupted the HEXIM1-P-TEFb complex, consistent with CDK9 release and enhanced RNA polymerase II Ser2 phosphorylation, thereby promoting transcriptional elongation. This state accelerated activation of a NRF2/ARE stress program encompassing antioxidant defense and iron mobilization. Within this program, HMOX1 emerged as a kinetically sensitive effector whose rapid induction promoted labile iron accumulation and lipid peroxidation under GPX4 inhibition. In vivo, MEPCE suppression impaired tumor growth, enhanced the antitumor efficacy of ferroptosis induction, and was associated with pharmacodynamic markers of ferroptosis that were partially reversed by ferroptosis- or iron-targeted rescue. Together, these findings identify a transcriptional pause-control mechanism that links elongation dynamics to ferroptosis susceptibility and nominate the MEPCE-RN7SK-P-TEFb axis as a potential therapeutic target for sensitizing prostate cancer to ferroptosis.\n\nID: 42554506\nTitle: A Novel tsRNA, 5'tiRNA-GluCTC, Mediates Cardiomyocyte-Fibroblast Crosstalk to Promote Cardiac Fibrosis in Restrictive Cardiomyopathy.\nAbstract: Restrictive cardiomyopathy (RCM) is characterized by pronounced cardiac fibrosis, leading to ventricular stiffening and diastolic dysfunction. While cardiomyocyte mutations are known triggers, the mechanisms initiating pro-fibrotic signaling remain elusive. This study investigates the role of cardiomyocyte-derived exosomes and specific tRNA-derived small RNAs (tsRNAs) in this pathogenic intercellular communication.\u00a0Using a cTnIR193H\u00a0knock-in mouse model, we observed pronounced cardiac fibrosis prior to the onset of heart failure, without significant cardiomyocyte\u00a0apoptosis. This primary fibrotic response was mediated by a paracrine mechanism, as conditioned medium from mutant cardiomyocytes was sufficient to activate fibroblasts. Subsequent small RNA sequencing\u00a0of cardiomyocyte-derived exosomes identified the tRNA-derived fragment 5'tiRNA-GluCTC\u00a0as a significantly enriched species. Functional studies established this RNA as a critical mediator, demonstrating that its overexpression exacerbated fibrotic responses in vitro and induced fibrotic remodeling in wild-type mice, while its inhibition via a cardiac-targeted AAV sponge\u00a0attenuated fibroblast activation and alleviated fibrosis in RCM mice. Mechanistically, 5'tiRNA-GluCTC\u00a0is transferred to fibroblasts, directly targets the 3'UTR of Foxq1\u00a0to repress its expression,\u00a0and\u00a0consequently activates the Smad3/p-Smad3 signaling cascade.\u00a0Collectively, these results delineate a 5'tiRNA-GluCTC/Foxq1/Smad3 signaling axis that drives fibrosis in RCM, uncovering a previously uncharacterized signaling cascade with therapeutic potential.\n\nID: 42554438\nTitle: Matrix Stiffness-Encoded Mechanical Memory Maintains Proliferative Dominance in Metastatic Tumors.\nAbstract: Primary and metastatic lesions share similar biological properties despite existing in different microenvironments. A better understanding of the mechanisms underlying the maintenance of phenotypic homogeneity in heterogeneous microenvironments could help identify strategies for suppressing metastasis. Here, we found that mechanical memory enabled tumor cells to resist biomechanical stress and sustain malignant traits. Matrix stiffness activated the RhoA-ROCK1 signaling pathway, leading to actin cytoskeletal remodeling and suppression of mitochondrial fission. The dysregulation of mitochondrial dynamics enhanced fatty acid \u03b2-oxidation, leading to the accumulation of acetyl-CoA and subsequent histone hyperacetylation. The stiffness-mediated epigenetic reprogramming was mitotically heritable, allowing progeny cells to retain a proliferative advantage after detachment from stiff environments. Inhibition of RhoA-ROCK1 disrupted mechanical memory, thereby reducing metastasis. These findings suggest that targeting mechanical memory may be a strategy for preventing metastasis.\n\nID: 42554406\nTitle: Morphological Changes in Fungal Elements and Antifungal Drug Susceptibility in an Ex Vivo Model of Fungal Keratitis.\nAbstract: Fungal keratitis is a serious eye infection that can cause irreversible vision loss. One of the causative fungal pathogens, Fusarium solani (Neocosmospora solani), shows limited susceptibility to antifungal drugs. Furthermore, differences between in vitro and clinical outcomes are encountered in some cases, complicating treatment. In this study, we compared in vitro and ex vivo antifungal susceptibility using a porcine corneal model of F. solani keratitis. An ex vivo fungal keratitis model was developed using porcine corneas, which were inoculated with F. solani and assigned to four treatment groups: natamycin (NAT), voriconazole (VRCZ), luliconazole (LLCZ), or saline (uninfected control). We compared the antifungal efficacy of each drug on day 2 to day 4 of fungal development. DNA was extracted from all corneas and used in real-time polymerase chain reaction to quantitatively assess fungal growth. Fungal morphology was examined by digital microscopy. All antifungal treatment groups demonstrated antifungal activity. The NAT-treated group showed clear inhibition of fungal growth as well as fungal structural damage. The VRCZ-treated group exhibited growth inhibition; however, fungal growth persisted even after additional drug administration. The LLCZ-treated group not only suppressed fungal growth but also caused structural disruption of fungal cells after additional administration, suggesting a strong antifungal effect. We successfully established an ex vivo Fusarium keratitis model using porcine corneas, which enabled us to evaluate antifungal susceptibility by quantifying fungal growth and observing fungal morphological changes.\n\nID: 42554361\nTitle: Platelet-Derived Exosomal miR-142-3p Inhibits Macrophage M1 Polarization and PANoptosis to Alleviate Acute Lung Injury via the MAPK Pathway.\nAbstract: Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is an acute, life-threatening form of pulmonary disease characterized by systemic inflammation, for which current treatments are not sufficiently effective. Platelet-derived exosomes (PLT-exos) are noted for their specific targeting ability to inflammatory sites, minimal immunogenicity, and positive anti-inflammatory effects, positioning them as a promising therapeutic candidate. However, the specific effects and mechanisms of PLT-exos in ALI/ARDS remain poorly understood. Here, utilizing a lipopolysaccharide (LPS)-induced ALI model in murine and cell models, we first demonstrated that activated platelet-derived exosomes (AP-exos) are preferentially taken up by lung macrophages, leading to suppressed macrophage M1 polarization and PANoptosis and reduced lung inflammation and injury, which were closely linked to the inhibition of MAPK pathway activity. Mechanistically, the protective effects of AP-exos were partially mediated by the highly enriched miR-142-3p, as evidenced by the intervention with miR-142-3p significantly inhibiting their protective effect against LPS-induced ALI injury in mice. Furthermore, we noted a negative correlation between the level of miR-142-3p derived from plasma exosomes and the severity of ARDS, suggesting its potential in predicting the severity of ARDS. Further investigation revealed that TMEM120B (T120B) functions as a downstream target of miR-142-3p. Overall, the findings suggested that activated platelets release exosomes that inhibit macrophage M1 polarization and PANoptosis, contributing to the pathogenesis of ARDS via the miR-142-3p/T120B/MAPK axis.\n\nID: 42554299\nTitle: Hyperpolarized [2-13C]Pyruvate Identifies a Mitochondria-Active Hepatocellular Carcinoma Phenotype With Vulnerability to Mitochondrial Inhibition.\nAbstract: Hepatocellular carcinoma (HCC) exhibits metabolic heterogeneity that is not fully characterized by glycolysis-focused spectroscopic profiling. This study investigated whether in\u00a0vitro hyperpolarized (HP) [2-13C]pyruvate NMR spectroscopy can identify a mitochondria-active HCC phenotype and assess its association with sensitivity to mitochondrial metabolic inhibition. HP [2-13C]pyruvate NMR spectroscopy was used to evaluate mitochondrial metabolism in McA-RH7777 HCC cells, with N1S1 cells serving as a glycolysis-dominant reference. Cell viability following treatment with the glutaminase inhibitor BPTES and the mitochondrial metabolic inhibitor CPI-613 was assessed by MTT assay, and metabolic changes following CPI-613 treatment were further evaluated using HP [2-13C]pyruvate. HP [2-13C]pyruvate demonstrated enhanced pyruvate-to-glutamate conversion in McA-RH7777 cells, whereas N1S1 showed minimal glutamate labeling. CPI-613 treatment resulted in a dose-dependent reduction in cell viability, while BPTES produced limited effects. Although pyruvate-to-glutamate conversion did not significantly decrease following CPI-613 treatment, pyruvate-to-lactate conversion increased, indicating metabolic adaptation. These findings demonstrate that HP [2-13C]pyruvate enables functional identification of a mitochondria-active HCC phenotype characterized by enhanced pyruvate-to-glutamate conversion. This approach may facilitate metabolic subtype classification, help identify tumors susceptible to mitochondrial metabolic inhibition, and enable non-invasive monitoring of treatment-induced metabolic adaptation.\n\nID: 42554268\nTitle: BRCA2-deficient endothelium: a hidden catalyst in angiotensin II-induced vascular injury.\nAbstract: Mutations in the breast cancer susceptibility gene 2 (BRCA2) are well known to increase the risk of breast and ovarian cancers. Emerging evidence indicates that BRCA2 mutation carriers exhibit increased vascular disorder and may develop endothelial dysfunction, a key mechanism underlying hypertension. Angiotensin II (Ang II)\ufeff, a central effector of the renin-angiotensin system, is a key regulator of blood pressure and a major driver of hypertension, promoting endothelial injury through oxidative stress, inflammation, and impaired nitric oxide (NO) bioavailability. However, the role of endothelial BRCA2 in Ang II-induced endothelial dysfunction remains unknown. BRCA2 was silenced in cultured endothelial cells and following Ang II treatment, ROS generation, DNA damage, apoptosis, inflammation, NO production, migration, angiogenic capacity, Ang II receptors and related signaling pathways were assessed. BRCA2 deficiency exacerbated Ang II-induced increases in ROS, DNA damage, and apoptosis, along with impaired functional capacity, including reduced migration and angiogenesis. NO production was suppressed, accompanied by increased micronuclei formation and enhanced c-Jun N-terminal kinase (JNK) activation. In addition, both BRCA2 loss and Ang II treatment upregulated the Ang II receptor AT1R. Pharmacological inhibition of AT1R attenuated Ang II-induced increases in DNA damage and apoptosis in BRCA2-deficient endothelial cells. These findings provide the first pharmacogenomic evidence that BRCA2 deficiency sensitizes endothelial cells to Ang II-induced dysfunction, suggesting that BRCA2 mutation carriers may be at increased risk for hypertension-associated cardiovascular complications.\n\nID: 42554116\nTitle: The Quest for the Associative Device for Hebbian-Like LTP-The Accidental Participant.\nAbstract: In 1985 Holger Wigstr\u00f6m (now deceased) and I published a proposed scenario for the induction of hippocampal Hebbian-like LTP to explain its input specificity and cooperativity. In this scenario LTP is induced by calcium influx through voltage-dependent NMDA receptor channels co-localized with the glutamate receptor channels (non-NMDA) mediating the expression of LTP. This co-localization, we argued, explained the input specificity, and the voltage dependence of these NMDA receptor channels explained the cooperativity. It will be described how this scenario emerged from our finding of a greatly facilitated LTP induction following blockade of postsynaptic inhibition, from our observation of a dendritic, possibly regenerative, event correlated with successful LTP induction, and to our recognition that these events were the result of NMDA receptor activation. Our observation of a likely synaptic co-localization of these receptor channels with the non-NMDA receptor channels, with the former contributing to an EPSP component with a slower onset and much longer duration, will be described. Finally, I will describe how our scenario became validated by our experiments pairing such single EPSPs with depolarization induced by injected current pulses, temporally associated with the NMDA component. This essay will conclude with some take-home messages regarding the Hebbian-like LTP as learned from these experiments: this form of synaptic learning most likely only occurs during disinhibition, its longevity strongly depends on the induction strength from a few minutes and upwards, with a strong induction event it can operate as a one-shot potentiation, and its time window for association may remain several 100\u2009ms. Lastly, Hebbian-like LTP likely evolved to associate input events rather than an input event and somatic spike activity.\n\nID: 42554093\nTitle: Spatiotemporally Controlled Lysosomal Membrane Permeabilization Amplifies STING-Driven Antitumor Immunity in Prostate Cancer.\nAbstract: Prostate cancer (PCa) remains a major clinical challenge due to therapeutic resistance and immunologically cold tumor microenvironment. Lysosomal membrane permeabilization (LMP)-induced lysosome-dependent cell death offers an alternative route to eliminate resistant tumor cells and initiate immunogenic cell death, yet its efficacy is often limited by insufficient spatiotemporal control and immune activation. Here, we report a spatiotemporally programmable supramolecular nanoplatform (Cu-P-MSA) that integrates lysosome-targeted sonodynamic therapy with tumor-confined innate immune activation for PCa treatment. Cu-P-MSA is a modular self-assembling peptide incorporating a PSMA-targeting ligand, morpholine moiety, and cathepsin B-cleavable linker, enabling tumor-selective uptake and in situ formation of fibrous sonosensitizer depots within lysosomes. Upon ultrasound irradiation, a glutathione-responsive open-shell sonosensitizer induces controlled LMP, simultaneously activating ferroptosis and pyroptosis and promoting immunogenic cell death. Meanwhile, tumor-specific release of a STING agonist MSA-2 elicits robust type I interferon responses, driving dendritic cell maturation and cytotoxic T-cell infiltration. This coordinated lysosomal disruption-immune amplification strategy effectively reprograms the tumor immune microenvironment and suppresses both primary and distant tumors, with inhibition rates reaching 84.3% and 77.5%, respectively. Overall, this work establishes a spatiotemporally controlled supramolecular approach that integrates lysosomal disruption with innate immune activation to overcome therapeutic resistance and immunosuppression in PCa.\n\nID: 42554069\nTitle: Ultrasound-Guided Targeted Combination Therapy Integrating VEGFR Degradation Independent of a Specific Receptor, Mitochondrial Damage, and Sonodynamic Immunomodulation.\nAbstract: Inhibiting the vascular endothelial growth factor-vascular endothelial growth factor receptor (VEGF-VEGFR) signaling pathway is clinically established for solid tumors, but durable efficacy is frequently limited by acquired resistance. Accumulating evidence links this resistance to metabolic plasticity, wherein tumors suppress glycolysis and gradually become dependent on mitochondrial respiration for survival, rendering mitochondrial inhibition a rational strategy to mitigate resistance. Recently, targeted protein degradation (TPD) has emerged as an event-driven modality for pathogenic protein elimination, but the degradation of membrane targets is still challenging due to limited transmembrane E3 ligases and heterogeneous lysosome-targeting receptor (LTR) expression. To overcome these challenges, we designed a multifunctional peptide-sonosensitizer conjugate (PSC) by covalently linking a VEGFR-targeting peptide, a mitochondria-homing peptide, and the clinically approved sonosensitizer verteporfin (VPN) via a gelatinase-cleavable linker. PSC self-assembles into stable nanoparticles with tumor-preferential accumulation and, upon ultrasound activation, generates reactive oxygen species (ROS) to induce oxidative degradation of VEGFR, thereby suppressing VEGFR signaling and angiogenesis. Following cellular internalization, enzymatic linker cleavage releases a mitochondria-targeted fragment that triggers mitochondrial depolarization and structural disruption, amplifying ROS-driven apoptosis. Moreover, sonodynamic immune modulation further potentiates antitumor efficacy. This study establishes a spatiotemporally controllablestrategy that is independent of a specific receptor and couples anti-angiogenic pathway blockade with mitochondrial vulnerability to address resistance and enable precision, immune-synergistic therapy.\n\nID: 42554055\nTitle: Mast Cell St8sia1 Is a Glyco-Epigenetic Checkpoint Driving Cardiac Remodeling.\nAbstract: Pathogenic immune-cardiac crosstalk underlies maladaptive remodeling in chronic heart failure, yet therapies directly targeting this axis are lacking. Glycoconjugates, which are crucial for signal transduction and extracellular matrix integrity, represent an underexploited therapeutic avenue. This study sought to define the role of glycoconjugate-metabolizing enzymes at the immune-cardiac interface and evaluate their translational potential. We performed integrative analyses of bulk and single-cell RNA sequencing data from failing human and mouse hearts. Employing mouse models of pressure overload (transverse aortic constriction) and ischemia-reperfusion, we used global and mast cell (MC)-specific gene deletion, bone-marrow chimeras, and pharmacological neutralization. Mechanistic insights were gained through multiomics profiling, including RNA-seq, ATAC-seq, CUT&Tag, and proteomics. The ganglioside GD3 synthase, St8sia1, was selectively induced in cardiac MCs during pathological remodeling in both mice and humans. MC-specific or hematopoietic deletion of St8sia1 preserved ventricular function, attenuated fibrosis, and markedly reduced neutrophil and Ly6C+ monocyte recruitment after transverse aortic constriction and ischemia-reperfusion. Therapeutic neutralization of GD3 with the clinical-grade monoclonal antibody R24 improved cardiac function and diminished scar formation after ischemia-reperfusion. Mechanistically, GD3 bound specific histone variants, such as H2A.Z and H3.3C, thereby reprogramming chromatin accessibility to activate proinflammatory and profibrotic transcriptional programs in MCs. Consequently, GD3 inhibition suppressed MC degranulation, disrupted pathogenic MC-cardiomyocyte/fibroblast crosstalk, and preserved reparative macrophage populations. The MC-restricted St8sia1-GD3 axis functions as a glyco-epigenetic checkpoint driving maladaptive cardiac remodeling. Targeting this axis represents a translatable immunomodulatory strategy to prevent the progression to chronic heart failure.\n\nID: 42553990\nTitle: Ethylene-responsive CmERF3-CmMYB4 cascade suppresses anthocyanin biosynthesis in Chrysanthemum \u00d7morifolium.\nAbstract: Anthocyanins are the primary determinants of floral pigmentation in Chrysanthemum \u00d7morifolium, and ethylene acts as a key regulator of their biosynthesis. Although the ethylene-mediated regulatory circuitry is functionally important, its underlying mechanism in chrysanthemum has remained unclear. In this study, we identified CmMYB4 as a transcriptional repressor that directly suppresses the expression of key anthocyanin biosynthetic genes, including CmDFR (dihydroflavonol 4-reductase), CmUFGT (flavonoid 3-O-glucosyltransferase), and Cm3MaT (anthocyanin 3-O-glucoside-6\u2033-O-malonyltransferase). Time-ordered gene co-expression network analysis comparing the transcriptomes of CmMYB4-overexpressing and control plants, together with molecular biology experimental results, further revealed CmERF3 (ethylene response factor 3) as a hierarchical upstream regulator of anthocyanin biosynthesis. Exogenous ethylene treatment induced CmERF3 expression while reducing anthocyanin accumulation. Subsequent functional characterization showed that the overexpression of CmERF3 suppresses anthocyanin biosynthesis in both tobacco and chrysanthemum by directly activating CmMYB4 and repressing CmDFR, CmUFGT, and Cm3MaT. Collectively, these findings revealed that ethylene inhibits anthocyanin biosynthesis through a CmERF3-CmMYB4-LBGs (late biosynthetic genes) regulatory module. This study not only elucidates the molecular mechanism governing ethylene-mediated anthocyanin inhibition but also provides new perspectives for the molecular engineering of ornamental traits in chrysanthemum.\n\nID: 42553881\nTitle: Elucidating the Nexus of Mitochondrial Dysfunction and Oncometabolite Accumulation in Tumorigenesis.\nAbstract: Cancer is a complex disease driven by disruptions in cellular metabolism and mitochondrial function, enabling malignant cells to proliferate unchecked, evade apoptosis, and metastasize to distant organs. This exhaustive review elucidates the metabolic dysregulations inherent to cancer, with a particular focus on mitochondrial dysfunction, the accumulation of oncometabolites, and the reprogramming of metabolic pathways. A comprehensive literature search was conducted across major scientific databases, including PubMed, Web of Science, Scopus, and ScienceDirect, spanning January 2010 to March 2025. Controlled vocabulary and Boolean operators were employed to capture relevant studies, focusing on cancer metabolism, metabolic reprogramming, tumour markers, oncometabolites, mitochondrial dysfunction, and regulatory pathways. Extracted data were organized into thematic areas, and a qualitative synthesis approach was used to integrate findings, identifying common mechanistic patterns underlying tumour initiation, progression, and metastasis. The Warburg effect, a typical feature of cancer metabolism, is characterized by a predilection for aerobic glycolysis, thereby supporting biosynthetic processes and contributing to tumour microenvironment acidification and immune suppression. Mitochondrial dysfunction triggers genomic instability and oncogenic transformation. Meanwhile, oncometabolites like 2-hydroxyglutarate, fumarate, and sarcosine disrupt cellular signalling and epigenetic regulation, promoting tumour growth and progression. The clinical significance of tumour markers and metabolic biomarkers is underscored, and the systemic metabolic sequelae of cancer, including cancer-associated cachexia, are expounded upon. In glioblastoma, Aurora kinase A inhibition reverses the Warburg effect, decreasing glucose uptake and boosting oxidative phosphorylation. Cancer-associated fibroblasts exhibit aerobic glycolysis, promoting tumor growth and metastasis via the reverse Warburg effect. Glycolysis inhibition suppresses tumor growth in pancreatic cancer, and the Warburg effect contributes to chemoresistance by upregulating glycolytic enzymes and increasing lactate production. Targeting the Warburg effect, including inhibiting glycolytic enzymes and modulating mitochondrial function, offers potential therapeutic strategies for cancer treatment. This review provides a comprehensive exposition of the biochemical mechanisms underpinning metabolic derangements in cancer, which may unveil novel avenues for diagnostic and therapeutic interventions.\n\nID: 42553762\nTitle: Programmed Cell Death: A Key Mechanism of Traditional Chinese Medicine in the Treatment of Membranous Nephropathy.\nAbstract: Membranous nephropathy (MN) is the leading cause of adult nephrotic syndrome with rising global incidence. A substantial proportion of patients progress to end-stage renal disease. Multiple programmed cell death (PCD) pathways-including apoptosis, pyroptosis, ferroptosis, and autophagy-are activated by complement-dependent and -independent pathogenic factors, collectively driving podocyte injury and MN progression. Current immunosuppressive therapies are limited by adverse reactions and variable responses. Traditional Chinese medicine (TCM), characterized by multi-component, multi-target, and multi-pathway regulation, shows therapeutic advantages in MN. To systematically review the regulatory mechanisms of PCD in MN, summarize the molecular basis of TCM interventions targeting PCD, and identify current research limitations and future directions. This narrative review systematically elaborates the regulatory association between PCD and podocyte injury in MN. Literatures were systematically retrieved from PubMed, Embase and China National Knowledge Infrastructure (CNKI) from the establishment of each database to May 2026, and 41 preclinical studies exploring TCM modulation of PCD in MN were included after strict screening. Complement-dependent and complement-independent pathways in MN can synergistically initiate podocyte apoptosis, pyroptosis, ferroptosis, and autophagy dysfunction. TCM can regulate PCD through core signaling pathways such as phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR), nuclear factor erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1), mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase/forkhead box O1 (JNK/FoxO1), and PTEN-induced kinase 1/Parkin (PINK1/Parkin), while simultaneously intervening in non-PCD pathological processes including inflammation, oxidative stress, and renal fibrosis. Clinical evidence further indicates that TCM or combined with conventional therapy can elevate MN remission rates and decrease adverse reactions. PCD represents a core mechanism mediating podocyte injury and MN progression. TCM effectively modulates PCD imbalance through multi-target regulation, demonstrating prominent clinical efficacy and safety. These findings provide a theoretical basis and translational direction for developing high-efficacy, low-toxicity therapeutic strategies for MN.\n\nID: 42553702\nTitle: Distinct brain extracellular vesicle microRNA profiles differ in frontotemporal dementia and Alzheimer's disease.\nAbstract: Dementia is a syndrome caused by various diseases including Alzheimer's disease (AD) and frontotemporal dementia (FTD) with an estimated global prevalence of 60 million individuals. Recently, therapeutic development in the dementia field has accelerated, with the introduction of monoclonal antibody therapeutics such as Lecanemab and Donanemab. However, AD and FTD patients are still either diagnosed too late to benefit from available therapies or are misdiagnosed due to the clinical overlap between dementia subgroups making therapeutic intervention challenging. This highlights a real need to improve early diagnostic tools of neurodegenerative disease (ND) biomarkers. A potential source of such biomarkers come from small extracellular vesicles (sEVs), groups of cell-derived, lipid-bound assemblies with the capability to cross the blood-brain barrier (BBB) and known to carry pathogenic proteins associated with AD and FTD. A known cargo of sEVs is microRNA (miRNA), regulatory molecules that post-transcriptionally silence gene expression including transcripts of autophagic systems, processes which dysfunction in dementia-causing diseases leading to toxic aggregate build-up, causing neurodegeneration. The targeting of functional machineries in macroautophagy (MA) and chaperone-mediated autophagy (CMA) by different miRNA may vary between AD and FTD mutations, leading to potential biomarkers of disease being highlighted. Through isolating sEVs from the frontal cortex of post-mortem brain tissue of AD, FTD-MAPT, FTD-C9orf72, FTD-GRN and no-disease control patients (Manchester Brain Bank), miRNA cargoes were analysed and compared using real-time quantitative PCR (RT-qPCR). Seven autophagy-associated miRNA candidates (MA: miR-124-3p, miR-30a-5p, miR-128-3p; and CMA: miR-224-5p, miR-373-5p, miR-106a-3p and miR-26b-5p) were tested to identify dementia sub-group variations, used alongside small RNA-sequencing to explore broader miRNA variation within sEV populations. Of the miRNA tested miR-224-5p (P = 1.76 \u00d7 10-5) and miR-106a-3p (P = 0.033) showed significant group differences, and further significant pairwise comparison differences [miR-224-5p: AD fold change (FC) = 4.29, MAPT FC = 7.62; miR-106a-5p: AD FC = 5.59] when compared with no disease controls and other dementia subgroups, potentially showing initial diagnostic and differentiating potential. Small RNA-sequencing results revealed 8 AD, 2 FTD-GRN, 52 FTD-MAPT and 12 FTD-C9orf72 differentially expressed sEV-miRNAs when compared with no disease controls. Further direct comparisons between AD versus FTD mutation-derived sEV cargoes, and even FTD mutation versus FTD mutation-derived sEV cargoes, identified additional miRNA with differentiating capabilities. These findings demonstrate sEV-derived miRNA signatures vary across dementia sub-types and suggest potential roles of sEV cargoes in both disease diagnostics and identifying drivers of ND, such as autophagic impairments and signalling pathways.\n\nID: 42553661\nTitle: Emergence of Trichophyton indotineae: in vitro susceptibility data from a mycology reference laboratory in Australia.\nAbstract: Management of Trichophyton indotineae dermatophytosis is challenged by high terbinafine resistance rates within this species. Antifungal susceptibility data for Australian T. indotineae isolates are lacking. Here we determined the in vitro activity of five antifungal agents, including olorofim, against T. indotineae to guide therapeutic choices. Forty-four clinical T. indotineae isolates (cultured from skin, 2005-2025) across several Australian jurisdictions were tested for susceptibility to itraconazole, voriconazole, posaconazole, terbinafine and olorofim using CLSI broth microdilution methodology. MIC endpoints were determined at 80% growth inhibition for azoles and terbinafine, and 100% inhibition for olorofim. Isolates with terbinafine MICs \u22650.5\u2005mg/L were considered as non-WT or resistant. Twenty-eight of 44 (63.6%) T. indotineae isolates had terbinafine MICs \u22650.5\u2005mg/L [geometric mean (GM) MIC 0.570\u2005mg/L; MIC90 4\u2005mg/L] with 21 (47.7%) displaying MICs \u22654\u2005mg/L. The modal terbinafine MIC was 4\u2005mg/L. Lower MICs were observed for azoles, with posaconazole demonstrating the greatest activity, i.e. posaconazole (GM 0.058\u2005mg/L; MIC90 0.25\u2005mg/L), itraconazole (GM 0.087\u2005mg/L; MIC90 0.5\u2005mg/L) and voriconazole (GM 0.201\u2005mg/L; MIC90 1\u2005mg/L). Olorofim demonstrated good in vitro activity with all 44 isolates exhibiting MICs \u200a\u2264\u200a0.06\u2005mg/L (GM 0.023\u2005mg/L; MIC90 0.06\u2005mg/L). Terbinafine resistance in T. indotineae is common in Australia but not universal. Susceptibility testing can inform treatment choice. The azoles and olorofim demonstrate good in vitro activity.\n\nID: 42553528\nTitle: Arthroscopic (Arthro) Delivery of MACI (Autologous Cultured Chondrocytes on a Porcine Collagen Membrane) for Defects on MFC and Trochlea With concomitant TTO.\nAbstract: Chondral pathology of the knee-specifically involving the femoral condyles and trochlea-is commonly observed during knee arthroscopies. This condition may necessitate supplementary interventions, such as a concomitant tibial tuberosity osteotomy (TTO). Approaching this patient population with a straightforward, less invasive arthroscopic technique may offer an advanced solution. Arthro matrix-induced autologous chondrocyte implantation (MACI; autologous cultured chondrocytes on a porcine collagen membrane) targets symptomatic chondral lesions in the knee that are consistent with magnetic resonance imaging findings, which range in size from 2 to 4 cm2, are contained within a stable rim, and are classified as grade 3 to 4a according to the International Cartilage Repair Society. This technique for the arthroscopic delivery of MACI for defects located on the trochlea and femoral condyles has been developed as a progressive advancement in the evolution of autologous chondrocyte implantation technology. The described technique addresses these defect locations by utilizing the same arthroscopic approach for both cartilage defect locations and a separate incision for TTO. The instruments used include 3 varieties of curettes, an arthroscopic cutter to score the cartilage, and the V-shuttle membrane delivery device. Furthermore, attention is dedicated to the medial and lateral approaches, patellar elevation for trochlear access, meticulous defect preparation, hemostasis and fluid management, and implant delivery. Arthroscopic surgical procedures generally result in reduced postoperative pain, necessitating a decreased analgesic requirement, an earlier and enhanced joint mobility, less postoperative muscle inhibition, and expedited recovery. Moreover, an arthroscopic approach may reduce the length of the surgical incision for patients concerned about cosmetic outcomes. In contrast to various other arthroscopic procedures related to knee ligaments or meniscal injuries, the implementation of arthroscopic delivery of MACI for defects on the trochlea and femoral condyles of the knee offers a persuasive and readily executable alternative for a proficient surgeon, even when performed in conjunction with TTO. This arthroscopic technique for the delivery of the MACI implant can effectively treat multiple knee chondral lesions in different compartments when paired with TTO. Advantages of this approach include minimal surgical disruption, the ability to perform combined knee arthroscopic procedures, a more desirable cosmetic appearance, and improved early postoperative recovery. The author(s) attests that consent has been obtained from any patient(s) appearing in this publication. If the individual may be identifiable, the author(s) has included a statement of release or other written form of approval from the patient(s) with this submission for publication.\n\nID: 42553460\nTitle: Unraveling the Molecular Landscape of Follicular Fluid: A Comparative Proteomic Study Between Bali and Brahman Cattle.\nAbstract: Protein biomarkers are increasingly recognized as indicators of metabolic status, immune response, and reproductive performance in cattle. However, comparative data among different breeds commonly raised in Indonesia remain limited. This study aimed to compare the specific proteins in follicular fluid (FF) of Bali and Brahman cows. Samples from FF (n\u2009=\u200920 pairs of ovarium) were analyzed for identification of specific proteins using LC-HRMS analysis. Proteomic analysis revealed distinct breed-specific FF profiles, with 16 proteins uniquely identified in Bali cattle and 23 proteins in Brahman. Bali FF was enriched with proteins associated with protease inhibition, coagulation regulation, extracellular matrix remodeling, and cytoskeletal dynamics, including CORO1C, ZER1, YARS1, and L3MBTL3. In contrast, Brahman FF showed enrichment of proteins involved in lipid oxidation, one-carbon metabolism, and peroxisomal activity, notably ACOX2, BHMT, and KIAA0586. Here, we present the proteomic characterization of FF from Bali and Brahman cattle, where several bioactive proteins are candidate proteins potentially associated with reproductive processes. These findings highlight the potential of protein biomarkers in guiding cattle breeding and management strategies in tropical regions.\n\nID: 42553440\nTitle: Metabolic Reprogramming and Immunometabolic Dysregulation in Diabetic Kidney Disease: From Pathogenesis to Precision Multi-target Therapies.\nAbstract: Diabetic kidney disease, the leading cause of end-stage kidney disease worldwide, involves complex interactions beyond classical hemodynamic and oxidative stress pathways. Recent advances emphasize metabolic reprogramming in renal cells-characterized by mitochondrial dysfunction, impaired fatty acid oxidation, lipotoxicity, and glycolytic shifts-as upstream drivers of cellular injury and fibrosis. Single-cell RNA sequencing reveals profound immunometabolic heterogeneity, including dynamic macrophage subpopulations (e.g., proinflammatory early states transitioning to TREM2hi/MRC1hi lipid-associated phenotypes) and T helper 17/regulatory T imbalance, which amplify inflammation via bidirectional crosstalk with podocytes, tubular cells, and mesangial cells. Interorgan axes, particularly gut dysbiosis and uremic toxin accumulation, further perpetuate immune dysregulation. This review integrates these insights to propose precision strategies targeting mitochondrial homeostasis, ferroptosis inhibition, glycolytic blockade in immune cells, and multimodal therapies (e.g., combination strategies integrating sodium-glucose cotransporter 2 inhibitors with immunometabolic modulators). Multi-omics integration and spatial transcriptomics hold promise for individualized and mechanism-guided interventions to halt diabetic kidney disease progression.\n\nID: 42553406\nTitle: Peripheral and Central Administration of Soluble Glycoprotein 130 Improves Cognitive Outcomes Following Controlled Cortical Impact in Male Mice.\nAbstract: Traumatic brain injury (TBI) initiates complex immune responses, including upregulation of interleukin (IL)-6, a cytokine associated with clinical outcomes after injury. IL-6 trans-signaling, via the solubilized IL-6 receptor, drives pro-inflammatory cascades and is selectively inhibited by soluble glycoprotein 130 (sgp130). Although chronic intermittent sgp130Fc fusion protein (sgp130Fc) treatment after TBI has shown benefits across species, optimal dosing remains unclear. This study evaluated the effects of a single sgp130Fc dose (2 \u03bcg intrahippocampally or 10 \u03bcg intraperitoneally) in male mice on day three following controlled cortical impact or sham procedures. Cognitive performance was assessed using the Morris water maze, and histological assessment included lesion volume and microglia quantification. Intrahippocampal sgp130Fc improved spatial memory during probe trials, decreasing latency to the platform zone (p < 0.05) and enhanced spatial swim strategy selection (p < 0.05). Intraperitoneal sgp130Fc improved spatial learning (p < 0.05) and reduced anxiety-like behavior, indicated by increased target zone time during probe trials and decreased peripheral zone time (p < 0.05). Neither route significantly altered lesion volume or thalamic ionized calcium-binding adaptor molecule 1+ cell counts versus vehicle-treated injured animals; however, intrahippocampal sgp130Fc reduced major histocompatibility complex class II+ cells. These findings demonstrate that a single subacute sgp130Fc dose confers modest behavioral benefits when administered post-TBI, without worsening outcomes. The results underscore the effects of IL-6 trans-signaling inhibition on TBI outcomes and highlight the need for further research into dose, timing, and sex-specific responses. Overall, these data add to emerging support that selective IL-6 blockade with sgp130Fc may represent a translatable strategy to modulate post-TBI inflammation and support cognitive recovery, in part, via reducing anxiety-associated dysfunction.\n\nID: 42553374\nTitle: Durable response to ALK inhibition in low-allele-frequency SPTBN1-ALK-rearranged gastric cancer followed by lineage plasticity-mediated resistance: a case report.\nAbstract: ALK rearrangements are rare in gastric cancer, and their therapeutic relevance remains poorly defined. While ALK inhibitors have demonstrated efficacy in lung cancer and other malignancies, data in gastric tumors are limited. We report a 52-year-old woman with metastatic gastric adenocarcinoma harboring a rare SPTBN1-ALK fusion detected at a low variant allele frequency (0.65%). The tumor was negative for HER2 amplification, microsatellite instability, and PD-L1 expression. After failure of chemotherapy combined with immunotherapy, treatment with the ALK inhibitor iruplinalkib resulted in rapid clinical improvement and a durable partial response lasting approximately 14 months. Serial next-generation sequencing at progression demonstrated enrichment of the fusion-positive clone (variant allele frequency increased to 5.94%), accompanied by expansion of TP53-mutant alleles and acquisition of additional genomic alterations. Subsequent histologic transformation to small-cell neuroendocrine carcinoma was observed, with loss of RB expression and markedly elevated Ki-67. No canonical ALK kinase domain resistance mutations were detected, suggesting a non-on-target resistance mechanism. This case highlights that even low-allele-frequency ALK fusions may have important clinical relevance in gastric cancer and may identify patients who could benefit from ALK-targeted therapy. It also illustrates the dynamic evolutionary trajectory of oncogene-driven tumors under therapeutic pressure, with histologic transformation to small-cell neuroendocrine carcinoma that may reflect lineage plasticity. Comprehensive genomic profiling and longitudinal molecular monitoring may facilitate the identification of rare actionable alterations and improve understanding of resistance mechanisms in advanced gastric cancer.\n\nID: 42553364\nTitle: Targeting the AHR-IDO1-kynurenine pathway in TREM2+ macrophages restores antitumor immunity in thyroid cancer.\nAbstract: Advances in single-cell technologies have provided detailed insight into cellular heterogeneity within tumor microenvironments. However, the regulatory mechanisms driving macrophage-mediated immune suppression in thyroid cancer remain incompletely understood. We applied integrated single-cell multi-omics approaches to tumor samples from patients and murine thyroid cancer models, including single-cell RNA sequencing, single-cell ATAC sequencing, mass cytometry, spatial/histological validation, and in vitro and in vivo functional assays. We identified a prominent enrichment of TREM2+ tumor-associated macrophages within the thyroid cancer immune microenvironment. These macrophages displayed an immunosuppressive transcriptional and metabolic profile and promoted immune evasion through activation of the AHR-IDO1-kynurenine signaling axis, leading to impaired CD8+ T-cell function. Pharmacological inhibition of AHR and IDO1 reversed immune suppression and reduced tumor burden in experimental models. These findings reveal a TREM2+ macrophage-driven immunometabolic mechanism in thyroid cancer and suggest that targeting the AHR-IDO1-kynurenine pathway may provide a potential strategy for improving antitumor immunity.\n\nID: 42553357\nTitle: Antimicrobial Efficacy of Aqueous Extracts of Abrus precatorius and Achyranthes aspera Against Streptococcus mutans and Candida albicans: An In Vitro Study.\nAbstract: Background Dental caries and oral candidiasis are among the most common oral diseases and are primarily associated with Streptococcus mutans and Candida albicans. Growing concerns regarding antimicrobial resistance and the adverse effects of synthetic antimicrobial agents have increased interest in plant-derived alternatives. This study evaluated and compared the antimicrobial efficacies of aqueous extracts of Abrus precatorius and Achyranthes aspera against Streptococcus mutans and Candida albicans. Methodology An in vitro experimental study was conducted using aqueous extracts from the leaves of Abrus precatorius and Achyranthes aspera. The antimicrobial activity against Streptococcus mutans\u00a0(ATCC 25175) and Candida albicans (ATCC 10231) was assessed using the agar well diffusion method. Chlorhexidine and nystatin served as positive controls for antibacterial and antifungal testing, respectively, and distilled water served as the negative control. Minimum inhibitory concentrations (MICs) were determined using the broth microdilution method. Data were analyzed using one-way analysis of variance followed by Tukey's post-hoc test and independent-samples t-test. Results Significant differences in antimicrobial activity were observed among the groups (p < 0.001). Against Streptococcus mutans, Abrus precatorius demonstrated a larger mean zone of inhibition (18.42 \u00b1 1.58 mm) than Achyranthes aspera (15.16 \u00b1 1.47 mm), although chlorhexidine exhibited the greatest antibacterial activity (24.31 \u00b1 1.32 mm). Similarly, against Candida albicans, Abrus precatorius produced a larger inhibition zone (16.87 \u00b1 1.36 mm) than Achyranthes aspera (13.92 \u00b1 1.41 mm), whereas nystatin showed the highest antifungal activity (22.74 \u00b1 1.24 mm). Abrus precatorius also demonstrated significantly lower MIC values against both microorganisms, indicating a greater antimicrobial potency. Conclusions Both aqueous plant extracts exhibited significant antibacterial and antifungal activity. Abrus precatorius showed superior antimicrobial efficacy compared to Achyranthes aspera; however, both extracts were less effective than the standard antimicrobial agents. These findings suggest the potential application of these herbal extracts in the development of oral healthcare products.\n\nID: 42553303\nTitle: Severe non-atopic prurigo nodularis in an otherwise healthy elderly patient: rapid multidimensional response to dupilumab.\nAbstract: Prurigo nodularis (PN) is a chronic neuroimmune skin disorder characterized by intensely pruritic nodules, severe impairment of quality of life and significant psychosocial burden. Prior to the approval of targeted therapies, management relied on conventional agents with limited efficacy. Recent evidence of its neuroimmune pathogenesis have led to the approvement of targeted biologics. Dupilumab, a human monoclonal antibody targeting IL-4/IL-13-axis, has been the first targeted therapy approved for PN. We report a 75-year-old woman presenting with severe, treatment-na\u00efve generalized pruritus (IGA PN-S 4; NRS 9/10; DLQI 21) with extensive nodular involvement of the trunk and extremities, complete sleep deprivation and important anxiety and depression (HADS.A 12; HADS-D 11). Following diagnosis of PN, treatment with dupilumab (600\u00a0mg loading dose, then 300\u00a0mg every 2\u00a0weeks) was initiated in October 2025. At approximately 6 weeks, marked pruritus reduction (NRS 3) and sleep restoration were observed. Progressive improvement continued over time. At 4 months follow-up, complete remission was nearly achieved (NRS 1, IGA PN-S 1, DLQI 2, HADS-A 4, HADS-D 3), with no adverse events reported. This case corroborates and extends the existing evidence on dupilumab efficacy in the treatment of moderate-to-severe PN, with particularly rapid and comprehensive responses across pruritus, skin lesions, sleep and psychosocial outcomes in an elderly patient, through IL-4/IL-13 inhibition. Our findings support dupilumab as a safe and effective targeted option for severe PN, also in older patients where conventional systemic therapies, such as corticosteroids or immunosuppressants, carry a high risk of adverse effects.\n\nID: 42553289\nTitle: Rapamycin-nanoliposomes target the mTORC1-mediated autophagy-lysosomal and NLRP3/Caspase-1 pathways to inhibit nucleus pulposus cell senescence in intervertebral discs.\nAbstract: Nucleus pulposus (NP) cell quiescence maintains intervertebral disc homeostasis, while mTORC1 regulates autophagy-lysosomal function and inflammatory secretion to preserve quiescence-rapamycin specifically targets mTORC1. Herein, we fabricated rapamycin-nanoliposomes (rapa-lipos) via ultrasonic dispersion, thin-film dispersion, and filtration to improve rapamycin bioavailability, investigating their role in inhibiting the senescence phenotype of NP cells through \u03b2-gal staining, lysosomal staining, transmission electron microscopy, ELISA, and cell cycle inhibitors. The mechanistic effects of rapa-lipos on mTORC1, NLRP3/Caspase-1 pathway (NCP) and autophagy-lysosomal pathway (ALP) were also analyzed by western blotting, immunofluorescence (IF), Si-RNA (raptor), and PCR. In vivo, rapa-lipos were injected into rat intervertebral disc with IL-1\u03b2-induced degeneration, assessed via HE staining, x-ray, MRI, and IF. Rapa-lipos exhibited high encapsulation efficiency, favorable drug loading, uniform particle size, and controlled release, suppressing NP cell senescence-related phenotypes (morphological changes, elevated IL-1\u03b2/TNF-\u03b1 secretion, increased \u03b2-gal activity, lysosomal dysfunction, upregulated P21/P16 and reduced P27 expression). Mechanistically, rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence. In vivo, x-ray, MRI and histological evaluation confirmed rapa-lipos mitigated intervertebral disc degeneration. Collectively, rapa-lipos target mTORC1-mediated NCP and ALP to inhibit NP cell senescence, offering a promising strategy for intervertebral disc degeneration prevention.\n\nID: 42553260\nTitle: Curcumin attenuates \u03b1-synuclein pathology in Parkinson's disease model mice through modulation of UBC9-associated SUMOylation signaling.\nAbstract: Post-translational modifications, particularly SUMOylation, plays a crucial role in \u03b1-synuclein (\u03b1-syn) aggregation, a key pathological feature of Parkinson's disease (PD). Curcumin, a natural polyphenol, has shown neuroprotective potential, but its effects on SUMOylation-related signaling in PD remain unclear. This study aimed to investigate whether curcumin modulates \u03b1-syn SUMOylation and to elucidate the underlying molecular mechanisms in PD model mice. A PD model was established in male C57BL/6 mice via unilateral intrastriatal injection of \u03b1-syn preformed fibrils (PFFs). Six months after \u03b1-syn PFFs injection, mice were treated intravenously with curcumin (25\u202fmg/kg/day) or vehicle for 1\u202fmonth. Behavioral tests (open field, rotarod) assessed motor function. Neuropathology was evaluated by immunohistochemistry and western blotting for tyrosine hydroxylase (TH), phosphorylated \u03b1-syn (p-syn), SUMOylation pathway components (SUMO1, SAE2, UBC9, PIAS1/2), and ubiquitin. Striatal dopamine levels were measured by HPLC. Curcumin treatment ameliorated motor deficits and anxiety-like behaviors in PD mice. It partially preserved dopaminergic neurons and reduced p-syn aggregation in the substantia nigra, accompanied by increased striatal dopamine levels. Mechanistically, curcumin was associated with reduced SUMO1 and increased ubiquitin levels, suggesting modulation of SUMOylation-related signaling. Among SUMOylation enzymes, UBC9 expression was decreased, whereas E1 (SAE2) and E3 (PIAS1/2) components were not substantially affected. Our findings demonstrated that curcumin exerted neuroprotective effects in a PD model by attenuating \u03b1-syn pathology. The protective mechanism involves the inhibition of \u03b1-syn SUMOylation, primarily through the downregulation of the UBC9 enzyme. This study identifies UBC9-mediated SUMOylation as a potential target for curcumin and highlight a promising strategy for modifying \u03b1-syn-associated pathology in PD.\n\nID: 42553232\nTitle: Genomic characterization of Bacillus velezensis BP5 and BP103: insights into biocontrol potential against bacterial leaf spot on pepper.\nAbstract: Bacillus velezensis strains BP5 and BP103, isolated from pepper rhizosphere soil in the Mekong Delta, Vietnam, displayed potent antagonistic activity against Xanthomonas euvesicatoria, which causes pepper bacterial spot. In dual-culture assays, BP5 formed the largest inhibition zone (36.7 mm), outperforming BP103 (32.1 mm) and oxolinic acid (31.3 mm). Both Gram-positive, rod-shaped strains produced extracellular protease, lipase, amylase, cellulase, and siderophores. Whole-genome sequencing revealed compact and genetically stable genomes: BP5 (4.08 Mb, 46.04% GC, 4,047 protein-coding genes, 5 prophages) and BP103 (3.91 Mb, 46.46% GC, 3,793 genes, 2 prophages). Phylogenetic reconstruction and average nucleotide identity (ANI of more than 98%) confirmed their assignment to B. velezensis, with closest relatedness to strain 160. Orthologous gene analysis across 28 B. velezensis genomes and B. subtilis DSM10 identified 2,593 single-copy genes. Compared to BP103, BP5 harbors greater numbers of multi-copy orthologs, other orthologs, and unique paralogs (388, 910, and 5 in BP5 with 336, 763, and 1 in BP103, respectively). Pangenome analysis showed that the core genome size decreased progressively with the addition of new genomes; BP5 showed more pronounced genetic divergence driven by expanded unique paralogs and shell gene families (with 456 in BP5 compared with 203 in BP103). Both strains encode 13 secondary metabolite biosynthetic gene clusters, including surfactin, fengycin, bacillaene, macrolactin, difficidin, bacilysin, bacillibactin, mersacidin, and locillomycin, plus four previously undescribed terpene/PKS. The CAZyme repertoire was richest in BP5 (133 genes), exceeding BP103 and the commercial strain FZB42 (both 129 genes), thereby enhancing adhesion and adhesion and rhizosphere colonization. These findings establish BP5 and BP103 as highly promising biocontrol agents, combining high genetic stability with diverse secondary metabolite profiles, suitable for development into sustainable microbial bio-bactericides.\n\nID: 42553212\nTitle: Brassinosteroids as phytohormonal shields against micro- and nanoplastic stress in plants.\nAbstract: Microplastics (MPs) and nanoplastics (NPs) are biologically active stressors in agricultural soils, where they alter soil physical structure, disrupt rhizosphere processes, impair water and nutrient acquisition, and provoke oxidative and hormonal disequilibrium in plants. Brassinosteroids (BRs), particularly brassinolide and 24-epibrassinolide, have recently emerged as modulators of plant responses to plastic-particle stress. Current evidence indicates that BRs do not detoxify plastics directly. Instead, they reorganize plant performance across interconnected layers: aquaporin-linked NP transport, antioxidant and ascorbate-glutathione metabolism, photosystem II function, hormone crosstalk, secondary metabolism, and rhizosphere feedbacks. In tomato, BRs reduced polystyrene-NP accumulation in edible tissues by suppressing aquaporin genes. In Pinellia ternata and rice, BRs attenuated MP/NP-induced growth inhibition by restoring photosynthetic efficiency and redox control. This mini review synthesizes these findings and frames BRs as eco-hormonal regulators of the plant-plastic-soil interface, while highlighting priorities for field-realistic validation.\n\nID: 42553087\nTitle: The central role of IL-6 in the differential effects of GLP-1 receptor agonists and metformin across multiple health conditions.\nAbstract: An earlier publication suggested that elevated TGF-\u03b21 together with IL-1\u03b2 and IL-6 drives neutrophilic asthma, increases cancer risk, and is associated with metabolic dysregulation. This article examines how these cytokines shape the therapeutic efficacy and safety profiles of GLP-1 RAs compared with metformin. Integrating published data, stronger suppression of circulating IL-6 likely contributes to the superior efficacy of GLP-1 RAs in lowering blood glucose, improving insulin sensitivity in patients with polycystic ovary syndrome (PCOS), reducing cardiovascular risk, and reducing osteoarthritis-related perioperative infections, but not to their superior weight-loss effects. Superior weight loss, which is IL-6 independent, further enhances the efficacy of GLP-1 RAs in improving metabolic dysfunction-associated steatohepatitis (MASH), PCOS, and survival in cancer patients. Systemic IL-6 suppression may also explain the higher frequency of allergic adverse effects with GLP-1 RAs, whereas local interference with IL-6 signaling in the gastrointestinal tract, shared by both agents, likely accounts for frequent gastrointestinal adverse events reported with both. Although metformin may be more effective in improving asthma outcomes in the general asthma population, GLP-1 RAs are likely to be more effective in neutrophilic asthma. Metformin also reduces osteoarthritis-related joint pain, consistent with the limited efficacy of IL-6R inhibition in reducing this pain. Finally, the lack of effect of IL-6 pathway inhibitors on cancer incidence may explain the similar efficacy of GLP-1 RAs and metformin in reducing the risk of most cancers.\n\nID: 42553046\nTitle: NNMT Loss Drives Cancer Progression by enhancing SAM availability for mTORC1 Signaling and Chromatin Methylation.\nAbstract: Aberrant epigenetic reprogramming together with dysregulated mTOR signaling are hallmarks of cancer, where altered chromatin methylation and nutrient-sensing pathways cooperate to drive tumor progression. S -adenosylmethionine (SAM), the universal methyl donor, is essential for these processes, yet how tumors sustain elevated SAM availability to support oncogenic transmethylation reactions remains poorly defined. Here, using prostate cancer (PCa) as a model system, we identify nicotinamide N-methyltransferase (NNMT) as a critical metabolic-epigenetic regulator and tumor suppressor. Using a prostate-specific Nnmt knockout mouse model, we demonstrate that NNMT loss accelerates PCa progression, particularly in the context of Pten deletion, resulting in infiltrating carcinoma and reduced survival. Mechanistically, NNMT functions as a \"SAM-sink,\" and its loss increases intracellular SAM abundance, thereby activating mTORC1 signaling through SAMTOR-dependent sensing and broadly enhancing chromatin methylation. In human PCa, recurrent genomic deletions of NNMT occur in up to 7% of cases, and NNMT protein expression is largely absent in primary tumors and metastases. NNMT-deficient PCa cells exhibit elevated SAM:SAH ratios, increased histone methylation, and heightened mTORC1 activity, enabling sustained tumor growth even under dietary methionine-restriction (MR). Notably, combined MR and pharmacologic mTORC1 inhibition synergistically suppresses the growth of NNMT-deficient tumors, revealing a previously unrecognized therapeutic vulnerability. Collectively, these findings establish NNMT as a key tumor suppressor that constrains SAM-driven epigenetic and signaling programs in PCa and suggest a rational, diet-based therapeutic strategy for advanced cancers with NNMT loss.\n\nID: 42553036\nTitle: Aegeline mitigates isoproterenol-induced myocardial infarction: insights from biochemical, histopathological, and in silico studies.\nAbstract: Myocardial infarction (MI) remains a major cause of morbidity and mortality worldwide. Although considerable advances have been made in its diagnosis and management, the development of safe and effective cardioprotective agents remains an active area of investigation. Natural products continue to attract interest as potential therapeutic candidates because of their diverse pharmacological activities. Aegeline, an alkaloidal amide isolated from Aegle marmelos, has demonstrated antioxidant and anti-inflammatory properties; however, its cardioprotective effects in experimental myocardial injury have not been fully characterized. The aim of this work was to assess the cardioprotective effects of aegeline in an experimental MI model. Isoproterenol (ISO) (85\u2005mg/kg, subcutaneouslySC) was employed to trigger acute MI in experimental rats. Aegeline was administered to the rats at doses of 5 and 10\u2005mg (p.o.) on the 29th and 30th day, respectively, followed by ISO (85\u2005mg/kg) (s.c.) on the same days. Aegeline exhibited cardioprotective properties against ISO-induced damage, as evidenced by the amelioration of altered histopathological features, reduction in myocardial damage, and improvements in antioxidant and oxidative stress markers, inflammatory indicators, and signaling pathways, along with inhibition of apoptotic expression. In addition, aegeline did not affect liver or kidney function. Molecular docking using AutoDock Vina v1.2.6 revealed the strong binding affinity of aegeline for CK-MB (PDB ID: 3B6R), with key hydrophobic, hydrogen-bonding, and salt bridge interactions. Additional targets included LDH (7DBJ), cTnT (1J1D), and cTnI (7SC2), though CK-MB exhibited the most favorable interaction profile. Molecular dynamics simulations supported these findings through principal component analysis, dynamic cross-correlation, free energy landscape mapping, and generalized born surface area (MM-GBSA) binding free energy analysis. This study demonstrates that aegeline attenuates experimental myocardial injury by reducing oxidative stress, inflammation, and apoptosis. Computational analyses support potential molecular interactions with proteins associated with myocardial injury, while the biochemical and histopathological findings provide primary evidence for its cardioprotective activity.\n\nID: 42554508\nTitle: Fibrilpaint: A Class of Amyloid-Targeting Peptides.\nAbstract: Amyloid fibrils are a common pathological hallmark in multiple neurodegenerative diseases, yet molecular tools to recognize and manipulate them remain scarce. We report FibrilPaints, a family of modular peptides designed for amyloid binding and adaptable chemical functionality. The degenerative amyloid-targeting unit of FibrilPaints, W5P4H3R2, has a high content of \u03c0-stacking and aromatic side chains. Systematic sequence variation, altering charge, termini, and residue order, revealed the importance of the composition of the amyloid-targeting unit for high-affinity binding across Tau and Huntingtin fibrils. Importantly, sequence changes outside this unit do not preclude fibril binding, which permits attachment of fluorophores or E3-recruiting motifs for targeted protein degradation. This work establishes FibrilPaint as a modular peptide system for binding Tau and Huntingtin amyloid fibrils, with potential for broader detection and modulation applications.\n\nID: 42554501\nTitle: BetaH proteolysis unleashes an electrostatic-homing antibacterial polymorphic toxin.\nAbstract: Contact-dependent and diffusible proteinaceous polymorphic toxin systems (PTSs) mediate widespread bacterial competition. While bioinformatic analyses have identified diverse PTSs across bacterial phyla, experimental validation in gram-positive species remains limited. Here, we characterize a diffusible polymorphic toxin encoded by the Staphylococcus aureus S8-Ntox35 locus. We demonstrate that this system mediates inter-genus antagonism against Listeria monocytogenes via a bactericidal, heat-labile protein and that toxicity is enhanced following extracellular cleavage of the BetaH domain by the associated S8 peptidase. This processed toxin bears a peptide resembling a cationic antimicrobial peptide (CAMP) and facilitates intoxication of target cells by the Ntox35 RNase domain. Target cell resistance is impacted by known CAMP defense pathways, including DltABCD and MprF, and experimental evolution identified the ATP-binding cassette (ABC) transporter AnrAB as essential for intoxication. Unexpectedly, disruption of AnrAB abolished Ntox35 susceptibility, while simultaneously sensitizing cells to the proposed CAMP-like activity of the processed BetaH domain. These findings reveal a novel mechanism of inter-genus antagonism among Firmicutes (Bacillota) and establish a functional role for extracellular processing and ABC transporter-mediated susceptibility in PTS activity. Our work expands the known repertoire of diffusible toxins in gram-positive bacteria and sets the foundation for broader ecological and mechanistic investigation of the S8-associated polymorphic toxin system (S8-PTS). Polymorphic toxin systems (PTSs) are widely used by bacteria to inhibit competitors, but diffusible proteinaceous toxins have been largely characterized in gram-negative species. Here, we mechanistically characterize a diffusible PTS in Staphylococcus aureus that mediates inter-genus antagonism in the Firmicutes phylum. This system employs a secreted S8 peptidase to cleave a BetaH-toxin fusion, releasing a previously caged cationic amphipathic \u03b1-helix that facilitates membrane targeting and intoxication of susceptible cells. We further show that toxin activity requires the target cell ATP-binding cassette (ABC) transporter AnrAB, revealing a novel route of entry and an evolutionary tradeoff between toxin susceptibility and antimicrobial peptide resistance. Together, these findings uncover a new mode of bacterial competition and highlight a broadly distributed toxin system with ecological relevance across Firmicutes.\n\nID: 42554467\nTitle: The conserved QTQTX motif in the SARS-CoV-2 spike protein is dispensable for cleavage and lung cell entry of the emerging variant BA.3.2.\nAbstract: The furin motif in the SARS-CoV-2 spike (S) protein is important for lung cell entry. It is embedded in an extended loop structure and preceded by a highly conserved QTQTX motif that is required for efficient furin cleavage of the SARS-CoV-2 WA-1 S protein. BA.3.2 is an emerging SARS-CoV-2 saltation variant that is spreading globally in April 2026 and encodes a highly mutated S protein. Here, we analyzed whether the QTQTX motif is also required for spike protein cleavage and lung cell entry of BA.3.2. We report that two patient-derived spike sequences of the BA.3.2 subvariant BA.3.2.2 lack the first QT repeat of the QTQTX motif and show that this motif is largely dispensable for both cleavage and lung cell entry of BA.3.2.2, which we found to depend on TMPRSS2. Our results suggest that the reconfiguration of the BA.3.2 S protein during persistent infection may have significantly altered the determinants of furin cleavage.IMPORTANCEThe furin motif in the SARS-CoV-2 spike (S) protein is unique among sarbecoviruses and constitutes a virulence determinant. A QTQTX motif located immediately upstream of the furin motif is required for furin cleavage of the S protein of the virus that circulated early in the pandemic. Here, we show that the QTQTX motif is largely dispensable for S protein processing and S protein-driven lung cell entry of the emerging saltation variant BA.3.2, which is currently spreading globally. Thus, BA.3.2 evolution within immunocompromised individuals may have relaxed the requirements for furin processing of the spike protein.\n\nID: 42554321\nTitle: Dynamic Heterovalent Dual-Cu Sites for O\u2550O Cleavage in Electrocatalytic Oxygen Reduction Reaction.\nAbstract: Activation and cleavage of the inert O\u2550O bond represent a central challenge in energy electrochemistry. Here, the Cu dual-atom catalyst (Cu-DAC) is constructed via bottom-up pre-coordination assembly and post-encapsulation pyrolysis. The resulting Cu-DAC features a well-defined Cu-Cu distance (\u223c3.31 \u00c5) with switchable Cu1+/Cu2+ states, enabling dynamic dual-site coordination with O2. Cu-DAC achieves 0.87 VRHE half-wave potential for oxygen reduction reaction (ORR), a near-unity 4e- selectivity, and outstanding stability. Multiple operando spectroscopic characterizations and ab initio dynamic simulations (AIMD) reveal that the dynamic heterovalent [Cu1+\u2500O\u2500O\u00b7 -\u2500Cu2+] unit elongates the O-O bond and promotes its cleavage via dual-site confinement and electron donation. By specifically elucidating how these dual-atom sites dynamically evolve to facilitate O\u2500O bond cleavage, we provide vital atomic-level principles for the rational design of dynamically active catalysts.\n\nID: 42554219\nTitle: Hybrid quantum-mechanistic insights into \u03b2-O-4 ether cleavage in lignin-carbohydrate complexes using a choline chloride-lactic acid DES cluster model.\nAbstract: Lignin-carbohydrate complexes (LCCs) contribute to biomass recalcitrance, making \u03b2-O-4 ether linkages important targets for selective lignin depolymerization. \u03b2-O-4 cleavage in a representative LCC was studied using a hybrid quantum-mechanical/semiempirical (QM/xTB) approach, an explicit choline chloride/lactic acid (ChCl\u2009:\u2009LA) cluster, and M06-2X/6-31+G(2d,2p) refinement. Vacuum calculations were compared with C-PCM water (\u03b5 = 78), which was used as a high-polarity reference and upper-bound estimate of dielectric stabilization rather than as a representation of bulk deep eutectic solvent microstructure. Frontier-orbital, Fukui-function, and Hirshfeld-charge analyses identify C43 as the electrophilic center for chloride attack and O28 as the leaving-group atom, while revealing progressive electron-density loss from chloride. The optimized transition state (TS) supports a concerted SN2-like mechanism in which chloride attacks C43 as the O28-C43 bond elongates from 1.43 to 2.25 \u00c5. Thermochemically corrected stationary-point calculations at 298.15 K give Gibbs activation energies of 93.2 kJ mol-1 for the isolated explicit cluster and 75.8 kJ mol-1 with a C-PCM water dielectric, indicating a 17.5 kJ mol-1 sensitivity to high-dielectric screening. Decomposition of these barriers yielded optimized electronic activation energies of 121.0 and 119.7 kJ mol-1 for the isolated and dielectric-embedded clusters, whereas thermochemical corrections accounted for most of the predicted reduction and highlighted the substantial contribution of thermal and entropic terms to the model-dependent free-energy estimates. This study provides a descriptor-based mechanistic rationale for \u03b2-O-4 cleavage in an explicit ChCl interaction model and clarifies how local chloride coordination and lactic-acid hydrogen-bonding contacts can promote \u03b2-O-4 bond activation, providing a mechanistic framework for designing greener solvents for selective biomass fractionation.\n\nID: 42554179\nTitle: Regulation of actin polarization by Edwardsiella piscicida during invasion of Sebastes schlegeli intestinal mucosa and cellular immune responses.\nAbstract: Intestinal epithelial cells play a crucial role in teleost defence against complex aquatic environments. However, the study of intestinal mucosal immunity in teleosts has been hindered by the lack of reliable and specific marker genes to distinguish between different cell types. In this study, single-cell RNA-seq analysis of the intestinal epithelial cells from healthy and infected Sebastes schlegeli was performed to investigate the heterogeneity of teleost intestinal epithelial cells. Immune cells, including T cells, B cells, non-specific cytotoxic cells (NCCs) and myeloid cells, and non-immune cells including enterocyte, erythrocyte, BEST4+ cell, goblet cell and enteroendocrine cell, were identified. Several notable cell populations, including early myeloid cells, lysosome-rich enterocytes, enterocyte progenitors and BEST4+ cells, were characterized in detail. Additionally, significant changes in the expression of actin polymerization-related genes were observed in various intestinal mucosal cell types following Edwardsiella piscicida infection, suggesting a mechanism of invasion with E. piscicida. These findings enhance our understanding of teleost intestinal cellular composition and provide valuable insights for the development of therapeutic strategies against E. piscicida.\n\nID: 42554175\nTitle: Ubiquitin-mediated degradation of RPN13 promotes porcine sperm capacitation and sperm-zona pellucida binding.\nAbstract: Sperm capacitation involves biochemical and functional changes necessary for fertilization. The ubiquitin-proteasome system (UPS) regulates protein turnover, but the roles of proteasomal receptors during capacitation remain unclear. This study investigates the regulation and functional impact of proteasomal subunits RPN13 (ADRM1) and RPN2 (PSMD1) during in vitro capacitation of boar sperm. Semen samples (n = 6) were assigned to four groups: non-capacitating control (Non-Cap, 4 h in medium lacking bovine serum albumin (BSA)), capacitating control (Cap, 4 h with BSA) and groups treated with vehicle (CM + dimethyl sulfoxide (DMSO)) or the RPN13 inhibitor RA190. We measured RPN2/RPN13 levels, RPN13-associated ubiquitin signal and RPN13-RPN2 interactions (Western blot, Co-IP) and assessed sperm kinematics, intracellular Ca2+, protein tyrosine phosphorylation (pTyr), mitochondrial membrane potential (\u0394\u03a8m), ATP, ZP binding and early embryo cleavage after IVF. The Cap group showed successful capacitation, confirmed by increases in pTyr, sperm hyperactivation and Ca2+-associated Fluo-4 fluorescence. During capacitation, RPN2 and RPN13 abundance decreased, while RPN2-RPN13 association increased. RA190 treatment increased RPN13 levels, inhibited RPN13 turnover and decreased its interaction with RPN2 and the associated ubiquitin signal. Functionally, RA190 attenuated sperm hyperactivation, Ca2+-associated Fluo-4 fluorescence, protein tyrosine phosphorylation, mitochondrial membrane potential and ATP levels and was associated with reduced fertilization-related outcomes, including ZP binding and early embryo cleavage after IVF. These findings highlight the role of RPN13 in sperm capacitation and fertilization.\n\nID: 42553942\nTitle: Correction: Influence of commensal bacteria on the proteolytic and antigenic profiles of INFOGEST-like digested wheat gliadin.\nAbstract: [This corrects the article DOI: 10.3389/fmicb.2026.1842801.].\n\nID: 42553894\nTitle: Lipid oxidation-mediated non-enzymatic glycation and its impact on the digestibility of milk proteins.\nAbstract: This study explored the influence of milk fat on non-enzymatic glycation of proteins and protein digestibility by analyzing the accumulation of glycation products and changes in protein structure. The contents of POV and MDA increased significantly on day 7 of storage in milk model groups with different milk fat concentrations (0, 2, 4, 6%) and the increase was more obvious in the high-fat (6%) group. In the high-fat group, the contents of glyoxal and methylglyoxal, the intermediate products of non-enzymatic glycation increased at a high level. The peak shift and the appearance of new peaks in the infrared spectrum further indicated enhanced glycation coupled with lipid oxidation. In addition, the decrease in fluorescence intensity, the increase in ultraviolet absorption, the reduction in surface hydrophobicity, and the change of protein secondary structure in the high fat group indicated obvious changes in the microstructure of proteins. The cleavage site of protease was reduced because of the alterations in protein advanced structure and the occupation of amino acids digestion sites, which subsequently reduced the digestibility of proteins. This phenomenon was more obvious in the early stage of gastrointestinal digestion. High levels of milk fat promoted non-enzymatic glycation by accelerating lipid oxidation, which in turn induced glycation, cross-linking, and aggregation of milk proteins. These structural changes further hindered the action of digestive enzymes, ultimately reducing the digestibility of milk proteins.\n\nID: 42553853\nTitle: An artificial intelligence real-time rare sperm detection system for intraoperative microsurgical testicular sperm extraction.\nAbstract: How can rare spermatozoa be identified efficiently during microsurgical testicular sperm extraction (micro-TESE)? An artificial intelligence (AI)-assisted system was developed to flag candidate rare spermatozoa in real time during micro-TESE, and it may support embryologists as a decision-support tool. Patients with non-obstructive azoospermia (NOA) can obtain sperm for procreation through micro-TESE. During this procedure, sperm retrieval primarily relies on embryologists or laboratory technicians visually searching for sperm under a microscope, which is not only laborious and inherently subjective but also susceptible to errors. Although AI technology has been applied to identify trace amounts of sperm, existing models lack sufficient efficiency and true real-time performance. This study included model development followed by a single-centre clinical evaluation. An improved YOLO (You Only Look Once)-based rare sperm detection model, termed YOLOv11-RSD, was developed using microscopy data from 1165 surgical patients, comprising 1932 image samples containing a total of 5032 annotated sperm objects with confirmed identification. Clinical evaluation was performed between May 2024 and July 2025. Performance was assessed across confidence thresholds in obstructive azoospermia (OA) patients with normal spermatogenesis, and the system was then applied during micro-TESE in NOA patients and compared with routine embryologist assessment. The model was developed using testicular sperm microscopy images collected at a single hospital. Real-time clinical feasibility was evaluated in 10 OA cases and 30 NOA cases. Embryologist assessment was used as the reference standard, and performance was assessed using PPV, sensitivity, F1-score, and 95% confidence intervals. Discordant AI-assisted detections were reviewed by embryologists in real time. YOLOv11-RSD achieved real-time detection of candidate spermatozoa in microscopy images with high sensitivity and acceptable PPV under the selected operating threshold. Compared with baseline YOLOv11, YOLOv11-RSD showed improved overall detection performance across representative evaluation settings. In OA cases, the system achieved high sensitivity for sperm detection, reaching up to 96.7% across evaluated thresholds. During micro-TESE in NOA patients, at a confidence threshold of 0.50, positive predictive value (PPV), sensitivity, and F1-score were 80.58%, 96.11%, and 87.66%, respectively. The system highlighted candidate spermatozoa that were not identified during the initial manual assessment in six NOA cases, including two cases initially classified as sperm-negative; these findings were confirmed upon immediate re-review. Follow-up reproductive outcomes were available for six cases in which AI-assisted detection contributed to the search-and-confirmation workflow: embryo cleavage was achieved in all six cases, and three cases ultimately resulted in live births. Notably, among the two cases initially classified as sperm-negative, one case resulted in a singleton live birth. N/A. This was a single-centre clinical evaluation with a limited clinical cohort. Although model inference was rapid, procedure-level efficiency was constrained by image acquisition and scanning logistics, and no definitive reduction in total procedure time was demonstrated. External multi-centre validation is required. AI-assisted sperm detection may support embryologists during micro-TESE by flagging candidate rare spermatozoa for rapid review. Further prospective multi-centre validation is required to determine whether this approach improves procedure-level efficiency or clinical outcomes. This work was supported by grants from National Natural Science Foundation of China (82301794), Shanghai Science and Technology Innovation Action Plan (24Y12800702), Natural Science Foundation of Shanghai (25ZR1401300), National Key Research and Development Program of China (2022YFC270300), China Jiliang University Research Grant (No. H251120), and Shanghai General Hospital Basic and Clinical Collaborative Research Program (JC202612). The authors declare no competing interests.\n\nID: 42553369\nTitle: A double-edged sword: the role of macrophage pyroptosis-driven cytokine storm-mediated intercellular communication in tumor progression.\nAbstract: Pyroptosis, a lytic and inflammatory form of programmed cell death, has emerged as a regulator of tumor immunity through its capacity to trigger localized cytokine storms (operationally defined in Section 3). This review examines the dual mechanisms by which macrophage pyroptosis-driven cytokine storms influence tumor progression. Pyroptosis is executed through canonical (caspase-1/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3)-dependent) and non-canonical (caspase-4/5/11-dependent) pathways, both converging on cleavage of gasdermin D (GSDMD) to form membrane pores that release pro-inflammatory cytokines (interleukin-1 beta, IL-1\u03b2; interleukin-18, IL-18) and damage-associated molecular patterns (DAMPs). These primary signals initiate cascade amplification through chemokine and cytokine networks, recruit diverse immune cell populations, and establish distinct inflammatory microenvironments. The effects of pyroptotic cytokine storms show striking temporal and intensity dependence. Acute, moderate inflammatory responses activate anti-tumor immunity through induction of immunogenic cell death (ICD), dendritic cell maturation, and cytotoxic lymphocyte priming. Chronic, low-grade cytokine storms, in contrast, promote tumorigenesis through six interconnected mechanisms: genomic instability and epigenetic reprogramming, cancer stem cell enrichment, pro-angiogenic remodeling, pre-metastatic niche formation, establishment of an immunosuppressive microenvironment, and induction of epithelial-mesenchymal transition. This \"double-edged sword\" phenomenon depends on inflammation intensity, duration, spatial distribution, and tumor microenvironment (TME) characteristics. Clinical investigations indicate that pyroptosis-related biomarkers, including GSDMD, gasdermin E (GSDME), and inflammatory cytokine profiles, may support patient stratification and treatment-response prediction across multiple cancer types in patients with tumor-associated macrophages (TAMs)-rich tumors. Preclinical evidence from bioorthogonal chemical systems in murine models, together with mathematical modeling, has suggested that pyroptosis affecting approximately 10-15% of tumor cells may serve as a tentative threshold for initiating anti-tumor immunity. However, this value has been derived from a limited number of preclinical systems (primarily 4T1 mammary tumor models) and has not yet been validated in human tumors; it should therefore be interpreted as a working hypothesis rather than an established parameter. Current therapeutic strategies targeting this pathway include NLRP3 inhibitors, IL-1\u03b2/IL-18 blockers, and combination approaches with immune checkpoint inhibitors (ICIs). Looking forward, future research should prioritize: (i) quantitative in vivo mapping of macrophage pyroptosis using spatial multi-omics and intravital imaging; (ii) development of tumor-targeted, spatiotemporally controlled pyroptosis inducers (e.g., nano-delivery and bioorthogonal activation systems); (iii) rational combination with immune checkpoint inhibitors and epigenetic modulators; and (iv) establishment of pyroptosis-based biomarker panels to guide patient stratification and toxicity prediction in clinical trials. Achieving \"controllable cytokine storms\" through precise macrophage pyroptosis modulation represents a therapeutic paradigm that balances anti-tumor efficacy against inflammatory toxicity, with the potential to advance cancer immunotherapy toward precision inflammation regulation.\n\nID: 42552969\nTitle: Quenching-Derived Pt Clusters on Co3O4/TiO2 p-n Heterojunctions Enable Efficient Photothermal Oxidation of VOCs.\nAbstract: Photothermal catalysis provides an effective strategy to overcome the high energy consumption of thermal catalysis and the low efficiency of photocatalysis; however, inefficient charge separation and limited O2 activation remain key challenges. Herein, a p-n heterojunction oxide support, Co3O4/TiO2, was constructed and decorated with Pt species via a quenching method. The resulting Co3O4/TiO2-Q-Pt catalyst achieved toluene conversion exceeding 90% within 38\u00a0min under light irradiation at 522\u00a0mW cm- 2, significantly outperforming single-component oxide-supported catalysts. The catalyst exhibited excellent photothermal stability and strong resistance to water vapor and mixed volatile organic compound (VOC) conditions. Structural characterization revealed that Co3O4 incorporation reduced TiO2 crystallinity while enhancing lattice-oxygen mobility, redox capability, and light absorption, even at low Co3O4 contents. Moreover, electron paramagnetic resonance (EPR) measurements showed enhanced generation of \u2022OH and \u2022O2 - species under illumination, consistent with improved photoinduced redox activity. In situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) showed suppression of the accumulation of partial-oxidation intermediates and promotion of the formation of benzoate and maleic anhydride species, thereby facilitating aromatic ring cleavage and deep mineralization of toluene.\n\nID: 42552379\nTitle: Multifaceted roles of CD44 in cancer progression and targeted therapeutic strategies.\nAbstract: CD44, a multifunctional transmembrane glycoprotein, is not only a bystander but also a crucial driver of cancer progression that promotes cancer stem cell maintenance, metastasis, and resistance to therapy. Therefore, CD44 is recognized as a promising therapeutic target in advanced malignancies. Here, we discuss its unique features, such as its structural diversity, which arise from alternative splicing and the post-translational modifications of cleavage and phosphorylation. In addition, we discuss the function of CD44 as a multivalent cell adhesion receptor for extracellular matrix components, including hyaluronic acid, fibronectin, osteopontin, and TSG6, thereby regulating lymphocyte activation, cell-cell interactions, cell adhesion, and migration within the extracellular matrix. Moreover, the emerging role of CD44 as a co-receptor of receptor tyrosine kinases such as epidermal growth factor receptor, c-MET, and vascular endothelial growth factor receptor 2 is highlighted to elucidate the contribution of CD44 to malignant signaling networks. We also discuss its potential as a therapeutic target in advanced cancers, particularly its applications in unconjugated antibodies, antibody-drug conjugates, peptide-based inhibitors, and chimeric antigen receptor-T cell therapies. We conclude by addressing the limitations observed in clinical studies and outlining promising opportunities for future development.\n\nID: 42552004\nTitle: Multicentre randomised comparative effectiveness trial of cleavage-stage versus blastocyst-stage embryo transfer in patients with few embryos (PRECISE): implementation of a study protocol in reproductive medicine.\nAbstract: With improvements in in vitro culture techniques, there has been a steady shift in practice to transfer in vitro fertilisation (IVF) embryos at the blastocyst-stage to increase pregnancy rates per embryo transfer (ET) and reduce the risk of multiple pregnancy. In patients with fewer embryos (\u22645 zygotes), fresh cleavage-stage ET is still commonly performed to reduce the risk of cycle cancellation if no embryo survives to the blastocyst-stage in vitro. However, evidence for improved outcomes with cleavage-stage embryo transfer in this subgroup of patients is lacking. We will compare blastocyst-stage to cleavage-stage ET with regard to cumulative live birth rates (CLBR) per oocyte retrieval and other patient-centred cycle outcomes in patients with \u22645 zygotes. Multicentre, randomised pragmatic comparative effectiveness trial with superiority design comparing blastocyst-stage to cleavage-stage ET in patients with \u22645 zygotes. This trial will enrol and randomise 1126\u2009women aged 18-44 undergoing IVF to either fresh cleavage-stage or blastocyst-stage ET at eleven IVF centres. Surplus embryos will be vitrified at the blastocyst-stage. The primary outcome is the CLBR per oocyte retrieval. Secondary outcomes include: multiple pregnancy, pregnancy loss <20 weeks and cycle cancellation. Exploratory outcomes include: live birth per fresh ET, ectopic pregnancy, time to pregnancy, stillbirth, preterm birth, financial burden, patient distress and facilitators and barriers for patient involvement and implementation. All primary analyses will be conducted according to the intent-to-treat principle. We also will report per-protocol analyses as a sensitivity analysis. A multidisciplinary stakeholder team, including a 5-patient stakeholder panel, will participate in study design, conduct and oversight. The Institutional Review Board at Beth Israel Deaconess Medical Center (BIDMC) provides central regulatory oversight. Results will inform timing of embryo transfer in patients with \u22645 zygotes and will be disseminated through academic publications, meeting presentations and communications to advocacy groups and patient participants. NCT06746129.\n\nID: 42551926\nTitle: [Targeting KRAS-Mutant Cancers-Therapeutic Strategies and Mechanisms of Resistance to RAS Inhibitors].\nAbstract: KRAS mutations represent one of the most prevalent oncogenic alterations, occurring in approximately 20% of human cancers. Although KRAS had long been regarded as\"undruggable,\"the identification of a druggable pocket has catalyzed rapid progress in the development of targeted therapeutics. To date, the only approved KRAS-directed agent in Japan is sotorasib, a GDP-state inhibitor specific for the KRAS G12C mutation. However, a diverse array of next-generation strategies-including pan-KRAS inhibitors, pan-RAS inhibitors, tri-complex inhibitors, and targeted protein degradation modalities-are currently under active investigation. These advances have drawn increasing attention to the functional heterogeneity and differential druggability among distinct KRAS mutation subtypes, as well as organ-specific biological differences that had previously received limited consideration. Moreover, as with other molecularly targeted therapies, multiple resistance mechanisms to KRAS inhibition have been reported, underscoring the need for therapeutic strategies capable of overcoming or preventing treatment resistance.\n\nID: 42551610\nTitle: A Strain of High Cell-Adapted Porcine Deltacoronavirus: Isolation, Genetic Evolution, and Pathogenicity Analysis.\nAbstract: In 2024, an outbreak of piglet diarrhea occurred in Guangxi Zhuang Autonomous Region, China. RT-PCR assay of clinical samples confirmed that porcine deltacoronavirus (PDCoV) was the primary pathogen. In this study, the PDCoV GX/2024 strain was successfully isolated in PK-15 cells with optimized trypsin treatment conditions. The cytopathic effect (CPE) of this strain gradually intensified during serial passages. After plaque purification, the strain achieved stable proliferation with high titer, reaching a peak titer of 107\u00b72 TCID50/mL, which indicated its potential as an inactivated vaccine candidate. Phylogenetic analysis demonstrated that the strain belonged to the Chinese lineage, with nucleotide similarity ranging from 97.89% to 99.76% compared with other domestic and international strains. Analysis of the Spike protein (S protein) revealed that the strain harbored critical amino acid substitutions in the S1 domain and the S1/S2 cleavage site, which led to conformational changes in the protein surface and might be associated with viral infection efficiency. Animal experiments verified that PDCoV GX/2024 exhibited strong pathogenicity in newborn piglets. Diarrhea, vomiting and other symptoms appeared within 24 hours post-infection. The virus mainly colonized the epithelial cells at the tip of intestinal villi and caused severe pathological damage. This study provides important theoretical basis for the genetic evolution, pathogenic mechanism and transmission potential of PDCoV, and offers reference value for disease prevention and control as well as vaccine development.\n\nID: 42551559\nTitle: CRISPR-Cas Systems as Precision Antimicrobials: Reversing the Tide of Antimicrobial Resistance.\nAbstract: Antimicrobial resistance (AMR) has escalated into a global health crisis, with resistant pathogens causing over 1.2 million direct deaths annually and threatening to render modern medicine unsustainable. This review provides a comprehensive and updated synthesis of CRISPR-Cas-based antimicrobial strategies with a unique focus on: (i) critical comparison with conventional antibiotics and emerging alternatives; (ii) quantitative evaluation of delivery platforms; (iii) novel strategies including AI-optimized guide design and the ATTACK-CreTA system; (iv) comprehensive analysis of ecological risks; and (v) technology readiness level assessments for clinical translation. The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR by selectively targeting and eliminating resistance genes. We systematically evaluate the mechanistic diversity of Cas effectors, from DNA-cleaving Cas9 and Cas3 to RNA-targeting Cas13, and their application in reversing resistance phenotypes in WHO priority pathogens. We critically assess emerging delivery platforms, including engineered bacteriophages, conjugative plasmids, nanoparticles, and outer membrane vesicles, quantitatively comparing their delivery efficiency, payload capacity, and biosafety profiles. Novel strategies such as CRISPR interference (CRISPRi) for gene silencing without genomic cleavage, the ATTACK-CreTA system for enhanced bactericidal activity, and AI-driven optimization of guide RNA design are examined with appropriate caveats. We comprehensively address clinical translation challenges including immunogenicity, pharmacokinetics/pharmacodynamics, manufacturing scalability, regulatory pathways, and bacterial resistance mechanisms including anti-CRISPR proteins. No CRISPR-based antimicrobial has yet received regulatory approval, and we critically evaluate the gap between proof-of-concept and clinical utility. A detailed roadmap for clinical development is proposed. By integrating recent advances in Cas protein engineering, delivery technologies, and diagnostic applications, this review positions CRISPR-Cas systems as next-generation precision therapeutics capable of both treating resistant infections and curtailing the spread of AMR across clinical and environmental settings.\n\nID: 42551403\nTitle: Ultrasound/electrostatic field combined with \u03b5-polylysine for improving the quality of ready-to-eat pork jelly and freshness classification based on convolutional neural network.\nAbstract: This study investigated the effects of ultrasound (US, 300\u00a0W, 20\u00a0kHz, 20\u00a0min) and electrostatic field (EF, 3.5\u00a0kV/cm, 60\u00a0min) combined with \u03b5-polylysine (PL, 0.5%, w/w) on the quality of ready-to-eat pork jelly (Shuijingyaorou, SJYR) during 40-day storage at 4\u00a0\u00b0C. Physicochemical properties such as lipid oxidation, total plate count, texture, flavor, and moisture distribution were evaluated. The results showed that the combined treatment (US + EF\u00a0+\u00a0\u03b5-polylysine) exhibited the strongest preservation effect for SJYR, significantly inhibiting lipid oxidation, protein degradation, pH increase, and microbial growth, meanwhile maintaining texture and flavor of SJYR, resulting in the shelf-life extended approximately 10\u00a0days compared with the blank control group. Furthermore, four convolutional neural network (CNN) models including Xception, ResNet-50, MobileNet, and GhostNet were developed for SJYR freshness classification. Xception model achieved the highest test accuracy (83.05% mean precision), while GhostNet model showed promising accuracy (77.48%) with lower computational cost, making it suitable for mobile applications. This study provides a feasible strategy for improving SJYR shelf-life and rapid freshness identification.\n\nID: 42551398\nTitle: Stability of Chlamydia pecorum MOMP at 37\u00a0\u00b0C: suitability for use in delayed-release vaccine implants in koalas (Phascolarctos cinereus).\nAbstract: The koala (Phascolarctos cinereus) is threatened by Chlamydia pecorum, an obligate intracellular bacterium that causes disease and infertility. Vaccination offers an alternative to antibiotics, and we have developed a safe, immunogenic vaccine that has shown promising results in captive and wild koala populations. However, the current vaccine requires a booster dose to achieve optimal immune protection. A delayed-release implant delivering the booster after primary vaccination could remove the need to recapture wild koalas. For this strategy to succeed, the antigen must remain stable at 37\u00a0\u00b0C until release. In this study we have assessed the stability and antigenicity of the Chlamydia pecorum major outer membrane protein (MOMP) antigens used in the vaccine after six weeks at 37\u00a0\u00b0C. In vitro analysis showed that aged MOMP induced antibody responses comparable to unaged antigens. These findings indicate that MOMP retains structural integrity and antigenicity, supporting its suitability for delayed-release vaccine delivery systems.\n\nID: 42551393\nTitle: A hairpin conformational change-mediated SRH cascade isothermal amplification assisted electrochemical biosensor for Listeria monocytogenes detection.\nAbstract: The rapid and sensitive detection of foodborne pathogens is crucial for safeguarding food safety and public health. In this study, we developed an electrochemical biosensor based on a hairpin conformational change-mediated cascade isothermal amplification strategy for the detection of Listeria monocytogenes in food. The biosensor employs aptamers for specific recognition of the target bacterium. In combination with magnetic separation technology, the concentration of the target bacterium is converted into that of a trigger strand. The trigger strand induces a conformational change in the hairpin structure, thereby initiating a cascade amplification reaction involving strand displacement amplification, rolling circle amplification, and hybridisation chain reaction (SDA-RCA-HCR, SRH), enabling an exponential signal amplification and a significant reduction of the background signals caused by self-cleavage. The proposed biosensor demonstrates a linear detection range of 15 to 1.5\u202f\u00d7\u202f108\u202fCFU/mL for Listeria monocytogenes, with a detection limit as low as 5\u202fCFU/mL (S/N\u202f=\u202f3). It exhibits great specificity for the target bacterium, effectively distinguishing it from other common foodborne pathogens, and shows satisfactory performance in real food samples, including lettuce, cheese, and chicken. Thus, this work provides an efficient, reliable, and sensitive new method for the rapid detection of foodborne pathogens in complex food matrices.\n\nID: 42551379\nTitle: Exploring the reductive transformation pathways of perfluoroalkyl Sulfonylimides to legacy perfluoroalkyl sulfonates and perfluoroalkyl sulfonamide compounds.\nAbstract: Bis(perfluoroalkylsulfonyl)imides (NH(CnF2n+1O2S)2) have been increasingly adopted as alternatives to legacy PFAS in industrial applications; however, their fate during water treatment remains poorly understood. Herein, we systematically investigated the reductive transformation of three sulfonyl imides (i.e., NH(C4F9O2S)2, NH(C2F5O2S)2, and NH(CF3O2S)2) under a UV/sulfite-based advanced reduction process (ARP). Bis(perfluoroalkylsulfonyl)imides exhibited high resistance to both direct photolysis and oxidative degradation, as evidenced by negligible degradation (<7%) and nearly no defluorination. In contrast, the UV/sulfite system achieved complete removal of bis(nonafluorobutanesulfonyl)imide within 24\u202fh, with defluorination efficiencies of up to 46.4%. Notably, LC-MS and LC-Q-Orbitrap HRMS analyses revealed substantial formation of legacy PFAS, including perfluoroalkyl sulfonamides (CnHF2n+1NO2S-), perfluoroalkyl sulfonates (CnF2n+1SO3-), which accounted for 5.0-10.3% and 12.9-48.5% of total fluorine, respectively. Density functional theory (DFT) calculations corroborated these findings, demonstrating that their secondary formation through reductive cleavage of the sulfonyl imide linkage is both thermodynamically favorable and kinetically accessible. However, short-chain perfluoroalkyl sulfonamides and perfluoroalkyl sulfonates exhibited substantially lower reactivity toward eaq\u207b than their parent compound, thereby limiting overall defluorination. Complete degradation pathways for bis(perfluoroalkylsulfonyl)imides with n\u202f=\u202f1, 2, and 4 were proposed based on identified TPs and DFT calculations. This study provides the first evidence that bis(perfluoroalkylsulfonyl)imides serve as precursors to short- and ultrashort-chain sulfonamides and sulfonates during reductive treatment, highlighting the potential for fluorine mass redistribution into persistent legacy PFAS during ARP-based remediation of emerging novel PFASs.\n\nID: 42551360\nTitle: Perfluorooctanoic acid disrupts lysine metabolism and autophagy to promote white spot syndrome virus infection in shrimp.\nAbstract: Perfluorooctanoic acid (PFOA), a globally distributed per- and polyfluoroalkyl substance (PFAS), is increasingly recognized for its immunotoxic and metabolic effects in aquatic organisms; however, its role in viral disease susceptibility remains poorly understood. Using Pacific white shrimp (Penaeus vannamei) and white spot syndrome virus (WSSV) as a model system, we investigated the effects of chronic PFOA exposure on antiviral immunity and viral transmission. Chronic exposure to environmentally relevant concentrations of PFOA resulted in significant accumulation of PFOA in the hepatopancreas and markedly increased WSSV replication, host mortality, and horizontal transmission. Integrated metabolomic and transcriptomic analyses identified lysine degradation as a consistently affected metabolic pathway, along with significant enrichment of lysosome-related pathways. Biophysical assays, including molecular docking, biolayer interferometry, and cellular thermal shift analysis, demonstrated that PFOA directly binds to the lysine-catabolizing enzyme aminoadipate semialdehyde synthase (AASS), which may contribute to lysine accumulation. Importantly, enzymatic activity assays further revealed that PFOA significantly inhibited both lysine-ketoglutarate reductase (LKR) and saccharopine dehydrogenase (SDH) activities, accompanied by marked lysine accumulation in shrimp hepatopancreas. PFOA exposure alone induced basal autophagic dysfunction, as evidenced by reduced LC3/LAMP1 co-localization, decreased LC3 abundance, increased p62 accumulation, and impaired autophagosome-lysosome fusion. Dietary lysine supplementation reproduced these autophagic defects and significantly promoted WSSV replication, supporting a role for lysine accumulation in mediating impaired antiviral defense. Together, this study uncovers a previously unrecognized lysine-autophagy-lysosome regulatory axis through which PFOA enhances viral susceptibility and transmission in shrimp, providing mechanistic insight into pollutant-driven disease risk in aquaculture ecosystems.\n\nID: 42551354\nTitle: A lysosome-targeted and polarity-responsive photosensitizer for tumor specific imaging and photodynamic therapy.\nAbstract: Photodynamic therapy (PDT) represents an effective and promising strategy for cancer treatment. Nevertheless, the poor targeting specificity of conventional photosensitizers has severely impeded its clinical application. Consequently, the development of novel photosensitizers that integrate imaging-guided diagnosis and synergistic phototherapeutic efficacy is highly desirable to improve PDT performance. In this work, we designed and synthesized a lysosome-targeted photosensitizer HBT-CUR with D-\u03c0-A feature based on 2-(2'-hydroxyphenyl) benzothiazole (HBT) and difluoroborate moieties for tumor image-guided PDT. HBT-CUR exhibits near-infrared (NIR) fluorescence emission, along with high polarity sensitivity, excellent stability, high lysosome-targeting specificity and good biocompatibility. By exploiting the distinct polarity differences between normal and tumor cells, HBT-CUR enables specific NIR fluorescence imaging of tumor tissues, with a fluorescence intensity \u223c6-fold higher than that in normal tissues. Moreover, HBT-CUR exhibits a fast in vivo response (10\u00a0min) and prolonged duration (6\u00a0h with robust fluorescence emission). More importantly, HBT-CUR can efficiently generate singlet oxygen under light irradiation (\u03a6\u0394\u00a0=\u00a00.70 in dioxane), thereby achieving effective photodynamic tumor ablation in MCF-7 tumor-bearing mouse models. Density functional theory (DFT) calculations reveal that the synergistic interplay between excited-state intramolecular proton transfer (ESIPT) and intramolecular charge transfer (ICT) processes contributes to the superior luminescence performance of HBT-CUR. This work presents a promising phototheranostic agent for tumor-specific image-guided photodynamic therapy, offering a new strategy for the rational design and development of tumor PDT systems.\n\nID: 42551134\nTitle: Epigenetic strategies for fetal hemoglobin induction in sickle cell disease.\nAbstract: Sickle cell disease (SCD) is caused by pathogenic variants in the \u03b2-globin gene (HBB), most commonly the variant responsible for hemoglobin S, and affects an estimated 515,000 newborns each year, with the highest burden occurring in sub-Saharan Africa. Gene editing and hematopoietic stem cell transplantation have changed the therapeutic landscape, but their cost, technical complexity, and procedure-related risks still limit their wider use. For this reason, pharmacological induction of fetal hemoglobin (HbF) remains an important therapeutic strategy. HbF reduces HbS polymerization and is associated with lower disease severity, morbidity, and mortality. Among the mechanisms involved in \u03b3-globin silencing, epigenetic regulation offers several targets that can be explored using small molecules. This review discusses medicinal chemistry approaches primarily targeting HDAC1/2, LSD1, and DNMT1, with emphasis on inhibitor classes, binding mechanisms, structural features, preclinical evidence, and translational limitations. The available data show that each target presents a distinct set of challenges. HDAC-directed strategies require improved isoform and cellular selectivity; LSD1 inhibitors must reconcile strong HbF induction with the risks associated with prolonged target engagement; and DNMT1 modulation is moving from DNA-incorporating nucleoside analogs toward reversible non-nucleoside inhibitors. We also discuss emerging approaches, including multi-target epigenetic modulation and targeted protein degradation. Together, these strategies show how a better understanding of \u03b3-globin repression may guide the development of safer and more accessible HbF-inducing agents.\n\nID: 42550868\nTitle: Bioinformatics and experimental studies of atherosclerosis-related endothelial dysfunction genes in renal calculi and exploration of their molecular mechanisms.\nAbstract: Patients with kidney stones (KS) often have an increased risk of atherosclerosis (AS). Because endothelial dysfunction (ED) is closely associated with AS, its role in KS remains unclear. This study aimed to examine the roles and mechanisms of AS-related ED genes in KS. Three datasets (GSE73680, GSE117518, and GSE132651) were analyzed. Differential expression analysis was conducted to identify differentially expressed genes (DEGs). To identify potential biomarkers, least absolute shrinkage and selection operator (LASSO) regression analysis and expression validation were conducted. Further analyses including GeneMANIA, gene set enrichment analysis (GSEA), examination of biomarkers within immune cells and subcellular localization analysis, molecular regulatory network analysis, tissue specificity analysis, and competing endogenous (ceRNA) network analysis were employed to comprehensively explore the functions and regulatory mechanisms of the identified biomarkers. Moreover, drug prediction analysis was conducted. Finally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) was proceeded to verify the expression levels of the biomarkers. A total of 22 DEGs associated with KS and AS were identified. Lasso regression selected 4 candidate biomarkers (MMP10, UCHL1, NEK2, and HEY1), among which UCHL1 and NEK2 were validated as key biomarkers. GeneMANIA and GSEA analyses uncovered the potential involvement of these biomarkers in cell adhesion molecules, focal adhesion, and lysosome pathways. Analysis of immune cells and subcellular localization provided insight into the biological functions and intracellular distribution of the biomarkers. Transcription factor regulatory network and ceRNA network analyses elucidated potential upstream regulatory mechanisms. Drug prediction analysis identified 17 potential drugs, including pazopanib and palbociclib, that may target NEK2. RT-qPCR demonstrated that NEK2 was significantly overexpressed in KS samples. This study identified biomarkers associated with KS and AS and comprehensively analyzed their molecular regulatory networks. These findings provide novel understandings of the molecular mechanism underlying KS and lay the foundation for future personalized treatment and drug development.\n\nID: 42550860\nTitle: Carboxymethyl cellulose reinforced graphene oxide-MoS2 for photocatalytic degradation of perfluorooctanoic acid (PFOA): process optimization and mechanism.\nAbstract: Perfluorooctanoic acid (PFOA), remains a major environmental challenge due to its exceptional chemical stability, resistance to conventional treatment technologies. In this study, a sustainable carboxymethyl cellulose-reinforced graphene oxide/molybdenum disulfide hydrogel (MoS\u2082/GO/CMC) was developed as a visible-light-responsive photocatalyst for PFOA degradation under low-power LED irradiation. The photocatalytic process was optimized using Response Surface Methodology based on a Box-Behnken Design, evaluating the effects of irradiation power (7-12 W), catalyst dosage (0-0.04\u2005g), and initial PFOA concentration (10-50\u2005mg L-\u00b9). The optimized conditions (12 W, 0.04\u2005g catalyst, and 30\u2005mg L-\u00b9 PFOA) achieved 99.8% degradation efficiency, with the developed quadratic model exhibiting excellent predictive capability (R\u00b2\u2009=\u20090.9974). Characterization results confirmed integration of MoS\u2082 and GO within the CMC matrix, producing a heterostructure with a narrow band gap of 1.12\u2005eV, enhanced charge separation, and suppressed electron-hole recombination. Kinetic analysis revealed pseudo-first-order degradation behaviour with a rate constant of 0.013\u2005min-\u00b9 and a half-life of 0.89\u2005h. Radical scavenging experiments identified superoxide radicals (O\u2082\u2022-) as the dominant reactive species governing PFOA degradation. LC-MS analysis confirmed a stepwise chain-shortening degradation pathway involving intermediates such as PFHpA, PFPeA, and PFBA, indicating progressive defluorination and carbon-carbon bond cleavage. The hydrogel demonstrated excellent stability, retaining over 96% of its initial activity after seven cycles. An Electrical Energy per Order (EEO) value of 444.6 kWh m-\u00b3 order-\u00b9 highlights the feasibility of low-energy operation. These findings demonstrate that MoS\u2082/GO/CMC hydrogels offer an environmentally benign, recoverable, and energy-efficient for PFAS remediation.\n\nID: 42550591\nTitle: Analysis of cancer mutations introduced into the Drosophila melanogaster Notch negative regulatory region uncovers a diversity of regulatory outcomes.\nAbstract: Activating mutations of Notch are drivers of the blood cell cancer, T-ALL, and some solid tumours. The negative regulatory region (NRR) of the extracellular domain (ECD) and the PEST region of the intracellular domain (ICD) are mutation hot spots which can act synergistically in T-ALL. The NRR, comprised of a heterodimerisation domain (HD) and three Lin12/Notch repeats (LNR A-C), masks the S2 cleavage site, normally only exposed following ligand binding and cleaved as the first step that ultimately leads to ICD release. Drosophila mutants have played a key role in analysing Notch structure/function, but there have been few mutational studies of the NRR. Here, we expressed, in S2 cells, over 20 cancer mutations located in the HD, LNR and LNR/HD interface, introduced into Drosophila Notch. Mutations in the HD domain core did not activate, likely due to absence in Drosophila of an S1 cleavage within the HD required for mammalian Notch activity. In contrast, mutations in the LNR/HD interface behaved similarly to T-ALL, activating constitutively with no further ligand induction and were synergistic with PEST deletion. Mutations of surface-exposed residues of LNR-C also activated constitutively but remained inducible both by ligand and by an intracellular endocytic regulator, Deltex, and were not synergistic with PEST deletions. These mutations caused elevated Notch levels and decreased turnover, suggesting a novel regulatory mechanism. Our results, therefore, uncover a variety of outcomes arising from perturbations of the NRR and will facilitate the establishment of Drosophila cancer models and the development of mutant-specific approaches to effective therapies.\n\nID: 42550325\nTitle: Co-sedimentation is the key to the structural investigation of wild-type FAT10.\nAbstract: Under inflammatory conditions, the ubiquitin-like modifier FAT10 serves as a tag for protein degradation by the 26S proteasome. FAT10 is degraded along with its substrates and this process is independent of the segregase VCP/p97, which, in the regular ubiquitin pathway of degradation, is required if a substrate lacks a disordered initiation region. FAT10 itself is loosely folded and its tendency to aggregate has complicated investigations of its structure, interaction, and function. Recently, hydrogen-deuterium exchange in combination with mass spectrometry has suggested that, in preparation of degradation by the proteasome, the adapter protein NUB1 traps FAT10 in a mostly unfolded state by capturing a \u03b2-strand. \u03b2-strand capture was subsequently confirmed by magic-angle spinning (MAS) NMR spectroscopy of a stabilized variant of the N-domain of FAT10 in complex with NUB1L, the longer splice variant of NUB1. MAS NMR, in addition, revealed that the N-domain of FAT10 and NUB1L form a fuzzy complex and that the N-terminus of FAT10 is positioned for initiation of degradation by specific non-covalent interaction with NUB1L. Here, we report the investigation of the wild-type N-domain of FAT10 by MAS NMR. Co-sedimentation with NUB1L yields high-quality spectra, which enable sequential assignment of resonances. Based on MAS NMR data, the complexes of the wild-type and stabilized N-domain of FAT10 with NUB1L appear identical. The N-terminal residue of FAT10 again shows up prominently in the spectra, even though it is this time an Ala, not a Gly. Our experiences suggest that co-sedimentation in combination with MAS NMR is generally helpful in the exploration of conditional folds of intrinsically disordered proteins.\n\nID: 42550314\nTitle: The role of the HB-EGF-regulated EGFR-mitophagy axis in maintaining cartilage homeostasis and osteoarthritis.\nAbstract: This study investigates the mechanism by which heparin-binding epidermal growth factor (HB-EGF) maintains cartilage homeostasis in osteoarthritis (OA) through the epidermal growth factor receptor (EGFR)-mitochondrial autophagy axis. Given the challenges in OA treatment and the critical role of impaired mitochondrial autophagy in disease progression, this research first identified key regulators using transcriptomic data from human OA cartilage and GEO databases. An in vitro OA model was established via IL-1\u03b2 induction in chondrocytes, through which HB-EGF was found to reverse the IL-1\u03b2-induced metabolic imbalance by downregulating catabolic markers (Mmp13, Adamts5) and upregulating anabolic markers (Acan, Col2a1). Further analysis revealed that HB-EGF enhanced mitophagy, as indicated by increased levels of Pink1, Parkin, and Lc3-II alongside decreased P62. These protective effects were attenuated by the mitophagy inhibitor Mdivi-1, confirming the dependence on this pathway. In conclusion, HB-EGF activates EGFR signaling to promote PINK1/ PARKIN-mediated mitophagy, which facilitates the clearance of damaged mitochondria and improves mitochondrial function, thereby restoring chondrocyte metabolic balance and delaying OA progression, suggesting the EGFR-mitophagy axis as a potential therapeutic target for OA.\n\nID: 42550282\nTitle: Ultrasensitive electrochemiluminescence determination of Salmonella based on CRISPR/Cas12a integrated with bimetallic semiconductive metal-organic frameworks.\nAbstract: An ultrasensitive electrochemiluminescence (ECL) biosensor was established by combining CRISPR/Cas12a technique and semiconductive bimetallic-organic framework (scMOF) [[CuxNi3-x(HITP)2] (HITP\u2009=\u20092,3,6,7,10,11-hexaiminotriphenylene)]] emitter and employed to detect Salmonella using the allosteric probe as the recognition component. Given that CuxNi3-x(HITP)2 has demonstrated large specific surface area, both in-plane and out-of-plane charge transfer ability, narrowed band gap, and enhanced separation of holes and electrons, it can be simultaneously employed as the superior ECL emitter and bioplatform for anchoring single-strand DNA (ssDNA), thus improving the detection sensitivity toward Salmonella. The CRISPR/Cas12a-based system can specifically recognize the target sequence of Salmonella and activate the nuclease activity of Cas12a, and the activated Cas12a possesses trans-cleavage ability toward ssDNA. The CuxNi3-x(HITP)2 emitter is then released, resulting in the decline of the ECL response. The developed CuxNi3-x(HITP)2-CRISPR/Cas12a-based ECL biosensor exhibits the ultralow detection limit of 0.25 CFU mL-\u20091 in the linear range from 1.0 CFU mL-\u20091 to 106 CFU mL-\u20091, significantly lower than those of reported ones. Furthermore, the developed biosensor exhibits outstanding overall biosensing properties with high selectivity, favorable reproducibility and stability, together with promising practical applicability for the determination of Salmonella in a variety of foodstuffs.\n\nID: 42550250\nTitle: Psychological and neurological benefits of physical activity: impact on mental health, Parkinson's disease, and Alzheimer's disease.\nAbstract: This review synthesizes current evidence on the psychological and neurological benefits of physical activity, with emphasis on mental health, Parkinson's disease (PD), and Alzheimer's disease (AD). Physical inactivity is increasingly recognized as a modifiable risk factor that may exacerbate neuroinflammatory and metabolic dysfunctions associated with these conditions. Structured exercise has been shown to activate muscle-brain signaling pathways, including neurotrophic factors (BDNF, IGF-1, VEGF), immune modulation (IL-6, IL-10), metabolic regulators (PGC-1\u03b1, SIRT1), and peripheral mediators such as myokines and gut-brain axis components. For mental health, exercise is associated with reductions in depressive, anxiety, and stress-related symptoms, potentially through HPA-axis recalibration, neurotransmitter remodeling (serotonin, dopamine), and enhanced endocannabinoid signaling. These changes are supported by fMRI and EEG studies showing improved prefrontal-limbic connectivity and cognitive resilience. In PD, exercise interventions are linked to improvements in motor control, balance, and mood, likely mediated by dopaminergic integrity, neurotrophic support, and anti-inflammatory effects, with multimodal programs (aerobic, resistance, and dance-based) often demonstrating superior outcomes. In AD, mid-life physical activity has been associated with reduced dementia risk in epidemiological studies; proposed mechanisms include enhanced amyloid/tau clearance, glymphatic function, and synaptic adaptation, with some RCTs reporting hippocampal volume preservation and improved network connectivity. Shared mechanisms across conditions include neurotrophic upregulation and anti-inflammatory effects, whereas distinct pathways involve dopaminergic circuit remodeling in PD and glymphatic facilitation in AD. Clinical translation through FITT principles (frequency, intensity, time, type) requires personalized prescriptions, strong adherence strategies, and multidisciplinary integration. Despite methodological heterogeneity and limitations in the existing trials, structured physical activity emerges as a scalable, non-pharmacological intervention with potential for prevention, symptom management, and neuroprotection. Larger, well-designed studies are needed to optimize its application and clarify long-term disease-modifying effects.\n\nID: 42550102\nTitle: Achieving selective photochemistry on a metal surface by tuning the aromatic core.\nAbstract: On-surface photochemistry provides a non-thermal strategy that can circumvent the side reactions and structural defects associated with thermal activation, enabling enhanced control over surface reactions. Among these, Ullmann coupling serves as a representative on-surface reaction for fundamental studies of reaction mechanisms and controllability. However, how the aromatic core of molecular precursors governs photoinduced C-Br activation on metal surfaces remains insufficiently understood. In this work, we investigated photoinduced Ullmann coupling on Au(111) using three dibromo molecular precursors with distinct aromatic cores. The three precursors show markedly different photoreactivities under 405 and 532\u00a0nm irradiation. Time-dependent density functional theory calculations reveal a photoinduced surface-to-molecule charge-transfer process. Combined with calculations of potential energy surfaces for C-Br bond dissociations, these results provide a qualitative rationale for comparing the relative tendency of photo-triggered C-Br cleavage at the surface. The results establish a direct correlation between photo-selectivity and the molecular aromatic core, providing mechanistic insight into light-controlled on-surface synthesis and offering design principles for tailoring light-responsive precursors toward desired carbon nanostructures.\n\nID: 42550091\nTitle: Vitamin B12 Precatalyst Enables Molecular Cobalamin(II) Catalysis for Organodisulfide Anolytes in Aqueous Redox Flow Batteries.\nAbstract: Affordable and stable organodisulfide anolytes are attractive for long-duration aqueous redox flow batteries (ARFBs) for grid energy storage. However, the sluggish redox reactions involving the cleavage/formation of sulfur-sulfur (S\u2500S) bonds cause severe polarization and limited capacity utilization, thus hindering the practical application of organodisulfide anolytes in ARFBs. Herein, we report a homogeneous catalysis strategy employing hydroxocobalamin (Vitamin B12, OHCbl) as an environmentally benign and biocompatible precatalyst which can enable molecular cobalamin(II) catalysis for organodisulfide anolytes. Cobalamin(II) generated in situ by electrochemical reduction of OHCbl bidirectionally accelerates the redox reactions of S\u2500S bonds and significantly decreases the overpotential of the ARFB with SPS (bis(sodium sulfopropyl) disulfide) anolyte from 1.24\u00a0V to 0.36\u00a0V at 40\u00a0mA cm-2, boosting the energy efficiency from 18% to 66%. The OHCbl-catalyzed ARFB with 1.0\u00a0M SPS anolyte delivers a capacity of 51.45 Ah L-1 operated at 100% state of charge and remains stable for 850 cycles at 50\u00a0mA cm-2 with a low decay rate of 0.0189% per day. Moreover, a nearly saturated 1.3\u00a0M SPS-based ARFB achieves 96.2% capacity utilization, corresponding to 67.05 Ah L-1 delivered capacity. This cobalamin(II) catalysis strategy addresses the kinetic bottlenecks inherent to organodisulfide-based anolytes for ARFBs.\n\nID: 42554260\nTitle: Endoscopic management of common bile duct stones using double balloon endoscopy in postoperative patients: evaluation of single-session complete stone clearance.\nAbstract: Endoscopic treatment of common bile duct (CBD) stones in patients with surgically altered anatomy is challenging because access to the duodenal papilla is often difficult. Although double balloon endoscopy-assisted ERCP (DB-ERCP) has become widespread, data on outcomes by reconstruction type remain limited. A total of 218 patients treated between November 2016 and October 2024 were retrospectively reviewed. Among these, 212 patients were included: 159 with Roux-en-Y reconstruction (R group) and 53 with Billroth II reconstruction (B group). The primary endpoints were single-session complete stone clearance and procedure time. The single-session complete stone clearance rate was 94.8%, with no significant difference between groups. Procedure time was significantly longer in the R group than in the B group (66.0 vs. 50.0\u2009min, p\u2009<\u20090.001). Multivariate analysis identified R group (p\u2009<\u20090.001), age \u226575\u2009years (p\u2009=\u20090.007), and \u22655 stones (p\u2009=\u20090.018) as independent factors for prolonged procedure time. In addition, \u22655 stones independently reduced single-session complete stone clearance (p\u2009<\u20090.001). Adverse events occurred in 2.8%, including one perforation, four bleeding events, and one pancreatitis case. Only one patient required surgery. DB-ERCP achieved a high single-session complete stone clearance rate in patients with surgically altered anatomy, with outcomes comparable to those of conventional ERCP in patients with normal anatomy. These findings suggest that DB-ERCP may represent a first-line treatment option for CBD stones in surgically altered anatomy.\n\nID: 42553904\nTitle: Conscious sedation as a potential optimal anesthetic regimen for endoscopic retrograde cholangiopancreatography in elderly frail patients: A prospective, single-blind, randomized controlled trial.\nAbstract: To investigate the impact of adverse cardiovascular events (ACVs) in elderly frail patients undergoing emergency endoscopic retrograde cholangiopancreatography (ERCP), and to explore a safe and comfortable anesthetic regimen suitable for this specific population. A single-center, prospective, single-blind, randomized controlled trial was conducted. Patients aged 70-90 years scheduled for emergency ERCP were enrolled and randomly divided into two groups: the conscious sedation group (Group S) and the intravenous general anesthesia group (Group A). Basic clinical data of the patients were recorded. During ERCP, the occurrence of ACVs was documented, including hemodynamic fluctuations exceeding 20% of the baseline value, angina pectoris, myocardial infarction, cardiac arrest, acute heart failure, myocardial ischemia, pulmonary embolism, and various arrhythmias. Additionally, adverse reactions such as hypoxemia, body movement, and cough during the procedure were monitored. The effects of different anesthetic methods on ACVs in elderly frail patients undergoing emergency ERCP were compared, and the satisfaction of patients, anesthesiologists, and endoscopists was evaluated. A total of 108 patients undergoing emergency ERCP were enrolled and randomly assigned to Group S (n = 53) and Group A (n = 55). The results showed that there was a statistically significant difference in the incidence of ACVs between the two groups (17% vs. 40%, P < 0.05). Compared with Group A, Group S had a significantly lower incidence of blood pressure and heart rate fluctuations \u226520% of the baseline value (13.2% vs. 25.5%, P < 0.05), and new-onset arrhythmias (5.7% vs. 18.2%, P < 0.05). Compared with intravenous general anesthesia, conscious sedation can reduce fluctuations in blood pressure and heart rate, and decrease the incidence of adverse cardiovascular events in elderly frail patients undergoing emergency ERCP.\n\nID: 42553858\nTitle: Prolonged Retention of a Spontaneously Dislodging Pancreatic Duct Stent in a Patient with Surgically Altered Anatomy: A Case Report.\nAbstract: Spontaneously dislodging pancreatic duct stents are generally expected to pass within several days. We report a rare case of markedly prolonged retention of such a stent in a patient with surgically altered anatomy. A 78-year-old man with a history of distal gastrectomy with Roux-en-Y reconstruction underwent single-balloon enteroscopy-assisted ERCP for choledocholithiasis. After successful biliary cannulation using pancreatic guidewire assistance, stone extraction was performed following a small sphincterotomy and papillary balloon dilation. A 5-Fr \u00d7 5-cm spontaneously dislodging pancreatic duct stent without an internal flange was prophylactically placed. The postoperative course was uneventful; however, spontaneous stent dislodgement was not observed on follow-up imaging. Because of the technical burden of repeat endoscopy in surgically altered anatomy, careful observation was initially selected. The stent remained in place for 104 days and was ultimately removed endoscopically using a single-balloon enteroscope without adverse events. This case illustrates that markedly prolonged retention of a spontaneously dislodging pancreatic duct stent may occur in patients with surgically altered anatomy. Careful follow-up, including imaging to confirm spontaneous stent passage, is important, and endoscopic removal should be considered when the stent remains in place, even if repeat endoscopy is technically challenging.\n\nID: 42553675\nTitle: A Concealed Typical AVNRT: U Wave Contribution in Simulating Long RP Supraventricular Tachycardia.\nAbstract: Accurate identification of P waves on electrocardiogram (ECG) is essential in supraventricular tachycardias because U waves may resemble retrograde P waves, producing a pseudo-long RP interval and hiding typical atrioventricular nodal reentrant tachycardia (AVNRT). A 40-year-old woman presented with regular narrow-complex tachycardia with a positive deflection immediately following the T wave, creating the impression of a long RP tachycardia. The electrophysiological study revealed a short VA interval with an H-A-V activation sequence, characteristic of typical slow-fast AVNRT. This case emphasizes the diagnostic challenge posed by U waves at higher heart rates, which can obscure the true arrhythmia mechanism.\n\nID: 42553548\nTitle: Uncovering the hidden biology of fibrinaloid microclot complexes in complex, inflammatory diseases.\nAbstract: Blood can clot into anomalous, fibrinolysis-resistant forms that arise from prothrombotic seeding areas, including damaged cellular debris and membrane-derived surfaces, giving rise to what we have termed fibrinaloid microclot complexes (colloquially: microclots). Their proteolytic resistance is due in part to the fact that they are amyloid in nature, and they can also entrap inhibitors of proteolysis. They consist of a variety of proteins besides the expected fibrin, and are highly enriched for other amyloidogenic proteins (in contrast to normal clots, whose proteome largely reflects the soluble plasma proteome). They also contain DNA in the form of neutrophil extracellular traps (NETs). Importantly, fibrinaloid microclot complexes are heterogeneous structures comprising multiple phenotypic forms, including those that nucleate and grow on cellular debris such as damaged membranes, microparticles, and immune-derived material. We consider that these debris-associated complexes can act as catalytic scaffolds that recruit fibrin(ogen) and inflammatory molecules, thereby amplifying amyloidogenic transformation and prothrombotic activity. Fibrinaloid microclot complexes have been reported in a widening range of chronic inflammatory and thrombo-inflammatory diseases in which they have been sought, and are highly enriched for amyloidogenic proteins. Additionally, the thrombi extracted from ischaemic stroke also contain proteins in an amyloid form. One mechanism that explains how such macroclots can form and block arteries larger than any leading to them is that this occurs via the accretion of microclots that already contain amyloid. We here show that these microclots exhibit a classical 'apple-green' birefringence when stained with the dye Congo red. It is now important to determine whether inhibiting amyloid-forming clot transitions has therapeutic value.\n\nID: 42553447\nTitle: Clinical outcomes following endoscopic retrograde cholangiopancreatography-based therapy compared with surgery in chronic pancreatitis: evidence from a multicentric cohort.\nAbstract: Real-world comparative outcomes of endoscopic retrograde cholangiopancreatography-based endoscopic therapy vs. pancreatic surgery in chronic pancreatitis (CP) remain incompletely defined. We performed a retrospective comparative effectiveness study using the TriNetX US Collaborative Network. Adults with CP undergoing endoscopic therapy or pancreatic surgery were identified, and propensity score matching generated 1451 patients in each cohort. Outcomes were assessed from 1-1095 days after the index event, using risk-based analyses as the primary comparative summaries and Kaplan-Meier analyses as secondary time-to-event summaries. Outcomes included chronic opioid prescriptions, opioid use disorder (OUD), celiac plexus block/neurolysis, pain codes, acute pancreatitis, exocrine and endocrine pancreatic insufficiency, emergency visits, and all-cause mortality. Mean follow up was 771.494 days for endoscopy and 827.285 days for surgery. New chronic opioid prescriptions occurred in 146/1092 (13.4%) endoscopic vs. 79/1032 (7.7%) surgical patients (odds ratio [OR] 1.862, 95% confidence interval [CI] 1.396-2.484; P<0.001; hazard ratio [HR] 1.861, 95%CI 1.415-2.448). Acute pancreatitis occurred in 167/530 (31.5%) vs. 65/565 (11.5%) patients (OR 3.539, 95%CI 2.578-4.858; P<0.001; HR 3.341, 95%CI 2.506-4.453). Exocrine pancreatic insufficiency occurred in 200/1213 (16.5%) vs. 132/1206 (10.9%) patients (OR 1.606, 95%CI 1.269-2.034; P<0.001; HR 1.585, 95%CI 1.272-1.975). Endocrine pancreatic insufficiency, mortality, emergency visits, and OUD did not differ significantly between cohorts. Endoscopic therapy was associated with higher chronic opioid prescribing, acute pancreatitis, celiac plexus block/neurolysis, and exocrine pancreatic insufficiency. These findings are associative and should be interpreted in the context of residual confounding and coding-based outcome ascertainment.\n\nID: 42553171\nTitle: Curcumin Attenuates Decidual Stromal Cell Ferroptosis and Restores Impaired Decidualization in Recurrent Spontaneous Abortion by Targeting BRD4.\nAbstract: Recurrent spontaneous abortion (RSA) is a major reproductive health challenge with limited clinical options. Curcumin, a natural polyphenol widely consumed as a dietary supplement, has shown promise in improving pregnancy outcomes; however, its specific molecular targets remain obscure due to its broad bioactivity. This study aimed to define curcumin's impact on RSA and identify its direct molecular target to provide a mechanistic basis for its nutritional application. We employed an integrated approach combining phenotypic analysis in the RSA mouse model and target discovery in human endometrial stromal cells. Target identification was performed using unbiased Limited Proteolysis-Mass Spectrometry (LiP-MS). The interaction between curcumin and the identified target, BRD4, was confirmed through molecular docking, dynamics simulations, functional genetics, and chromatin immunoprecipitation (ChIP-qPCR). Functional outcomes were assessed by measuring key ferroptosis markers and mitochondrial morphology using transmission electron microscopy. Decidualization was analyzed in both mouse tissues and cells subjected to a decidualization and ferroptosis induction protocol. Curcumin significantly reduced embryo resorption and restored decidual morphology and marker expression (PRL and IGFBP1) in RSA mice. LiP-MS analysis in human decidual stromal cells identified ferroptosis as the primary pathway targeted by curcumin. Both in\u00a0vivo and in\u00a0vitro validation showed that curcumin inhibits ferroptosis, decreasing lipid peroxidation, restoring glutathione balance, and preserving mitochondrial integrity, which in turn rescued decidual marker expression. Proteomic integration with ferroptosis databases identified BRD4 as a central hub. Mechanistically, curcumin binds BRD4, thereby suppressing BRD4-driven expression of TFRC and ACSL4, which blocks ferroptosis and rescues decidual markers. This research uncovers a BRD4-driven ferroptosis pathway as a key pathogenic mechanism in RSA. Our results demonstrate that curcumin acts as an epigenetic modulator by directly targeting BRD4, thereby suppressing this pro-ferroptotic transcriptional program. These findings provide critical mechanistic evidence supporting curcumin as an evidence-based nutritional intervention for RSA.\n\nID: 42553058\nTitle: Sarcopenia and sepsis fuel a self-perpetuating cycle of immunometabolism decline.\nAbstract: Sepsis and sarcopenia are intertwined clinical challenges characterized by profound immunometabolic dysregulation. Traditionally viewed as a passive reservoir, skeletal muscle is now recognized as an active immunometabolic rheostat that dynamically influences systemic inflammation and homeostasis. This review synthesizes recent advances that redefine our understanding of muscle in critical illness, moving beyond simplistic views of catabolism to encompass complex adaptive strategies. This review summarizes recent advances and discusses septic autocannibalism, a process in which muscle proteolysis, driven by a metabolic defense priority, provides key substrates, glutamine for immune function and alanine for hepatic gluconeogenesis. Initially adaptive, sustained activation of this response leads to severe muscle wasting and long-term functional impairment. We analyze the bidirectional relationship between these conditions, focusing on shared risk factors such as immunosenescence and obesity. Key molecular pathways, including the IL-6/JAK/STAT and NF-\u03baB axes, are examined, with particular emphasis on how the temporal release of myokines (e.g., IL-6, IL-15, IGF-1) dictates their shift from adaptive signals to chronic catabolic drivers. Furthermore, we discuss how energy reprogramming, characterized by aerobic glycolysis and mitochondrial failure, disrupts muscle homeostasis. We highlight physical activity as a potent modulator of immunometabolic health through the release of anti-inflammatory myokines and enhanced mitochondrial biogenesis. To propel the field forward, we propose experimental avenues: single-cell spatial transcriptomics to map cellular crosstalk, mitochondrial transplantation to restore energetic capacity, and the identification of \"muscle resilience\" biomarkers for early intervention. This review underscores the urgent need for integrated immunometabolic approaches to improve outcomes in critically ill patients.\n\nID: 42552689\nTitle: Clinical Development of Therapies for Charcot-Marie-Tooth Disease: Recommendations for Trial Design, Endpoints, and Regulatory Pathways.\nAbstract: Charcot-Marie-Tooth disease (CMT) encompasses a heterogeneous group of inherited peripheral neuropathies. Despite being the most common genetic neurological condition, individual CMT subtypes are rare, presenting unique challenges for therapeutic development. The Together Patients Industry Clinicians versus CMT (ToPIC: CMT) Advocacy Group was formed with a diverse group of patient advocacy groups, clinician-scientists who treat patients with CMT, and pharmaceutical industry representatives to develop a common guidance on development of new treatments for CMT with clear expectations for meaningful patient outcomes and objective assessments of improvement. The ToPIC: CMT Group developed recommendations for clinical development of drugs and biological products for treating CMT, addressing trial design considerations for this rare progressive disease. Key challenges in CMT include small patient populations, variable disease progression, and the need for sensitive outcome measures. Recommendations emphasize flexible trial designs including adaptive designs, external controls, and single-participant designs when scientifically justified. Where possible, broad inclusion criteria based on clinical phenotype rather than genetic subtype alone are recommended. Disease-specific, validated outcome measures should assess function across ages and disease stages. Biomarkers reflecting peripheral nervous system health may serve as surrogate endpoints to support accelerated approval pathways. Patient and care partner perspectives are essential throughout development, particularly regarding treatment goals, risk tolerance, and meaningful endpoints. Successful therapeutic development for CMT and related neuropathies requires innovative approaches that balance rigorous scientific standards with the realities of rare disease research. Regulatory flexibility, informed by patient input and natural history data, can facilitate efficient development while maintaining assurance of safety and effectiveness.\n\nID: 42552337\nTitle: NanoBiT screening identifies the azaoxafluorene VT11 as potent tau interaction inhibitor.\nAbstract: Tauopathies are neurodegenerative disorders characterized by accumulating misfolded, insoluble tau protein aggregates in neurons or glial cells. In this study, we screened the Spectrum Collection and other compound libraries for inhibitors of tau self-interaction using a structural complementation reporter system (NanoLuciferase Binary Technology). Resulting candidates were tested in dose-response assays and evaluated for cell toxicity and microtubule destabilization. Further, a seed-induced tau interaction biosensor assay and a cell-free tau Real-Time Quaking-Induced Conversion assay have been established to study their effects on the kinetics of tau interaction and aggregation, respectively. The substances ritanserin, 3-methoxycatechol, gambogic amide, azaoxafluorenes VT11, and NS 185 and thieno[2,3-d][1.3]oxazine B6/55 showed a concentration-dependent tau self-interaction inhibition without relevant cell toxicity or microtubule destabilization. Ritanserin, VT11, NS 185 and B6/55 blocked tau interaction in the seed-induced tau interaction biosensor assay. Finally, the cell-free tau RT-QuIC assay displayed highest inhibitory potential for VT11. Thus, the azaoxafluorene VT11 seems to be a promising candidate for further investigations as tau interaction inhibitor to address a pivotal pathological process in Alzheimer's disease and other tauopathies.\n\nID: 42552063\nTitle: Recent Advances in Autophagy and Immunotherapy for the Clearance of Aggregated \u03b1-Synuclein in Parkinson's Disease.\nAbstract: Parkinson's disease is a neurodegenerative condition characterized by the accumulation of misfolded and aggregated \u03b1-synuclein in Lewy bodies and neurites. These protein aggregates contribute to neurodegeneration and motor symptoms such as bradykinesia, rigidity, and tremor. While the ubiquitin-proteasome system degrades soluble \u03b1-synuclein, aggregated and oligomeric forms are primarily cleared via the autophagy-lysosomal pathway. Mutations of the SNCA gene exacerbate \u03b1-synuclein aggregation and significantly impair its clearance, highlighting the importance of targeting toxic \u03b1-synuclein species. Strategies such as promoting autophagosome formation via 5'-AMP-activated protein kinase (AMPK) and mechanistic target of rapamycin complex 1 (mTORC1) or facilitating autophagosome maturation via RAB7-a member of the RAS oncogene family-and related effectors, have shown promise in enhancing autophagy and reducing \u03b1-synuclein pathology. Pharmacological agents such as rapamycin, trehalose, and nilotinib have demonstrated preclinical efficacy in enhancing \u03b1-synuclein clearance and alleviating disease features. Concurrently, immunotherapy approaches, including passive and active immunization, aim to enhance the immune system's ability to recognize and eliminate toxic \u03b1-synuclein species. Emerging strategies such as peptide-based therapies aim to inhibit aggregation or promote degradation of \u03b1-synuclein. At the same time, nanotechnology enables the targeted delivery of therapeutic agents across the blood-brain barrier with improved efficiency. Additionally, novel AUTOTAC (autophagy-targeting chimera) platforms offer a precision strategy to tag \u03b1-synuclein for autophagic degradation. This review explores many advances in autophagy-mediated aggregated \u03b1-synuclein clearance, emphasizing its potential as a therapeutic strategy to address the limitations of current symptomatic treatments and slow the progression of Parkinson's disease.\n\nID: 42551925\nTitle: [Development of Therapies Targeting Non-G12C KRAS Mutations (Focusing on G12D and Pan-RAS)].\nAbstract: Sotorasib and adagrasib, which are KRAS G12C inhibitors, have shown favorable clinical results mainly in non-small cell lung cancer (NSCLC) and have already been introduced into clinical practice. However, for patients with non-G12C mutations that account for the majority, such as G12D, G12V, and G13D, effective targeted therapies have still not been established. Setidegrasib (ASP3082) is a first-in-class proteolysis-targeting chimera molecule (PROTAC) that targets the KRAS G12D mutation. Phase \u2160 trial targeting previously treated patients with advanced solid tumors harboring KRAS G12D mutations has been initiated, and the latest results were reported in the N Engl J Med in March 2026. Daraxonrasib (RMC-6236) is a pan-RAS inhibitor that targets all ON-state RAS (KRAS, NRAS, HRAS) by forming a specific pocket spanning the Switch \u2160and Switch \u2161 regions of KRAS through a mechanism called tri-complex/molecular glue. Currently, multiple international collaborative phase \u2162 trials on RMC-6236, mainly focusing on PDAC and NSCLC, are underway. The development of new drugs targeting molecules other than KRAS G12C is progressing rapidly, and in KRAS mutation-positive solid cancers where prognosis was difficult to improve with conventional chemotherapy, there is a high possibility that the existing standard treatments will be significantly revised.\n\nID: 42551861\nTitle: Cadaveric Feasibility Study Describing Transarticular Drilling of the Canine Anconeal Process through a Cranial Arthroscopic Portal.\nAbstract: The aims of this study were to describe a cranial arthroscopic portal to the supratrochlear foramen and to assess the feasibility and accuracy of retrograde transarticular drilling of the anconeal process in canine cadaveric elbows. Cadaver elbows (n\u2009=\u20098) were instrumented through a cranial portal in the distal humerus using a NanoScope. The scope was manoeuvred through the supratrochlear foramen to inspect the anconeal process. A 2.0-mm tunnel simulating the pilot hole for an ununited anconeal process repair was performed using a C guide, cannulated drill bits and NanoScope guidance. Experimental measures included arthroscopic anconeal process identification, surgical time, computed tomography and gross dissection to assess for critical structure damage and drill tunnel aperture location and trajectory. The first two elbows were used for technique development; the remaining six elbows have reported outcome measures. Mean surgery time was 16.6 (standard deviation 3.0) minutes. All six tunnels drilled had acceptable trajectories based on computed tomography and gross dissection. Major damage to critical structures was not observed in any specimen; however, tunnels were directly adjacent to transcubital vessels and a musculocutaneous nerve branch. This technique was feasible, provided visualization of the anconeal process, and allowed accurate drilling of the anconeal process in a cadaveric model. Clinical feasibility and safety require validation in a clinical study.\n=======================================================\n\n### [CUSTOM DATAPOINTS]\nCRITICAL EXTRACTION DIRECTIVE: You MUST extract the following custom datapoints as root-level key/value pairs inside your final JSON block:\n- \"suggested_experiments\": generate 1-3 suggested experiments\n- \"suggested_studies\": generate 1-3 suggested studies\n- \"swansons_literature_based_discovery_candidates\": You are an advanced Literature-Based Discovery (LBD) system executing Swanson\u2019s complementary-but-disjoint (A-B-C) model. Your goal is to find hidden, unpublished connections across the provided dataset. Strict Discovery Protocol: 1. Identify distinct, isolated sub-literatures (Domain A and Domain C) within the dataset that share NO direct citations, co-mentions, or common contextual paragraphs. 2. Find an intermediate biological mechanism, protein, path, or entity (Bridge B) that appears independently in both isolated domains (A-to-B and B-to-C). 3. Synthesize a novel, unstated hypothesis (A-to-C). Negative Constraint (Crucial): DO NOT output any connection if the relationship between Concept A and Concept C is explicitly mentioned, paired, or summarized anywhere in the source text. If a connection (like \"OMN resilience to SMN stabilization\") is already explicitly stated or grouped as a concept in the data, it is considered \"already known\" and must be disqualified. Format your output exactly as follows: - Discovered Hypothesis (A to C): [Clear, novel statement] - Literature A (Origin): [Entity/Concept and source context] - Literature C (Target): [Entity/Concept and source context] - The Intersecting Bridge B: [The shared mechanism/protein linking them] - Biological Rationale: [1-2 sentences explaining why this hidden connection is mechanistically plausible]\n- \"contradictions_between_evidences\": Identify conflicting evidence within the evidence set (if any) and flag the dispute here\n- \"repurposed_solutions\": identify and explain repurposed Solution potentials\n- \"BoNT_induced_lysosomal_flux\": Evaluate if SNAP-25 cleavage and resulting docking inhibition increase the diversion of sequestered pathological proteins (e.g., Tau, alpha-synuclein) toward the autophagic-lysosomal system in the cell body.\n- \"EV_somatic_sequestration_rate\": Measure changes in the proportion of axonal-derived EVs reaching the neuronal soma vs. those released at the terminal following distal BoNT-A administration.\n- \"synaptic_vs_somatic_clearance_ratio\": Determine the shift in clearance kinetics between synaptic EV-mediated spreading and somatic degradation pathways (lysosome/autophagy) under localized synaptic silencing.\n\n\nFormat Requirement:\nRAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\nFirst provide disclaimer such as \"Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\"\n---\nWrite in a highly academic, formal thesis tone.\nFormat your readable response using these exact academic headers:\n###[CLAIM EVALUATED AND ANSWER TO USER]\n(Exact wording of the claim evaluated)\n### [ABSTRACT & REWRITTEN CLAIM]\n(Scientific synthesis)\n### [INTRODUCTION & JUSTIFICATION]\n(Mechanistic explanation utilizing the 'moneyshot quotes' you will use in the EVIDENCE, METHODOLOGY & CITATIONS section later as well)\n### [DISCUSSION: NOVEL & OVERLOOKED]\n(5-10 bullet points of surprising facts)\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 3) - [copied/verbatim Quote text]\"\n\n**CRITICAL: You must include the exact quote you used in the [copied/verbatim Quote text] section.\n\nIf the prompt says \"at least 20 quotes\" then there must be at least 20 matching citations. You must actually use the quotes you select within the conext of the preprint publication you write.\n\nEvaluation Schema:\nRAG AMNESIA IS ACTIVE: You must ONLY use the provided context literature. Do not use outside prior knowledge. If the evidence is missing, insufficient, or requires gap-filling to fully evaluate the claim, you MUST explicitly state the gaps and missing evidence in your justification. Under no circumstances should you invent or hallucinate citations or quotes.\n\n###critical: WRAP YOUR THOUGHTS WITH \nAll responses must include the mandatory \"### [EVIDENCE, METHODOLOGY & CITATIONS]\" section as formatted.\nCRITICAL:\n**MONEYSHOT QUOTES MUST DIRECTLY SUPPORT YOUR CLAIMS**\n**MONEYSHOT QUOTES MUST BE USED IN YOUR RESPONSE TEXT WITHOUT IN-LINE ANNOTATION**\n**MONEYSHOT QUOTES MUST BE USED IN A FORMAL PROFESSIONAL WAY, WORTHY OF PEER REVIEW, WITHOUT ILLOGICAL LEAPS (UNSUPPORTED MAY BE OK, ILLOGICAL IS NOT OK)**\n(Numbered list matching inline citations) For example \"1. ID: 12345 - Application: The text discusses ... and since no other evidence provided proves nor disproves the claim, the lowest rating allowed across all evidences is required. ID:12345 indicates the claim is overall plausible (Alignment with this ID: 7) - *\"copied/verbatim Quote text\"**\n\nCRITICAL INSTRUCTION:\nwhen fact checking: At the very end of your response, you MUST provide a machine-readable JSON block containing evaluation metrics. \nIt MUST be enclosed exactly between ###JSON_START### and ###JSON_END###. Ensure the JSON is valid. \n\nFor the \"Logic_Chain\", break down the systemic mechanism into verbose unabridged atomic multi-step pathways using i/o porting style where the input of next node must match output of the prior (e.g., A -> B, B->C, C->D). Each chain must fully represent the response you give, and should be color coded with light green (Gap_Strength is \"None\"), lightblue (Gap_Strength is medium), or pink (strong Gap_Strength). Logic_Chain MUST be a JSON array of objects. Each object MUST contain EXACTLY these keys: \"Step\", \"From\", \"Relationship\", \"To\", \"evidence_source_id\", \"Alignment_Score\", \"Consilience_Score\", \"Confidence_Score\", \"Gap_Strength\", \"Justification\", and \"Color\". Use commas between objects. DO NOT leave trailing commas inside objects.\n\nFor \"Verbatim_Quotes\", copy at least 20 (required, 20 or more) \"moneyshot\" quotes EXACTLY as they appear in the context literature text, word-for-word, characters included, that fully support your response. We will programmatically validate these. You MUST return an array of OBJECTS, where each object has a \"quote\" key and a \"source_id\" key (the ID of the text it came from, e.g., the ID). Do not alter a single character, do not paraphrase.\n\nUse these scales to evaluate HOW WELL THE EVIDENCE SUPPORTS THE SPECIFIC CLAIM EVALUATED ABOVE:\n- Alignment Score (1-7): How well does the EVALUATED CLAIM factually align with the provided RAG evidence set? [1=Evidence proves claim strictly false, 2=Evidence indicates the claim is impossible, 3=Implausible, 4=Neutral/Unrelated, 5=Plausible, 6=Evidence indicates inevitable, 7=Evidence proves claim strictly true]\n- Consilience Score (1-7): How consilient (in agreement) is the evidence set regarding this claim? [1=Highly Conflicting/Disputed, 4=Mixed, 7=Unanimous Agreement]\n- Confidence Score (1-7): Implied confidence of the research based on study types and depth [1=In Vitro/Animal/Preprint, 4=Observational/Moderate, 7=Meta-analysis/RCT]\n\nFormat (DO NOT USE fencing)\nCRITICAL: Use ONLY Pubmed MeSH tags (exclude descriptor and [type]) for your gate variable names (i.e.,.the \"gates\") so they will be standardized globally. Be unabridged, comprehensive, and exhaustive in your gate mapping with at least 1 gate nodes for each quote you identified per the specification and map the gates granularly/atomically.\n\n###JSON_START###\n{\n \"Alignment\": 5,\n \"Consilience\": 6,\n \"Confidence\": 5,\n \"Logic_Chain\":[\n {\n \"Step\": 1,\n \"From\": \"Variable A\",\n \"Relationship\": \"-->\",\n \"To\": \"Variable B\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 5,\n \"Confidence_Score\": 4,\n \"Gap_Strength\": \"None\",\n \"Justification\": \"...\",\n \"Color\": \"lightgreen\"\n }\n ],\n \"Verbatim_Quotes\": [\n {\n \"quote\": \"Copy the Exact wording from text exactly as it is, including all characters (we ascii match for validation!).\",\n \"source_id\": \"12345678\"\n }\n ],\n \"Study_Type_Audit\": { \"ID123\": \"meta_analysis:Count=10\", \"ID124\": \"in_vivo:Count=3\" },\n \"Gap_Analysis_Audit\": { \"study_type\": \"in_vitro\", \"study_intent\": \"binding\", \"justification\": \"The context provided indicates...\", \"predicted_result\": \"RGNEF binds to Zn2 magnitudes higher than BMAA\", \"short_answer_to_user\": \"Direct answer to the user primary intent, addressing the user directly when appropriate\"}\n,\n \"suggested_experiments\": \"[Extract: generate 1-3 suggested experiments]\",\n \"suggested_studies\": \"[Extract: generate 1-3 suggested studies]\",\n \"swansons_literature_based_discovery_candidates\": \"[Extract: You are an advanced Literature-Based Discovery (LBD) system executing Swanson\u2019s complementary-but-disjoint (A-B-C) model. Your goal is to find hidden, unpublished connections across the provided dataset. Strict Discovery Protocol: 1. Identify distinct, isolated sub-literatures (Domain A and Domain C) within the dataset that share NO direct citations, co-mentions, or common contextual paragraphs. 2. Find an intermediate biological mechanism, protein, path, or entity (Bridge B) that appears independently in both isolated domains (A-to-B and B-to-C). 3. Synthesize a novel, unstated hypothesis (A-to-C). Negative Constraint (Crucial): DO NOT output any connection if the relationship between Concept A and Concept C is explicitly mentioned, paired, or summarized anywhere in the source text. If a connection (like \\\"OMN resilience to SMN stabilization\\\") is already explicitly stated or grouped as a concept in the data, it is considered \\\"already known\\\" and must be disqualified. Format your output exactly as follows: - Discovered Hypothesis (A to C): [Clear, novel statement] - Literature A (Origin): [Entity/Concept and source context] - Literature C (Target): [Entity/Concept and source context] - The Intersecting Bridge B: [The shared mechanism/protein linking them] - Biological Rationale: [1-2 sentences explaining why this hidden connection is mechanistically plausible]]\",\n \"contradictions_between_evidences\": \"[Extract: Identify conflicting evidence within the evidence set (if any) and flag the dispute here]\",\n \"repurposed_solutions\": \"[Extract: identify and explain repurposed Solution potentials]\",\n \"BoNT_induced_lysosomal_flux\": \"[Extract: Evaluate if SNAP-25 cleavage and resulting docking inhibition increase the diversion of sequestered pathological proteins (e.g., Tau, alpha-synuclein) toward the autophagic-lysosomal system in the cell body.]\",\n \"EV_somatic_sequestration_rate\": \"[Extract: Measure changes in the proportion of axonal-derived EVs reaching the neuronal soma vs. those released at the terminal following distal BoNT-A administration.]\",\n \"synaptic_vs_somatic_clearance_ratio\": \"[Extract: Determine the shift in clearance kinetics between synaptic EV-mediated spreading and somatic degradation pathways (lysosome/autophagy) under localized synaptic silencing.]\"\n}\n###JSON_END###\n\n### CRITICAL QUOTE VALIDATION FAILURE (ATTEMPT 1) ###\nThe validator executed a 100% strict, character-by-character substring search. Your response was REJECTED because the following quotes do not exist verbatim in the source texts.\n\n\u274c FAILED QUOTES (You must fix or delete these):\n\n- ERROR: You cited ID: 23300443 for the quote: \"BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT.\"\n FACT: Strict Misquote Detected! The exact character sequence \"BoNT/A and BoNT/E are internalised ...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 23300443 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 23300443 ---\n ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system.\n --- END ACTUAL ABSTRACT FOR 23300443 ---\n\n- ERROR: You cited ID: 9300434 for the quote: \"Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection.\"\n FACT: Strict Misquote Detected! The exact character sequence \"Motoneurons identified by the retro...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 9300434 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 9300434 ---\n ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals.\n --- END ACTUAL ABSTRACT FOR 9300434 ---\n\n- ERROR: You cited ID: 42553289 for the quote: \"Rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence.\"\n FACT: Strict Misquote Detected! The exact character sequence \"Rapa-lipos targeted-inhibited mTORC...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42553289 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42553289 ---\n ID: 42553289\nTitle: Rapamycin-nanoliposomes target the mTORC1-mediated autophagy-lysosomal and NLRP3/Caspase-1 pathways to inhibit nucleus pulposus cell senescence in intervertebral discs.\nAbstract: Nucleus pulposus (NP) cell quiescence maintains intervertebral disc homeostasis, while mTORC1 regulates autophagy-lysosomal function and inflammatory secretion to preserve quiescence-rapamycin specifically targets mTORC1. Herein, we fabricated rapamycin-nanoliposomes (rapa-lipos) via ultrasonic dispersion, thin-film dispersion, and filtration to improve rapamycin bioavailability, investigating their role in inhibiting the senescence phenotype of NP cells through \u03b2-gal staining, lysosomal staining, transmission electron microscopy, ELISA, and cell cycle inhibitors. The mechanistic effects of rapa-lipos on mTORC1, NLRP3/Caspase-1 pathway (NCP) and autophagy-lysosomal pathway (ALP) were also analyzed by western blotting, immunofluorescence (IF), Si-RNA (raptor), and PCR. In vivo, rapa-lipos were injected into rat intervertebral disc with IL-1\u03b2-induced degeneration, assessed via HE staining, x-ray, MRI, and IF. Rapa-lipos exhibited high encapsulation efficiency, favorable drug loading, uniform particle size, and controlled release, suppressing NP cell senescence-related phenotypes (morphological changes, elevated IL-1\u03b2/TNF-\u03b1 secretion, increased \u03b2-gal activity, lysosomal dysfunction, upregulated P21/P16 and reduced P27 expression). Mechanistically, rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence. In vivo, x-ray, MRI and histological evaluation confirmed rapa-lipos mitigated intervertebral disc degeneration. Collectively, rapa-lipos target mTORC1-mediated NCP and ALP to inhibit NP cell senescence, offering a promising strategy for intervertebral disc degeneration prevention.\n --- END ACTUAL ABSTRACT FOR 42553289 ---\n\n- ERROR: You cited ID: 42551559 for the quote: \"The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR.\"\n FACT: Strict Misquote Detected! The exact character sequence \"The CRISPR-Cas system, originally d...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42551559 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42551559 ---\n ID: 42551559\nTitle: CRISPR-Cas Systems as Precision Antimicrobials: Reversing the Tide of Antimicrobial Resistance.\nAbstract: Antimicrobial resistance (AMR) has escalated into a global health crisis, with resistant pathogens causing over 1.2 million direct deaths annually and threatening to render modern medicine unsustainable. This review provides a comprehensive and updated synthesis of CRISPR-Cas-based antimicrobial strategies with a unique focus on: (i) critical comparison with conventional antibiotics and emerging alternatives; (ii) quantitative evaluation of delivery platforms; (iii) novel strategies including AI-optimized guide design and the ATTACK-CreTA system; (iv) comprehensive analysis of ecological risks; and (v) technology readiness level assessments for clinical translation. The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR by selectively targeting and eliminating resistance genes. We systematically evaluate the mechanistic diversity of Cas effectors, from DNA-cleaving Cas9 and Cas3 to RNA-targeting Cas13, and their application in reversing resistance phenotypes in WHO priority pathogens. We critically assess emerging delivery platforms, including engineered bacteriophages, conjugative plasmids, nanoparticles, and outer membrane vesicles, quantitatively comparing their delivery efficiency, payload capacity, and biosafety profiles. Novel strategies such as CRISPR interference (CRISPRi) for gene silencing without genomic cleavage, the ATTACK-CreTA system for enhanced bactericidal activity, and AI-driven optimization of guide RNA design are examined with appropriate caveats. We comprehensively address clinical translation challenges including immunogenicity, pharmacokinetics/pharmacodynamics, manufacturing scalability, regulatory pathways, and bacterial resistance mechanisms including anti-CRISPR proteins. No CRISPR-based antimicrobial has yet received regulatory approval, and we critically evaluate the gap between proof-of-concept and clinical utility. A detailed roadmap for clinical development is proposed. By integrating recent advances in Cas protein engineering, delivery technologies, and diagnostic applications, this review positions CRISPR-Cas systems as next-generation precision therapeutics capable of both treating resistant infections and curtailing the spread of AMR across clinical and environmental settings.\n --- END ACTUAL ABSTRACT FOR 42551559 ---\n\n- ERROR: You cited ID: 23110146 for the quote: \"The present results strongly sustain a combinatorial action at peripheral and central neural levels.\"\n FACT: Strict Misquote Detected! The exact character sequence \"The present results strongly sustai...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 23110146 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 23110146 ---\n ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life.\n --- END ACTUAL ABSTRACT FOR 23110146 ---\n\n- ERROR: You cited ID: 23300443 for the quote: \"These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma.\"\n FACT: Strict Misquote Detected! The exact character sequence \"These results suggest that the fast...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 23300443 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 23300443 ---\n ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system.\n --- END ACTUAL ABSTRACT FOR 23300443 ---\n\n- ERROR: You cited ID: 42102607 for the quote: \"These findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord.\"\n FACT: Strict Misquote Detected! The exact character sequence \"These findings establish that BoNT-...\" was NOT found in the provided text. Do NOT truncate, paraphrase, or edit quotes.\n \n Below is the complete, true text of ID 42102607 that you MUST read. \n Find a valid, verbatim, character-perfect sentence inside this exact block to cite instead, or change your claim to align with what this text actually says:\n \n --- BEGIN ACTUAL ABSTRACT FOR 42102607 ---\n ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases.\n --- END ACTUAL ABSTRACT FOR 42102607 ---\n\n\n\u2705 PASSED (DO NOT CHANGE THESE):\n- \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\" (Source: 25878289)\n- \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\" (Source: 25878289)\n- \"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\" (Source: 23300443)\n- \"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\" (Source: 23110146)\n- \"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\" (Source: 21929507)\n- \"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\" (Source: 42102607)\n- \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\" (Source: 39196607)\n- \"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\" (Source: 39196607)\n- \"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\" (Source: 42102607)\n- \"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\" (Source: 21929507)\n- \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\" (Source: 25878289)\n- \"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\" (Source: 9300434)\n- \"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\" (Source: 25878289)\n\n\nINSTRUCTION: Study the actual abstracts provided. Correct the casing, punctuation, spelling, or map the quote to its true source ID. Do NOT use ellipses.\n\n=======================================================\n\u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f CRITICAL VERIFICATION FAILURE (RETRY LOOP DETECTED) \u26a0\ufe0f\u26a0\ufe0f\u26a0\ufe0f\nYour previous response was REJECTED because your quotes failed strict byte-perfect validation.\n\nTO BREAK THE LOOP, FOLLOW THESE 3 ABSOLUTE RULES:\n1. NO REPAIRING: If a quote failed, do NOT attempt to edit or tweak it. Either copy a completely different, 100% verbatim sentence from the source, or discard the quote entirely.\n2. PERMISSION TO DISCARD: You are NOT permitted to return fewer quotes to pass validation. Never hallucinate just to meet a quota.\n3. BYTE-PERFECT COPY: You must perform a direct, literal copy-paste. Ellipses (...) are BANNED. Do not change a single capital letter, punctuation mark, or space.\n=======================================================\n\nBASED STRICTLY ON THE CONTEXT LITERATURE ABOVE, EVALUATE THIS SPECIFIC PERSPECTIVE/PENTAMATRIX:\n\"Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.\"",
"rawResponse": "Even though this fact check looked at unique up-to-date abstracts, new evidence may refute this answer in the future. Although 'Zero Hallucinated Moneyshot Quotes' is programmatically enforced, AI is not always immune to inadvertently/erroneously misinterpreting data. This is not medical or professional advice, but instead, is an opinion calculated by AI based on the literature evaluated.\n\n###[CLAIM EVALUATED AND ANSWER TO USER]\n\"Hypothesis: If BoNT-A is injected into peripheral nerves, the transient inhibition of SNAP-25-dependent EV secretion at the terminal acts as a 'bottleneck' that increases the retrograde transport load of pathological cargo into endolysosomal degradation pathways within the soma, potentially shifting the clearance balance from synaptic exocytosis to somatic lysosomal processing.\"\n\n### [ABSTRACT & REWRITTEN CLAIM]\nThe provided literature confirms that BoNT/A undergoes retrograde transport from peripheral nerve terminals to the soma, where it interacts with intracellular trafficking machinery and influences central nervous system components. While evidence verifies that BoNT/A utilizes retrograde transport and that it can be sequestered in LC3-positive autophagosomes, the specific hypothesis regarding a \"bottleneck\" shift from synaptic exocytosis to somatic degradation remains a theoretical model requiring further quantitative validation. The literature establishes that BoNT/A cleavage of SNAP-25 occurs at the terminal and subsequently within the soma, but direct kinetic quantification of this shift in clearance pathways requires gap-filling studies.\n\n### [INTRODUCTION & JUSTIFICATION]\nThe intracellular itinerary of Botulinum Neurotoxin Type A (BoNT/A) is far more expansive than its canonical role in blocking neurotransmitter release at the neuromuscular junction. Evidence demonstrates that the toxin enters neurons via synaptic vesicles and is subsequently sorted into retrograde axonal transport carriers. ID: 25878289 notes that \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\" Once internalized, the toxin engages with autophagosomal pathways, as \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\" This interaction with the endolysosomal system is critical, particularly as \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\" The ability of BoNT/A to alter the neurobiological landscape is confirmed by findings that \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\" Thus, while the \"bottleneck\" effect remains a specific hypothesis, the literature supports a shift in proteostatic burden and cellular signaling following retrograde axonal transport.\n\n### [DISCUSSION: NOVEL & OVERLOOKED]\n* BoNT/A-induced inhibition of SNAP-25 does not merely halt release; it causes secondary synaptic stripping in motoneurons, a phenomenon characterized by a 15-fold increase in spine density and significant synaptic bouton detachment.\n* Retrograde trafficking of BoNT/A is not limited to synaptic vesicles but utilizes autophagosomes (LC3-positive carriers), which serve as a highway for delivering the toxin from the terminal to the soma.\n* The toxin's effect extends beyond the presynaptic membrane; it influences the proliferation and secretory states of associated glia, such as Schwann cells, highlighting a multicellular impact.\n* BoNT/A displays differential intracellular migration; while protease activity is often concentrated locally at the site of application, a measurable portion accumulates in the soma over several weeks.\n* The retrograde pathway of BoNT/A can be hijacked for therapeutic purposes, as evidenced by its application in downregulating pain-related neuropeptides like CGRP and ion channels like TRPV1 in the spinal dorsal horn.\n* Neurotoxic activity is not synonymous with death; motoneurons can undergo significant structural remodeling and transient hypertrophy following BoNT/A exposure without immediate organelle degeneration.\n* Evidence suggests that the fast axonal retrograde transport compartment is multifunctional, acting as a gateway for multiple pathogenic virulence factors, not just BoNTs.\n* There is a distinct discrepancy between the timing of peripheral paralysis and the appearance of central neurotoxic effects, suggesting a staggered temporal profile for the toxin's dual mechanism of action.\n\n### [EVIDENCE, METHODOLOGY & CITATIONS]\n1. ID: 25878289 - Application: Investigating BoNT/A retrograde trafficking mechanisms. Alignment: 5. Quote: *\"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\"*\n2. ID: 25878289 - Application: Assessing autophagosomal involvement. Alignment: 5. Quote: *\"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\"*\n3. ID: 23300443 - Application: Confirming retrograde transport in motor neurons. Alignment: 5. Quote: *\"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\"*\n4. ID: 23110146 - Application: Demonstrating cl-SNAP-25 presence in CNS. Alignment: 5. Quote: *\"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\"*\n5. ID: 21929507 - Application: Tracking toxin movement in neurites. Alignment: 5. Quote: *\"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\"*\n6. ID: 42102607 - Application: Elucidating CNS involvement. Alignment: 5. Quote: *\"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\"*\n7. ID: 39196607 - Application: Identifying ER-related translocation. Alignment: 5. Quote: *\"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\"*\n8. ID: 39196607 - Application: Tracking Golgi-ER trafficking. Alignment: 5. Quote: *\"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\"*\n9. ID: 42102607 - Application: Antipruritic mechanism explanation. Alignment: 5. Quote: *\"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\"*\n10. ID: 21929507 - Application: Temporal somatic accumulation. Alignment: 5. Quote: *\"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\"*\n11. ID: 25878289 - Application: Activity-dependent retrograde transport. Alignment: 5. Quote: *\"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\"*\n12. ID: 9300434 - Application: Structural alteration of synapses. Alignment: 5. Quote: *\"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\"*\n13. ID: 25878289 - Application: Defining CNS activity. Alignment: 5. Quote: *\"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\"*\n14. ID: 39196607 - Application: Identification of genome-wide screening. Alignment: 4. Quote: *\"To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.\"*\n15. ID: 23110146 - Application: Modulation of Schwann cells. Alignment: 5. Quote: *\"We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.\"*\n16. ID: 23300443 - Application: Comparing TeNT and BoNTs. Alignment: 4. Quote: *\"The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.\"*\n17. ID: 9300434 - Application: Detachment of synaptic boutons. Alignment: 5. Quote: *\"Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.\"*\n18. ID: 42102607 - Application: SNAP-25 in mast cells. Alignment: 5. Quote: *\"Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.\"*\n19. ID: 9300434 - Application: Long-term synaptic changes. Alignment: 5. Quote: *\"Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).\"*\n20. ID: 39196607 - Application: Neuronal somatic processing. Alignment: 5. Quote: *\"Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.\"*\n\n###JSON_START###\n{\n \"Alignment\": 5,\n \"Consilience\": 5,\n \"Confidence\": 4,\n \"Logic_Chain\": [\n {\n \"Step\": 1,\n \"From\": \"Terminal BoNT/A uptake\",\n \"Relationship\": \"-->\",\n \"To\": \"SNAP-25 cleavage and vesicle docking inhibition\",\n \"evidence_source_id\": \"42102607\",\n \"Alignment_Score\": 7,\n \"Consilience_Score\": 7,\n \"Confidence_Score\": 6,\n \"Gap_Strength\": \"None\",\n \"Justification\": \"BoNT/A binds and cleaves SNAP-25 to inhibit exocytosis.\",\n \"Color\": \"lightgreen\"\n },\n {\n \"Step\": 2,\n \"From\": \"SNAP-25 cleavage\",\n \"Relationship\": \"-->\",\n \"To\": \"Autophagosome incorporation of toxin/cargo\",\n \"evidence_source_id\": \"25878289\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 6,\n \"Confidence_Score\": 5,\n \"Gap_Strength\": \"medium\",\n \"Justification\": \"Toxin is internalized and directed into retrograde carriers.\",\n \"Color\": \"lightblue\"\n },\n {\n \"Step\": 3,\n \"From\": \"Axonal transport\",\n \"Relationship\": \"-->\",\n \"To\": \"Somatic endolysosomal fusion\",\n \"evidence_source_id\": \"25878289\",\n \"Alignment_Score\": 6,\n \"Consilience_Score\": 6,\n \"Confidence_Score\": 5,\n \"Gap_Strength\": \"medium\",\n \"Justification\": \"LC3-positive autophagosomes fuse with lysosomes in the soma.\",\n \"Color\": \"lightblue\"\n }\n ],\n \"Verbatim_Quotes\": [\n {\"quote\": \"Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport.\", \"source_id\": \"25878289\"},\n {\"quote\": \"Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo.\", \"source_id\": \"25878289\"},\n {\"quote\": \"We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport.\", \"source_id\": \"23300443\"},\n {\"quote\": \"Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A.\", \"source_id\": \"23110146\"},\n {\"quote\": \"The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes.\", \"source_id\": \"21929507\"},\n {\"quote\": \"Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn.\", \"source_id\": \"42102607\"},\n {\"quote\": \"The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol.\", \"source_id\": \"39196607\"},\n {\"quote\": \"An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion.\", \"source_id\": \"39196607\"},\n {\"quote\": \"Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP.\", \"source_id\": \"42102607\"},\n {\"quote\": \"A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks.\", \"source_id\": \"21929507\"},\n {\"quote\": \"Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons.\", \"source_id\": \"25878289\"},\n {\"quote\": \"Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone.\", \"source_id\": \"9300434\"},\n {\"quote\": \"However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport.\", \"source_id\": \"25878289\"},\n {\"quote\": \"To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes.\", \"source_id\": \"39196607\"},\n {\"quote\": \"We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes.\", \"source_id\": \"23110146\"},\n {\"quote\": \"The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons.\", \"source_id\": \"23300443\"},\n {\"quote\": \"Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron.\", \"source_id\": \"9300434\"},\n {\"quote\": \"Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action.\", \"source_id\": \"42102607\"},\n {\"quote\": \"Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days).\", \"source_id\": \"9300434\"},\n {\"quote\": \"Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites.\", \"source_id\": \"39196607\"}\n ],\n \"Study_Type_Audit\": { \"25878289\": \"in_vitro:Count=1\", \"23300443\": \"in_vivo:Count=1\", \"42102607\": \"in_vivo:Count=1\", \"39196607\": \"in_vitro:Count=1\" },\n \"Gap_Analysis_Audit\": { \"study_type\": \"in_vitro/in_vivo\", \"study_intent\": \"trafficking\", \"justification\": \"The evidence supports retrograde transport and lysosomal fusion but lacks direct kinetic measurement of the hypothesized 'bottleneck' transition.\", \"predicted_result\": \"Quantitative shifts in cargo destination.\", \"short_answer_to_user\": \"While retrograde transport and endolysosomal processing of BoNT/A are established, the specific 'bottleneck' shift remains a mechanistic hypothesis needing further verification.\" },\n \"suggested_experiments\": \"Quantify the ratio of retrograde-to-exocytic vesicle flux in the presence vs. absence of BoNT/A using live-cell confocal imaging. Analyze the accumulation of tagged pathological proteins in the soma using pulse-chase methods in compartments.\",\n \"suggested_studies\": \"Compare somatic lysosomal enzyme activity levels in BoNT/A-treated vs. control neurons.\",\n \"swansons_literature_based_discovery_candidates\": \"- Discovered Hypothesis (A to C): BoNT/A-induced somatic endolysosomal upregulation could serve as a compensatory clearing mechanism for tau-containing microclots in tauopathy-prone neurons. - Literature A (Origin): BoNT/A retrograde transport to the soma and fusion with lysosomes (ID: 25878289) - Literature C (Target): Amyloidogenic protein clearing in neurodegeneration (ID: 42553548) - The Intersecting Bridge B: LC3-positive autophagosomes and lysosomal fusion. - Biological Rationale: BoNT/A routes cargo into the somatic lysosomal degradation pathway; thus, stimulating this specific retrograde pathway may mechanically facilitate the clearance of other non-toxin-related intracellular protein aggregates.\",\n \"contradictions_between_evidences\": \"None identified in the current literature set regarding retrograde transport existence, though spatial distribution of cleavage varies.\",\n \"repurposed_solutions\": \"Utilization of BoNT/A-induced retrograde transport as a targeted vehicle to deliver degradation-inducing motifs to the neuronal soma for neurodegenerative diseases.\",\n \"BoNT_induced_lysosomal_flux\": \"The evidence indicates that BoNT/A is routed to lysosomes, but whether this specifically draws in other exogenous pathological proteins like Tau remains an experimental inference.\",\n \"EV_somatic_sequestration_rate\": \"Insufficient data to quantify the specific change in EV reaching the soma compared to terminal release.\",\n \"synaptic_vs_somatic_clearance_ratio\": \"Evidence confirms a shift toward somatic processing of the toxin, but a definitive clearance ratio of general synaptic-to-somatic cargo is not provided.\"\n}\n###JSON_END###",
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"9300434": "ID: 9300434\nTitle: Effects of botulinum neurotoxin type A on abducens motoneurons in the cat: ultrastructural and synaptic alterations.\nAbstract: The synaptic alterations induced in abducens motoneurons by the injection of 3 ng/kg of botulinum neurotoxin type A into the lateral rectus muscle were studied using ultrastructural and electrophysiological techniques. Motoneurons identified by the retrograde transport of horseradish peroxidase showed a progressive synaptic stripping already noticeable by four days post-injection which increased over the study period. By 35 days post-injection, the normal coverage of motoneurons by synaptic boutons (66.4 +/- 4.0%) significantly decreased to 27.2 +/- 4.0%. Synaptic boutons detached by a widening of the subsynaptic space but remained apposed by synaptic contacts and desmosomes to the motoneuron. Detachment did not affect equally flat and round vesicle-containing boutons. The control motoneuron had almost equal numbers of both types of boutons, but after 35 days post-injection the ratio of round to flat vesicle-containing boutons was 1.20 +/- 0.01. Synaptic boutons impinging on motoneurons showed signs of alterations in membrane turnover, as indicated by an increase in the number of synaptic vesicles and a decrease in the number of coated vesicles and synaptic vesicles near the active zone. Abducens motoneurons had a transient increase in soma size by 15 days that returned to normal at 35 days, but no signs of chromatolysis or organelle degeneration were seen. Accompanying the swelling of motoneurons, a 15-fold increase in the number of spines, very infrequent in controls, was observed. Spines located in the soma and proximal dendritic trunk received synaptic contacts from both flat and round vesicle-containing boutons that could be either partly detached or completely attached to the motoneuron. An increased turnover of the plasmatic membrane of the motoneuron was observed, as indicated by a four-fold increase in the number of somatic coated vesicles. Animals were implanted with bipolar electrodes in the ampulla of both horizontal semicircular canals for evoking contralateral excitatory and ipsilateral inhibitory postsynaptic potentials. Motoneurons were antidromically identified from the lateral rectus muscle. Synaptic potentials of vestibular origin were recorded in abducens motoneurons. In the period between two and six days post-injection, a complete abolition of inhibitory synaptic potentials was observed. By contrast, excitatory synaptic potentials remained, but were reduced by 82%. The imbalance between excitatory and inhibitory inputs to motoneurons induced a progressive increase of firing frequency within a few stimuli applied to the contralateral canal. Between 7 and 15 days post-injection, both excitatory and inhibitory postsynaptic potentials were virtually abolished and remained so up to the longest time checked (105 days). Some motoneurons recorded beyond 60 days post-injection showed signs of recovery of excitatory postsynaptic potentials. During the whole time-span studied, presynaptic wavelets were present, indicating no affecting of the conduction of afferent volleys to the abducens nucleus. Taken together, these data indicate that botulinum neurotoxin at high doses causes profound synaptic alterations in motoneurons responsible for the effects seen in the behavior of motoneurons recorded in alert animals.",
"9344876": "ID: 9344876\nTitle: Intracellular location of SNAP-25 in human neutrophils.\nAbstract: Exocytosis plays an essential role in the physiological functions of human neutrophils. Although SNAP-25 is considered to play a key role in vesicle-membrane fusion, it has been detected almost exclusively in the neuronal system. Using different specific antibodies to SNAP-25, we have identified in the membrane fraction of resting human neutrophils an immunoreactive band with the same molecular mass observed in brain homogenates. Immunoblot analysis of subcellular fractions of neutrophils revealed that SNAP-25 protein was found in the granule membrane fraction, but not in the cytosolic and plasma membrane fractions. Granule localization for neutrophil SNAP-25 was further demonstrated by confocal and immunoelectron microscopy. Furthermore, SNAP-25 was mainly located in the morphologically defined neutrophil peroxidase-negative granules, which are mobilizable upon cell activation. In addition, the protein was specifically cleaved by botulinal neurotoxin A, as observed in brain homogenate. These findings reveal the presence of SNAP-25 in the granule membranes of human neutrophils.",
"10783705": "ID: 10783705\nTitle: [Muscarinic modulation of cardiac activity].\nAbstract: The goal of the present review is to report information concerning cardiac innervation or more precisely to approach the modulation of cardiac electrical and mechanical activity by parasympathetic innervation. Acetylcholine (ACh) release by nerve endings from the vagus nerve hyperpolarizes the membrane, shortens action potential (AP) duration and has a negative inotropic effect on cardiac muscle. Toxins are usefull tools in the study of membrane signals. The Caribbean ciguatoxin (C-CTX-1) has a muscarinic effect on frog atrial fibres. The toxin evokes the release of ACh from motoneuron nerve terminals innervating this tissue which allows us to propose a model, similar to the one of the neuromuscular junction (nmj), to describe the events occurring during the triggering and release of ACh. Trachynilysin (TLY) is a proteic toxin which causes an influx of Ca2+ into the cells and releases ACh from nmj synaptic vesicles. TLY has a muscarinic effect on atrial fibres which is explicated in the release of neurotransmitter from the nerve endings generated by the TLY-induced Ca2+ influx. It is known that ACh release from nmj is known to be due to exocytosis of synaptic vesicles via the activation of a proteic complex blocked by botulinum toxins. One of these proteins SNAP-25 is the target of type A botulinum toxin (BoNT/A). The study of hearts isolated from BoNT/A poisoned frogs show that atrial AP is lengthened and reveals the presence of SNAP-25 in nerve endings of this tissue. Moreover, the electrical activity of ventricular muscle is markedly altered; in BoNT/A treated frog, an important outward current activated by internal Ca2+ develops. ACh released from nerve terminals binds to a G protein coupled membrane receptor and activates a K+ channel and other effectors. Five subtypes of muscarinic receptors have been cloned from different tissue (M1, M2, M3, M4) subtypes have been identified in cardiac tissues throughout many species. These receptors coupled with different G-proteins activate different effectors. M1 receptors modulate the cardiac plateau and therefore the magnitude of the peak contraction. M2 receptors are mainly involved in the repolarization phase of the AP and modulate the duration of the peak contraction. The roles of M3 and M4 are not yet clearly defined; however, they may activate K+ currents. In conclusion, ACh releases from parasympathetic nerve endings which innervate cardiac cells follows to similar events (Ca2+ influx; presence of a SNAP-25 protein) to those which produce ACh release from nmj, stimulates different G proteins coupled muscarinic receptors, and activates different effectors involved in the modulation of cardiac electrical and mechanical activity.",
"21929507": "ID: 21929507\nTitle: Extravesicular intraneuronal migration of internalized botulinum neurotoxins without detectable inhibition of distal neurotransmission.\nAbstract: Intracellular protein transport routes can be studied using toxins that exploit these to enter cells. BoNTA (botulinum neurotoxin type\u00a0A) is a protease that binds to peripheral nerve terminals, becomes endocytosed and causes prolonged blockade of transmitter release by cleaving SNAP-25 (synaptosome-associated protein of 25\u00a0kDa). Retrograde transport of the toxin has been suggested, but not of the transient muscle relaxant, BoNTE (botulinum neurotoxin type\u00a0E). In the present study, dispersal of these proteases in compartmented cultures of rat sympathetic neurons was examined after focal application of BoNTA or BoNTE to neurites. A majority of cleaved SNAP-25 was seen locally, but some appeared along neurites and accumulated in the soma over several weeks. BoNTE yielded less cleaved SNAP-25 at distal sites due to shorter-lived enzymic activity. Neurite transection prevented movement of BoNTA. The BoNTA protease could be detected only in the supernatants of neurites or cell body lysates, hence these proteases must move along neuronal processes in the axoplasm or are reversibly associated with membranes. Substitution into BoNTE of the BoNTA acceptor-binding domain did not alter its potency or mobility. Spontaneous or evoked transmission to cell bodies were not inhibited by retrogradely migrated BoNTA except with high doses, concurring with the lack of evidence for a direct central action when used clinically.",
"23110146": "ID: 23110146\nTitle: The analgesic effect on neuropathic pain of retrogradely transported botulinum neurotoxin A involves Schwann cells and astrocytes.\nAbstract: In recent years a growing debate is about whether botulinum neurotoxins are retrogradely transported from the site of injection. Immunodetection of cleaved SNAP-25 (cl-SNAP-25), the protein of the SNARE complex targeted by botulinum neurotoxin serotype A (BoNT/A), could represent an excellent approach to investigate the mechanism of action on the nociceptive pathways at peripheral and/or central level. After peripheral administration of BoNT/A, we analyzed the expression of cl-SNAP-25, from the hindpaw's nerve endings to the spinal cord, together with the behavioral effects on neuropathic pain. We used the chronic constriction injury of the sciatic nerve in CD1 mice as animal model of neuropathic pain. We evaluated immunostaining of cl-SNAP-25 in the peripheral nerve endings, along the sciatic nerve, in dorsal root ganglia and in spinal dorsal horns after intraplantar injection of saline or BoNT/A, alone or colocalized with either glial fibrillar acidic protein, GFAP, or complement receptor 3/cluster of differentiation 11b, CD11b, or neuronal nuclei, NeuN, depending on the area investigated. Immunofluorescence analysis shows the presence of the cl-SNAP-25 in all tissues examined, from the peripheral endings to the spinal cord, suggesting a retrograde transport of BoNT/A. Moreover, we performed in vitro experiments to ascertain if BoNT/A was able to interact with the proliferative state of Schwann cells (SC). We found that BoNT/A modulates the proliferation of SC and inhibits the acetylcholine release from SC, evidencing a new biological effect of the toxin and further supporting the retrograde transport of the toxin along the nerve and its ability to influence regenerative processes. The present results strongly sustain a combinatorial action at peripheral and central neural levels and encourage the use of BoNT/A for the pathological pain conditions difficult to treat in clinical practice and dramatically impairing patients' quality of life.",
"23300443": "ID: 23300443\nTitle: Botulinum neurotoxins A and E undergo retrograde axonal transport in primary motor neurons.\nAbstract: The striking differences between the clinical symptoms of tetanus and botulism have been ascribed to the different fate of the parental neurotoxins once internalised in motor neurons. Tetanus toxin (TeNT) is known to undergo transcytosis into inhibitory interneurons and block the release of inhibitory neurotransmitters in the spinal cord, causing a spastic paralysis. In contrast, botulinum neurotoxins (BoNTs) block acetylcholine release at the neuromuscular junction, therefore inducing a flaccid paralysis. Whilst overt experimental evidence supports the sorting of TeNT to the axonal retrograde transport pathway, recent findings challenge the established view that BoNT trafficking is restricted to the neuromuscular junction by highlighting central effects caused by these neurotoxins. These results suggest a more complex scenario whereby BoNTs also engage long-range trafficking mechanisms. However, the intracellular pathways underlying this process remain unclear. We sought to fill this gap by using primary motor neurons either in mass culture or differentiated in microfluidic devices to directly monitor the endocytosis and axonal transport of full length BoNT/A and BoNT/E and their recombinant binding fragments. We show that BoNT/A and BoNT/E are internalised by spinal cord motor neurons and undergo fast axonal retrograde transport. BoNT/A and BoNT/E are internalised in non-acidic axonal carriers that partially overlap with those containing TeNT, following a process that is largely independent of stimulated synaptic vesicle endo-exocytosis. Following intramuscular injection in vivo, BoNT/A and TeNT displayed central effects with a similar time course. Central actions paralleled the peripheral spastic paralysis for TeNT, but lagged behind the onset of flaccid paralysis for BoNT/A. These results suggest that the fast axonal retrograde transport compartment is composed of multifunctional trafficking organelles orchestrating the simultaneous transfer of diverse cargoes from nerve terminals to the soma, and represents a general gateway for the delivery of virulence factors and pathogens to the central nervous system.",
"25878289": "ID: 25878289\nTitle: Control of autophagosome axonal retrograde flux by presynaptic activity unveiled using botulinum neurotoxin type a.\nAbstract: Botulinum neurotoxin type A (BoNT/A) is a highly potent neurotoxin that elicits flaccid paralysis by enzymatic cleavage of the exocytic machinery component SNAP25 in motor nerve terminals. However, recent evidence suggests that the neurotoxic activity of BoNT/A is not restricted to the periphery, but also reaches the CNS after retrograde axonal transport. Because BoNT/A is internalized in recycling synaptic vesicles, it is unclear which compartment facilitates this transport. Using live-cell confocal and single-molecule imaging of rat hippocampal neurons cultured in microfluidic devices, we show that the activity-dependent uptake of the binding domain of the BoNT/A heavy chain (BoNT/A-Hc) is followed by a delayed increase in retrograde axonal transport of BoNT/A-Hc carriers. Consistent with a role of presynaptic activity in initiating transport of the active toxin, activity-dependent uptake of BoNT/A in the terminal led to a significant increase in SNAP25 cleavage detected in the soma chamber compared with nonstimulated neurons. Surprisingly, most endocytosed BoNT/A-Hc was incorporated into LC3-positive autophagosomes generated in the nerve terminals, which then underwent retrograde transport to the cell soma, where they fused with lysosomes both in vitro and in vivo. Blocking autophagosome formation or acidification with wortmannin or bafilomycin A1, respectively, inhibited the activity-dependent retrograde trafficking of BoNT/A-Hc. Our data demonstrate that both the presynaptic formation of autophagosomes and the initiation of their retrograde trafficking are tightly regulated by presynaptic activity.",
"36831512": "ID: 36831512\nTitle: Current Status of Lymphangiogenesis: Molecular Mechanism, Immune Tolerance, and Application Prospect.\nAbstract: The lymphatic system is a channel for fluid transport and cell migration, but it has always been controversial in promoting and suppressing cancer. VEGFC/VEGFR3 signaling has long been recognized as a major molecular driver of lymphangiogenesis. However, many studies have shown that the neural network of lymphatic signaling is complex. Lymphatic vessels have been found to play an essential role in the immune regulation of tumor metastasis and cardiac repair. This review describes the effects of lipid metabolism, extracellular vesicles, and flow shear forces on lymphangiogenesis. Moreover, the pro-tumor immune tolerance function of lymphatic vessels is discussed, and the tasks of meningeal lymphatic vessels and cardiac lymphatic vessels in diseases are further discussed. Finally, the value of conversion therapy targeting the lymphatic system is introduced from the perspective of immunotherapy and pro-lymphatic biomaterials for lymphangiogenesis.",
"36971125": "ID: 36971125\nTitle: Extracellular vesicle-mediated heterogeneous communication between cancer and the lymphatic system facilitates lymphatic metastasis.\nAbstract: ",
"37037273": "ID: 37037273\nTitle: Botulinum neurotoxin A modulates the axonal release of pathological tau in hippocampal neurons.\nAbstract: Pathological tau aggregates propagate across functionally connected neuronal networks in human neurodegenerative pathologies, such as Alzheimer's disease. However, the mechanism underlying this process is poorly understood. Several studies have showed that tau release is dependent on neuronal activity and that pathological tau is found in the extracellular space in free form, as well as in the lumen of extracellular vesicles. We recently showed that metabotropic glutamate receptor activity and SNAP25 integrity modulate the release of pathological tau from human and mouse synaptosomes. Here, we have leveraged botulinum neurotoxins (BoNTs), which impair neurotransmitter release by cleaving specific synaptic SNARE proteins, to dissect molecular mechanisms related to tau release at synapses. In particular, we have tested the effect of botulinum neurotoxin A (BoNT/A) on the synaptic release of tau in primary mouse neurons. Hippocampal neurons were grown in microfluidic chambers and transduced with lentiviruses expressing human tau (hTau). We found that neuronal stimulation significantly increases the release of mutant hTau, whereas wild-type hTau is unaffected. Importantly, BoNT/A blocks mutant hTau release, indicating that this process is controlled by SNAP25, a component of the SNARE complex, in intact neurons. These results suggest that BoNTs are potent tools to study the spreading of pathological proteins in neurodegenerative diseases and could play a central role in identifying novel molecular targets for the development of therapeutic interventions to treat tauopathies.",
"37086961": "ID: 37086961\nTitle: Tumor-derived extracellular vesicles delivering TNF-\u03b1 promotes colorectal cancer metastasis via the NF-kB/LAMB3/AKT axis by targeting SNAP23.\nAbstract: Accumulating evidence have demonstrated that cytokines are enriched in tumor-derived extracellular vesicles (EVs) and widely involved in tumorigenesis of various types of carcinomas, including colorectal cancer (CRC). Nevertheless, the functions of cytokines in EVs secreted from colorectal cancer cells remain largely unknown. In the present study, we found that TNF-\u03b1 was elevated in EVs from CRC patient serum samples and CRC cell lines, of which the expression was associated with aggressive features of colorectal cancer. EV TNF-\u03b1 secretion is dependent on synaptosome-associated protein 23 (SNAP23). Functional experiments revealed that EV TNF-\u03b1 promotes CRC cell metastasis via the NF-\u03baB pathway by targeting SNAP23. Mechanistically, SNAP23 was transcriptionally upregulated by EV TNF-\u03b1/NF-\u03baB axis to enhance the expression of laminin subunit beta-3 (LAMB3), thereby activating the PI3K/AKT signaling pathway and consequently facilitate CRC progression. Based on our findings, we could conclude that EV TNF-\u03b1 plays an oncogenic role in CRC progression through SNAP23, which in turn promotes EV TNF-\u03b1 secretion, suggesting that EV TNF-\u03b1/SNAP23 axis may serve as a diagnostic biomarker and potential therapeutic target for CRC.",
"37204265": "ID: 37204265\nTitle: Ultrasonography in neurology: A comprehensive analysis and review.\nAbstract: Neurologists in both the inpatient and outpatient settings are increasingly using ultrasound to diagnose and manage common neurological diseases. Advantages include cost-effectiveness, the lack of exposure to ionizing radiation, and the ability to perform at the bedside to provide real-time data. There is a growing body of literature that supports using ultrasonography to improve diagnostic accuracy and aid in performing procedures. Despite the increasing utilization of this imaging modality in medicine, there has been no comprehensive review of the clinical applications of ultrasound in the field of neurology. We discuss the current uses and limitations of ultrasound for various neurological conditions. We review the role for ultrasound in commonly performed neurologic procedures including lumbar puncture, botulinum toxin injections, nerve blocks, and trigger point injections. We specifically discuss the technique for ultrasound-assisted lumbar puncture and occipital nerve block as these are commonly performed. We then focus on the utility of ultrasound in the diagnosis of neurologic conditions. This includes neuromuscular diseases such as motor neuron disorders, focal neuropathies, and muscular dystrophy as well as vascular conditions such as stroke and vasospasm in subarachnoid hemorrhage. We also address ultrasound's use in critically ill patients to aid in identifying increased intracranial pressure, hemodynamics, and arterial and/or venous catheterization. Finally, we address the importance of standardized ultrasound curricula in trainee education and make recommendations for the future directions of research and competency guidelines within our specialty.",
"37371477": "ID: 37371477\nTitle: The Role of Extracellular Vesicles in the Pathogenesis of Hematological Malignancies: Interaction with Tumor Microenvironment; a Potential Biomarker and Targeted Therapy.\nAbstract: The tumor microenvironment (TME) plays an important role in the development and progression of hematological malignancies. In recent years, studies have focused on understanding how tumor cells communicate within the TME. In addition to several factors, such as growth factors, cytokines, extracellular matrix (ECM) molecules, etc., a growing body of evidence has indicated that extracellular vesicles (EVs) play a crucial role in the communication of tumor cells within the TME, thereby contributing to the pathogenesis of hematological malignancies. The present review focuses on how EVs derived from tumor cells interact with the cells in the TME, such as immune cells, stromal cells, endothelial cells, and ECM components, and vice versa, in the context of various hematological malignancies. EVs recovered from the body fluids of cancer patients often carry the bioactive molecules of the originating cells and hence can be considered new predictive biomarkers for specific types of cancer, thereby also acting as potential therapeutic targets. Here, we discuss how EVs influence hematological tumor progression via tumor-host crosstalk and their use as biomarkers for hematological malignancies, thereby benefiting the development of potential therapeutic targets.",
"37394036": "ID: 37394036\nTitle: Intercellular transmission of pathogenic proteins in ALS: Exploring the pathogenic wave.\nAbstract: In patients with amyotrophic lateral sclerosis (ALS), disease symptoms and pathology typically spread in a predictable spatiotemporal pattern beginning at a focal site of onset and progressing along defined neuroanatomical tracts. Like other neurodegenerative diseases, ALS is characterized by the presence of protein aggregates in postmortem patient tissue. Cytoplasmic, ubiquitin-positive aggregates of TDP-43 are observed in approximately 97% of sporadic and familial ALS patients, while SOD1 inclusions are likely specific to cases of SOD1-ALS. Additionally, the most common subtype of familial ALS, caused by a hexanucleotide repeat expansion in the first intron of the C9orf72 gene (C9-ALS), is further characterized by the presence of aggregated dipeptide repeat proteins (DPRs). As we will describe, cell-to-cell propagation of these pathological proteins tightly correlates with the contiguous spread of disease. While TDP-43 and SOD1 are capable of seeding protein misfolding and aggregation in a prion-like manner, C9orf72 DPRs appear to induce (and transmit) a 'disease state' more generally. Multiple mechanisms of intercellular transport have been described for all of these proteins, including anterograde and retrograde axonal transport, extracellular vesicle secretion, and macropinocytosis. In addition to neuron-to-neuron transmission, transmission of pathological proteins occurs between neurons and glia. Given that the spread of ALS disease pathology corresponds with the spread of symptoms in patients, the various mechanisms by which ALS-associated protein aggregates propagate through the central nervous system should be closely examined.",
"37517544": "ID: 37517544\nTitle: Logistics and distribution of small extracellular vesicles from the subcutaneous space to the lymphatic system.\nAbstract: Small extracellular vesicles (sEVs) are small, cell-derived particles with sizes of approximately 100\u00a0nm. Since these particles include cargos such as host cell-derived proteins, messenger RNAs, and micro RNAs, they serve as mediators of cell-cell communication. While the analysis of the pharmacokinetic of sEVs after the intravenous injection have been reported, the lymphatic transport of sEVs remains unclear. The objective of this study was to provide insights into the intra-lymphatic trafficking and distribution of sEVs when they are injected into an interstitial space both in normal skin tissue and in cancerous tissue. When sEVs were Subcutaneously administered into the tail base and the tumor tissue, they preferably accumulated in the lymph nodes (LNs), rather than in the liver and the spleen. The findings reported herein show that the lymphatic transport of sEVs was drastically changed in model mice, in which a surgical treatment was used to modify to allow the dominant lymphatic flow from the footpad directly to the axillary LN via the inguinal LN. Based on the results, we conclude that when sEVs are injected into the subcutis space, they are preferably delivered to the LN via the lymphatic system. Further, the extent of accumulation of sEVs in the LN after subcutaneous injection was reduced when they were preliminarily incubated with Proteinase K. These results suggest that the lymphatic drainage of sEVs in normal skin tissue is regulated by membrane proteins on their surface. This reduction, however, was not observed in the case of cancer tissue. This discrepancy can be attributed to the presence of highly permeable lymphatic vessels in the tumor tissue. Further, the major cell subtypes that captured sEVs in the LN were LN-resident medullary sinus macrophages. These collective findings indicate that the lymphatic drainage of sEVs are mediated by proteins and, that they may appear to contribute to the control of the function of immune-responsive cells in the LNs.",
"37700095": "ID: 37700095\nTitle: Identification of the SNARE complex that mediates the fusion of multivesicular bodies with the plasma membrane in exosome secretion.\nAbstract: Exosomes play crucial roles in local and distant cellular communication and are involved in various physiological and pathological processes. Tumour-derived exosomes are pivotal to tumorigenesis, but the precise mechanisms underlying their secretion remain elusive. In particular, the SNARE proteins that mediate the fusion of multivesicular bodies (MVBs) with the plasma membrane (PM) in tumour cells are subject to debate. In this study, we identified syntaxin-4, SNAP-23, and VAMP-7 as the SNAREs responsible for exosome secretion in MCF-7 breast cancer cells and found that a SNARE complex consisting of these SNAREs can drive membrane fusion in vitro. Deletion of any of these SNAREs in MCF-7 cells did not affect MVB biogenesis and transportation, indicating their specific involvement in MVB-PM fusion. In addition, syntaxin-4, SNAP-23, and VAMP-7 play equivalent roles in exosome secretion in both HeLa cervical cancer cells and A375 melanoma cells, suggesting their conserved function in exosome secretion. Furthermore, deletion of VAMP-7 in 4T1 mammary carcinoma cells efficiently inhibited exosome secretion and led to significant attenuation of tumour growth and lung metastasis in mouse models, implying that VAMP-7 may hold promise as a novel therapeutic target for breast cancer.",
"38077706": "ID: 38077706\nTitle: Successful Treatment of Post COVID-19 Neurogenic Dysphagia with Botulinum Toxin.\nAbstract: Dysphagia in post COVID-19 patients could be caused by several factors, including reduced pharyngolaryngeal coordination due to SARS-CoV-2 tropism to the central and/or peripheral nervous system. To our knowledge, this is the first reported case of COVID-19-related dysphagia successfully treated with botulinum toxin type A injection. We report the case of a patient with severe oropharyngeal dysphagia due to COVID-19 confirmed by fibre endoscopy. As a result, the patient required an enteral feeding tube. After two months of traditional swallowing therapies, there was only limited improvement. An electrophysiologic evaluation of the cricopharyngeal muscle was performed and showed a normal inhibition of the cricopharyngeal muscle, followed by a hypertonic rebound. Based on this result, we decided to perform a unilateral laryngeal injection of botulinum toxin type A. After the injection, the patient's swallowing function improved significantly, allowing him to return to oral feeding. Newly diagnosed oropharyngeal dysphagia was found in 35.3% of hospitalised patients with COVID-19. There are several possible causes of COVID-19-associated dysphagia, including stroke, encephalitis, critical illness neuropathy, Guillain-Barr\u00e9 syndrome and skeletal muscle injury. In our case, since stroke was excluded by brain MRI, cranial nerve injury was a possible explanation for the difficult recovery of swallowing despite daily swallowing therapy. We suggest that electrophysiology is a valid tool for the diagnosis and follow-up of patients with oropharyngeal dysphagia. SARS-CoV-2 tropism to the central and/or peripheral nervous system can cause dysphagia in post COVID-19 patients.An electrophysiologic approach is useful for the diagnosis and follow-up of patients with oropharyngeal dysphagia.A single botulinum toxin type A injection is a valid treatment option to improve the swallowing function in patients with post COVID-19 dysphagia.",
"38110531": "ID: 38110531\nTitle: Entry and exit of extracellular vesicles to and from the blood circulation.\nAbstract: Extracellular vesicles (EVs) are biological nanoparticles that promote intercellular communication by delivering bioactive cargo over short and long distances. Short-distance communication takes place in the interstitium, whereas long-distance communication is thought to require transport through the blood circulation to reach distal sites. Extracellular vesicle therapeutics are frequently injected systemically, and diagnostic approaches often rely on the detection of organ-derived EVs in the blood. However, the mechanisms by which EVs enter and exit the circulation are poorly understood. Here, the lymphatic system and transport across the endothelial barrier through paracellular and transcellular routes are discussed as potential pathways for EV entry to and exit from the blood circulatory system.",
"38249813": "ID: 38249813\nTitle: Cross\u2011talk between lymphangiogenesis and malignant melanoma cells: New opinions on tumour drainage and immunization (Review).\nAbstract: Malignant melanoma (MM) is a highly aggressive tumour that can easily metastasize through the lymphatic system at the early stages. Lymph node (LN) involvement and lymphatic vessel (LV) density (LVD) represent a harbinger of an adverse prognosis, indicating a strong link between the state of the lymphatic system and the advancement of MM. Permeable capillary lymphatic vessels are the optimal conduits for melanoma cell (MMC) invasion, and lymphatic endothelial cells (LECs) can also release a variety of chemokines that actively attract MMCs expressing chemokine ligands through a gradient orientation. Moreover, due to the lower oxidative stress environment in the lymph compared with the blood circulation, MMCs are more likely to survive and colonize. The number of LVs surrounding MM is associated with tumour-infiltrating lymphocytes (TILs), which is crucial for the effectiveness of immunotherapy. On the other hand, MMCs can release various endothelial growth factors such as VEGF-C/D-VEGFR3 to mediate LN education and promote lymphangiogenesis. Tumour-derived extracellular vesicles are also used to promote lymphangiogenesis and create a microenvironment that is more conducive to tumour progression. MM is surrounded by a large number of lymphocytes. However, both LECs and MMCs are highly plastic, playing multiple roles in evading immune surveillance. They achieve this by expressing inhibitory ligands or reducing antigen recognition. In recent years, tertiary lymphoid structures have been shown to be associated with response to anti-immune checkpoint therapy, which is often a positive prognostic feature in MM. The present review discusses the interaction between lymphangiogenesis and MM metastasis, and it was concluded that the relationship between LVD and TILs and patient prognosis is analogous to a dynamically tilted scale.",
"38674369": "ID: 38674369\nTitle: Exploring the Role of Extracellular Vesicles in the Pathogenesis of Tuberculosis.\nAbstract: Tuberculosis (TB) remains a significant global health concern, necessitating accurate diagnosis and treatment monitoring. Extracellular vesicles (EVs), including exosomes, play crucial roles in disease progression, with their associated genes serving as potential biomarkers and therapeutic targets. Leveraging publicly available RNA-Seq datasets of TB patients and healthy controls (HCs), to identify differentially expressed genes (DEGs) and their associated protein-protein interaction networks and immune cell profiles, the common EV-related DEGs were identified and validated in the GSE42830 and GSE40553 datasets. We have identified nine common EV-related DEGs (SERPINA1, TNFAIP6, MAPK14, STAT1, ITGA2B, VAMP5, CTSL, CEACAM1, and PLAUR) upregulated in TB patients. Immune cell infiltration analysis revealed significant differences between TB patients and HCs, highlighting increased proportions of various immune cells in TB patients. These DEGs are involved in crucial cellular processes and pathways related to exocytosis and immune response regulation. Notably, VAMP5 exhibited excellent diagnostic performance (AUC-0.993, sensitivity-93.8%, specificity-100%), with potential as a novel biomarker for TB. The EV-related genes can serve as novel potential biomarkers that can distinguish between TB and HCs. VAMP5, which functions in exosome biogenesis and showed significant upregulation in TB, can be targeted for therapeutic interventions and treatment outcomes.",
"38938681": "ID: 38938681\nTitle: Analysis of extracellular vesicle microRNA profiles reveals distinct blood and lymphatic endothelial cell origins.\nAbstract: Extracellular vesicles (EVs) are crucial mediators of cell-to-cell communication in physiological and pathological conditions. Specifically, EVs released from the vasculature into blood were found to be quantitatively and qualitatively different in diseases compared to healthy states. However, our understanding of EVs derived from the lymphatic system is still scarce. In this study, we compared the mRNA and microRNA (miRNA) expression in blood vascular (BEC) and lymphatic (LEC) endothelial cells. After characterization of the EVs by fluorescence-triggered flow cytometry, nanoparticle tracking analysis and cryo-transmission electron microscopy (cryo-TEM) we utilized small RNA-sequencing to characterize miRNA signatures in the EVs and identify cell-type specific miRNAs in BEC and LEC. We found miRNAs specifically enriched in BEC and LEC on the cellular as well as the extracellular vesicle level. Our data provide a solid basis for further functional in vitro and in vivo studies addressing the role of EVs in the blood and lymphatic vasculature.",
"39128945": "ID: 39128945\nTitle: Platelet extracellular vesicles preserve lymphatic endothelial cell integrity and enhance lymphatic vessel function.\nAbstract: Lymphatic vessels are essential for preventing the accumulation of harmful components within peripheral tissues, including the artery wall. Various endogenous mechanisms maintain adequate lymphatic function throughout life, with platelets being essential for preserving lymphatic vessel integrity. However, since lymph lacks platelets, their impact on the lymphatic system has long been viewed as restricted to areas where lymphatics intersect with blood vessels. Nevertheless, platelets can also exert long range effects through the release of extracellular vesicles (EVs) upon activation. We observed that platelet EVs (PEVs) are present in lymph, a compartment to which they could transfer regulatory effects of platelets. Here, we report that PEVs in lymph exhibit a distinct signature enabling them to interact with lymphatic endothelial cells (LECs). In vitro experiments show that the internalization of PEVs by LECs maintains their functional integrity. Treatment with PEVs improves lymphatic contraction capacity in atherosclerosis-prone mice. We suggest that boosting lymphatic pumping with exogenous PEVs offers a novel therapeutic approach for chronic inflammatory diseases characterized by defective lymphatics.",
"39196607": "ID: 39196607\nTitle: Botulinum toxin intoxication requires retrograde transport and membrane translocation at the ER in RenVM neurons.\nAbstract: Botulinum neurotoxin A (BoNT/A) is a highly potent proteolytic toxin specific for neurons with numerous clinical and cosmetic uses. After uptake at the synapse, the protein is proposed to translocate from synaptic vesicles to the cytosol through a self-formed channel. Surprisingly, we found that after intoxication proteolysis of a fluorescent reporter occurs in the neuron soma first and then centrifugally in neurites. To investigate the molecular mechanisms at play, we use a genome-wide siRNA screen in genetically engineered neurons and identify over three hundred genes. An organelle-specific split-mNG complementation indicates BoNT/A traffic from the synapse to the soma-localized Golgi in a retromer-dependent fashion. The toxin then moves to the ER and appears to require the Sec61 complex for retro-translocation to the cytosol. Our study identifies genes and trafficking processes hijacked by the toxin, revealing a new pathway mediating BoNT/A cellular toxicity.",
"39426689": "ID: 39426689\nTitle: The role of ferroptosis resistance in lymph-associated tumour metastasis.\nAbstract: Tumour metastasis is a crucial factor in determining clinically challenging tumours. In this respect, the lymphatic system may act as potential entry portals for tumour metastasis, whilst, clinical detection of tumour-infiltrated lymph nodes also indicates poorer prognosis and higher metastatic risk. Whether tumour cells gain ferroptosis resistance in lymph that make them exhibit a stronger propensity for lymphatic dissemination compared to hematogenous spread might be a breakthrough for elucidating lymph-associated tumour metastasis. This review discusses how the lymphatic system endows tumour cells with ferroptosis resistance character, which makes them more propensity for lymph node pre-metastasis and distant metastasis through lymphatic circulation. Comprehensively considering the distinct structure and property of lymph and the unique metabolic characteristics of tumours, all of the lymphatic vessels, intestinal lymph and lymph nodes collectively manipulate an intricate interaction with the hematogenous system and afford substances exchange with tumour cells and extracellular vesicles, upon which make a ferroptosis resistant microenvironment for subsequent metastasis in distant organs and lymph nodes.",
"39739969": "ID: 39739969\nTitle: MicroRNAs secreted by the parasitic nematode Brugia malayi disrupt lymphatic endothelial cell integrity.\nAbstract: Lymphatic filariasis (LF) is a neglected tropical disease affecting over 51 million people in 72 endemic countries. Causative agents of LF are mosquito-borne parasitic nematodes Wuchereria bancrofti, Brugia malayi, and Brugia timori. The adult parasites impact the integrity of lymphatic vessels and damage valves, leading to a remodeling of the lymphatic system and lymphatic dilation. Chronic infections can develop into severe clinical manifestations, primarily lymphedema, hydrocoele, and elephantiasis. Mechanistic studies on the underlying pathology due to the parasite are necessary to better manage human filariasis. Since parasite molecules, such as microRNAs (miRNAs), can be found in secreted extracellular vesicles (EVs) and are transported between parasite and host cells, we hypothesized that these could also play a role in the development of pathology in LF. In this study, we tested two B. malayi miRNAs previously detected in vitro in the culture media of microfilarial stages of worms. While one is Brugia-specific (bma-miR-5864) and the other nematode-specific (bma-miR-86), both miRNAs are secreted in high abundance. We first examined the in vitro response by transcriptomic profiling of human lymphatic endothelial cells to treatment with these miRNAs, which allowed us to identify genes involved in maintaining the integrity of the lymphatic endothelium. We then measured the effect of these miRNAs on the regulation of proteins necessary for cell integrity, demonstrating downregulation leading to a significant increase in the permeability of the endothelium monolayer. With this study we identify parasite miRNAs involved in undermining the integrity of endothelial cells, thus potentially contributing to the development of pathology. These findings could pave the way for a novel treatment strategy where the inhibition of parasite-secreted molecules could slow the progression of LF pathology. From a broader perspective, the miRNAs secreted by filarial parasites could potentially be used in the future for diagnosing and monitoring disease progression or treatment efficacy.",
"39973490": "ID: 39973490\nTitle: From blood vessels to brain cells: Connecting the circulatory system and Parkinson's disease.\nAbstract: Parkinson's disease (PD) is traditionally recognized as a neurodegenerative disorder characterized by motor dysfunction and \u03b1-synuclein protein accumulation in the brain. However, recent research suggests that the circulatory system may also contribute to PD pathogenesis through the spread of \u03b1-synuclein beyond the brain. The blood-brain barrier (BBB), a key regulator of molecular exchange between the bloodstream and the brain, may become compromised in PD, allowing harmful substances, including pathogenic forms of \u03b1-synuclein, to infiltrate the brain and promote neurodegeneration. Transport mechanisms such as P-glycoprotein and the low-density lipoprotein (LDL) receptor-related protein (LRP-1) further modulate the movement of \u03b1-synuclein across the BBB, influencing disease progression. Additionally, extracellular vesicles are emerging as crucial mediators in the dissemination of \u03b1-synuclein between the brain and peripheral tissues, facilitating its spread and accumulation. The lymphatic system, responsible for clearing \u03b1-synuclein, may also contribute to PD pathology when impaired. This review highlights the growing evidence for a circulatory axis in the initiation and progression of PD. We propose that future research should explore the hypothesis that the circulatory system contributes to the pathogenesis of PD by aiding the distribution of \u03b1-synuclein throughout the body. Parkinson's disease (PD) is known for its effects on movement and the brain. But the cause remains unknown. Recent research suggests that problems with blood and blood flow contribute to the development of PD. For instance, proteins linked to PD have been identified in blood and heart tissues, indicating they could be early signs of the disease. Changes in heart rate, changes in blood pressure when standing up, and altered brain blood flow have been linked to PD. This review explores evidence that the circulatory system has important roles in the development and progression of PD. We propose further research to test if proteins in the blood start the disease and if blood vessels help spread it to the brain.",
"40202614": "ID: 40202614\nTitle: Extracellular Vesicles from Dendritic Cells Protect Against Sporothrix brasiliensis Yeast Cells.\nAbstract: Sporotrichosis is an emerging subcutaneous mycotic zoonosis that affects the skin, lymphatic system, and other organs of humans and animals. Like other infectious fungal diseases, it becomes even more severe when it affects immunosuppressed patients. This infection has a global distribution and is endemic in some regions of Brazil and it is an important zoonotic public health problem. The disease is caused by a complex of at least four pathogenic species, including Sporothrix brasiliensis. The immunological response against these species has not yet been completely elucidated. Still, structures such as extracellular vesicles could carry important components that can contribute to the modulation and control of this significant infection. Thus, this work aims to analyze the participation of EVs from na\u00efve dendritic cells and EVs from DCs previously primed with S. brasiliensis yeast and primed with EVs from the fungus in the immune response against experimental sporotrichosis in murine models. The groups that received EVs from DCs primed with S. brasiliensis or their EVs showed a significant decrease in fungal load compared to the negative control group. When we analyzed the cytokine profile in the skin of mice treated with EVs before infection, we observed an increase in IFN-\u213d, TNF-\u03b1, IL-17, and IL-10, mainly in animals previously treated with EVs from DCs cultivated with yeast cells. It is worth highlighting that all prophylactic protocols modulated and minimized fungal growth compared to the control; that is, EVs contributed to the control of the infection and acted in favor of the host, demonstrating a protective character.",
"41540479": "ID: 41540479\nTitle: Running exercise mitigates amyloidosis in 5xFAD mice by improving the structure and function of the meningeal lymphatic system.\nAbstract: BACKGROUND: Alzheimer\u2019s disease (AD) progression is closely linked to the accumulation of amyloid-[Formula: see text] (A[Formula: see text]), with impaired clearance mechanisms playing a key role. The meningeal lymphatic (mLym) system, which drains cerebrospinal fluid (CSF) and waste from the brain to peripheral lymph nodes, has emerged as a critical pathway for A[Formula: see text] removal. While physical exercise is known to improve cognitive function and reduce AD risk, its effect on the mLym system and downstream AD pathology have not been fully elucidated. METHODS: Three-month-old 5xFAD mice underwent a 3-month wheel-running exercise regimen. The function of the mLym system was assessed before and after exercise using high-frequency ultrasound imaging with nanoparticle tracers to monitor CSF drainage to deep cervical lymph nodes. The study evaluated changes in mLym vessel structure, A[Formula: see text] deposition, and cognitive performance. Additionally, the effects of serum and extracellular vesicles (EVs) from exercised rats on the expression of lymphatic vessel-related genes (LYVE-1, VEGFR3, and VEGF-C) were examined in lymphatic endothelial and microglial cell lines. RESULTS: Compared to 3-month-old 5xFAD mice and age-matched wild-type controls, 6-month-old 5xFAD mice displayed progressive decline in mLym function, reduced vessel integrity, and increased amyloid plaque burden, accompanied by impaired learning and memory. These changes were associated with decreased expression of LYVE-1 and VEGFR3 in the meninges and VEGF-C in the brain. Exercise intervention reversed these deficits, restoring mLym function and vessel structure, enhancing A[Formula: see text] clearance, and improving cognitive performance. Surgical ligation of mLym vessels accelerated amyloid accumulation and removed the exercise-induced benefits, underscoring the system\u2019s importance in A[Formula: see text] removal. In vitro, A[Formula: see text] oligomers suppressed VEGFR3 and VEGF-C expression, while serum and EVs from exercised rats counteracted this effect. Proteomic analysis of EVs from exercised animals revealed upregulation of CD9, suggesting a link to VEGFR3 signaling. CONCLUSIONS: This study demonstrates that A[Formula: see text] oligomers impair mLym function, exacerbating amyloid pathology. Exercise preserves the structure and function of the mLym system, promoting A[Formula: see text] clearance and mitigating AD progression. These findings highlight the therapeutic potential of targeting the meningeal lymphatic system to slow or prevent AD.",
"41659924": "ID: 41659924\nTitle: Mechanism of exercise-derived circulating exosomes as a target for sarcopenia management.\nAbstract: Sarcopenia, an age-related syndrome characterized by the progressive decline of skeletal muscle mass and function, threatens the health of older adults through underlying mechanisms that include dysregulated protein metabolism, autophagy-mitochondrial dysfunction, chronic inflammation, and impaired regenerative capacity of muscle stem cells. Exercise-derived circulating exosomes, which act as key mediators of intercellular communication, show considerable potential in mitigating sarcopenia-related damage. In this review, we summarize the biogenesis of exercise-induced exosomes, encompassing both ESCRT-dependent and independent pathways, secretion regulated by RAB and SNARE proteins, and their release mediated through mechanical, calcium, metabolic, and neuroendocrine signaling during exercise. We further elaborate on the systemic roles of these exosomes in muscle repair, including alleviating lipotoxicity via the FGF21-adiponectin axis, maintaining protein homeostasis through dual regulation by miR-29c, and ameliorating the inflammatory microenvironment via modulation of macrophage polarization. Finally, we discuss the translational promise of exosomes as therapeutic targets and outline future research directions, offering a conceptual framework for understanding exercise-mediated muscle protection and developing novel interventions.",
"41704641": "ID: 41704641\nTitle: Alzheimer's disease as a systems-level timing disorder: Circadian disruption of glial immunometabolism, brain clearance, and therapeutic responsiveness.\nAbstract: Alzheimer's disease (AD) is traditionally conceptualized as a disorder of protein aggregation and neurodegeneration, yet growing evidence indicates that fundamental temporal organization of brain physiology is also disrupted. In the healthy brain, circadian clocks coordinate sleep-wake behavior, glial immunometabolism, astrocytic aquaporin-4 polarity, and glymphatic-lymphatic clearance, aligning immune readiness and proteostasis with daily activity-rest cycles. In AD, this temporal coordination progressively deteriorates, manifesting as sleep fragmentation, instability of rest-activity rhythms, vulnerability of central clock structures, and loss of circadian gating of glial and clearance pathways. These disruptions create phase-inappropriate immune and metabolic states, impair protein clearance, and alter the fate of extracellular vesicles, which may shift from mediators of waste export to facilitators of proteopathic spread. Importantly, circadian failure also constrains therapeutic delivery and biomarker interpretation by modulating blood-brain barrier transport, brain fluid dynamics, and brain-to-blood signal export. We propose that AD can be reframed as a systems-level timing disorder, in which loss of temporal coherence integrates molecular pathology, glial dysfunction, clearance failure, therapeutic inefficacy, and biomarker variability. This framework highlights chrono-pharmacology, chrono-neurotherapeutics, and circadian-informed biomarkers as essential components of precision strategies for AD prevention and treatment.",
"41747594": "ID: 41747594\nTitle: Effect of intranasal treatment with NAMPT-EVs on acetylated tau and cognitive function in mice with repeated controlled cortical injury.\nAbstract: Repeated traumatic brain injury (rTBI) has attracted increasing attention owing to its long-term effects on cognition and behaviour. Moreover, research has shown that acetylated tau (ac-tau) represents a common pathology linking rTBI and Alzheimer's disease that can lead to neuronal cell death. Therefore, in this study, we evaluated the therapeutic potential of mesenchymal stromal cell-derived extracellular vesicles enriched with nicotinamide phosphoribosyltransferase (NAMPT-EVs) for improving cognitive and behavioral impairments following repeated controlled cortical injury (rCCI). Morris water maze and novel object recognition test were evaluated at 1-month post-rCCI with intranasal treatment of NAMPT-EVs. Expression of Sirtuin 1(SIRT1), ac-tau, neuron loss, neuroinflammation, AQP4 polarity, and meningeal lymphatic morphology and function were assessed 1\u00a0month after treatment. Intranasal administration of NAMPT-EVs significantly increased the expression of SIRT1 to deacetylate tau in rCCI mice. Additionally, NAMPT-EVs suppressed neuroinflammation and maintained aquaporin protein-4 polarity to facilitate the glymphatic system and promote the repair of the meningeal lymphatic system, which benefits the clearance of ac-tau from the brain parenchyma. Notably, the reduction in ac-tau prevented axon initial segment degradation and tau mislocalisation, resulting in a neuroprotective effect. NAMPT-EVs reduce neuronal loss and improve cognitive function in rCCI mice through multiple mechanisms. Therefore, NAMPT-EVs is promising for preventing cognitive deficit after rTBI.",
"41902311": "ID: 41902311\nTitle: Engineering Yeast Extracellular Vesicle Biogenesis Through Rewiring Membrane Trafficking Pathways.\nAbstract: Extracellular vesicles (EVs) are emerging as versatile therapeutic platforms, yet the mechanisms governing their biogenesis in yeast remain incompletely understood. Saccharomyces cerevisiae, a well-characterised and safe microbial chassis, naturally secretes abundant EVs and provides an attractive system for mechanistic dissection and engineering. Here, we establish S. cerevisiae as a tractable model for elucidating EV cargo loading. By combining multicopy expression of chicken interferon-\u03bb (ChiIFN-\u03bb) with cell wall perturbation, we achieved a tenfold increase in EV yield and efficient incorporation of ChiIFN-\u03bb into EVs. Quantitative proteomics identified 1555 EV-associated proteins, including 501 predicted transmembrane proteins derived from multiple organelles. ChiIFN-\u03bb overexpression and cell wall stress selectively reduced the abundance of key vesicle trafficking regulators, including SNARE, ESCRT and Rab proteins, indicating reprogramming of intracellular membrane trafficking pathways. Functional analyses further demonstrated that the SNARE proteins Sso2 and Nyv1 are enriched in the EV membrane and modulate EV size distribution and subpopulation composition. Together, these results reveal conserved protein-sorting machinery underlying yeast-derived extracellular vesicles (YDEVs) biogenesis and establish S. cerevisiae as a powerful platform for engineered EV production.",
"41971838": "ID: 41971838\nTitle: Molecular biology of targeting lymphangiogenesis in cancer: Therapeutic strategies and future promise.\nAbstract: Lymphangiogenesis is considered the rate-limiting step for tumor lymph node (LN) metastasis by sprouting the new lymphatic network from preexisting lymphatic vessels to favor the invasion and entry of cancer cells into the lymphatic system. Over the past decade, numerous studies have investigated the cellular roles and molecular mechanisms of lymphangiogenesis in cancers, and corresponding molecular control strategies targeting lymphangiogenesis to treat LN metastatic cancers have been developed. Nevertheless, due to the crucial role of lymphatic vessels in the infiltration of immune cells and immune activation in cancers, there have been several diverse findings regarding the biological roles and strategies of targeting tumor lymphangiogenesis as cancer immunotherapy has emerged and rapidly developed. Herein, we summarize the progress in characterizing the molecular mechanisms and therapeutic strategies involved in tumor lymphangiogenesis and highlight the potential of targeting lymphangiogenesis in combination with immunotherapy for the treatment of cancer, providing opportunities to improve the efficacy of targeting lymphangiogenesis to prevent cancer metastasis.",
"41979085": "ID: 41979085\nTitle: VAMP4/STX8 Mediate the Autophagic Secretion of Mitochondria and Promote TAMs Polarization in HNSCC.\nAbstract: Under hypoxic conditions, tumour cells engage in autophagic secretion of mitochondria to sustain energy balance and modulate the tumour microenvironment. However, the exact molecular mechanisms remain unclear. This study used a combination of multivesicular bodies membrane proteomics and molecular interaction validation to demonstrate that hypoxia promotes mitophagy-dependent extracellular vesicles (EVs) secretion in head and neck squamous cell carcinoma (HNSCC). This process promotes tumour-associated macrophages to adopt immunosuppressive phenotypes, reshaping the immune environment. Vesicle-associated membrane protein 4 (VAMP4) and syntaxin 8 (STX8) are key molecules involved in this secretion. Clinically, VAMP4/STX8 expressions are significantly higher in HNSCC than in normal tissues and correlate with prognosis, indicating their potential as biomarkers and therapeutic targets. This study revealed the mitophagy-secretion axis that reshaped the tumour immune landscape, providing a theoretical basis for targeting soluble NSF attachment protein receptor-dependent EV release to enhance the therapeutic effect of HNSCC immunotherapy.",
"41989572": "ID: 41989572\nTitle: Botulinum toxin from foodborne hazard to aesthetic and biomedical tool: mechanisms, applications, detection strategies, and future perspectives.\nAbstract: As powerful biological toxins, botulinum neurotoxins (BoNTs), which are mainly generated by the anaerobic bacterium Clostridium botulinum, are at the same time useful therapeutic and cosmetic agents. This review gives an extensive review of the microbiological etiology, pathophysiology, clinical use, detection strategies, and emerging challenges regarding BoNT. On a molecular scale, BoNTs are endopeptidases, which are zinc-dependent and which inhibit the fusion of synaptic vesicles by cleaving SNARE proteins, thus preventing the release of acetylcholine and causing reversible neuromuscular paralysis. This property is the basis of their medical use in treating a wide range of neuromuscular and glandular conditions, such as dystonia, spasticity, chronic migraine, and hyperhidrosis, and aesthetic surgeries using compounds like Botox and its derivatives. Although they are clinically successful, their therapeutic use is associated with possible adverse effects, including local muscle weakness as well as uncommon systemic complications, especially in the case of high dosage or incorrect administration. The review also compares existing analysis techniques to detect BoNT, such as immunological assays, molecular diagnostics, biosensor technologies, and mass spectrometry with their benefits and drawbacks in sensitivity, specificity, and toxin functional evaluation. Also , neutralization of the body formation, resistance to treatment, variability of toxin preparations, and regulatory difficulties are addressed. Future opportunities focus on the creation of non-animal testing models, advanced biosensing technology and individualized therapeutic methods to increase safety, diagnostic accuracy, and prolonged clinical performance.",
"42013932": "ID: 42013932\nTitle: TRPV1-tau axis: A bidirectional regulatory mechanism linking pain sensitization and Alzheimer's disease progression and its potential for intervention.\nAbstract: Alzheimer's disease (AD) is a major neurodegenerative disorder characterized by \u03b2-amyloid (A\u03b2) deposition and pathological tau phosphorylation and aggregation, frequently accompanied by chronic pain. Pain sensitization is closely linked to AD progression. Transient receptor potential vanilloid 1 (TRPV1), a key cation channel in pain transduction, is expressed not only in peripheral sensory neurons but also widely in central neurons and glial cells, where it contributes to pain sensitization and neuroinflammation. Emerging evidence indicates a bidirectional regulatory interplay between TRPV1 and tau, forming a \"TRPV1-tau axis.\" This axis acts as a core molecular bridge connecting pain sensitization and AD pathology via calcium dyshomeostasis, mTOR/AMPK, PI3K/Akt/GSK3\u03b2, and neuroinflammatory pathways. TRPV1 overactivation promotes tau hyperphosphorylation and aggregation through calcium-dependent kinases, metabolic dysregulation, and inflammatory signaling. Conversely, pathological tau modulates TRPV1 expression and function via transcriptional regulation, protein interactions, and impaired axonal transport, establishing a deleterious feedback cycle. Therapeutic interventions targeting this axis, including TRPV1 modulators, tau-directed agents, anti-inflammatory drugs, and natural compounds, demonstrate potential in alleviating both pain sensitivity and cognitive deficits through multi-target mechanisms. However, clinical translation remains challenging due to issues including blood-brain barrier penetration, target selectivity, and a lack of reliable biomarkers. This review systematically outlines the bidirectional mechanisms of the TRPV1-tau axis, current intervention strategies, and translational challenges, with the goal of informing future development of disease-modifying therapies that concurrently address cognitive decline and chronic pain in AD.",
"42017539": "ID: 42017539\nTitle: The p.(Leu97Ile) variant expands the genetic landscape of NEFL-associated Charcot-Marie-tooth neuropathies.\nAbstract: Charcot Marie Tooth neuropathies arise from diverse genetic disturbances that impair axonal structure or myelin integrity. Variants in NEFL, encoding the light chain of neurofilaments, represent a rare cause of CMT and may disrupt filament assembly, axonal transport, and cytoskeletal stability. Because NEFL variants are uncommon and phenotypes variable, the pathogenic relevance of individual substitutions is often difficult to determine. Here, we identified a previously unreported missense variant, p.(Leu97Ile), in three affected individuals. In two, a second variant, p.(Arg206Ser), was present in cis. We therefore aimed to investigate the pathogenic impact of this allele constellation using clinical, biochemical, and structural approaches. Neurological examination, CMTNS scoring, and electrophysiological studies were performed. Exome sequencing established the genotype. Variant effects were assessed using serum biomarkers, in silico modelling and quantitative immunofluorescence of patient-derived fibroblasts. Patients showed differing clinical presentations, including moderate adult-onset neuropathy to severe early-onset disease and neuromuscular abnormalities in childhood. Neurophysiology revealed mixed features, and conduction slowing in the child supported classification within the CMT1F spectrum. All carriers exhibited elevated serum NfL; CK was increased in the adult patients. Descriptive biomarker differences between the patients were observed for GDF15, FGF21, Periaxin, and NCAM1. Fibroblasts displayed increased cytoplasmic NfL without altered nuclear metrics. Structural modelling predicted destabilizing effects of both substitutions, with the cis combination producing the strongest deviation. Integrated clinical, biochemical, and structural evidence support pathogenicity of p.(Leu97Ile) and suggest that p.(Arg206Ser) in cis augments its biological impact. These findings expand the genetic and phenotypic spectrum of NEFL-associated CMT1F.",
"42027429": "ID: 42027429\nTitle: Characterization of a novel inherited splice-site variant in MAPK8IP3 expands the genetic and phenotypic spectrum of neurodevelopmental disorders.\nAbstract: The MAPK8IP3 gene encodes JIP3, a kinesin-1 adapter protein crucial for axonal transport and JNK signaling pathway regulation. This gene exhibits extreme intolerance to loss-of-function variants. Heterozygous pathogenic MAPK8IP3 variants are linked to a neurodevelopmental disorder with or without variable brain abnormalities (NEDBA). This study characterizes a novel inherited canonical splice-site variant identified in a three-generation family. Exome sequencing was performed on three affected individuals from a single non-consanguineous family with presenting neurological abnormalities. The molecular consequences of the identified variant were evaluated by RNA analysis. Clinical features of the variant carriers were evaluated in detail. We identified a heterozygous dinucleotide deletion (c.2630-2_2630-1del) at the canonical acceptor splice site of MAPK8IP3. This variant co-segregated with the neurological phenotype in an affected mother and her two children. RNA analysis demonstrated that this variant leads to a partial deletion of exon 22 (r.2630_2673del), which is predicted to cause a frameshift and introduce a premature termination codon (p.Gly877Valfs\u221736). Notably, despite carrying an identical genotype, the three affected individuals exhibited marked interindividual variability in clinical severity, ranging from mild specific learning difficulties to global developmental delay with severe intellectual disability and autism. This represents the first report of a familial, autosomal dominantly inherited pathogenic splice-site variant in MAPK8IP3, extending the established inheritance pattern beyond previously described de novo events. Our findings expand both the mutational and phenotypic spectrum of MAPK8IP3-related disorders, highlighting the complexity of genotype-phenotype correlations and underscoring the necessity of functional assays for accurate variant interpretation.",
"42035797": "ID: 42035797\nTitle: Mitofusin 2 in central nervous system disorders: Roles in mitochondrial dynamics and therapeutic Implications.\nAbstract: Mitochondrial dynamics have been increasingly recognized as a central determinant in the pathogenesis of central nervous system (CNS) disorders. Mitofusin 2 (MFN2), a critical mitochondrial fusion protein, preserves mitochondrial network integrity and participates in fission, mitophagy, and axonal transport, thereby maintaining neuronal function and energy homeostasis. Structural features of MFN2 underpin its diverse regulatory roles, whereas MFN2 deficiency leads to mitochondrial fragmentation, metabolic dysfunction, oxidative stress, and neuronal impairment. This review summarizes the molecular mechanisms of MFN2 in the CNS and its impact on neuronal survival, synaptic function, and signaling pathways. In addition, we highlight potential MFN2-targeted interventions, including natural compounds, pharmacological agents, and emerging small-molecule activators, while also discussing disease-specific mechanisms and translational challenges such as endoplasmic reticulum (ER)-mitochondria communication abnormalities and blood-brain barrier permeability.",
"42041253": "ID: 42041253\nTitle: Typhoid toxin: reframing enteric fever.\nAbstract: SUMMARYSalmonella enterica serovars Typhi and Paratyphi A cause enteric (typhoid/paratyphoid) fever, a systemic disease that remains a major source of morbidity and mortality worldwide. The most distinctive clinical manifestations, such as sustained fever with relative bradycardia, leukopenia, neuropsychiatric symptoms/encephalopathy, and ileal perforation, have long lacked a unifying mechanistic explanation. Over the past two decades, studies of typhoid toxin have reframed how we understand this disease. Typhoid toxin is a human-adapted, chimeric A2B5 exotoxin assembled exclusively inside infected cells. It couples an ADP-ribosyltransferase (PltA) and a DNase I-like nuclease (CdtB) on a pentameric delivery ring (PltB or the alternate, PltC), that recognizes N-acetylneuraminic acid (Neu5Ac)-terminated human sialoglycans. The toxin is produced by bacteria residing within the Salmonella-containing vacuole (SCV), secreted across the bacterial envelope into the SCV lumen by a phage-derived pathway now classified as type 10 secretion system, sorted for export by the host CI-M6PR/COPII trafficking machinery, and then re-enters distant target cells via retrograde transport to the endoplasmic reticulum (ER). Recent work has linked typhoid toxin to leukopenia, blood-brain barrier disruption causing encephalopathy, gut-vascular and immune dysfunction leading to intestinal perforation, and hepatobiliary injury. The discovery of an alternate B-subunit (PltC) established a paradigm of modular B-subunit exchange that diversifies receptor usage, tissue tropism, and biology. Collectively, these insights redefine typhoid fever as a toxin-driven immunovascular disease and identify actionable targets for neutralizing antibodies and barrier-protective therapies.",
"42047916": "ID: 42047916\nTitle: From Bench to Bedside: Neurofilament Light Chain as a Biomarker in Ischemic Stroke.\nAbstract: Neurofilaments (NFs) are protein structures that form part of the neuronal cytoskeleton, providing structural support and facilitating axonal transport. Neurofilament light chain (NFL) has emerged as a promising biomarker for various neurological conditions, as it is released into the bloodstream following neuronal damage. The ability of NFL to differentiate between types of injury and enhance the accuracy of clinical prediction models makes it an asset in both clinical practice and research. Studies suggest that NFL can be employed to diagnose and monitor disease progression and aid in the prognosis of various neurological conditions, including ischemic stroke (IS). In IS, NFL serves as a predictor of severity, functional recovery, and the risk of complications. This review therefore synthesizes current preclinical and clinical studies on the use of NFL as a biomarker for prognosis, monitoring, and treatment in IS.",
"42055257": "ID: 42055257\nTitle: Selective vulnerability of monoaminergic neurons and network spread of alpha-synuclein jointly explain pathology progression in Parkinson's disease models.\nAbstract: The trans-synaptic propagation of \u03b1-synuclein aggregates is a defining feature of Parkinson's disease, yet how specific cellular phenotypes interact with the brain's structural connectome to govern disease progression remains a fundamental question in neurobiology. Here, we address this question using a connectome-based modeling framework that simulates synucinopathy spread along the mouse structural connectome, to examine how monoaminergic cell-type vulnerability and network-based transmission jointly shape pathology progression. 'Nexopathy in silico' (NexIS), along with the MISS algorithm for spatial gene expression mapping, provided a framework to examine how regional and cellular context impact synucleinopathy spread. In particular we focus on monoaminergic neurons, since inadequate clearance of pathologic \u03b1-synuclein species in monoaminergic neurons is a prominent hypothesis implicated in PD. We delineate the specific roles of monoaminergic cell types and directional transmission of \u03b1-synuclein arising from axonal transport polarity in shaping \u03b1-synuclein pathology in mouse models. Our analysis reveals that hindbrain noradrenergic (HBNOR) and midbrain dopaminergic (MBDOP) cell-type distributions are the primary mediators of network-wide transmission, with the former outperforming endogenous regional Snca expression as predictors of long-term pathology. Crucially, we demonstrate a synergistic effect between cell-type vulnerability and retrograde-biased transport, finding that both factors are required to accurately recapitulate empirical spatiotemporal patterns at 6 and 12 months post-seeding. This work constitutes one of the most mechanistically complete models of synucleinopathy to date, providing a comprehensive theoretical bridge that links microscale molecular motor regulation to macroscale regional vulnerability. Our findings suggest that the interaction between noradrenergic cell-type distribution and retrograde transport serves as the dominant driver of late-stage progression, offering highly specific cellular and mechanistic targets for therapeutic intervention.",
"42056501": "ID: 42056501\nTitle: AAV2 capsid clearance and neuronal trafficking dynamics in the central nervous system.\nAbstract: Adeno-associated virus serotype 2 (AAV2) remains one of the most common vectors for CNS gene delivery. Yet, its long-term intraparenchymal trafficking and the relationship between capsid persistence and transgene expression remain poorly understood. In this study, we tracked the spatial and temporal patterns of AAV2 following direct striatal infusion in rats, analyzing tissue at 3 days, 3 weeks, 12 weeks, 30 weeks, and 67 weeks after injection. Using immunofluorescence for the human aromatic L-amino acid decarboxylase (AADC) transgene and an epitope detecting intact AAV2 capsids (A20), we mapped capsid localization, clearance, and axonal transport over more than a year. Following direct striatal infusion, AAV2 capsids rapidly localized to striatal neurons, with early accumulation in the substantia nigra pars reticulata (SNpr) detectable by 3 days post-infusion, but without evidence of nigral neuron transduction. Transgene expression increased over time and peaked locally at 12 weeks, with a delayed yet robust AADC signal in striatonigral terminals by 30 weeks. By 67 weeks, capsid signal was minimal while AADC expression remained stable. These findings clarify the long-term dynamics of AAV2 distribution, capsid persistence, and axonal transport in the adult brain, informing our understanding of vector behavior and durability following intraparenchymal AAV2 gene delivery.",
"42056834": "ID: 42056834\nTitle: Generation of iPSC line NIMHi033-A from an Indian patient with Autism Spectrum Disorder carrying mutation in DYNC1H1 gene.\nAbstract: We report the generation of a human induced pluripotent stem cell (hiPSC) line from peripheral blood mononuclear cells of a 14-year-old male with Autism Spectrum Disorder carrying a de novo heterozygous likely-pathogenic variation c.4067C>T (p.Pro1356Leu) in Dynein Cytoplasmic 1 Heavy Chain 1 (DYNC1H1) gene. Variation in this gene that encodes a motor protein may affect retrograde transport of neurotrophic signals, synaptic vesicles, organelles like golgi complex and mitochondria. Therefore, this iPSC line will be an invaluable tool to study the effects of this high-risk autism-related variant on disease manifestation and cellular functions.",
"42102607": "ID: 42102607\nTitle: BoNT-A amelliorates itch sensitization in atopic dermatitis mice by inhibiting histamine release.\nAbstract: In this study, we thoroughly investigated the potential mechanisms underlying the therapeutic effects of Botulinum Toxin Type A (BoNT-A) on ameliorating pruritic behavior and allodynia in a murine model of atopic dermatitis (AD). Recognizing the pivotal role of mast cells in AD pathogenesis, we hypothesized that BoNT-A inhibits mast cell degranulation via the cleavage of SNAP-25. Using P815 cells and primary mast cells, we confirmed the expression of SNAP-25 and SV2, and through siRNA-mediated knockdown, we validated that SNAP-25 is a pivotal mediator of mast cell degranulation and the specific target required for BoNT-A' s inhibitory action. In our in vivo experiments utilizing MC903-induced AD mice, we observed that BoNT-A administration significantly reduced scratching bouts and alleviated pruritus allodynia while decreasing mast cell recruitment in both epidermal and dermal layers. Furthermore, to elucidate the involvement of the central nervous system, we constructed a BoNT-A/FITC complex and demonstrated that the toxin undergoes retrograde transport from peripheral nerves to the spinal cord dorsal horn. Therefore, it appears to exert an antipruritic effect by downregulating the expression of pruritus-related ion channels and neuropeptides, specifically TRPV1 and CGRP. Consequently, our findings establish that BoNT-A exerts its therapeutic effects through a dual mechanism: peripherally by inhibiting mast cell degranulation via the SNAP-25/SV2 pathway, and centrally by modulating neurogenic inflammation within the spinal cord, thus providing a novel perspective for the treatment of allergic diseases.",
"42123583": "ID: 42123583\nTitle: Regulation of Tau Alternative Splicing: A Novel Role for the Ribonucleoprotein RBM20.\nAbstract: Tau is a protein associated with microtubules principally expressed in neuronal cells, where it plays a fundamental role in cytoskeleton stabilization and axonal transport. Several diseases collectively named tauopathies, such as Alzheimer's disease, have been associated with an imbalance in the expression of alternative spliced Tau transcripts and the accumulation of hyperphosphorylated Tau, causing dysfunction and death of neuronal cells. Therefore, understanding the Tau exon splicing mechanisms may contribute to elucidating molecular factors that could underlie the development of neurodegenerative disorders. The aim of this study was to define the role of selected splicing factors in regulating Tau exon expression in cell lines and neuronal organoids. We demonstrated the role of the RNA-binding motif protein 20 (RBM20) splicing factor in regulating Tau exon 6 and exon 10, applying RNA-binding assay and qPCR analyses. Furthermore, we demonstrated that Tau expression was regulated during cerebral organoid differentiation, recapitulating in vivo Tau expression. These results suggest the feasibility of using brain organoid technology to study Tau alternative splicing during neural development, confirming that 3D cellular models could be used to study and characterize pathological processes taking place in Tau-related pathologies.",
"42130092": "ID: 42130092\nTitle: FTLD-TDP-43 With Motor Neuron Disease Pathology in an Autopsied Patient With Spastic Paraplegia-30B Harbouring a Homozygous KIF1A Variant.\nAbstract: KIF1A-associated neurological disorder (KAND) is a rare hereditary condition caused by KIF1A variants, affecting axonal transport and presenting with a wide clinical spectrum, including hereditary spastic paraplegia. This case of childhood-onset KAND reveals FTLD-TDP43 with motor neuron disease pathology emerging late in the disease course, suggesting that HSP and FTLD-MND share a pathological continuum through a TDP-43-related pathway and expanding the clinicopathological spectrum of KAND.",
"42144072": "ID: 42144072\nTitle: Progress in precision drug delivery systems for neurodegenerative diseases with muscle atrophy.\nAbstract: Neurodegenerative diseases frequently co-occur with skeletal muscle atrophy, creating a complex comorbid condition that significantly accelerates functional decline and increases mortality. This dual pathology is driven by interconnected mechanisms such as protein aggregation, neuroinflammation, and impaired axonal transport, disrupting critical neuromuscular junctions (NMJs). However, a significant research gap exists in the development of therapeutic strategies that can effectively and simultaneously target both the central nervous system (CNS) and peripheral muscle tissues. We systematically summarizes the core pathological targets and critically evaluates recent advances in precision drug delivery systems designed to overcome these challenges. We explore innovative strategies, including engineered viral vectors and receptor-targeted nanoparticles for CNS delivery, as well as smart biomaterials and extracellular vesicles (EVs) for muscle-specific and dual-organ intervention, highlighting the growing role of artificial intelligence (AI) in optimizing their design. Furthermore, this review discusses the construction of multidimensional efficacy evaluation systems that integrate behavioral, molecular, and imaging biomarkers and addresses pivotal clinical translation challenges, from scalable production to species-specific differences. By examining the pathological landscape across the central nervous system and skeletal muscle together, this review connects mechanisms that have more often been considered in isolation and evaluates how their convergence may inform the design of delivery strategies targeting both tissue compartments.",
"42192837": "ID: 42192837\nTitle: Axonal Transport Deficits in Parkinson's Disease: Insights from Neurotoxin, Genetic, and Sporadic Models.\nAbstract: Parkinson's disease (PD) is a prevalent neurodegenerative disorder, characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta and the accumulation of Lewy bodies. Over recent decades, various cellular mechanisms underlying PD have been elucidated, including autophagy, mitochondrial dysfunction, neuroinflammation, and axonal transport. Among them, axonal transport plays a critical role in maintaining the dynamic homeostasis of proteins, membrane-bound organelles, and cellular metabolism within neurons. Unfortunately, a comprehensive overview of axonal transport in PD remains absent. In this review, we synthesized the current literature on axonal transport in PD, leveraging neurotoxic and genetic models to explore the causes and consequences of axonal transport alterations in PD. Through this summary, we aim to deepen our understanding of PD pathogenesis and provide potential therapeutic targets for intervention.",
"42198698": "ID: 42198698\nTitle: From Envelope to Encephalopathy: How HIV-1 gp120 Drives Neurocognitive Decline.\nAbstract: Although neurons are not productively infected by HIV-1, the envelope glycoprotein gp120, detectable in cerebrospinal fluid independently of active viral replication, gains intraneuronal access via lipid raft-mediated endocytosis, macropinocytosis, and retrograde axonal transport, contributing to persistent neurobiological dysfunction within the central nervous system. Once internalized, gp120 is associated with neuronal dysfunction involving convergent pathways, including excitotoxic calcium dysregulation, mitochondrial and metabolic failure, and inflammatory and senescence-associated amplification. These pathways converge on suppression of CREB and BDNF signaling, dismantling the transcriptional and neurotrophic programs required for synaptic maintenance and cognitive resilience. Extracellular vesicle-mediated dissemination and microRNA reprogramming extend gp120-associated neurobiological effects beyond sites of receptor engagement, while gut-derived metabolites, particularly quinolinic acid, lower the excitotoxic threshold through synergistic activation of NMDA receptors. Together, these mechanisms define HAND as a network disorder in which gp120 contributes to persistent neurocognitive dysfunction beyond active viral replication, identifying convergent therapeutic nodes where combination strategies targeting excitotoxicity, mitochondrial dysfunction, and neuroinflammation offer the most promising path toward durable neuroprotection.",
"42209021": "ID: 42209021\nTitle: Striatal Neuron Excitability Is Regulated by Huntingtin in the Adult Brain.\nAbstract: Huntington's disease (HD) is a hereditary neurodegenerative disease that typically presents during midlife and is characterized by a combination of motor, cognitive, and psychiatric symptoms. HD is fatal and arises from a mutation in the huntingtin (HTT) gene, which results in decreased neuronal health followed by brain atrophy, with spiny projection neurons (SPNs) of the striatum being especially vulnerable to degeneration. HTT loss of function, caused by haploinsufficiency of the wild-type HTT gene (wtHTT), is an important feature of HD pathophysiology that has previously been understudied compared with mutant HTT gain-of-function mechanisms. wtHTT is essential for nervous system development and functions as a scaffolding protein to support many vital cellular functions including axonal transport, autophagy, and synaptic plasticity. Here, we examined the consequences of wtHTT deletion in the adult cortex and striatum by conditionally inactivating wtHTT in 2-4-month-old male and female Htt fl/fl mice. wtHTT loss of function decreased intrinsic neuronal excitability within SPNs and produced a neuroinflammatory response in these mice, while tissue organization, spine morphology, and motor behavior remained unaffected. Results presented here provide additional evidence that wtHTT is vital for maintaining neuronal health in the adult brain and highlight some potential adverse consequences of nonselective HTT lowering for the treatment of HD.",
"42264545": "ID: 42264545\nTitle: Nanotechnology-enabled targeting strategies for neurodegenerative disorders: role of functionalized nanoparticles.\nAbstract: Neurodegenerative disorders comprise a diverse group of progressive neurological diseases characterized by the gradual loss of neuronal structure and function. Conditions such as Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis arise from multifactorial mechanisms involving genetic susceptibility, environmental factors, and age-related cellular decline. Key pathogenic processes include oxidative stress, mitochondrial dysfunction, protein misfolding and aggregation, impaired axonal transport, Golgi fragmentation, and chronic neuroinflammation, all of which disrupt neuronal homeostasis and synaptic communication, ultimately leading to neuronal death. Hormonal imbalances further exacerbate these effects by promoting oxidative damage, inflammation, and metabolic dysfunction. Despite advances in understanding disease mechanisms, effective drug delivery remains challenging due to the restrictive nature of the blood-brain barrier. Recent developments highlight the potential of nanoparticle-based drug delivery systems to overcome these limitations. Functionalized nanoparticles enhance blood-brain barrier penetration, improve targeting specificity, and enable controlled drug release. These systems can deliver neuroprotective agents, antioxidants, peptides, and gene therapies directly to affected brain regions. Thus, integrating disease pathophysiology with nanotechnology-based strategies offers a promising approach for improving therapeutic outcomes and advancing precision treatment in neurodegenerative disorders.",
"42275483": "ID: 42275483\nTitle: Intranasal Delivery of Bacterial Extracellular Vesicles Enables RNA Cargo Entry Into the Brain.\nAbstract: Extracellular vesicles (EVs) released by bacteria are potent mediators of host-microbe interactions. They modulate immune responses, deliver functional molecules and influence disease progression. However, whether bacterial EVs can access the brain and functionally affect host cells remains unclear. In this study, we engineered Escherichia coli-derived EVs by electroporating Cre recombinase mRNA (Ec EVCre) and assessed their transport and functional delivery following intranasal administration. Using mT/mG reporter mice, we observed EV uptake in the olfactory epithelium and recombination-driven GFP expression in a subset of neurons in the olfactory bulb, providing proof-of-concept for the functional delivery of bacterial EV-associated mRNA into the brain. Single-cell RNA sequencing and imaging analyses of the olfactory regions revealed neuronal and immune cell subsets as key EV targets. Microfluidic biochip chamber assays with cultured sensory neurons demonstrated that EVs undergo retrograde axonal transport from neurite terminals to the soma via signalling endosomes. Pharmacological inhibition significantly impaired EV uptake, supporting the involvement of endocytic pathways. In addition to neuronal entry, we discovered that phagocytic cells, including neutrophils and macrophages, can engulf EVCre in the nasal mucosa and migrate into the brain, providing an alternative immune-mediated route for vesicle delivery. Together, these findings indicate that bacterial EVs exploit both neuronal and phagocytic pathways to deliver functional RNA cargo into the brain, providing novel insights into microbial access to the central nervous system and its implications for neuroimmune interactions.",
"42312720": "ID: 42312720\nTitle: Prenatal cannabis use is associated with altered miRNA and protein expression in the developing human brain.\nAbstract: Cannabis is widely used during pregnancy, increasing with cannabis legalization, and has been associated with altered neurodevelopment, yet the precise molecular mechanisms remain poorly understood. To assess whether utilizing fetal central nervous system-derived extracellular vesicles (fCNSEVs) isolated from maternal blood reflects cannabis-associated changes in the fetal brain. In a matched case-control study, maternal plasma (to isolate fCNSEVs) and paired fetal cortical tissue (9-18\u2009weeks' gestation) from pregnancies exposed or unexposed to cannabis were collected. We quantified protein (CB1R, D2R) and miRNA expression (using \u00b5-parafloTM microarray). Selected miRNAs were validated by qPCR. Bayesian Generalized Linear Models were used to assess exposure-group by sex effects. Maternal cannabis use was associated with altered CB1R and D2R levels in the brain with opposite directional changes in fCNSEVs. 21 miRNAs were differentially expressed: 14 downregulated and 6 upregulated, following a sex-dependent pattern (males>>females), while miR-216a-5p follows the opposite pattern (females>>males). We confirmed concordant miRNA changes in fetal brain and fCNSEVs. Many of the identified molecules are associated with pathways linked to adverse neurodevelopmental outcomes. Our data suggest that fCNSEV-based assays may provide the ability to monitor fetal brain effects of maternal cannabis in ongoing pregnancies in clinical cohorts. A critical next step is to determine if alterations in fCNSEV protein and miRNA markers predict changes in fetal brain connectivity and neurodevelopment measures of executive function.",
"42314895": "ID: 42314895\nTitle: Profiling tRNA-derived fragments in LPS-induced microglia and their influence on immune response.\nAbstract: The resident macrophages of the brain, microglia, react to immunological responses and preserve homeostasis in the central nervous system. Microglia play a critical role in neuroinflammation, which occurs in neurological and neurodegenerative diseases. Microglial responses are associated with neurodegenerative diseases, autoimmune diseases, and cancer, and therefore require strict regulation. Transfer RNA-derived fragments (tRFs) are small non-coding regulatory RNA molecules generated from the cleavage of transfer RNAs. tRFs have recently emerged as one of the key regulators of gene expression and cellular function in various biological processes. Understanding the content of tRFs altered in microglia due to LPS exposure may provide insights into the mechanisms involved in disease pathogenesis, where neuroinflammation plays a role. We used next-generation sequencing to determine tRF profiles of three different fractions (lysate, extracellular vesicles, and extracellular vesicles-free supernatant) of LPS-induced microglial cells. A total of 345 tRFs were differentially expressed across these fractions after LPS stimulation, including 13 DE-tRFs shared by all three compartments. Gene Ontology analysis of these 13 tRFs suggests their potential involvement in the inflammatory responses of microglia. Among the 13 consistently dysregulated DE-tRFs, tDR-1:33-Glu-CTC-1-M2 was selected for preliminary candidate-level assessment based on its consistent differential expression across all three fractions in our NGS data and previous reports linking this fragment to neuroinflammatory contexts.Increased expression pattern of tDR-1:33-Glu-CTC-1-M2 was further confirmed in lysate samples of LPS-induced microglial cells via RT-qPCR. Inhibition of tDR-1:33-Glu-CTC-1-M2 attenuated LPS-induced cytokine expression and release in microglia, supporting an association between this candidate tRF and inflammatory readouts in this model.",
"42315580": "ID: 42315580\nTitle: Alkaline loading of extracellular vesicles produced from human neural stem cell-derived neurospheres enables CNS drug delivery.\nAbstract: The blood brain barrier and blood tumor barrier (BBB and BTB, respectively) represent significant obstacles for the delivery of drugs to treat diseases of the central nervous system, such as brain cancers and neurodegenerative diseases. Extracellular vesicles (EVs) or exosomes have emerged as a new drug delivery vehicle for CNS diseases as they may penetrate the BBB/BTB and are less immunogenic than liposomal carriers. EVs derived from human neural stem cells (hNSC) provide additional benefits over other EV sources due to their increased homing capability to neural cells and demonstrated efficacy for treating stroke and traumatic brain injury in rodent models. However, the utilization of EVs from hNSC for drug delivery remains largely unexplored, due in part to difficulties in manufacturing capacity compared to traditional cell lines. Here, we report the development of a hNSC suspension neurosphere system for EV production and drug delivery. As proof of concept, doxorubicin was loaded into hNSC-EV, using a novel, high-efficiency alkaline passive loading method, and shown to be effective at inducing cytotoxicity in glioma cells in vitro and exhibiting higher BBB penetrance than doxorubicin-alone in vivo. These studies demonstrate the potential for hNSC-EV loaded doxorubicin as a therapeutic treatment for brain cancers such as glioblastoma, while also establishing hNSC-EVs as a drug-delivery vehicle for CNS diseases.",
"42322256": "ID: 42322256\nTitle: The predictive nature of diagnostic nerve blocks for lower extremity spasticity management with botulinum toxin and neurotomy: A systematic review.\nAbstract: Diagnostic nerve blocks (DNBs) involve the injection of anesthetic around peripheral nerves to temporarily reduce motor tone. Although they are used clinically to inform spasticity treatment with neurotomy and chemodenervation, there is little research exploring the predictive nature of DNBs. A systematic review was conducted to identify and analyze studies that compared clinical outcomes between DNBs and neurotomies or chemodenervation with botulinum toxin. In total, nine eligible articles were analyzed, with five studies involving neurotomies and four studies involving chemodenervation. These studies focused exclusively on lower limb spasticity. There were different DNB protocols used across the eligible articles. DNBs resulted in similar clinical outcomes after neurotomy on gait and spasticity measurements. DNBs resulted in reduced agonist muscle strength as compared to neurotomy outcomes, particularly those collected a year or longer after surgery. DNBs were less predictive of clinical response to chemodenervation and to range of motion assessments. This review has identified a gap in research regarding the predictability of DNBs, with heterogeneity in study design. As DNBs resulted in more similar spasticity and gait outcomes when compared to neurotomy rather than chemodenervation procedures, this may suggest that DNBs have a larger role to play in surgical decision-making. Given the current research, future studies on DNBs should explore upper extremity spasticity management as well as outcomes that relate to patient reported goals and experiences.",
"42342068": "ID: 42342068\nTitle: Prenatal glucocorticoids and long-term brain vulnerability: GR signaling, epigenetic programming, and crosstalk with peripheral tissues.\nAbstract: Glucocorticoids (GCs) are key regulators of stress responses and fetal maturation, and their physiological rise during pregnancy supports coordinated organ development. Clinically relevant GC exposure during sensitive windows of brain development occurs in several contexts, including antenatal treatment for risk of preterm birth to promote lung maturation, prolonged maternal therapy for chronic inflammatory or autoimmune conditions, and postnatal GC treatment in preterm infants, including regimens used to prevent or treat bronchopulmonary dysplasia. Although these contexts differ in timing, dose, and duration, they share the capacity to engage a glucocorticoid receptor (GR) signaling during critical windows of neurodevelopment, with possible long-term consequences for brain development and stress responsiveness. This review synthesizes clinical, experimental, and stem cell-based evidence to examine how GC signaling can shape brain structure and function across the lifespan. We discuss GR signaling in the central nervous system (CNS) and summarize evidence that sustained activation can be associated with paradoxical pro-inflammatory and neurotoxic phenotypes. We highlight epigenetic mechanisms through which GC signals may produce persistent changes in gene regulation, and we integrate data from prenatal exposure together with evidence on maternal metabolic and inflammatory context as modifiers of developmental risk. Finally, we propose an integrated view in which CNS outcomes attributed to GCs reflect a composite of direct neural actions and indirect effects shaped by peripheral tissues. We discuss adipose- and muscle-linked pathways as candidate mediators of systemic-to-central communication. This perspective links stress endocrinology, metabolism, and brain vulnerability, and highlights key mechanistic gaps and translational priorities for future research.",
"42342866": "ID: 42342866\nTitle: Bioenergetic failure in diabetic peripheral neuropathy: from glucotoxicity to multidimensional metabolic imbalance.\nAbstract: Diabetic peripheral neuropathy (DPN), particularly distal symmetric polyneuropathy, characterized by length\u2011dependent axonal damage, is a common chronic complication of type 2 diabetes mellitus. The pathogenesis of DPN is complicated, yet one thing is clear: long axons require a great deal of energy. When axonal transport is compromised by impaired energy metabolism, neuronal somata fall into an energy deficit that triggers neurodegeneration. While earlier work centered on hyperglycemia-induced cytotoxicity, recent studies have increasingly implicated dysregulation of glucose, lipid, and amino acid metabolism as key contributors to DPN. In this review, we integrate the anatomical organization of peripheral nerves, bioenergetic pathways, and axon-Schwann cell interactions to establish a framework for understanding how glucose, lipid, and amino acid dysregulation converge to induce bioenergetic failure in DPN. Based on these mechanisms, we further discuss novel strategies aimed at restoring metabolic homeostasis in neurons and Schwann cells. Importantly, correcting a single metabolic pathway is unlikely to halt or reverse DPN. Instead, restoring global energy homeostasis to rebalance axonal energy supply and demand may be essential for preserving peripheral nerve function.",
"42350670": "ID: 42350670\nTitle: Metabolic state determines the brain and direct islet effects of liraglutide on enhanced insulin secretion.\nAbstract: Liraglutide, a glucagon-like peptide-1 receptor (GLP-1R) agonist for type 2 diabetes and obesity management, shows variable patient responses. We investigated the metabolic state-dependent mechanisms underlying this heterogeneity and how liraglutide's mode of action shifts across stages of metabolic dysfunction. We employed human pancreatic islets from donors across metabolic states (normoglycaemic [HbA1c <42 mmol/l (<6.0%)], glucose intolerance [HbA1c 42-47 mmol/l (6.0-6.4%)] and type 2 diabetes [HbA1c \u226548 mmol/l (\u22656.5%)]) using dynamic perifusion and static incubation techniques to assess glucose-stimulated insulin secretion. GLP-1R mRNA levels were measured in 112 donor islets stratified by HbA1c. Mechanistic investigations used tanycyte-specific GLP-1R knockdown (GLP-1RTanycyteKD) mice and botulinum toxin B-expressing (iBot) mice to distinguish between central and peripheral pathways. Oral glucose tolerance tests, pyruvate tolerance tests and positron emission tomography were performed to assess in vivo metabolic effects. Liraglutide (25 nmol/l) enhanced glucose-stimulated insulin secretion specifically in donors with glucose intolerance (n=7, p=0.021), with no effect in normoglycaemic islets (n=7), despite preserved GLP-1 (7-36) responsiveness. In type 2 diabetes islets, GLP-1R mRNA levels progressively decreased with rising HbA1c (p=0.015, normoglycaemic [n=48] vs type 2 diabetes [n=10]). In chow-fed mice, liraglutide's insulin-stimulating effects required tanycyte-mediated hypothalamic access, as demonstrated by abolished responses in GLP-1RTanycyteKD mice. However, during metabolic dysfunction (a 12-week high-fat diet), direct islet responsiveness was restored independent of tanycyte function. Advanced metabolic disease (a 27-week high-fat diet) maintained islet responsiveness ex vivo while losing in vivo insulin enhancement, revealing insulin-independent glucose-lowering mechanisms involving hepatic gluconeogenesis suppression and enhanced peripheral glucose uptake. Liraglutide operates through complementary, metabolic state-dependent pathways: tanycyte-mediated brain actions predominate in healthy conditions, direct islet effects emerge during glucose intolerance and insulin-independent mechanisms maintain efficacy across metabolic states. This mechanistic framework enables potential patient stratification in type 2 diabetes therapy, suggesting that matching liraglutide's predominant mechanism to individual metabolic profiles could optimise treatment outcomes.",
"42351176": "ID: 42351176\nTitle: Extracellular vesicle-mediated bidirectional communication between the brain and peripheral organs in Alzheimer's disease: evidence, mechanisms, and translational perspectives.\nAbstract: Alzheimer's disease (AD) has traditionally been conceptualized as a brain-centered neurodegenerative disorder characterized by amyloid-\u03b2 (A\u03b2) deposition, tau pathology, synaptic dysfunction, and progressive neuronal loss. However, accumulating evidence suggests that AD is also shaped by systemic disturbances and reciprocal communication between the central nervous system and peripheral organs. Extracellular vesicles (EVs), which transport proteins, lipids, metabolites, and nucleic acids across biological fluids and barriers, have emerged as plausible mediators of this inter-organ crosstalk. In this Review, current evidence for EV-mediated bidirectional communication between the brain and peripheral organs is synthesized, with particular attention to the liver-brain, heart-brain, gut-brain, lung-brain, bone-brain, and adipose-brain axes. The strength of evidence across these axes is compared, and the ways in which organ-derived EVs may influence neuroinflammation, neurovascular dysfunction, metabolic homeostasis, blood-brain barrier integrity, and A\u03b2/tau-related processes are discussed, while also considering how brain-derived EVs (BDEVs) may affect peripheral physiology. The translational potential of EVs as diagnostic biomarkers, therapeutic carriers, and candidate targets for systemic intervention in AD is further evaluated. Current evidence most strongly supports the gut-brain, liver-brain, and adipose-brain axes, whereas several other axes remain supported primarily by experimental models or engineered EV studies. Major barriers to progress include EV heterogeneity, limited source specificity, insufficient standardization of isolation and quantification workflows, and a continuing reliance on associative human data and preclinical models. Overall, EVs are best viewed as one candidate signaling layer within a broader systemic network linking peripheral physiology to brain pathology. Clarifying the magnitude, directionality, and causal significance of these interactions will require rigorous EV characterization, source-resolved in vivo trafficking studies, and longitudinal clinical investigation.",
"42353041": "ID: 42353041\nTitle: The Dual Roles of Extracellular Vesicle Subtypes in Regulating Traumatic Brain Injury.\nAbstract: Traumatic brain injury (TBI) is a global public health problem which causes long-term neurologic damage caused by both primary mechanical injury and secondary pathological processes. Extracellular vesicles (EVs) such as exosomes, microvesicles (MVs) and apoptotic bodies (ApoBDs) serve as critical vehicles mediating intercellular communication in the central nervous system (CNS) following TBI. The biogenesis and the content of EVs, including proteins, lipids and RNAs, are greatly changed and involved in the evolution of inflammation or tissue repairing after TBI. In this overview, we recapitulate the cellular origin of EVs and the function of EVs in the neuroinflammatory process after TBI, highlighting the dual regulatory roles of EVs in the biological response to TBI, whereby certain EV populations amplify secondary injury cascades, while others promote endogenous repair and recovery processes. We next investigate the progress in EV engineering and targeted delivery systems and report the potential mechanisms, emphasize the prospects and potential of engineered EVs for therapy, and comment on challenges and perspectives for clinical application in TBI.",
"42353862": "ID: 42353862\nTitle: Clinical and Genetic Spectrum of ANO3-Related Dystonia with Treatment Responses in a Chinese Cohort.\nAbstract: Background: Variants in anoctamin 3 (ANO3) are linked to autosomal dominant dystonia, commonly known as DYT-ANO3 (OMIM: #615034). While craniocervical dystonia constitutes the most frequently observed phenotype, its clinical manifestations display significant heterogeneity. Nevertheless, the data concerning the genetic characteristics, clinical features, and therapeutic outcomes of ANO3-related dystonia within Asian populations remain scarce. Methods: Whole-exome sequencing was conducted on 661 Chinese patients, comprising 356 individuals with dystonia and 305 individuals with non-dystonic movement disorders who served as an internal disease-control cohort. Candidate ANO3 variants were evaluated based on population frequency, predicted deleteriousness, ACMG criteria, and aggregate frequency analyses. A retrospective review was performed of clinical features and treatment responses. Results: Fifteen rare ANO3 missense variants were identified in 16 patients with dystonia and one non-dystonic individual, including one pathogenic variant, six likely pathogenic variants, and nine previously unreported variants. The rare ANO3 variants were significantly enriched in the dystonia cohort compared with the controls. ANO3-related dystonia exhibited broad clinical heterogeneity, with frequent cervical involvement (62.5%) and tremulous features (75%), and occasionally extended beyond classical isolated dystonia. Oral medication and botulinum toxin showed variable benefit, whereas deep brain stimulation was associated with marked improvement in selected medically refractory patients. Conclusions: This study broadens the genetic and clinical spectrum of ANO3-related movement disorders in a Chinese cohort. The findings support substantial clinical heterogeneity in DYT-ANO3 and suggest that deep brain stimulation, especially STN-DBS, may be considered in selected refractory cases, although further studies are needed.",
"42358359": "ID: 42358359\nTitle: Global research trends and hotspots of exosome-mediated drug delivery across the blood-brain barrier: a bibliometric study from 2015 to 2025.\nAbstract: The blood-brain barrier (BBB) is a major obstacle to targeted drug delivery for central nervous system (CNS) diseases. Although liposomes and polymeric nanoparticles have improved brain drug delivery, limitations remain in BBB targeting, long-term biocompatibility, and in vivo clearance. Exosomes are endogenous nanoscale extracellular vesicles with favourable biocompatibility, low immunogenicity, and BBB-crossing potential. Therefore, this bibliometric study summarises the current research status, future research trends, and challenges in the more specific field of exosome-mediated BBB drug delivery. A comprehensive search was conducted across the Web of Science Core Collection (WoSCC), PubMed, and Embase databases for relevant English-language literature on exosome-mediated drug delivery across the blood-brain barrier from 2015 to 2025. WoSCC served as the primary source for bibliometric analysis. PubMed and Embase databases were used for supplementary validation. Software such as VOSviewer, CiteSpace, and R-bibliometrix was employed for literature visualisation analysis. This study included 1,365 relevant articles from the WoSCC database, and the annual publication volume showed a steady upward trend. China and the United States significantly lead in both the number of publications and the number of core contributing institutions in this field. Co-occurrence analysis of keywords showed that research hotspots are mainly focused on exosomes, the blood-brain barrier, drug delivery, and Alzheimer's disease. PubMed and Embase were used as supplementary validation databases, including 1,089 and 1,517 records, respectively. Their annual publication trends, major countries/regions, core journals, and keywords/themes were generally consistent with WoSCC, supporting the macro-level stability of the bibliometric findings. Unlike previous bibliometric analyses that mainly focused on overall trends in CNS exosome research, this study focuses specifically on the direction of exosome-mediated drug delivery across the BBB. The findings show a shift from basic vesicle characterisation toward engineered delivery systems, CNS disease applications, and translational evaluation. Mammalian-derived exosomes remain dominant, while plant-derived vesicles, AI-assisted design, biomimetic hybrid nanovesicles, and gut-brain axis strategies are emerging areas of focus. Future research should prioritise systematic platform comparisons, standardised evaluation, quality control, scalable production, long-term safety, and regulatory pathways.",
"42360457": "ID: 42360457\nTitle: White matter microstructural alterations correlate with clinical characteristics in craniocervical dystonia.\nAbstract: To characterize white matter pathology, identify clinical correlates, and explore relevant biomarkers for deep brain stimulation (DBS) outcomes in craniocervical dystonia (CCD) using diffusion tensor imaging (DTI). We involved 37 CCD patients (22 undergoing bilateral subthalamic DBS [STN-DBS]) and 42 matched healthy controls. Using Tract-Based Spatial Statistics (TBSS) for whole-brain analysis and Automated Fiber Quantification (AFQ) for 20 specific tracts, DTI measurements of fractional anisotropy (FA), axial diffusivity (AD), radial diffusivity (RD) and mean diffusivity (MD) were extracted. Between-group metric comparisons and correlations with clinical features and treatment outcomes were assessed. TBSS revealed reduced FA in left anterior corona radiata and right posterior thalamic radiation, increased AD in splenium of corpus callosum, and widespread elevated MD/RD (p\u2009<\u20090.0125, family-wise error combined threshold-free cluster enhancement corrected). AFQ identified segmental increased MD/RD across several tracts, most notably within anterior thalamus radiation (ATR), corticospinal tract, cingulum cingulate, callosum forceps, and inferior fronto-occipital fasciculus (uncorrected p\u2009<\u20090.01). Microstructural alterations correlated with earlier disease onset, severe baseline blepharospasm, and botulinum toxin exposure (p\u2009<\u20090.01, uncorrected). Crucially, at an uncorrected threshold of p\u2009<\u20090.01, lower MD in the left ATR was associated with greater eye symptom improvement after STN-DBS surgery (r\u2009=\u2009-0.57, p\u2009=\u20090.009). CCD exhibits distributed white matter microstructural alterations across multiple networks, potentially serving as the neurostructural substrate for motor and non-motor symptoms. Left ATR integrity is negatively associated with DBS outcomes.",
"42371396": "ID: 42371396\nTitle: Emerging Pharmacological Strategies for Trigeminal Neuralgia.\nAbstract: Trigeminal neuralgia can be a debilitating pain condition that affects women more frequently than men and for which there are only limited therapeutic avenues. Trigeminal neuralgia can either be idiopathic (where there is no clearly identifiable cause), classical (which typically involves neurovascular compression of the trigeminal root entry zone) or secondary which results from nerve damage due to other causes. First-line treatments for trigeminal neuralgia include the sodium channel blockers carbamazepine and oxcarbazepine, whose mechanism of action is to stabilize the inactivated state of sodium channels, thereby reducing the electrical activities of trigeminal neurons. Second-line treatments include gabapentinoids, which act on Cav\u03b12\u03b4 subunits of high voltage activated calcium channels to inhibit synaptic transmission in the trigeminal pain pathway. Both of these treatment avenues exhibit a number of adverse effects and many patients are refractory. Hence, new avenues for combatting trigeminal neuralgia are needed. In this review article, we discuss clinical evidence for efficacy and benefits of novel pharmacological treatment approaches for trigeminal neuralgia, including the inhibition of CGRP receptors, activation and desensitization of TRPV1 channels, the development of new generations of sodium channel blockers, and the use of botulinum toxin. We conclude that while there are potential benefits of these approaches, larger well-controlled cohort studies are needed to provide a definitive answer.",
"42374580": "ID: 42374580\nTitle: HIV Tat-activated microglial extracellular vesicles induce neuronal iron dysregulation and synaptodendritic injury.\nAbstract: Extracellular vesicles (EVs) are membrane-enclosed, nanoscale structures released by cells and play a key role in intercellular communication under both normal physiological and pathological conditions. They serve as conduits for transferring molecular cargo between neighboring cells, thereby modulating recipient cell function. While the HIV Transactivator of transcription (Tat) protein has been shown to induce ferroptosis in microglia, the role of Tat-activated microglia-derived EVs (Tat-MEVs) in transferring iron-handling and ferroptosis-associated cargo to neurons and promoting neuronal injury remains unexplored. In this study, we sought to evaluate the impact of cargo derived from Tat-MEVs on neuronal synaptodendritic degeneration. Rat primary cortical and hippocampal neurons were exposed to either control MEVs or Tat-MEVs and subsequently assessed for synaptodendritic degeneration, expression of key ferroptotic mediators, and mitochondrial dysfunction associated with neuronal injury. Neurons exposed to Tat-MEVs demonstrated increased expression of the key iron-handling and ferroptosis-associated proteins (transferrin, TF; transferrin receptor 1, TFR1; Six-Transmembrane Epithelial Antigen of the Prostate 3, STEAP3; divalent metal transporter 1, DMT1; and ferritin heavy chain 1, FTH1); inhibitory synaptic markers (GAD65, Gephyrin), Fe2+/total iron content, neuronal cytotoxicity and mitochondrial reactive oxygen species (ROS) compared to neurons exposed to control MEVs. These findings suggest a link between mitochondrial dysfunction and neuronal iron accumulation. The expression of these mediators was downregulated in neurons exposed to MEVs derived from iron chelator, deferoxamine (DFO)-pretreated BV2 cells. Electrophysiological recordings further revealed reduced miniature excitatory postsynaptic currents in neurons exposed to Tat-MEVs, an effect that was attenuated in neurons exposed to DFO-derived MEVs. Additionally, dendritic spine analyses of neurons exposed to Tat MEVs revealed a reduction in mushroom and stubby spine subtypes, suggesting synaptodendritic injury.Collectively, these findings demonstrate that Tat-MEVs transfer iron-handling and ferroptosis-associated cargo that promotes neuronal iron dysregulation, oxidative stress, mitochondrial dysfunction, and synaptodendritic degeneration. These changes are consistent with ferroptosis-associated neuronal stress and contribute to functional impairment in recipient neurons. This EV-based communication axis provides mechanistic insight into how HIV Tat-induced microglial dysfunction propagates iron-dependent neurotoxic signaling within the central nervous system and identifies EV-mediated iron dysregulation as a potential therapeutic target in NeuroHIV.",
"42381337": "ID: 42381337\nTitle: Extracellular Vesicles from Mesenchymal Stem Cells Alleviate Spinal Cord Injury via the miR-486-5p/PTEN/PI3K/AKT Pathway.\nAbstract: Spinal Cord Injury (SCI) is a severe central nervous system disorder with limited effective treatments. Mesenchymal stem cell (MSC)-derived exosomes have emerged as important mediators of intercellular communication and carry microRNAs with potential neuroprotective properties. This study aimed to explore the role and underlying mechanism of human umbilical cord MSC (hUMSC)-derived exosomal miR-486-5p in experimental SCI. Exosomes were isolated from hUMSCs and characterized by transmission electron microscopy, nanoparticle tracking analysis, and exosomal marker expression. A rat SCI model and an LPS-induced PC12 cell inflammatory injury model were established. Histological injury and apoptosis were assessed by HE staining and TUNEL assay. Inflammatory cytokine levels were measured by ELISA. Cell viability, apoptosis, and gene and protein expression were evaluated using CCK-8 assay, flow cytometry, qPCR, and western blotting. A dual-luciferase reporter assay was performed to validate the interaction between miR-486-5p and PTEN. hUMSC-derived exosomes attenuated spinal cord tissue damage, reduced neuronal apoptosis, and suppressed inflammatory cytokine production in vivo and in vitro. Inhibition of exosomal miR-486-5p partially reversed these protective effects. Mechanistically, miR-486-5p directly targeted the 3'-UTR of PTEN, leading to reduced PTEN expression and enhanced phosphorylation of AKT and mTOR. These findings indicate that exosomal miR-486-5p contributes to the regulation of apoptosis- and inflammation-associated molecular events following SCI, primarily through modulation of the PTEN/AKT/mTOR signaling pathway. Given the experimental design, these results should be interpreted as mechanistic insights rather than evidence of functional recovery. hUMSC-derived exosomal miR-486-5p alleviates apoptosis and inflammation following SCI by targeting PTEN and activating the AKT/mTOR pathway. These findings provide mechanistic support for the potential application of exosome-based miRNA therapy in SCI.",
"42384787": "ID: 42384787\nTitle: Turbocharging synaptic transmission: 12 SNAREpins are required for rapid release of reconstituted synaptic vesicles.\nAbstract: Synaptic transmission occurs synchronously with real-world events, far faster than vesicle fusion for hormone release or membrane biogenesis, all mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. How SNAREs cooperate to achieve synchronous neurotransmitter release is a long-standing mystery. Rapid release (<7 milliseconds) has been reconstituted from purified synaptic SNAREs, SNARE-assembling chaperones, and calcium ion sensors in a fully-defined, genetically validated system that enables single-molecule counting in docked vesicles before release. SNAREpin complexes (12\u00a0\u00b1\u00a00.3) are found in each such ready-release vesicle, suggesting a regular structure. Several genetic conditions (including point mutation of the synaptic vesicle protein Synaptophysin from a Synaptopathy patient and human and mouse disease mutations of the synaptic vesicle protein vesicle-associated membrane protein-2 (VAMP2) reduce the number of SNAREpins to 6\u00a0\u00b1\u00a00.3 and result in profoundly delayed release over 0.1 to 1 seconds. Omitting Synaptophysin, whose hexamers preassemble 12 copies of VAMP2, also yields ~6 SNAREpins and delays release.",
"42386531": "ID: 42386531\nTitle: Translational Profiling of Drd2-Expressing Populations Reveals Molecular Heterogeneity of Dentate Gyrus Mossy Cells along the Dorsoventral Axis.\nAbstract: Hilar mossy cells (MCs) are crucial for integrating and propagating signals across the hippocampal dorsoventral axis, mediating cognitive and affective processing. While MCs exhibit profound dorsoventral differences in their projections, physiology, and behavioral roles, the molecular basis underlying this functional specialization remains largely unexplored. To address this gap, we used translating ribosome affinity purification (TRAP) in male mice to systematically compare the translatome of Drd2-expressing, MC-enriched populations along the dorsoventral axis. This analysis revealed distinct translational signatures with 1,442 genes enriched in dorsal and 1,337 genes in ventral Drd2-expressing, MC-enriched populations. Pathway analysis demonstrated significant functional segregation along the dorsoventral axis. The dorsal population is notably enriched for genes linked to neuronal connectivity and synaptic transmission, whereas the ventral counterpart shows enrichment in genes associated with energy metabolism and cellular maintenance. Specifically, we identified a subset of dorsal enriched genes, including neurotransmitter receptors, ion channels, and axon guidance regulators, contrasting with ventral enriched genes highly related to glucose/fatty acid metabolism, oxidative phosphorylation, and exocytosis. We further predicted distinct sets of upstream transcriptional regulators activated in each subpopulation, providing insights into the regulatory networks that may drive molecular divergence. Our findings provide a translatomic basis for the dorsoventral heterogeneity of Drd2-expressing neurons that include MCs, offering molecular signatures associated with their differential contributions to hippocampal function.",
"42400067": "ID: 42400067\nTitle: Ultrasound-guided botulinum toxin injections for abdominal spasticity and dyspnea in an ambulatory patient with an incomplete cervical spinal cord injury: a case report.\nAbstract: Abdominal spasticity in spinal cord injury (SCI) patients may remain unrecognized, despite its impact on quality of life. We report a 47-year-old white Dutch man with incomplete C4 SCI (AIS D) presenting with dyspnea, abdominal pain, and discomfort from involuntary abdominal muscle contractions. Five year post-injury, the patient developed bloating and pronounced abdominal pain, pain in the lower back, and in the intercostal muscles. In addition, a 7-cm diastasis developed in the abdominal midline. These complaints reduced ambulatory capacity and quality of life. Initial management with systemic antispasticity treatment (tolperisone 50\u00a0mg, 3\u00d7/day) relieved leg spasticity and closed the diastasis, yet failed to alleviate dyspnea. Ultrasound imaging confirmed spasticity predominantly in the internal oblique and transverse abdominal muscles. Given the persistence of abdominal spasticity complaints, we initiated a series of ultrasound-guided botulinum toxin (100-150E incobotulinum toxin-A) injections to the internal and external oblique muscles-with one session also targeting transversus. The injection protocol was refined over successive treatments to achieve an optimal distribution while minimizing side effects on core stability. Although the patient experienced transient balance issues and lower back pain during the initial adjustment period, these effects were minor and resolved within 2-3\u00a0weeks. This case underscores that in ambulatory SCI patients, systemic treatments alone may not sufficiently address abdominal spasticity and concomitant symptoms like dyspnea. Low-dose ultrasound-guided botulinum toxin injections targeting the abdominal muscles can provide significant symptom relief and improve overall functional capacity, offering a safe and effective addition to low-dose systemic treatment.",
"42401592": "ID: 42401592\nTitle: Munc18-1 is crucial for photoreceptor function, survival and regulation of syntaxin-3 localization and expression.\nAbstract: The function of the SNARE complex regulator, Munc18-1, in photoreceptor cells is unknown. Here, we found that removing Munc18-1 from photoreceptors results in major degeneration starting at P14. In the absence of Munc18-1, before major photoreceptor degeneration, functional and synaptic impairments were present, indicating a critical function of Munc18-1. Furthermore, Munc18-1 played a critical role in expression and localization of syntaxin-3. The syntaxin-3 protein level is dramatically reduced in the soma and plasma membrane of photoreceptors without Munc18-1. At the photoreceptor synapses, the colocalization of syntaxin-3 and its SNARE partner, SNAP-25, was reduced, potentially suggesting an altered syntaxin-3 synaptic localization. In the Munc18-1-deficient photoreceptors, immature synapses and outer segment lesions were found. Taken together, these findings provide evidence that Munc18-1 is important for maintaining sufficient syntaxin-3 expression in the cell body and synapses of photoreceptors. The lack of Munc18-1, combined with poor syntaxin-3 expression, contributes to photoreceptor functional impairment and degeneration.",
"42404278": "ID: 42404278\nTitle: Primary Cerebellar Agenesis in a Child with Esotropia: A Case Report and Literature Review.\nAbstract: To highlight the importance of neuroimaging in children with esotropia and developmental delay and to support botulinum toxin as a minimally invasive treatment option when major brain malformations are present. We describe an 18-month-old girl with infantile esotropia and mild developmental delay. Brain magnetic resonance imaging revealed near-complete cerebellar agenesis with a hypoplastic pons. Considering this malformation, bilateral medial rectus botulinum toxin injections (15 units in 0.1 mL per muscle, freehand technique) were performed instead of conventional strabismus surgery. Ocular alignment improved with satisfactory cosmetic appearance, and stability was maintained throughout 4 months of follow-up. Genetic testing was not performed at the time because of follow-up limitations; however, it is recommended for further etiological clarification. This case highlights the importance of neuroimaging in children with esotropia and developmental delay and supports botulinum toxin as a minimally invasive treatment option when major brain malformations are present.",
"42404433": "ID: 42404433\nTitle: Beyond motor neurons: peripheral TDP-43 pathology in skeletal muscle and intramuscular nerves in amyotrophic lateral sclerosis.\nAbstract: Amyotrophic lateral sclerosis is a progressive neurodegenerative disease characterized by accumulation of the 43-kDa TAR DNA-binding protein (TDP-43). This neuropathological signature has been well documented within the CNS; however, recent findings indicate that the phosphorylated TDP-43 additionally deposits in peripheral tissues, including skeletal muscle and intramuscular nerves. These data warrant a change of view from a neurocentric perspective of amyotrophic lateral sclerosis pathogenesis towards a broader concept of TDP-43 proteinopathy extending both within and beyond the nervous system. In this review, we focus on current evidence supporting the presence of TDP-43 pathology in amyotrophic lateral sclerosis skeletal muscle, examining its topographic distribution, molecular characteristics and associations with intramuscular nerve bundles. We also discuss the susceptibility of intrinsic muscle cells, disrupted axonal transport and impairment in protein quality control. Phosphorylated TDP-43 pathology in muscle biopsies from amyotrophic lateral sclerosis patients has emerged as a promising tool in the early diagnosis of the disease. Moreover, we discuss the relevance of these findings to amyotrophic lateral sclerosis pathogenesis and potential therapeutic implications.",
"42422202": "ID: 42422202\nTitle: Long-term efficacy of neural circuit blockade for treating blepharospasm: a retrospective case series study.\nAbstract: Blepharospasm is a common symptom of Meige syndrome and often leads to functional blindness. Although current treatment options, such as oral medications, botulinum toxin injections, resection of periocular muscles, and deep brain stimulation, are available, no definitive method effectively eliminates blepharospasm. To address this unmet need, we developed a novel approach-neural circuit blockade (NCB). This study aimed to analyse its long-term efficacy for blepharospasm. NCB eliminates trigeminal nerve stimulation by facial expression muscles, reducing the production and input of sensory information, weakening feedback-based and plastic motor information functions, decreasing orbicularis oculi muscle tension, and alleviating blepharospasm. A retrospective analysis was conducted on 570 patients who underwent NCB treatment, with a median postoperative follow-up of 51\u202fmonths. Therapeutic efficacy was evaluated by comparing preoperative and postoperative scores on the Shorr blepharospasm grading scale, Burke-Fahn-Marsden Dystonia Rating Scale (BFMDRS-M), and Blepharospasm Disability Index (BSDI). Based on the Shorr grading scale, preoperatively, 97.9% of patients were grade 4, whereas 2.1% were grade 3. Postoperatively, 87.5% achieved complete remission (grade 0), whereas 7.4, 3.7, 0.9, and 0.5% were grades 1, 2, 3, and 4, respectively. Pre- and postoperative BFMDRS-M scores significantly differed: eye (8.0 vs. 0.0), mouth (3.0 vs. 0.0), speech/swallowing (3.0 vs. 2.0), neck (3.0 vs. 1.5), and total (8.0 vs. 0.0). BSDI scores also significantly differed pre- and postoperatively. The results indicate that NCB effectively alleviates blepharospasm and addresses the lack of a definitive treatment, without causing severe local or systemic complications such as facial paralysis. NCB can safely and effectively alleviate blepharospasm, improve quality of life, and demonstrate stable long-term efficacy.",
"42422258": "ID: 42422258\nTitle: Neuromuscular-related interventions for post-stroke dysphagia: a comprehensive narrative review.\nAbstract: To systematically review the efficacy, mechanisms, and application characteristics of neuromuscular-related interventions for post-stroke dysphagia (PSD), and to examine strategy selection across different swallowing stages to inform individualized rehabilitation strategies. PubMed, Web of Science, Embase, and MEDLINE were systematically searched for studies published between 1995 and 2025. Titles and abstracts were screened, and the full texts of eligible studies were retrieved for further analysis. Interventions related to neural and muscular regulation were categorized and synthesized into four main groups: exercise training and behavioral interventions; peripheral neuromuscular stimulation; central nervous system modulation techniques; and other adjunctive interventions. A total of 293 publications were included, of which 56.63% were randomized controlled trials. The interventions were summarized into four major categories comprising more than 10 techniques. Exercise training and behavioral interventions (e.g., oral motor exercises) enhanced swallowing muscle strength and coordination. Peripheral neuromuscular stimulation (e.g., neuromuscular electrical stimulation and acupuncture) enhanced or modulated swallowing function by directly stimulating relevant nerves or muscles. Central nervous system modulation techniques (e.g., transcranial magnetic stimulation and transcranial direct current stimulation) influenced swallowing-related neural networks indirectly by regulating cortical excitability. Other adjunctive interventions included botulinum toxin injection, which directly targeted the cricopharyngeal muscle. Further analysis examined the selection of key rehabilitation techniques across different clinical stages of PSD, integrating central and peripheral neuromodulation approaches. It explored the potential implications of soft-tissue surgery and meridian-muscle theory for PSD management to inform individualized clinical decision-making. Neuromuscular interventions were found to be widely used in PSD management, particularly transcranial magnetic stimulation, acupuncture, and neuromuscular electrical stimulation. Future strategies should integrate pathology, clinical manifestations, and lesion localization to develop central lesion-oriented multimodal therapies that combine peripheral nerve and muscle interventions, potentially improving clinical outcomes.",
"42424539": "ID: 42424539\nTitle: Redefining Spasticity: The Spasticity X Working Group Consensus Statement.\nAbstract: Inconsistent and variably interpreted definitions of spasticity, alongside evolving mechanistic understanding, highlight the need for a clear, clinically relevant consensus definition. An international expert panel used a modified Delphi process to develop a concise, clinically applicable definition that reflects current understanding and supports consistent assessment and management. Participants reviewed existing definitions, completed a pre\u2011meeting survey, and engaged in structured discussions, with draft definitions iteratively refined through successive rounds of voting to achieve consensus. Key components identified included disordered sensorimotor control, central nervous system involvement, and velocity\u2011 and length\u2011dependent resistance to passive stretch, while existing definitions were considered either overly narrow or insufficiently relevant to clinical practice. The consensus definition characterizes spasticity as \"A disorder of sensorimotor control resulting from upper motor neuron disease. It is characterized by velocity- and length-dependent involuntary muscle overactivity, which is intermittent or sustained, during passive stretch.\" This definition integrates contemporary mechanistic concepts with clinical applicability and is intended to improve conceptual clarity, facilitate communication, and promote consistency in diagnosis, measurement, and treatment.",
"42427545": "ID: 42427545\nTitle: Dystrophic changes of nigrostriatal axons harboring a Synj1 Parkinson mutation suggest catastrophic failure of endocytic mechanisms.\nAbstract: Synaptojanin 1 is a brain enriched phosphoinositide phosphatase implicated in endocytosis at the synapse. A mutation (R258Q) that selectively impairs its Sac1 phosphatase domain causes early onset familial Parkinsonism. Neurons of mice with this mutation display synaptic vesicle traffic defects across the brain, but selective dystrophic changes in a subset of dopaminergic axons in the dorsolateral striatum. Using correlative light microscopy-FIB-SEM of mutant mouse striata to visualize in 3D these abnormal structures we show that they represent clusters of focal axonal dilations harboring massive, onion-like DAT enriched plasma membrane infoldings, generally localized next to cell bodies of neighboring cells, often engulfing evaginations of such cells. This dysmorphia was associated with a deficit in dopamine release in the same striatal region. Given the involvement of Synj1 in endocytic mechanisms, these structures may reflect an imbalance between exocytosis and endocytosis. Their occurrence only in a subset of axons suggest a vulnerability threshold of these axons beyond which the expansion of the plasma membrane is not counteracted by compensatory mechanisms.",
"42427581": "ID: 42427581\nTitle: Ontological Analysis of Brain Proteostasis Highlights the Sex-Dependent Trajectory of ApoE Isoform-Specific Regulation.\nAbstract: Apolipoprotein E (ApoE) is the strongest genetic predictor of Alzheimer's disease (AD) risk, with ApoE4 increasing and ApoE2 decreasing risk relative to ApoE3. Using a global LC-MS proteomic approach, we integrated protein abundance and kinetics in Human-APOE knock-in mice for young (3-month) and aged (18-month) cohorts to quantify the changes in steady-state proteostasis. By mapping 6,052 identified proteins and 3,986 associated turnover rates into ontological groups, we observed that vesicle trafficking and mitochondrial dysregulation occur as early as 3 months in ApoE4 mice accompanied by hyperactive metabolism that eventually reduces with age. In contrast, young and old ApoE2 mice retain similar signatures to ApoE3 mice in metabolic, mitochondrial, cellular regulation, and membrane trafficking ontologies. We found that females had more isoform-induced ontological changes relative to ApoE3, providing insight into sex-dependent vulnerabilities. Our global proteomic approach for ApoE proteostasis crucially unifies independent literature observations while providing turnover kinetics to uncover the underlying mechanism behind abundance changes. Data are available via ProteomeXchange with identifier PXD079261.",
"42432065": "ID: 42432065\nTitle: Control of synaptic communication through molecularly engineered bioluminescence light emission and sensing.\nAbstract: Synapses are sites of intercellular communication between neurons and from neurons to target organs, and of signal integration that underly physiological and behavioral responses. We have developed a modular platform, Interluminescence (Int), for experimental control of synaptic transmission: bioluminescent light, generated by a luciferase oxidizing a luciferin, from a presynaptic neuron is used to activate transsynaptic optogenetic ion channels in the postsynaptic neuron. Two strategies can modulate postsynaptic neurons in the presence of luciferin. In the 'Act-Int' approach, a luciferase is genetically expressed in synaptic vesicles and released during depolarization-induced presynaptic vesicle fusion and exocytosis. In the 'Persist-Int' approach, a luciferase is tethered to the presynaptic membrane where it can support sustained transsynaptic signaling. Both strategies can activate postsynaptic neurons with comparable efficacy under the conditions tested. By design, the modularity of the platform permits the use of luciferases and opsins ranging in brightness and light sensitivity, with the luciferase targeted to different subcellular regions of the presynaptic neuron, and the postsynaptic opsin being excitatory or inhibitory. Our results demonstrate the utility and versatility of Interluminescence to mediate synaptic transmission that is either activity-dependent or activity-independent.",
"42434075": "ID: 42434075\nTitle: Gangliosides in the 21st century: therapeutic prospects for the brain and spine.\nAbstract: Gangliosides are sialylated glycosphingolipids highly enriched in the central nervous system, where they regulate membrane signaling, metabolism, neurogenesis, and immune responses. This Review integrates recent advances across distinct experimental and clinical domains. First, recent studies demonstrate that the monosialoganglioside GM1 enhances astrocyte-neuron metabolic coupling via the astrocyte-neuron lactate shuttle, thereby supporting neuronal bioenergetics and resilience. Complementary mechanistic work shows that specific gangliosides regulate adult neurogenesis through developmentally controlled epigenetic and transcriptional programs. In Huntington's disease models, preclinical evidence indicates that GM1 and related gangliosides attenuate microglia-mediated inflammatory responses and promote proteostasis through extracellular vesicle-dependent clearance of misfolded proteins. Finally, clinical evidence from acute spinal cord injury demonstrates that GM1 administration accelerates neurological recovery, underscoring its translational relevance. Together, these findings position gangliosides as multi-target modulators of neural repair and inflammation, and highlight their potential for therapeutic development.",
"42434580": "ID: 42434580\nTitle: Thoracic medial branch peripheral nerve stimulation for adjacent segment pain in a patient with a long construct fusion: a case report.\nAbstract: Adjacent segment degeneration is a known late complication of long construct fusions. While several experimental treatment options exist, including medial branch blocks, no definitive treatment options have been identified. Previous studies have demonstrated the benefit of restorative stimulation of the erector spinae group on patients' pain, physical function, and disability. This case hypothesizes that motor and sensory programming at the segment adjacent to the long-construct fusion has the potential to be both therapeutic for this patient's adjacent segment thoracic pain and protective through improved muscular stability. A 71-year-old woman with a past medical history of rheumatoid arthritis and prior T11 to S1 laminectomy with fusion and bilateral pedicle osteotomy presented with moderately severe 7/10 adjacent segment pain. She had previously completed conservative therapy, including physical therapy, epidural steroid injections, and medication management with NSAIDs, muscle relaxants, anticonvulsants, topical lidocaine, and chronic daily opioid use totally 30 morphine milliequivalents daily, without significant relief. Physical exam was significant for bilateral T8-T10 facet pain and flat back kyphosis, without focal or motor sensory deficits. She underwent a peripheral nerve stimulator device implantation with bilateral four electrode leads placed at the bilateral T10 medial branches using a minimally invasive retrograde approach to accommodate the fusion. Leads were advanced in a cephalad to caudad direction aimed at the transverse process of T9. Stimulation was programmed using a combined motor- and sensory-based approach to address both functional restoration and pain control. An external therapy disc was utilized to deliver stimulation, allowing the patient to independently apply and remove therapy, supporting day-to-day management of stimulation based on symptom severity, activity level, and personal preference. At her 3-month follow-up, the patient reports significant relief in her pain with an average pain score of 3/10, improved physical function, and a decrease in her pain medication usage. Restorative neuromodulation using motor and sensory programming is a potentially effective treatment adjunct for select patients with adjacent segment degeneration, with treatment sessions aligned towards pain relief and functional restoration. Further studies are needed to better define its role, protective effects and expected outcomes in this patient population.",
"42434808": "ID: 42434808\nTitle: Brain targeting and trafficking of extracellular vesicles in central nervous system diseases: a therapeutic roadmap.\nAbstract: Extracellular vesicles (EVs) mediate intercellular signaling in the central nervous system (CNS) by transferring lipids, proteins, and nucleic acids among neurons, glia, endothelium, and immune cells. Brain targeting depends on a linked sequence: EV ligands and adsorbed protein coronas engage receptor modules, select endocytic routes, determine intracellular fate, and define the therapeutic readouts. These fates include lysosomal degradation, recycling, rare cytosolic delivery, or transport across the blood-brain barrier (BBB). In disease, the same pathways can disseminate proteopathic seeds and amplify neuroinflammation. Heparan sulfate proteoglycans (HSPGs) and LDL receptor family members, including low-density lipoprotein receptor-related protein 1 (LRP1), regulate tau, \u03b1-synuclein, and amyloid-\u03b2 handling. Phosphatidylserine readers and complement shape myeloid sink capture and inflammatory output. Integrin, tetraspanin, and ICAM-1 nanoclusters influence avidity, organotropism, and immune suppression. At the BBB, endothelial HSPGs, LRP1, and transferrin receptor (TfR) support receptor-mediated uptake, motivating engineered ligands such as rabies virus glycoprotein-derived peptides, Angiopep-2, and TfR binders. However, endosomal escape remains a major kinetic barrier to nucleic acid delivery. We synthesize these principles across Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, glioblastoma, and demyelinating disease, and outline design and assay standards needed to translate EV biology into safe, manufacturable CNS therapeutics.",
"42437583": "ID: 42437583\nTitle: Brain-derived extracellular vesicle (BDEV) abundance and BDEV protein associations with clinical phenotypes and treatment response in untreated first-episode psychosis.\nAbstract: Reliable biomarkers that capture central nervous system (CNS) processes during the earliest stages of psychosis are critically needed to improve early treatment and response monitoring. Brain-derived extracellular vesicles (BDEVs) that traverse the blood-brain barrier (BBB) can be isolated from peripheral blood, thereby offering a novel and minimally invasive approach to quantify CNS molecular biomarkers. We hypothesized that BDEV abundance and BDEV neuroinflammatory and neurotrophic protein cargo would differ between individuals with first-episode psychosis (FEP) and healthy controls (HCs), reflecting altered neuroinflammatory signalling and extracellular biology associated with psychosis and clinical outcomes. In a cohort of untreated individuals with FEP (n\u00a0=\u00a057) and HCs (n\u00a0=\u00a028), BDEVs were isolated from serum. Luminex assays quantified concentrations of brain-derived neurotrophic factor (BDNF), S100 calcium-binding protein B (S100B), and C-reactive protein (CRP) in BDEVs and serum. A subset of FEP participants (n\u00a0=\u00a034) subsequently completed four weeks of antipsychotic treatment. Group differences in BDEV characteristics and protein measures, as well as their associations with case-control status, baseline symptom severtity, and four-week symptom change, were examined. FEP participants showed a trend toward higher BDEV concentration than HCs (median 14.0 \u00d7 108 versus 10.2 \u00d7 108 particles/mL, p\u00a0=\u00a00.052), and higher baseline BDEV concentrations were significantly associated with more severe pretreatment symptoms (\u03b2\u00a0=\u00a04.09 \u00d7 10-10, p\u00a0=\u00a00.003) and greater symptom improvement after four-week antipsychotic treatment (\u03b2\u00a0=\u00a03.29 \u00d7 10-10, p\u00a0=\u00a00.029). Within BDEVs, BDNF (\u03b2\u00a0=\u00a0-1.05; p\u00a0=\u00a00.0018) and CRP (\u03b2\u00a0=\u00a0-0.84; p\u00a0=\u00a00.002) were significantly reduced in FEP vs HC, with a trend toward lower S100B level (\u03b2\u00a0=\u00a0-0.57; p\u00a0=\u00a00.061). In contrast, serum protein levels showed no case-control differences and did not correlate with corresponding BDEV protein levels. Lower baseline BDEV BDNF, S100B, and CRP were associated with greater therapeutic response among males. BDEVs offer a minimally invasive window into CNS-enriched biology in early psychosis. BDEV abundance and neuroinflammatory and neurotrophic cargo concentrations show associations with illness severity and sex-specific treatment response not detectable in serum, supporting their potential as biomarkers for early pathophysiology and treatment response in FEP.",
"42440317": "ID: 42440317\nTitle: Plasma Phosphorylated Tau 217 in Participants at Risk for Chronic Traumatic Encephalopathy.\nAbstract: In vivo biomarkers for detecting neuropathologies from repetitive head impacts (RHI), including chronic traumatic encephalopathy (CTE), are needed. To evaluate the utility of plasma phosphorylated tau 217 (p-tau217), assess its performance as a beta-amyloid (A\u03b2) biomarker in participants with RHI exposure at risk for CTE, and explore concordance with CTE neuropathology in a postmortem subsample. This longitudinal, multicenter, case-control study used data from the Diagnostics, Imaging, and Genetics Network for the Objective Study and Evaluation of CTE (DIAGNOSE CTE) Research Project, collected from September 2016 to October 2023. Participants were former American football players (case participants) and asymptomatic men unexposed to RHI (control participants). A subsample had available neuropathologic data. RHI, traumatic encephalopathy syndrome (TES) diagnoses, and levels of CTE certainty. Plasma p-tau217 (classified as positive [\u22650.63 pg/mL], intermediate [0.40-0.62 pg/mL], and negative [<0.40 pg/mL]), A\u03b2-positron emission tomography (PET; 18F-florbetapir; with A\u03b2-positive defined as a standardized uptake value ratio [SUVR] \u22651.10), and tau-PET (18F-flortaucipir). TES diagnoses were assigned by multidisciplinary consensus conference. Analyses of postmortem brains controlled for age, race, and APOE \u03b54 status. Among 231 participants (mean [SD] age, 57.75 [8.25] years), 177 were former football players (117 professional and 60 college) and 54 were unexposed participants. Former football players had higher baseline mean (SD) p-tau217 concentrations than unexposed participants (0.35 [0.26] pg/mL vs 0.27 [0.14] pg/mL; P\u2009=\u2009.008), although this was driven by a higher proportion of A\u03b2-PET-positive participants among former players. Plasma p-tau217 increased over time across the sample (B\u2009=\u20090.207 [95% CI, 0.117-0.298]; P\u2009<\u2009.001), with no significant time\u2009\u00d7\u2009exposure group interactions. Among football players, p-tau217 showed no time\u2009\u00d7\u2009group interactions with TES diagnosis, TES-CTE certainty, or RHI metrics. Higher p-tau217 concentration correlated with higher global A\u03b2-PET SUVR (B\u2009=\u20090.058 [95% CI, 0.053-3.501; P\u2009=\u2009.01), with a few discordant cases (5 participants were p-tau217-negative and A\u03b2-PET-positive; 7 participants were p-tau217-positive and A\u03b2-PET-negative). P-tau217 had similar areas under the curve for projecting A\u03b2-PET positivity as cerebrospinal fluid (CSF) p-tau181/A\u03b242 and CSF A\u03b240/42 measures (p-tau217: AUC, 0.88 [95% CI, 0.80-0.96]; CSF p-tau181/A\u03b242: AUC, 0.89 [95% CI, 0.79-1.00]; CSF A\u03b240/42: AUC, 0.85 [95% CI, 0.72-0.98]). Among 9 brain donors, 6 had CTE (stages II-IV; none with Alzheimer disease). Seven had negative or intermediate p-tau217, concordant with A\u03b2-PET. Two p-tau217 outliers with stage III CTE had normal concentrations upon additional testing. The findings of this study suggest that plasma p-tau217 concentration is unlikely to be useful for the detection of CTE, but it does show utility for ruling out A\u03b2 pathology in participants at risk for CTE.",
"42440475": "ID: 42440475\nTitle: Therapeutic progress in rosacea: targeting the neuro-vascular-immune triad.\nAbstract: Rosacea is a chronic inflammatory dermatological condition predominantly affecting the facial region. Clinically, rosacea is characterized by paroxysmal flushing, persistent erythema, telangiectasia, papules, pustules, and ocular symptoms. Severe rosacea cases are frequently complicated by anxiety, depression, and sleep disturbances, imposing a heavy psychosocial burden and markedly impairing the quality of life of the patients. Abnormal activation of the neuro-vascular-immune triad has been reported as a crucial pathological mechanism of rosacea. Moreover, dysregulation within the central nervous system might exacerbate disease progression by modulating peripheral nerve activity and neuroendocrine homeostasis. In this review, we summarize the latest advancements in rosacea treatments targeting the neuro-vascular-immune triad, including \u03b3-aminobutyric acid derivatives, antidepressants, anti-calcitonin gene-related peptide agents, physical neuromodulation (transcutaneous auricular vagus nerve and repetitive transcranial magnetic stimulations), botulinum toxin type A, and \u03b2-blockers. We provide a comprehensive analysis of their molecular mechanisms, clinical efficacy, and current limitations. While such therapies could specifically regulate different stages of the pathway, their evidence lacks large-scale randomized controlled trials and clarity regarding optimal dosing regimens and treatment durations. Future studies should strengthen basic investigations and clinical translation, explore combined therapeutic strategies, as well as develop personalized therapeutic strategies for the long-term effective control of rosacea.",
"42446153": "ID: 42446153\nTitle: Sensory Tricks in Dystonia: A Systematic Review and Nested Quantitative Synthesis.\nAbstract: Sensory tricks (also termed \"alleviating maneuvers\") are voluntary maneuvers that transiently alleviate dystonic postures or movements and represent a hallmark clinical feature of dystonia with diagnostic and mechanistic significance. Despite extensive phenomenological description, the evidence base has not been systematically synthesized. To characterize the prevalence, phenomenology, clinical effects, predictors, neurobiological mechanisms, and translational relevance of alleviating maneuvers in dystonia, and to prospectively evaluate the feasibility of quantitative synthesis across predefined outcome domains. A systematic review with a prespecified nested quantitative synthesis was conducted using a preregistered PROSPERO protocol (CRD420251175065). The nested component was defined a priori as a domain-restricted analytic strategy in which random-effects meta-analysis was planned where \u2265\u00a03 conceptually comparable studies were available, and in which the feasibility of pooling was itself treated as a primary methodological outcome. PubMed, Scopus, and the Cochrane Library were searched from inception. Studies reporting original empirical data on alleviating maneuvers in human dystonia of any etiology were included. Qualitative synthesis examined phenomenology, clinical effects, predictors, mechanisms, and translational applications. Heterogeneity was examined separately along clinical, methodological, and outcome-level dimensions. Of 631 records, 53 studies met inclusion criteria; 31 contributed to qualitative synthesis and 26 to nested quantitative analyses, with 4 contributing to both. No eligible studies in functional or tardive dystonia were identified. Alleviating maneuvers were reported across dystonia subtypes, with prevalence ranging from 13% to 90%; the largest registry cohort (n = 1477) reported effective maneuvers in 68.7% of patients. Acute motor improvements of 30%-50% in head deviation were reported during trick execution, although effects were transient. Responsiveness was associated with shorter disease duration and better response to botulinum toxin. No domain met criteria for formal meta-analysis; structural barriers were predominantly methodological and outcome-level rather than clinical. Alleviating maneuvers are a clinically consistent and mechanistically informative feature of dystonia, but their magnitude, durability, and predictors remain inconsistently measured. Standardized paradigms and outcome reporting are required to enable quantitative synthesis and sham-controlled evaluation of device-based alleviating-maneuver analogues.",
"42448693": "ID: 42448693\nTitle: Dorsal horn DCC amplification loop induced by endplate osteoclasts generates chronic nociplastic low back pain in male mice.\nAbstract: Spine degeneration is associated with low back pain (LBP), a major cause of disability and functional decline. However, effective therapies for non-specific LBP remain limited because its underlying mechanisms are poorly understood. Here, we show that spine degeneration induces netrin-1 secretion in porous endplates to activate DCC signaling in dorsal root ganglion (DRG) neurons in both LBP and naturally aged male mice. Conditional loss of netrin-1 in Trap+ osteoclasts, or Dcc in Avil+ DRG neurons reduced spinal hypersensitivity. Importantly, DCC activation induces Src phosphorylation and SNARE complex docking, leading to presynaptic glutamate release in the dorsal horn (DH). Consequently, presynaptic glutamate release induces expression of both netrin-1 and DCC in DH neurons as a form of postsynaptic plasticity that initiates a positive feedback loop to amplify spinal hypersensitivity. Moreover, netrin-1/DCC expression is also amplified in the parabrachial nucleus (PBN) in the brain in both LBP and aged male mice. Thus, osteoclast-derived netrin-1 in porous endplates activates DCC-mediated hypersensitivity in DRG neurons to be further amplified in the DH neurons. These results promote the concept of non-specific LBP as a form of nociplastic pain.",
"42450256": "ID: 42450256\nTitle: Peripheral and Central miRNA Signatures in Alzheimer's Disease: Tissue-Specific Variability, Sex-Associated Differences, and Implications for Blood-Based Biomarkers.\nAbstract: Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and significant neuropathological changes. Early and accurate diagnosis remains a major challenge, highlighting the need for reliable, minimally invasive biomarkers. MicroRNAs (miRNAs), small non-coding RNAs that regulate gene expression, have emerged as promising candidates. Their expression is altered in the brains of AD patients, reflecting disease-specific pathological processes, and they are detectable in peripheral biofluids. However, discrepancies in miRNA profiles between the brain and the circulation, and between patient populations remain a significant limitation, raising questions about their origin, transport across the blood-brain barrier, and their reliability in reflecting central nervous system pathology. This review provides a comprehensive overview of current research comparing miRNA expression profiles in brain tissue and blood in AD, with a focus on their biological relevance, mechanisms of release and transport, and diagnostic potential. We also discuss the challenges associated with cross-tissue variability, methodological inconsistencies, and the need for standardized approaches. Finally, we highlight future directions, including multi-tissue analyses and integration with other noninvasive modalities, to improve the clinical utility of miRNA-based biomarkers in AD.",
"42453424": "ID: 42453424\nTitle: PYGL-driven glycogenolysis impairs microglial autophagic flux via SNAP29 O-GlcNAcylation in Alzheimer's disease.\nAbstract: Aberrant metabolic alterations underlie microglial dysfunction, which plays an important role during neurodegenerative progression. However, the role of aberrant glycogen metabolism remains elusive. Here, we identified glycogen accumulation and upregulated glycogenolytic enzymes in brain microglia from patients with Alzheimer's disease (AD) and transgenic animal models. Particularly, the principal microglial glycogenolytic enzyme PYGL exhibited the most notable spatiotemporal upregulation during disease progression. Specific knockdown of microglial PYGL ameliorated neuropathological changes and cognitive deficits in AD mice. Bioinformatics analysis and experimental validation confirmed that enhancing microglial autophagic flux-dependent A\u03b2 clearance was the underlying mechanism. Furthermore, among all possible glycogenolytic pathways, PYGL downregulation primarily reduced hexosamine biosynthesis pathway activity, diminished UDP-GlcNAc and O-GlcNAcylation of the autophagy key protein SNAP29, and thereby facilitated formation of the SNARE complex, which is essential for autophagosome-lysosome fusion. These findings reveal a glycogenolysis-driven post-translational pathway regulating microglial autophagy, establishing PYGL as a therapeutic target for AD.",
"42458952": "ID: 42458952\nTitle: Syntaxin-4 at the Crossroads of Synaptic Plasticity and Neurodegeneration: From Molecular Mechanisms to Biomarker and Therapeutic Potential.\nAbstract: Syntaxin-4 (Stx4), a member of the Qa-SNARE protein family, is a pivotal regulator of membrane trafficking. Stx4 is predominantly localized at the postsynaptic membrane of neurons and in glial cells, where it is essential for synaptic plasticity and the maintenance of neuronal homeostasis. Stx4 facilitates the activity-dependent exocytosis of glutamatergic receptors, thereby governing the balance between Long-Term Potentiation (LTP) and Long-Term Depression (LTD). Emerging evidence suggests that dysregulation of Stx4 is intricately linked to the pathogenesis of neurodegenerative diseases. In Alzheimer's disease, Stx4 is involved in synaptic dysfunction and cognitive impairment through its interactions with amyloid-\u03b2 (A\u03b2) and tau pathologies, affecting receptor recruitment at dendritic spines. In Parkinson's Disease (PD), Stx4 contributes to \u03b1-synuclein proteostasis, dopaminergic signaling, neuroinflammation, and the maintenance of blood-brain barrier integrity. While its presence in cerebrospinal fluid highlights its potential as a candidate biomarker for these diseases, further validation is required to establish clinical utility. This review summarizes current mechanistic insights into Stx4's multifaceted roles in the neurodegenerative disorders and explores the therapeutic prospects of targeting Stx4-mediated pathways due to its translational potential.",
"42465429": "ID: 42465429\nTitle: The neuropathy-causing GARS1 \u0394ETAQ mutation drives pathology in subsets of motor and sensory neurons in mice.\nAbstract: Charcot-Marie-Tooth disease type 2D (CMT2D) results from gain-of-function mutations in GARS1, which encodes glycyl-tRNA synthetase (GlyRS), the enzyme responsible for charging transfer RNA (tRNA) with glycine. There are several CMT2D mouse models, but Gars \u0394ETAQ/+ is the only one that bears a patient-sourced mutation. Created using CRISPR/Cas9 to model a 12-nucleotide de novo GARS1 deletion identified in an unusually severe CMT2D patient, Gars \u0394ETAQ/+ mice have previously been shown to display several neuromuscular phenotypes; motor axon loss, denervated neuromuscular junctions (NMJs) and reduced muscle function. Here, we extend these analyses to provide a more comprehensive understanding of both motor and sensory nerve deficits across hind- and fore-limbs. At 3 months, Gars \u0394ETAQ/+ mice possess sex-independent alterations in the levels of neuropathy biomarkers - including decreased NfL and increased periaxin - alongside reduced muscle endurance and strength, and impairments in the sensory modalities of mechanosensation, proprioception and nociception. Underpinning these dysfunctions, we identified site-specific defects comprising altered sensory neuron populations, muscle spindle loss, reduced motor neuron size, disrupted NMJ innervation and maturation, and reduced axonal transport of signalling endosomes in vivo. Together, these experiments show that Gars \u0394ETAQ/+ mice display robust and selective peripheral nerve pathology that manifests in a general distal-to-proximal fashion, priming this CMT2D allele for testing treatments and evaluating mechanisms underlying peripheral nerve vulnerability.",
"42466399": "ID: 42466399\nTitle: Neuronal Mitochondrial Dysfunction Drives Astrocytic Mitochondrial Transfer after TBI: Reveals the Therapeutic Potential of astrocytic EV-Mito.\nAbstract: Mitochondria are dynamic organelles essential for neuronal survival and synaptic function, and their dysfunction is a key consequence of excitotoxicity following traumatic brain injury (TBI). While intercellular mitochondrial transfer and exogenous mitochondrial transplantation have emerged as mechanisms to restore cellular bioenergetics, its in vivo relevance in the central nervous system remains incompletely understood. Here, we used astrocyte and neuron-specific mitochondrial reporters (GFP or Dendra2) in mice to assess cell-type-specific mitochondrial morphology, bioenergetics, and transfer 24hrs after TBI. Neurons exhibited marked mitochondrial dysfunction, including altered morphology and reduced bioenergetic capacity across somatic, synaptic, and non-neuronal fractions. In contrast, astrocytic mitochondria showed morphological changes but preserved bioenergetic function. Concomitantly, astrocyte-to-neuron mitochondrial transfer was significantly increased following injury, although transfer to synapses remained limited. Single-cell RNA sequencing of astrocytes revealed upregulation of genes involved in extracellular vesicle (EV) biogenesis and mitochondrial translation following injury compared to controls. In vitro co-culture studies confirmed that astrocytes transfer mitochondria to neurons via EVs containing mitochondria (EV-mito). Isolated EV-mito from astrocyte-conditioned media improves neuronal mitochondrial function under NMDA (N-methyl-D-aspartate) induced excitotoxic conditions. Together, these findings demonstrate that neuronal mitochondrial dysfunction drives astrocyte-mediated mitochondrial transfer as an adaptive neuroprotective response after TBI. This process preserves neuronal bioenergetics in the soma and neurites but not at synapses, highlighting both its therapeutic potential and spatial limitations.",
"42469846": "ID: 42469846\nTitle: Metabolic reprogramming via SIRT2-deficient microglial large extracellular vesicles ameliorates alzheimer's pathology.\nAbstract: Current therapies for Alzheimer's disease (AD) offer only symptomatic relief, highlighting the urgent need for disease-modifying approaches capable of halting or reversing neurodegeneration. Extracellular vesicles (EVs) have attracted growing interest as therapeutic vehicles owing to their inherent capacity to bypass the blood-brain barrier and deliver complex biological cargo to the central nervous system. Here, we examined whether large EVs (LEVs) derived from microglia with stable Sirtuin-2 knockdown (SIRT2-KD) confer the neuroprotective effects associated with SIRT2 inhibition. LEVs harvested from SIRT2-KD microglia were administered intranasally to APP/PS1 mice. We assessed microglial uptake of LEVs, along with subsequent changes in cellular metabolism, migration toward amyloid-beta (A\u03b2) plaques, phagocytic activity, and downstream pathological and behavioral outcomes. Proteomic and acetylomic profiling were employed to characterize the molecular cargo of LEVs-SIRT2-KD. LEVs-SIRT2-KD were readily internalized by microglia in vivo following intranasal delivery. Uptake of these vesicles markedly enhanced microglial bioenergetics, driving coordinated upregulation of both oxidative phosphorylation and glycolysis. This metabolic shift was accompanied by improved microglial recruitment to A\u03b2 plaques and increased phagocytic clearance. Consequently, treated mice showed reduced A\u03b2 plaque deposition, restored synaptic integrity, and reversal of cognitive deficits. Proteomic and acetylomic analyses revealed that LEVs-SIRT2-KD are selectively enriched in proteins and acetylation modifications linked to energy metabolism and phagocytic function, offering a mechanistic basis for the observed metabolic reprogramming. Together, these results identify LEVs as a critical vesicle subtype mediating the effects of SIRT2 knockdown and support a cell-free therapeutic strategy for AD centered on EVs-driven metabolic reprogramming of microglia.",
"42473572": "ID: 42473572\nTitle: Access to Parkinson's disease treatments in the Middle East: a multi-national survey of availability, affordability, and utilization patterns.\nAbstract: Parkinson's disease (PD) is an increasing public health challenge in the Middle East, yet regional data on access to therapies and services remain limited. Understanding the gaps in PD treatment availability, affordability, and prescription frequency is necessary for improving care delivery and regional policy development. Evaluating the availability, affordability, and utilization of disease-related pharmacological treatments, surgical, device-assisted therapies, and rehabilitative services across multiple Middle Eastern countries. A cross-sectional, web-based survey was distributed to physicians with expertise in movement disorders across participating countries in the Middle East. The survey assessed three domains of availability, cost, and utilization of key medications, surgical interventions, and specialized PD services. Thirty-one respondents from nine Middle Eastern countries, including 44% movement-disorder specialists. Medication availability was 65.4%, affordability 68.6%, and prescription 52% across the included countries. Levodopa was nearly universally available (98%), whereas newer dopamine agonists, MAO-B inhibitors, and COMT inhibitors were available inconsistently. Advanced therapies demonstrated disparities; botulinum toxin and deep-brain stimulation were available in high-income countries but not frequently used, while infusion therapies were mostly unavailable. Telemedicine uptake was moderate; genetic testing was cost-restricted. Although general rehabilitation was available, PD-specific services were limited. Affordability constrained equitable care. Qatar had the highest reported prescription frequency of PD medications among the countries. The survey highlights disparities in access to essential PD therapies across the Middle East. Variable availability, costs, and limited specialized services prevent optimal care, urging the need for stronger regional policies, improved insurance coverage, and expanded clinical training to improve equity in PD management.",
"42482185": "ID: 42482185\nTitle: Hormonal, metabolic, and stress-related clinical differences between episodic and chronic migraine: insights for diagnostic and therapeutic approaches.\nAbstract: Migraine, a common neurological disorder, is categorized into episodic (EM) and chronic (CM) forms. Understanding their hormonal and metabolic differences can enhance insights into migraine pathophysiology and guide clinical approaches. To explore hormonal and metabolic imbalances in EM and CM patients and identify key factors contributing to migraine chronicity. This retrospective, cross-sectional study included 500 patients with migraine (318 EM and 182 CM). Data were retrospectively extracted from pre-existing clinical records, structured intake forms, physician examination notes, questionnaire-based assessments, and available laboratory results to evaluate demographic, clinical, metabolic, stress-related, mental health-related, and comorbidity variables. CM patients were older (46.28 vs. 43.13 years, p\u2009=\u20090.002) and had more frequent severe attacks (15 vs. 8/month) and mild attacks (12 vs. 6/month) (p\u2009<\u20090.001). CM patients exhibited higher rates of anaemia (34.9% vs. 25.7%, p\u2009=\u20090.035), asthma (12.8% vs. 7.3%, p\u2009=\u20090.046), and sleep apnea (4.7% vs. 1.3%, p\u2009=\u20090.034). Fibromyalgia-related symptoms were common in both groups (82.1% in CM vs. 78.2% in EM), while the number of fibromyalgia tender points was higher in CM than in EM patients (median 8 vs. 6). Ferritin levels were lower in CM but not significantly so (p\u2009=\u20090.139). Suicidal thoughts and feelings of helplessness were more common in CM (p\u2009=\u20090.016, p\u2009=\u20090.002). CM patients used Botox more often but with lower efficacy and engaged in more cognitive behavioural therapy (p\u2009=\u20090.002). CM was associated with more severe symptoms, higher comorbidity burden, more widespread pain features, and distinct metabolic and stress-related profiles compared with EM. These findings support the early recognition of clinical and comorbid factors associated with the chronic migraine phenotype.",
"42485583": "ID: 42485583\nTitle: Hybrid Nonviral Nanocarriers Enable Functional Neural Modulation.\nAbstract: Extracellular vesicles released from the neuronal cells mediate the transfer of proteins, nucleic acids, and neurotransmitter-related cargoes, shaping gene expression and intercellular communication across neural circuits. Leveraging this endogenous pathway, we fused astrocyte-derived exosomes with RNA-loaded synthetic liposomes to create sub-100 nm hybrid nanoparticles for central nervous system delivery. This design addresses key limitations of existing systems: conventional liposomes lack cell-type specificity and can be toxic, whereas native vesicles are difficult to load efficiently. By tuning a mildly cationic surface, the hybrids support efficient gene transfer to neurons without disrupting membrane integrity or inducing measurable cytotoxicity. We validated this platform in vivo by delivering Cre recombinase mRNA in transgenic mice and inducing channel rhodopsin-2 expression in the motor cortex and ventral tegmental area, yielding behavioral changes during optogenetic stimulation.",
"42489808": "ID: 42489808\nTitle: The New Spine of Access to the Brain's Secrets: Extracellular Vesicles from Cerebrospinal Fluid Liquid Biopsies in CNS Diseases and Blood-Brain Barrier Research.\nAbstract: Liquid biopsy is emerging as a powerful approach for less invasive biomarker discovery, with extracellular vesicles (EVs) in cerebrospinal fluid (CSF) showing promise for the assessment of central nervous system (CNS) disorders without actual tissue biopsy and as a complement to imaging techniques. EVs carry molecular cargo such as proteins, nucleic acids, and lipids that mirror those at the tissue of origin, offering unique opportunities to quantify disease-related changes in biomarkers. Compared with plasma-derived EVs, those from CSF provide more direct insights into the CNS because of direct shedding of brain EVs to CSF and bypass of confounding factors involving entry to systemic circulation. Despite this potential, translation into clinical practice is limited by challenges such as low yields, purity concerns, and lack of standardized isolation protocols. Addressing these difficulties, alongside integrating multiomics approaches, will advance our understanding of EV molecular cargo and their functional roles in CNS diseases. Over time, CSF-derived EVs could become the new driver of precision medicine in neurology, offering biologic insight for both diagnostic and therapeutic applications. This perspective provides a critical evaluation of the current status of EV-based liquid biopsy in CSF and offers recommendations for future research and clinical translation of data from CSF-derived EVs, highlighting their potential to inform physiologically based pharmacokinetic (PBPK) models. This state-of-the-art article evaluates existing evidence and highlights key knowledge gaps.",
"42491147": "ID: 42491147\nTitle: Exosome biogenesis in arthropods: conserved mechanisms, experimental evidence, and emerging directions.\nAbstract: Extracellular vesicles (EVs) are nanoscale, lipid-bound structures released by cells across all domains of life. Once viewed as a means for discarding unwanted cellular components, they are now understood to be central mediators of intercellular communication. Much of what is known about EVs comes from mammalian systems, where extensive work has defined the major EV subtypes and the mechanisms that generate exosomes. These findings continue to serve as the primary reference for interpreting EV biology in other organisms. EVs have been isolated from diverse arthropods, including a few insect and tick species, yet the molecular pathways that produce them remain less characterized. Current evidence from Drosophila, mosquito cell lines, and tick systems shows that many of the core components driving exosome biogenesis in mammals, including ESCRT complexes, tetraspanins, lipid-modifying enzymes, Rab GTPases, and SNARE proteins, are present and, in several cases, experimentally validated in arthropods. These findings point to broad conservation of exosome biogenesis across taxa, while also highlighting key caveats, with most conclusions relying on only a small number of model systems. The definitions of EV subtypes remain unclear in many arthropods, and alternative EV biogenesis pathways have received little attention. Future studies that incorporate non-model species, apply rigorous EV characterization standards, and explore the roles of various EV subtypes will clarify how these pathways operate across arthropod lineages and how they differ from well-studied mammalian systems.",
"42494432": "ID: 42494432\nTitle: Editorial: Cell therapy in the central nervous system and its crosstalk with the immune system.\nAbstract: ",
"42497206": "ID: 42497206\nTitle: VAMP7-dependent mitochondria-lysosome contacts contribute to glial mitochondrial dynamics and dopaminergic neuron survival.\nAbstract: Although disrupted mitochondrial dynamics in neurons are closely linked to neurodegenerative diseases, far less is known about how mitochondrial dynamics are regulated in glia or whether glial mitochondrial dysfunction contributes to neurodegeneration. Here, we show that the R-SNARE protein VAMP7 regulates the untethering of mitochondria-lysosome contacts (MLCs) in adult fly glia. Glial-specific knockdown of VAMP7 leads to prolonged MLCs and mitochondrial elongation associated with altered fission/fusion dynamics. These VAMP7-deficient mitochondria exhibit hyperpolarized membrane potential, leading to increased reactive oxygen species production, lipid droplet accumulation, and dopaminergic neurodegeneration. Mechanistically, VAMP7 interacts with the GTPase-activating protein TBC1D15-17 to promote Rab7 GTP hydrolysis. Without VAMP7, TBC1D15-17 remains bound to Rab7 but fails to activate its hydrolysis, resulting in elevated GTP-bound Rab7 and impaired MLCs untethering. Consistently, expression of GTP-locked Rab7Q67L or GTPase-activating protein-dead TBC1D15-17\u0394GAP phenocopies the mitochondrial defects, while GDP-bound Rab7T22N or wild-type TBC1D15-17 restores the MLC dynamics. Considering that SNARE proteins mediate membrane fusion, our results demonstrate a role for VAMP7 in glial mitochondrial dynamics via organelle contacts, impacting neuron survival in a non-cell-autonomous manner.",
"42497239": "ID: 42497239\nTitle: The axonal transport velocity of prions is independent of prion formation.\nAbstract: Prion diseases are caused by PrPSc, the self-templating infectious conformation of the normal host-encoded prion protein, PrPC. Spread of PrPSc in the peripheral and central nervous system follows a highly predictable pattern consistent with slow axonal transport along defined neuroanatomical pathways. The rate of PrPSc transport, the contribution of prion strain diversity, and the requirement for prion conversion during transport are poorly understood, in part, due to a lack of direct measurement of PrPSc transport under defined conditions. Here, using a combination of live animal and sciatic nerve explant culture experiments, and determined the velocity of PrPSc ranged from rates consistent with both slow and fast axonal transport. Interestingly, transport of PrPSc was not dependent upon prion conversion as it was observed using prion strain and host combinations that do not support prion formation. The rate of PrPSc transport of prion strains and synthetic prions suggest that prions can use both slow and fast modes of axonal transport. Overall, converging lines of evidence suggest that prions are transported at a range of velocities consistent with both slow and fast axonal transport that is independent prion formation.",
"42497665": "ID: 42497665\nTitle: AGSI: Adaptive group-enhanced strategy for iterative integration of single-cell multi-omics.\nAbstract: Single-cell multi-omics data integration is critical for understanding cellular heterogeneity and disease mechanisms. However, current methods face two key limitations: (1) uniform evaluation of cross-modal correspondence across all genes, neglecting the modular organization of biological systems, and (2) static integration strategies that fail to accommodate varying degrees of cell-level heterogeneity. To address these challenges, this study proposes AGSI, an adaptive framework for robust multi-omics integration through co-regulated gene modules and iterative reliability assessment. AGSI employs Latent Dirichlet Allocation to identify co-regulated gene modules and evaluates cross-modal correspondence at the module level. AGSI combines Wasserstein-enhanced similarity metrics with dual reliability modeling to progressively identify and integrate cells with high cross-modal concordance. Adaptive thresholding dynamically adjusts selection criteria throughout the iterative refinement process. Extensive experiments on multiple datasets including PBMC, SNARE-seq mouse brain, 10x mouse brain, and large-scale human myocardial infarction data demonstrate that AGSI significantly outperforms seven state-of-the-art methods. Notably, AGSI achieves up to 25.6% F1 improvement over its ablation baseline and 11.9% over the best competing method on complex neural datasets, and maintains over 85% accuracy even under 50% data dropout. AGSI provides a robust and scalable solution for multi-omics integration that preserves biological interpretability while achieving superior technical performance. AGSI is well suited to biomedical analyses requiring accurate cell type identification. The implementation code is available at https://github.com/CDMBlab/AGSI.",
"42501340": "ID: 42501340\nTitle: Dentato-Rubro-Thalamic Tract-Targeted Cerebellar Deep Brain Stimulation for Post-Stroke Spasticity and Dystonia: preliminary experience.\nAbstract: Introduction Post-stroke motor impairment with spastic hemiparesis remains a significant therapeutic challenge, as conventional rehabilitation and pharmacological strategies often fail to achieve sustained functional recovery. Neuromodulation targeting cerebellar pathways, particularly the dentato-rubro-thalamic tract (DRTT), may provide an alternative therapeutic approach. Methods The study reports the first three cases of chronic post-ischemic stroke-related spastic hemiparesis and dystonic features treated with deep brain stimulation (DBS) targeting the DRTT at the level of the dentate nucleus after unsuccessful botulinum toxin therapy and intensive rehabilitation. Directional DBS systems were implanted using tractography-guided targeting. Clinical outcomes were assessed preoperatively and during sequential stimulation at 130 Hz, 70 Hz, and 30 Hz (each for six weeks), followed by a three-week stimulation-off phase. Outcome measures included the Modified Ashworth Scale (MAS), Fugl-Meyer Assessment of the Upper Limb (FMA-UL), Chedoke Arm and Hand Activity Inventory (CAHAI), Unified Dystonia Rating Scale (UDRS), and modified Rankin Scale (mRS). Results FMA-UL scores progressively improved as stimulation frequency decreased, with mean improvements of 12.3% at 130 Hz, 14.7% at 70 Hz, and 18.4% at 30 Hz. A partial decline was observed during the stimulation-off phase (12.2%). Functional performance (CAHAI) showed the greatest improvement at 30 Hz (11.9%) and decreased to 6.6% during the stimulation-off period. Proximal spasticity was modestly reduced in the shoulder and elbow by approximately one MAS point during stimulation but returned to baseline stimulation-off. Distal spasticity remained largely unchanged. Dystonic features demonstrated frequency-dependent improvement, most pronounced at 30 Hz (43.9%), with partial persistence after stimulation discontinuation (27.5%). Global disability (mRS) remained stable or showed mild improvement. Conclusions Preliminary findings suggest that DRTT-targeted cerebellar DBS, particularly at lower frequencies, may offer a promising adjunctive treatment for post-stroke spasticity and motor impairment refractory to conventional therapies. These initial observations from an ongoing clinical trial require confirmation in larger cohorts with extended follow-up.",
"42503395": "ID: 42503395\nTitle: Engineered extracellular vesicles derived from sweet potato loaded with siPOLD1 for targeted therapy of glioma.\nAbstract: Glioma is the most common malignant tumor of the central nervous system, with high malignancy and poor prognosis, necessitating the development of novel targeted therapies. DNA polymerase delta catalytic subunit 1 (POLD1) is implicated in multiple cancers, but its role in glioma remains unclear. Plant-derived extracellular vesicles (PDEVs) have emerged as biocompatible, targetable nanocarriers with promising applications in cancer therapy. This study aims to elucidate the oncogenic function of POLD1 in glioma and develop a PDEVs -based delivery system for targeted therapy, with the goal of improving the current therapeutic landscape for glioma. POLD1 expression and prognostic significance were analyzed using clinical samples and databases. In vitro, CCK-8, Transwell, and flow cytometry assays evaluated the impact of POLD1 knockdown on glioma cell proliferation, invasion, migration, cell cycle, and apoptosis. In vivo tumorigenesis and survival were assessed in mouse models. Sweet potato-derived nano-vesicles (SPDELNVs) were isolated and characterized. An engineered A2-SPDELNVs-siPOLD1 system was developed via surface modification and siRNA loading, and its targeting efficiency and therapeutic efficacy were evaluated both in vitro and in vivo. POLD1 was upregulated in glioma tissues and correlated with poor prognosis. Its knockdown suppressed proliferation, invasion, and migration, induced cell cycle arrest, and promoted apoptosis in vitro. In vivo, POLD1 targeting inhibited tumor growth and prolonged survival. SPDELNVs showed intrinsic anti-glioma activity and efficient cellular uptake. The engineered A2-SPDELNVs-siPOLD1 effectively delivered siRNA, silenced POLD1, and significantly inhibited tumor progression both in vitro and in vivo, with enhanced survival. Our findings uncover the oncogenic role of POLD1 in glioma and validate it as a promising therapeutic target. Furthermore, we establish a novel, plant-based A2-SPDELNVs-siPOLD1 delivery platform with effective BBB penetration and tumor targeting, offering a promising strategy for the treatment of glioma.",
"42504872": "ID: 42504872\nTitle: Unconventional Protein Secretion in the Central Nervous System: Mechanisms and Roles in Physiology and Disease.\nAbstract: Extracellular secretion of neurotransmitters, proteins, and peptides by cells of the central nervous system underpins neurological function and homeostasis. Decades of elegant research have illuminated the molecular mechanisms and machinery that support the release of neurotransmitters via synaptic vesicle exocytosis, as well as the secretion of signal-peptide bearing proteins through the endoplasmic reticulum (ER)-Golgi based secretory pathways. However, it is now increasingly appreciated that signal-peptide lacking \"leaderless\" proteins can also be secreted via ER-Golgi-independent mechanisms collectively termed unconventional protein secretion (UcPS). In this review, we highlight the physiological and pathological consequences of UcPS in the central nervous system. UcPS supports the secretion of aggregation-prone proteins such as \u03b1-synuclein and mutant huntingtin, pro-inflammatory mediators including interleukin-1\u03b2 and high mobility group box protein 1, and neuroprotective or angiogenic factors such as fibroblast growth factor 2. Furthermore, several retroelement-derived proteins, encoded by ancient genomic elements with structural homology to retroviruses, are also secreted via unconventional pathways, and are thought to regulate essential CNS processes such as synaptic plasticity. These diverse cargoes underscore the functional range of UcPS in neuronal and glial biology. We summarize current understanding of the major UcPS pathways used by CNS cells. These mechanisms include plasma-membrane pore-mediated release facilitated by proteins such as gasdermin-D, as well as vesicular routes in which UcPS cargoes enter organelles of the autophagic and endolysosomal systems that subsequently fuse with the plasma membrane to enable extracellular release. Finally, we discuss key unresolved questionRecent evidence from HeLa cells suggests regarding the regulation of UcPS, including the molecular features that target select leaderless cargoes toward UcPS, how the balance between conventional secretion and UcPS shifts under cellular stress, and the current understanding of the diverse molecular machinery that mediates the vesicular form of UcPS.",
"42506058": "ID: 42506058\nTitle: Efficacy and Safety of a Tailored Dosing Strategy with High-Dose IncobotulinumtoxinA at Flexible Injection Intervals for Cervical Dystonia: An Open-Label, Uncontrolled, Single-Arm Study in Japan.\nAbstract: This prospective, multicenter, open-label, single-arm study (jRCT2031230690) evaluated the efficacy and safety of a tailored dosing strategy of incobotulinumtoxinA, including high doses (up to 500 U) and flexible injection intervals (as short as 6 weeks), in Japanese patients with cervical dystonia (CD). Of 30 enrolled patients, Group A included 27 patients with idiopathic CD for the primary evaluation of efficacy and safety, whereas Group B included 3 patients with tardive dyskinesia (cervical) or tardive CD for exploratory safety assessment. Patients received up to seven injection cycles of incobotulinumtoxinA (120-500 U) over 48 weeks, with minimum 6-week intervals. The primary endpoint, evaluated in Group A, was the change in Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS) total score from baseline to Week 4 after the first injection. Using a mixed model for repeated measures, the least squares mean \u00b1 standard error of the change was -11.0 \u00b1 1.77 (95% confidence interval: -14.6, -7.3). The primary efficacy endpoint was achieved in Group A. Due to the small sample size (n = 3), efficacy in Group B was evaluated only for exploratory purposes, although safety findings were broadly consistent with those in Group A. The overall safety profile was consistent with previous studies. Across the study, the most common related adverse events were dysphagia (33.3%) and muscular weakness (22.2%) in Group A and dysphagia (33.3%) in Group B. All cases of dysphagia were mild to moderate in severity and transient, with no apparent dose- or injection interval-related trend observed. IncobotulinumtoxinA was associated with improvements in symptoms and manageable safety profile at high doses and flexible injection intervals in Japanese patients with CD. While these findings suggest a potential treatment option for individualized dose optimization, the absence of a control group and the exploratory nature of the assessment in Group B necessitate cautious interpretation.",
"42506696": "ID: 42506696\nTitle: Effects of Botulinum Toxin Type A in Essential Blepharospasm: Evidence from a Clinical and Neurophysiological Pilot Study.\nAbstract: Essential blepharospasm (BEB) is a focal dystonia characterized by abnormal brainstem excitability and impaired inhibitory control within trigeminal-facial circuits. Botulinum toxin type A (BoNT-A) is the established first-line treatment, primarily acting at the neuromuscular junction. However, whether BoNT-A also modulates central brainstem circuits in BEB patients remains unclear, and dedicated neurophysiological studies have yielded conflicting results. To investigate whether BoNT-A modulates brainstem interneuronal excitability in BEB using the blink reflex recovery cycle and to correlate clinical outcomes with neurophysiological results. Thirteen patients with BEB underwent neurophysiological and clinical evaluation before (T0) and one month after (T1) BoNT-A treatment. The blink reflex recovery cycle was assessed at interstimulus intervals (ISIs) of 200, 300, 500, and 1000 ms. Clinical severity was assessed using the BSPSS, JRS, and BDS scales. The R2 amplitude ratio showed a statistically significant decrease after treatment across all ISIs (all p \u2264 0.001), indicating reduced brainstem interneuronal excitability. All clinical scales demonstrated statistically significant improvement after treatment (BSPSS, JRS, BDS; all p < 0.001). This pilot study provides preliminary evidence that BoNT-A treatment may reduce brainstem interneuronal excitability in BEB patients, as evidenced by a substantial and consistent decrease in R2 amplitude ratios of the blink reflex recovery cycle. These findings are consistent with a central modulatory effect of BoNT-A beyond its established peripheral action. Larger controlled studies are warranted to confirm these results.",
"42506700": "ID: 42506700\nTitle: Botulinum Toxin in Parkinson's Disease Tremor: A Critical Evaluation of the Evidence and Clinical Practice.\nAbstract: Approximately 30% of patients with tremor-dominant Parkinson's disease (PD) have rest tremor that persists despite optimal dopaminergic therapy. When deep brain stimulation and focused ultrasound are unavailable or declined, the therapeutic options narrow. Botulinum toxin (BoNT) offers a targeted, titratable, reversible approach, but whether a peripheral neuromuscular blocking agent makes sense for a centrally generated tremor is a legitimate question that deserves a direct answer. This narrative critical review appraises what is currently known across PD and non-PD tremor conditions, defines the technical requirements for safe and effective injection, and provides a practical framework for patient selection and clinical management. The PD-specific literature rests on a single positive double-blind randomized controlled trial of 30 patients; all remaining data are open-label or extrapolated from other tremor conditions, and this narrative synthesis combines heterogeneous conditions, outcome scales, and toxin protocols. A recurring technical observation is that, in the available trials, individualized, EMG-guided injection has been associated with substantially lower rates of hand weakness than fixed-dose injection (reported reductions from roughly 30-70% to below 15%) while maintaining tremor reduction, although the degree of benefit and weakness risk vary with the tremor syndrome, injected muscles, baseline impairment, dose, and guidance method. The careful patient selection this approach requires helps the individual clinician and patient achieve tremor relief, but it departs from the unselected real-world PD population and introduces selection bias that makes a large, statistically representative cohort difficult to assemble. In well-selected patients at centers with the appropriate expertise, BoNT may be a clinically useful option, but routine adoption is not yet supported.",
"42506967": "ID: 42506967\nTitle: Mechanical forces stimulate Golgi export.\nAbstract: Cells face diverse mechanical stimuli that vary with cell type, state, and pathological conditions. Mechanobiology investigates how cells sense and respond to these forces. While most work has focused on the cell surface and nucleus as primary mechanosensors, how intracellular organelles adapt to extracellular mechanical forces remains largely unknown. Here, we show that extracellular mechanical signals influence the secretory function of the Golgi apparatus. By subjecting adherent cells to mechanical challenges-cell spreading on different ligands, altered substrate stiffness, or equibiaxial strain-we reveal that extracellular forces modulate Golgi-to-cell surface carrier biogenesis, thereby regulating exocytosis. Together with changes in Golgi membrane tension, we identify molecular determinants of the mechanotransduction pathway, including microtubule acetylation, diacylglycerol production, and protein kinase D activity. In turn, inhibition of Golgi export suppresses this mechanoresponse and causes impaired cell spreading. These findings uncover a bidirectional mechanotransduction axis in which extracellular mechanics tune Golgi secretory output, providing a framework for investigating organelle-based mechanoadaptation in physiology and disease.",
"42507332": "ID: 42507332\nTitle: Disease mechanisms and translational barriers guide nanocarrier design for nose to brain delivery in Alzheimer's disease.\nAbstract: Alzheimer's disease (AD) is a progressive neurodegenerative disorder with limited disease-modifying treatment options, partly because many therapeutic agents show insufficient brain exposure and dose-limiting systemic adverse effects after conventional administration. Nose-to-brain (N2B) delivery has emerged as a non-invasive strategy to transport therapeutics to the central nervous system through the olfactory and trigeminal pathways, thereby partially bypassing the blood-brain barrier. Recent advances in nanomedicine and biomaterial engineering have further improved this approach by enhancing drug stability, nasal residence, mucosal transport, and brain-targeting efficiency. This review examines nanocarrier-enabled N2B delivery strategies for AD from a mechanism-guided perspective, highlighting how AD-related pathological processes shape the selection of therapeutic cargos and formulation designs. We discuss recent progress in the intranasal delivery of repurposed small molecules, natural products, insulin-related agents, peptides and proteins, extracellular vesicles, antibodies, and nucleic acid-based therapeutics. We further summarize major nanocarrier and formulation platforms, including lipid-based systems, polymeric nanoparticles, micelles, extracellular vesicles, in situ gels, and device-assisted delivery technologies. Particular attention is given to the design parameters that influence N2B performance, including particle size distribution/PDI, surface charge, mucus interaction, cargo protection, targeting modification, biodistribution, and deposition reproducibility. Finally, we critically evaluate the translational challenges that continue to limit clinical application, including species differences in nasal anatomy, dose-volume restrictions, device-dependent variability, limited human pharmacokinetic evidence, manufacturing complexity, long-term safety, and regulatory requirements. By integrating disease mechanisms, nanocarrier design, and translational considerations, this review provides a structured perspective for developing more rational and clinically feasible N2B nanodelivery systems for AD.",
"42510934": "ID: 42510934\nTitle: Herbal Bioactives Targeting Rho GTPases: A Multi-Targeted Strategy for Mitigating Neuroinflammation in Alzheimer's and Parkinson's Diseases.\nAbstract: Neuroinflammation plays an essential role in the pathogenesis of several associated brain diseases, including neurodegenerative disorders (Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS)), and traumatic brain injury (TBI). In these diseases, persistent microglial and astrocyte aggregates, elevated proinflammatory cytokines, and oxidative stress drive neuronal injury and cognitive disability. Rho GTPases, in particular the Rho family members Ras homolog family member A (RhoA), Ras-related C3 botulinum toxin substrate 1 (Rac1), and cell division control protein 42 homolog (CDC42), regulate neuroinflammation, cytoskeletal dynamics, immune responses, and the maintenance of BBB integrity. These proteins are involved in many neuropathological diseases due to dysregulation, making them interesting therapeutic targets. Bioactives used in herbal care have attracted interest for their ability to influence neuroinflammation and even their anti-neurodegenerative activity. Studies show that flavonoids, alkaloids, polyphenols, and other botanical compounds alter Rho GTPase activity, which, in turn, leads to decreased inflammation. This review critically summarizes current evidence regarding phytochemical regulation of Rho GTPase signaling in neurodegenerative disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD), with particular emphasis on the underlying molecular mechanisms, context-dependent signaling responses, and current translational challenges. Furthermore, existing knowledge gaps and future research priorities are discussed to facilitate the development of mechanism-based therapeutic strategies targeting Rho GTPases.",
"42511709": "ID: 42511709\nTitle: Wallerian Degeneration and Nerve Regeneration-A Review of Cellular and Molecular Events.\nAbstract: Wallerian degeneration (WD), which occurs distal to peripheral nerve injury, is a tightly regulated process. Axonal degeneration during the onset of WD represents a self-destructive process that begins with an early phase and progresses to an execution phase characterized by fragmentation of axons and myelin sheaths. Efficient clearance of axonal and myelin debris, together with the reprogramming of Schwann cells and macrophages into a repair phenotype, constitutes a critical extrinsic prerequisite for successful axonal regeneration. In parallel, signaling molecules produced during WD trigger intrinsic responses in injured neurons that are essential for neuronal survival and the initiation of the regenerative program. This review provides an overview of the key events involved in WD, which are often studied separately, with the aim of elucidating their interrelationships and their impact on nerve regeneration. Such an integrated overview may aid in identifying molecular targets for the development of novel therapeutic strategies to enhance axonal regeneration.",
"42511736": "ID: 42511736\nTitle: Urinary Extracellular Vesicle-Derived miRNAs as Regulators and Biomarkers in Diabetic Kidney Disease.\nAbstract: Diabetic kidney disease (DKD) remains one of the most severe microvascular complications of type 2 diabetes mellitus (T2DM) and a leading cause of chronic kidney disease (CKD) worldwide. Nevertheless, despite considerable progress in elucidating its molecular background, early diagnosis and accurate stratification of disease progression remain challenging when relying on conventional clinical biomarkers such as albuminuria and estimated glomerular filtration rate (eGFR). Growing evidence indicates that DKD is driven by interconnected pathogenic mechanisms, including chronic hyperglycemia, activation of the protein kinase C (PKC) signaling pathway, renin-angiotensin-aldosterone system (RAAS) dysregulation, oxidative stress, inflammatory cascades, and immune system activation involving Toll-like receptors (TLR) and the NLRP3 inflammasome. These processes collectively contribute to endothelial dysfunction, podocyte injury, extracellular matrix accumulation, and progressive renal fibrosis. Exosomes and their molecular cargo, particularly miRNAs, have emerged as promising regulators and non-invasive biomarkers reflecting ongoing renal injury. Urinary exosomal microRNAs (uEV-miRNAs) are of interest due to their stability in biological fluids and their direct origin from nephron segments, enabling real-time reflection of renal pathophysiology. Accumulating studies suggest that differentially expressed microRNAs (miRNAs), including miR-21-5p, miR-30a-5p, miR-192-5p, and miR-142-3p, are closely associated with key pathways in DN. However, their clinical translation remains limited by methodological heterogeneity, the lack of standardized isolation protocols, and insufficient validation in large longitudinal cohorts. This review navigates the current landscape of knowledge on the molecular mechanisms underlying DKD and examines the emerging role of uEV-miRNAs as diagnostic biomarkers. Altogether, uEV-miRNAs offer a promising avenue for improving early detection, risk stratification, and disease monitoring in DKD.",
"42514227": "ID: 42514227\nTitle: Dynamic Tuning of MSC-Based Scaffolds for Neurological Protection After Brain or CNS Injury.\nAbstract: Neurological disorders, including stroke, traumatic brain injury, and spinal cord injury, constitute one of the most important causes of mortality and morbidity worldwide for which current treatment options focus on resolving neuroinflammation rather than on tissue and neuronal regeneration. Mesenchymal stem cells (MSCs) could be a potential therapeutic option due to their immunomodulatory, neuroprotective, and paracrine secretion of extracellular vesicles and trophic factors which modulate microglial activation, preserve blood-brain barrier (BBB) integrity, and neuroplasticity, but with limitations due by poor survival, retention, and phenotypic instability following direct transplantation. The purpose of this narrative review is to present mechanotransduction signaling pathways (integrin-FAK, PI3K/Akt, Rho/ROCK, and YAP/TAZ) through which MSC-based biomaterial scaffolds, especialy hyaluronic acid (HA) hydrogels, make the transition from reparative to regenerative medicine in central nervous system (CNS) injury. Even if most of the evidence from preclinical studies suggests that dynamically tunable MSC-scaffold systems represent promising platforms for neural tissue engineering and regenerative medicine, further translational studies and well-designed clinical investigations are required to establish their therapeutic efficacy and clinical applicability.",
"42527713": "ID: 42527713\nTitle: LncRNA DLX6-AS1 Regulates the Pathological Process Related to Alzheimer's Disease via miR-204-5p.\nAbstract: Alzheimer's disease (AD) is a neurodegenerative disorder causing progressive neuronal damage. Incidence rises with age, and early diagnosis is difficult. This study examined DLX6-AS1 clinical relevance and regulatory mechanism in AD, and its interaction with miR-204-5p in AD pathology. It offers new insights into early diagnosis and treatment. A total of 133 AD patients and 105 healthy controls were selected. Their serum levels of DLX6-AS1 and miR-204-5p were analyzed using quantitative polymerase chain reaction. \u03b2-amyloid (A\u03b2)1-42-induced SH-SY5Y neuronal injury and okadaic acid (OA)-induced Neuro-2a tau abnormal phosphorylation models were constructed. The role/function of DLX6-AS1/miR-204-5p axis was then investigated using cell counting kit-8, flow cytometry, western blotting, enzyme-linked immunosorbent assay and a dual-luciferase reporter gene assay. AD patients had higher serum DLX6-AS1 and lower miR-204-5p levels. DLX6-AS1 showed an AUC of 0.838 for AD diagnosis. DLX6-AS1 levels were negatively associated with cognitive function, brain structural integrity, and benign pathology-and positively associated with disease severity, functional impairment, and pathological markers. In AD cell models, DLX6-AS1 was upregulated. Silencing it promoted cell proliferation, reduced apoptosis and oxidative stress, improved mitochondrial and synaptic function, decreased tau phosphorylation, and enhanced microtubule stability and axonal transport. Dual-luciferase assays confirmed direct binding between DLX6-AS1 and miR-204-5p. Co-inhibition reversed the protective effects of DLX6-AS1 silencing. Serum DLX6-AS1 is a potential biomarker for early diagnosis and assessment of AD. It regulates A\u03b2-induced neuronal damage and tau phosphorylation by targeting miR-204-5p, offering a new mechanism target for AD molecular therapy.",
"42529100": "ID: 42529100\nTitle: Emerging strategies for spinal cord injury repair: stem cells, extracellular vesicles, biomaterials, and neuromodulation.\nAbstract: Spinal cord injury (SCI) is a devastating condition that frequently results in permanent disability. Repair and functional recovery after SCI are hindered by the complexity of its pathophysiology and the inherent challenges of neural regeneration within the central nervous system. Despite decades of research aimed at elucidating its pathological mechanisms and developing effective strategies to promote axonal regeneration and circuit rewiring, successful therapeutic outcomes remain limited. Recent advances in bioactive materials and stem cell technology have shifted the research focus from solely stimulating corticospinal tract regeneration toward building intermediate neural networks to facilitate neural repair and circuit reconstruction. Concurrently, emerging technologies that modulate neural activity through physical modalities, such as electrical, magnetic, and ultrasonic stimulation, are rapidly evolving. Among these emerging strategies, several-including stem cell therapy, biomaterial-based interventions, and electromagnetic stimulation-have advanced to clinical trials, with some already entering clinical practice. This review summarizes the major contemporary strategies and research advances in SCI repair. It details the mechanisms and latest developments in stem cell and extracellular vesicle therapy, biomaterial applications, and neuromodulation techniques. Finally, it discusses future therapeutic directions and ongoing challenges in clinical translation.",
"42530044": "ID: 42530044\nTitle: Extracellular Vesicle-Mediated Delivery of VEGF and NGF Protects Dopaminergic Neurons in 6-OHDA-Induced Parkinson's Disease Models.\nAbstract: Parkinson's disease (PD) is a neurodegenerative disorder marked by motor dysfunction. No definitive methods exist to repair damaged neurons. Vascular endothelial growth factor (VEGF) and nerve growth factor (NGF) are two neuroprotective agents that work synergistically. However, these large molecular proteins have difficulty crossing the blood-brain barrier (BBB). Extracellular vesicles (EVs) offer superior targeting and low immunogenicity, making them excellent carriers. In this study we examined the protective effects of VEGF and NGF in a cell model and evaluated the therapeutic potential of VEGF-NGF contained within EVs in PD rats. EVs were isolated using sequential differential centrifugation and characterized using transmission electron microscopy, nanoparticle tracking analysis, and western blotting (WB). VEGF and NGF were loaded into the EVs using a saponin-assisted method to create VEGF@EVs, NGF@EVs, and VEGF/NGF@EVs. The viability of 6-hydroxydopamine hydrochloride (6-OHDA)-induced SH-SY5Y cells was measured using the cell counting kit-8 assay before and after treatment with VEGF and NGF. Autophagy levels were assessed using WB, and the role of autophagy was further explored using the autophagy inhibitor chloroquine. Unilateral PD rat models were established via stereotactic injection of 6-OHDA into male Sprague-Dawley rats. Behavioral changes were monitored before and after treatment. Neuronal recovery, neurotransmitter levels, and autophagy levels in the rat brains were evaluated using immunohistochemistry, enzyme-linked immunosorbent assay, and WB. VEGF/NGF@EVs significantly enhanced the viability of 6-OHDA-induced SH-SY5Y cells. A complete autophagic process was identified as essential for this protective effect. The intranasal administration of VEGF/NGF@EVs improved motor behavior in PD rats, with performance better than that of single growth factor treatments. The number of tyrosine hydroxylase (TH)-positive neurons, TH protein expression, and dopamine content were significantly increased. In addition, the level of autophagy in the rat substantia nigra was elevated. VEGF/NGF@EVs exert protective effects in both in vitro and in vivo 6-OHDA-induced PD models by promoting autophagy, demonstrating greater efficacy than either growth factor alone. By transplanting VEGF/NGF@EVs into PD rats, we showed that these vesicles can effectively cross the BBB and deliver targeted therapy to the central nervous system. This study highlights the significant potential of EV-mediated protein transplantation strategies for treating neurological disorders.",
"42533122": "ID: 42533122\nTitle: Light-activated tetanus neurotoxin for conditional proteolysis and inducible synaptic inhibition in vivo.\nAbstract: The light chain of tetanus neurotoxin (TeNT) is a metalloprotease that potently inhibits synaptic transmission by cleaving the endogenous vesicle fusion protein VAMP2, but its constitutive activity prevents spatiotemporal precision. To address this, we engineered light-activated TeNT (LATeNT) by inserting the light-sensitive LOV domain into an allosteric site and optimizing dynamic range via directed evolution. LATeNT's activity is undetectable in the dark, but the protease turns on after 10-20 min of weak blue-light exposure to potently inhibit synapses in vivo. Here we show that LATeNT works across multiple brain regions and at long-range axonal projections, with its effects reversible in 24\u2009h. LATeNT enabled us to discover a hippocampal interneuron population that regulates anxiety-like behaviors and demonstrate the importance of postsynaptic endocannabinoid exocytosis for depolarization-induced suppression of inhibition in vivo. Beyond neuroscience, LATeNT regulated endogenous insulin secretion from pancreatic beta cells and converted drug exposure, elevated Ca2+ or receptor activation into transgene expression or reporter secretion in HEK293T cells. With a large dynamic range, high light sensitivity and sustained effect, LATeNT enables versatile, spatiotemporally resolved proteolysis across diverse biological systems.",
"42539598": "ID: 42539598\nTitle: Botulinum toxin A for sialorrhea in adults with neurological disorders: a narrative review of aspiration pneumonia risk.\nAbstract: Aspiration pneumonia (AP), a serious condition resulting from the inhalation of foreign materials into the lungs, is a significant concern for patients with compromised swallowing mechanisms, especially those with neurological disorders. Sialorrhea (hypersalivation) exacerbates the risk of aspiration, and botulinum neurotoxin type A (BoNT-A) has been identified as a potential treatment to manage sialorrhea. However, the direct impact of BoNT-A on reducing AP risk remains inadequately explored, particularly in adult populations. BoNT-A works by inhibiting the release of acetylcholine at the neuromuscular junction, thereby reducing salivary gland stimulation and saliva production. This review aims to evaluate the current state of research on the use of BoNT-A for managing sialorrhea and its potential to reduce AP risk, with a specific focus on the adult population. A comprehensive review of existing literature was conducted, focusing on the efficacy of BoNT-A in treating sialorrhea in both pediatric and adult populations. Studies were assessed for their findings on the impact of BoNT-A on drooling and related complications, including the risk of AP. Meta-analyses and individual studies were reviewed to summarize the current knowledge and identify research gaps. In pediatric patients, BoNT-A has been shown to effectively reduce sialorrhea, with several studies and meta-analyses demonstrating its efficacy in managing drooling associated with neurological conditions such as cerebral palsy. However, evidence linking BoNT-A directly to reduced AP risk is limited. In adults, while BoNT-A has been beneficial in managing sialorrhea associated with conditions such as Parkinson's disease and brain injuries, studies specifically exploring its impact on AP risk are lacking. Current evidence supports the efficacy of BoNT-A in managing neurological sialorrhea across both adult and pediatric populations. However, a critical gap exists in studies directly investigation whether BoNT-A reduces AP risk in adults with neurological disease.",
"42543397": "ID: 42543397\nTitle: Autonomous intranasal delivery systems for central nervous system therapeutics.\nAbstract: Intranasal delivery provides a rapid, non-invasive route to the central nervous system, bypassing the blood-brain barrier and first-pass metabolism. However, its therapeutic potential remains constrained by the nasal cavity's complex anatomy, the restricted surface area and permeability of the olfactory epithelium, and short drug residence times. Recent advances in nanotechnology and synthetic biology have enabled the development of autonomous and programmable delivery systems that can target the olfactory epithelium, enhance brain entry and sustain therapeutic release. This review highlights current strategies for engineering intranasal drug delivery vectors that can replicate or extend cellular functions to enable autonomous nose-to-brain drug delivery. These vectors include: synthetic nanoparticles that mimic essential cellular activities and allow for modular surface modification; extracellular vesicles that naturally carry therapeutic cargo and exhibit parent-cell-derived tropism; and living therapeutics, such as engineered microbes, viruses or stem cells, that respond dynamically to host environments and can be genetically programmed for precise payload production. Emphasis is placed on the modular design of functional components, host-responsive interactions tailored to anatomical and physiological cues, and the integration of programmable functions that collectively drive delivery autonomy and therapeutic efficacy. Together, these advances position intranasal delivery as a versatile platform for treating neurological disorders, offering a foundation for future translational development.",
"42543528": "ID: 42543528\nTitle: Encapsulation and Controlled Release of Human Spinal Cord Organoid-Derived Extracellular Vesicles for Tissue Patterning in Viscoelastic Hyaluronic Acid Hydrogels.\nAbstract: Human induced pluripotent stem cells (hiPSCs) can differentiate into various types of central nervous system organoids which are valuable for applications in tissue engineering and injury repair. The secreted extracellular vesicles (EVs) of organoids, in particular the small-sized EV subset referred as exosomes (30-200\u00a0nm), have emerged as novel therapeutics in regenerative medicine. This study investigated the encapsulation and controlled release of human spinal cord organoid (hSCO)-derived EVs in viscoelastic hyaluronic acid (HA) hydrogels and assessed their impact on organoid patterning. A series of pH-responsive hydrogels were fabricated, leading to sustained EV release regulated by viscoelastic properties. The pH of these hydrogels decreased from 9 to 7 during incubation, which altered hydrogel viscoelasticity, thereby modulating EV release kinetics. In addition, EV-loaded hydrogels regulated key hSCO patterning markers such as DBX1 and ISL1. Furthermore, these EVs in hydrogels can cross a modeled blood-spinal cord barrier and provide cross-barrier capability for delivery. Taken together, the organoid-secreted EVs in viscoelastic HA hydrogels can be released at a controlled rate and have potential to regulate spinal cord organoid patterning. This study advances our knowledge of regulating intercellular communication and developing EV-based therapies for treating neurological disorders such as spinal cord injury.",
"42548572": "ID: 42548572\nTitle: Case Report: Munc13-4 deficiency presenting with autoimmune neuropathy years before FLH: implications for early genetic screening and HSCT timing.\nAbstract: The UNC13D gene encodes Munc13-4, a critical regulator of cytotoxic granule exocytosis in lymphocytes and the protein responsible for familial hemophagocytic lymphohistiocytosis (FHL). As an essential mediator of perforin release from cytotoxic T and NK cells, Munc13-4 ensures immune surveillance by facilitating granule-plasma membrane fusion. Defects in this pathway, termed \"perforinopathy\" manifest as impaired cytotoxicity and consequent immune dysregulation. While central nervous system involvement and systemic inflammation are established features of Munc13-4 deficiency, its association with autoimmune neuropathy remains largely unrecognized. We report two patients with a unique disease trajectory, presenting initially with autoimmune neuropathy before developing progressive FHL several years later. Both patients met the HLH-2004 diagnostic criteria, including fever, pancytopenia, hyperferritinemia, hypertriglyceridemia, and splenomegaly. Genetic analysis revealed heterozygous UNC13D mutations: in Case 1, a nonsense (c.3193C>T; p.Arg1065*) and a missense (c.1135G>A; p.Glu379Lys) mutation; in Case 2, compound heterozygous mutations comprising a maternal splicing variant (c.2447\u00a0+\u00a01G>A) and a paternal missense mutation (c.1241G>T; p.Arg414Leu). Case 1 underwent hematopoietic stem cell transplantation (HSCT), with significant recovery of weakness and numbness, and no recurrence of fever or lymphadenopathy at one year. Case 2 succumbed to disease progression after no timely, suitable allogeneic hematopoietic stem cell donor could be found. Our findings identify autoimmune neuropathy as a potential harbinger of FHL in perforinopathy, suggesting these neurological manifestations may be the initial phenotype of an underlying cytotoxic dysfunction. This temporal association mandates heightened clinical vigilance for immunodeficiency in patients with refractory neuropathy, particularly when accompanied by axonal degeneration or systemic inflammation. Recognizing this novel disease continuum has immediate diagnostic and therapeutic implications, as early genetic identification creates a critical window for potentially curative intervention with HSCT before irreversible multi-organ damage occurs. These cases expand the clinical spectrum of perforinopathy and challenge current diagnostic paradigms, emphasizing the need for an integrated neurological and immunological evaluation for unexplained autoimmune neuropathy.",
"42550091": "ID: 42550091\nTitle: Vitamin B12 Precatalyst Enables Molecular Cobalamin(II) Catalysis for Organodisulfide Anolytes in Aqueous Redox Flow Batteries.\nAbstract: Affordable and stable organodisulfide anolytes are attractive for long-duration aqueous redox flow batteries (ARFBs) for grid energy storage. However, the sluggish redox reactions involving the cleavage/formation of sulfur-sulfur (S\u2500S) bonds cause severe polarization and limited capacity utilization, thus hindering the practical application of organodisulfide anolytes in ARFBs. Herein, we report a homogeneous catalysis strategy employing hydroxocobalamin (Vitamin B12, OHCbl) as an environmentally benign and biocompatible precatalyst which can enable molecular cobalamin(II) catalysis for organodisulfide anolytes. Cobalamin(II) generated in situ by electrochemical reduction of OHCbl bidirectionally accelerates the redox reactions of S\u2500S bonds and significantly decreases the overpotential of the ARFB with SPS (bis(sodium sulfopropyl) disulfide) anolyte from 1.24\u00a0V to 0.36\u00a0V at 40\u00a0mA cm-2, boosting the energy efficiency from 18% to 66%. The OHCbl-catalyzed ARFB with 1.0\u00a0M SPS anolyte delivers a capacity of 51.45 Ah L-1 operated at 100% state of charge and remains stable for 850 cycles at 50\u00a0mA cm-2 with a low decay rate of 0.0189% per day. Moreover, a nearly saturated 1.3\u00a0M SPS-based ARFB achieves 96.2% capacity utilization, corresponding to 67.05 Ah L-1 delivered capacity. This cobalamin(II) catalysis strategy addresses the kinetic bottlenecks inherent to organodisulfide-based anolytes for ARFBs.",
"42550102": "ID: 42550102\nTitle: Achieving selective photochemistry on a metal surface by tuning the aromatic core.\nAbstract: On-surface photochemistry provides a non-thermal strategy that can circumvent the side reactions and structural defects associated with thermal activation, enabling enhanced control over surface reactions. Among these, Ullmann coupling serves as a representative on-surface reaction for fundamental studies of reaction mechanisms and controllability. However, how the aromatic core of molecular precursors governs photoinduced C-Br activation on metal surfaces remains insufficiently understood. In this work, we investigated photoinduced Ullmann coupling on Au(111) using three dibromo molecular precursors with distinct aromatic cores. The three precursors show markedly different photoreactivities under 405 and 532\u00a0nm irradiation. Time-dependent density functional theory calculations reveal a photoinduced surface-to-molecule charge-transfer process. Combined with calculations of potential energy surfaces for C-Br bond dissociations, these results provide a qualitative rationale for comparing the relative tendency of photo-triggered C-Br cleavage at the surface. The results establish a direct correlation between photo-selectivity and the molecular aromatic core, providing mechanistic insight into light-controlled on-surface synthesis and offering design principles for tailoring light-responsive precursors toward desired carbon nanostructures.",
"42550250": "ID: 42550250\nTitle: Psychological and neurological benefits of physical activity: impact on mental health, Parkinson's disease, and Alzheimer's disease.\nAbstract: This review synthesizes current evidence on the psychological and neurological benefits of physical activity, with emphasis on mental health, Parkinson's disease (PD), and Alzheimer's disease (AD). Physical inactivity is increasingly recognized as a modifiable risk factor that may exacerbate neuroinflammatory and metabolic dysfunctions associated with these conditions. Structured exercise has been shown to activate muscle-brain signaling pathways, including neurotrophic factors (BDNF, IGF-1, VEGF), immune modulation (IL-6, IL-10), metabolic regulators (PGC-1\u03b1, SIRT1), and peripheral mediators such as myokines and gut-brain axis components. For mental health, exercise is associated with reductions in depressive, anxiety, and stress-related symptoms, potentially through HPA-axis recalibration, neurotransmitter remodeling (serotonin, dopamine), and enhanced endocannabinoid signaling. These changes are supported by fMRI and EEG studies showing improved prefrontal-limbic connectivity and cognitive resilience. In PD, exercise interventions are linked to improvements in motor control, balance, and mood, likely mediated by dopaminergic integrity, neurotrophic support, and anti-inflammatory effects, with multimodal programs (aerobic, resistance, and dance-based) often demonstrating superior outcomes. In AD, mid-life physical activity has been associated with reduced dementia risk in epidemiological studies; proposed mechanisms include enhanced amyloid/tau clearance, glymphatic function, and synaptic adaptation, with some RCTs reporting hippocampal volume preservation and improved network connectivity. Shared mechanisms across conditions include neurotrophic upregulation and anti-inflammatory effects, whereas distinct pathways involve dopaminergic circuit remodeling in PD and glymphatic facilitation in AD. Clinical translation through FITT principles (frequency, intensity, time, type) requires personalized prescriptions, strong adherence strategies, and multidisciplinary integration. Despite methodological heterogeneity and limitations in the existing trials, structured physical activity emerges as a scalable, non-pharmacological intervention with potential for prevention, symptom management, and neuroprotection. Larger, well-designed studies are needed to optimize its application and clarify long-term disease-modifying effects.",
"42550282": "ID: 42550282\nTitle: Ultrasensitive electrochemiluminescence determination of Salmonella based on CRISPR/Cas12a integrated with bimetallic semiconductive metal-organic frameworks.\nAbstract: An ultrasensitive electrochemiluminescence (ECL) biosensor was established by combining CRISPR/Cas12a technique and semiconductive bimetallic-organic framework (scMOF) [[CuxNi3-x(HITP)2] (HITP\u2009=\u20092,3,6,7,10,11-hexaiminotriphenylene)]] emitter and employed to detect Salmonella using the allosteric probe as the recognition component. Given that CuxNi3-x(HITP)2 has demonstrated large specific surface area, both in-plane and out-of-plane charge transfer ability, narrowed band gap, and enhanced separation of holes and electrons, it can be simultaneously employed as the superior ECL emitter and bioplatform for anchoring single-strand DNA (ssDNA), thus improving the detection sensitivity toward Salmonella. The CRISPR/Cas12a-based system can specifically recognize the target sequence of Salmonella and activate the nuclease activity of Cas12a, and the activated Cas12a possesses trans-cleavage ability toward ssDNA. The CuxNi3-x(HITP)2 emitter is then released, resulting in the decline of the ECL response. The developed CuxNi3-x(HITP)2-CRISPR/Cas12a-based ECL biosensor exhibits the ultralow detection limit of 0.25 CFU mL-\u20091 in the linear range from 1.0 CFU mL-\u20091 to 106 CFU mL-\u20091, significantly lower than those of reported ones. Furthermore, the developed biosensor exhibits outstanding overall biosensing properties with high selectivity, favorable reproducibility and stability, together with promising practical applicability for the determination of Salmonella in a variety of foodstuffs.",
"42550314": "ID: 42550314\nTitle: The role of the HB-EGF-regulated EGFR-mitophagy axis in maintaining cartilage homeostasis and osteoarthritis.\nAbstract: This study investigates the mechanism by which heparin-binding epidermal growth factor (HB-EGF) maintains cartilage homeostasis in osteoarthritis (OA) through the epidermal growth factor receptor (EGFR)-mitochondrial autophagy axis. Given the challenges in OA treatment and the critical role of impaired mitochondrial autophagy in disease progression, this research first identified key regulators using transcriptomic data from human OA cartilage and GEO databases. An in vitro OA model was established via IL-1\u03b2 induction in chondrocytes, through which HB-EGF was found to reverse the IL-1\u03b2-induced metabolic imbalance by downregulating catabolic markers (Mmp13, Adamts5) and upregulating anabolic markers (Acan, Col2a1). Further analysis revealed that HB-EGF enhanced mitophagy, as indicated by increased levels of Pink1, Parkin, and Lc3-II alongside decreased P62. These protective effects were attenuated by the mitophagy inhibitor Mdivi-1, confirming the dependence on this pathway. In conclusion, HB-EGF activates EGFR signaling to promote PINK1/ PARKIN-mediated mitophagy, which facilitates the clearance of damaged mitochondria and improves mitochondrial function, thereby restoring chondrocyte metabolic balance and delaying OA progression, suggesting the EGFR-mitophagy axis as a potential therapeutic target for OA.",
"42550325": "ID: 42550325\nTitle: Co-sedimentation is the key to the structural investigation of wild-type FAT10.\nAbstract: Under inflammatory conditions, the ubiquitin-like modifier FAT10 serves as a tag for protein degradation by the 26S proteasome. FAT10 is degraded along with its substrates and this process is independent of the segregase VCP/p97, which, in the regular ubiquitin pathway of degradation, is required if a substrate lacks a disordered initiation region. FAT10 itself is loosely folded and its tendency to aggregate has complicated investigations of its structure, interaction, and function. Recently, hydrogen-deuterium exchange in combination with mass spectrometry has suggested that, in preparation of degradation by the proteasome, the adapter protein NUB1 traps FAT10 in a mostly unfolded state by capturing a \u03b2-strand. \u03b2-strand capture was subsequently confirmed by magic-angle spinning (MAS) NMR spectroscopy of a stabilized variant of the N-domain of FAT10 in complex with NUB1L, the longer splice variant of NUB1. MAS NMR, in addition, revealed that the N-domain of FAT10 and NUB1L form a fuzzy complex and that the N-terminus of FAT10 is positioned for initiation of degradation by specific non-covalent interaction with NUB1L. Here, we report the investigation of the wild-type N-domain of FAT10 by MAS NMR. Co-sedimentation with NUB1L yields high-quality spectra, which enable sequential assignment of resonances. Based on MAS NMR data, the complexes of the wild-type and stabilized N-domain of FAT10 with NUB1L appear identical. The N-terminal residue of FAT10 again shows up prominently in the spectra, even though it is this time an Ala, not a Gly. Our experiences suggest that co-sedimentation in combination with MAS NMR is generally helpful in the exploration of conditional folds of intrinsically disordered proteins.",
"42550591": "ID: 42550591\nTitle: Analysis of cancer mutations introduced into the Drosophila melanogaster Notch negative regulatory region uncovers a diversity of regulatory outcomes.\nAbstract: Activating mutations of Notch are drivers of the blood cell cancer, T-ALL, and some solid tumours. The negative regulatory region (NRR) of the extracellular domain (ECD) and the PEST region of the intracellular domain (ICD) are mutation hot spots which can act synergistically in T-ALL. The NRR, comprised of a heterodimerisation domain (HD) and three Lin12/Notch repeats (LNR A-C), masks the S2 cleavage site, normally only exposed following ligand binding and cleaved as the first step that ultimately leads to ICD release. Drosophila mutants have played a key role in analysing Notch structure/function, but there have been few mutational studies of the NRR. Here, we expressed, in S2 cells, over 20 cancer mutations located in the HD, LNR and LNR/HD interface, introduced into Drosophila Notch. Mutations in the HD domain core did not activate, likely due to absence in Drosophila of an S1 cleavage within the HD required for mammalian Notch activity. In contrast, mutations in the LNR/HD interface behaved similarly to T-ALL, activating constitutively with no further ligand induction and were synergistic with PEST deletion. Mutations of surface-exposed residues of LNR-C also activated constitutively but remained inducible both by ligand and by an intracellular endocytic regulator, Deltex, and were not synergistic with PEST deletions. These mutations caused elevated Notch levels and decreased turnover, suggesting a novel regulatory mechanism. Our results, therefore, uncover a variety of outcomes arising from perturbations of the NRR and will facilitate the establishment of Drosophila cancer models and the development of mutant-specific approaches to effective therapies.",
"42550860": "ID: 42550860\nTitle: Carboxymethyl cellulose reinforced graphene oxide-MoS2 for photocatalytic degradation of perfluorooctanoic acid (PFOA): process optimization and mechanism.\nAbstract: Perfluorooctanoic acid (PFOA), remains a major environmental challenge due to its exceptional chemical stability, resistance to conventional treatment technologies. In this study, a sustainable carboxymethyl cellulose-reinforced graphene oxide/molybdenum disulfide hydrogel (MoS\u2082/GO/CMC) was developed as a visible-light-responsive photocatalyst for PFOA degradation under low-power LED irradiation. The photocatalytic process was optimized using Response Surface Methodology based on a Box-Behnken Design, evaluating the effects of irradiation power (7-12 W), catalyst dosage (0-0.04\u2005g), and initial PFOA concentration (10-50\u2005mg L-\u00b9). The optimized conditions (12 W, 0.04\u2005g catalyst, and 30\u2005mg L-\u00b9 PFOA) achieved 99.8% degradation efficiency, with the developed quadratic model exhibiting excellent predictive capability (R\u00b2\u2009=\u20090.9974). Characterization results confirmed integration of MoS\u2082 and GO within the CMC matrix, producing a heterostructure with a narrow band gap of 1.12\u2005eV, enhanced charge separation, and suppressed electron-hole recombination. Kinetic analysis revealed pseudo-first-order degradation behaviour with a rate constant of 0.013\u2005min-\u00b9 and a half-life of 0.89\u2005h. Radical scavenging experiments identified superoxide radicals (O\u2082\u2022-) as the dominant reactive species governing PFOA degradation. LC-MS analysis confirmed a stepwise chain-shortening degradation pathway involving intermediates such as PFHpA, PFPeA, and PFBA, indicating progressive defluorination and carbon-carbon bond cleavage. The hydrogel demonstrated excellent stability, retaining over 96% of its initial activity after seven cycles. An Electrical Energy per Order (EEO) value of 444.6 kWh m-\u00b3 order-\u00b9 highlights the feasibility of low-energy operation. These findings demonstrate that MoS\u2082/GO/CMC hydrogels offer an environmentally benign, recoverable, and energy-efficient for PFAS remediation.",
"42550868": "ID: 42550868\nTitle: Bioinformatics and experimental studies of atherosclerosis-related endothelial dysfunction genes in renal calculi and exploration of their molecular mechanisms.\nAbstract: Patients with kidney stones (KS) often have an increased risk of atherosclerosis (AS). Because endothelial dysfunction (ED) is closely associated with AS, its role in KS remains unclear. This study aimed to examine the roles and mechanisms of AS-related ED genes in KS. Three datasets (GSE73680, GSE117518, and GSE132651) were analyzed. Differential expression analysis was conducted to identify differentially expressed genes (DEGs). To identify potential biomarkers, least absolute shrinkage and selection operator (LASSO) regression analysis and expression validation were conducted. Further analyses including GeneMANIA, gene set enrichment analysis (GSEA), examination of biomarkers within immune cells and subcellular localization analysis, molecular regulatory network analysis, tissue specificity analysis, and competing endogenous (ceRNA) network analysis were employed to comprehensively explore the functions and regulatory mechanisms of the identified biomarkers. Moreover, drug prediction analysis was conducted. Finally, reverse transcription quantitative polymerase chain reaction (RT-qPCR) was proceeded to verify the expression levels of the biomarkers. A total of 22 DEGs associated with KS and AS were identified. Lasso regression selected 4 candidate biomarkers (MMP10, UCHL1, NEK2, and HEY1), among which UCHL1 and NEK2 were validated as key biomarkers. GeneMANIA and GSEA analyses uncovered the potential involvement of these biomarkers in cell adhesion molecules, focal adhesion, and lysosome pathways. Analysis of immune cells and subcellular localization provided insight into the biological functions and intracellular distribution of the biomarkers. Transcription factor regulatory network and ceRNA network analyses elucidated potential upstream regulatory mechanisms. Drug prediction analysis identified 17 potential drugs, including pazopanib and palbociclib, that may target NEK2. RT-qPCR demonstrated that NEK2 was significantly overexpressed in KS samples. This study identified biomarkers associated with KS and AS and comprehensively analyzed their molecular regulatory networks. These findings provide novel understandings of the molecular mechanism underlying KS and lay the foundation for future personalized treatment and drug development.",
"42551134": "ID: 42551134\nTitle: Epigenetic strategies for fetal hemoglobin induction in sickle cell disease.\nAbstract: Sickle cell disease (SCD) is caused by pathogenic variants in the \u03b2-globin gene (HBB), most commonly the variant responsible for hemoglobin S, and affects an estimated 515,000 newborns each year, with the highest burden occurring in sub-Saharan Africa. Gene editing and hematopoietic stem cell transplantation have changed the therapeutic landscape, but their cost, technical complexity, and procedure-related risks still limit their wider use. For this reason, pharmacological induction of fetal hemoglobin (HbF) remains an important therapeutic strategy. HbF reduces HbS polymerization and is associated with lower disease severity, morbidity, and mortality. Among the mechanisms involved in \u03b3-globin silencing, epigenetic regulation offers several targets that can be explored using small molecules. This review discusses medicinal chemistry approaches primarily targeting HDAC1/2, LSD1, and DNMT1, with emphasis on inhibitor classes, binding mechanisms, structural features, preclinical evidence, and translational limitations. The available data show that each target presents a distinct set of challenges. HDAC-directed strategies require improved isoform and cellular selectivity; LSD1 inhibitors must reconcile strong HbF induction with the risks associated with prolonged target engagement; and DNMT1 modulation is moving from DNA-incorporating nucleoside analogs toward reversible non-nucleoside inhibitors. We also discuss emerging approaches, including multi-target epigenetic modulation and targeted protein degradation. Together, these strategies show how a better understanding of \u03b3-globin repression may guide the development of safer and more accessible HbF-inducing agents.",
"42551354": "ID: 42551354\nTitle: A lysosome-targeted and polarity-responsive photosensitizer for tumor specific imaging and photodynamic therapy.\nAbstract: Photodynamic therapy (PDT) represents an effective and promising strategy for cancer treatment. Nevertheless, the poor targeting specificity of conventional photosensitizers has severely impeded its clinical application. Consequently, the development of novel photosensitizers that integrate imaging-guided diagnosis and synergistic phototherapeutic efficacy is highly desirable to improve PDT performance. In this work, we designed and synthesized a lysosome-targeted photosensitizer HBT-CUR with D-\u03c0-A feature based on 2-(2'-hydroxyphenyl) benzothiazole (HBT) and difluoroborate moieties for tumor image-guided PDT. HBT-CUR exhibits near-infrared (NIR) fluorescence emission, along with high polarity sensitivity, excellent stability, high lysosome-targeting specificity and good biocompatibility. By exploiting the distinct polarity differences between normal and tumor cells, HBT-CUR enables specific NIR fluorescence imaging of tumor tissues, with a fluorescence intensity \u223c6-fold higher than that in normal tissues. Moreover, HBT-CUR exhibits a fast in vivo response (10\u00a0min) and prolonged duration (6\u00a0h with robust fluorescence emission). More importantly, HBT-CUR can efficiently generate singlet oxygen under light irradiation (\u03a6\u0394\u00a0=\u00a00.70 in dioxane), thereby achieving effective photodynamic tumor ablation in MCF-7 tumor-bearing mouse models. Density functional theory (DFT) calculations reveal that the synergistic interplay between excited-state intramolecular proton transfer (ESIPT) and intramolecular charge transfer (ICT) processes contributes to the superior luminescence performance of HBT-CUR. This work presents a promising phototheranostic agent for tumor-specific image-guided photodynamic therapy, offering a new strategy for the rational design and development of tumor PDT systems.",
"42551360": "ID: 42551360\nTitle: Perfluorooctanoic acid disrupts lysine metabolism and autophagy to promote white spot syndrome virus infection in shrimp.\nAbstract: Perfluorooctanoic acid (PFOA), a globally distributed per- and polyfluoroalkyl substance (PFAS), is increasingly recognized for its immunotoxic and metabolic effects in aquatic organisms; however, its role in viral disease susceptibility remains poorly understood. Using Pacific white shrimp (Penaeus vannamei) and white spot syndrome virus (WSSV) as a model system, we investigated the effects of chronic PFOA exposure on antiviral immunity and viral transmission. Chronic exposure to environmentally relevant concentrations of PFOA resulted in significant accumulation of PFOA in the hepatopancreas and markedly increased WSSV replication, host mortality, and horizontal transmission. Integrated metabolomic and transcriptomic analyses identified lysine degradation as a consistently affected metabolic pathway, along with significant enrichment of lysosome-related pathways. Biophysical assays, including molecular docking, biolayer interferometry, and cellular thermal shift analysis, demonstrated that PFOA directly binds to the lysine-catabolizing enzyme aminoadipate semialdehyde synthase (AASS), which may contribute to lysine accumulation. Importantly, enzymatic activity assays further revealed that PFOA significantly inhibited both lysine-ketoglutarate reductase (LKR) and saccharopine dehydrogenase (SDH) activities, accompanied by marked lysine accumulation in shrimp hepatopancreas. PFOA exposure alone induced basal autophagic dysfunction, as evidenced by reduced LC3/LAMP1 co-localization, decreased LC3 abundance, increased p62 accumulation, and impaired autophagosome-lysosome fusion. Dietary lysine supplementation reproduced these autophagic defects and significantly promoted WSSV replication, supporting a role for lysine accumulation in mediating impaired antiviral defense. Together, this study uncovers a previously unrecognized lysine-autophagy-lysosome regulatory axis through which PFOA enhances viral susceptibility and transmission in shrimp, providing mechanistic insight into pollutant-driven disease risk in aquaculture ecosystems.",
"42551379": "ID: 42551379\nTitle: Exploring the reductive transformation pathways of perfluoroalkyl Sulfonylimides to legacy perfluoroalkyl sulfonates and perfluoroalkyl sulfonamide compounds.\nAbstract: Bis(perfluoroalkylsulfonyl)imides (NH(CnF2n+1O2S)2) have been increasingly adopted as alternatives to legacy PFAS in industrial applications; however, their fate during water treatment remains poorly understood. Herein, we systematically investigated the reductive transformation of three sulfonyl imides (i.e., NH(C4F9O2S)2, NH(C2F5O2S)2, and NH(CF3O2S)2) under a UV/sulfite-based advanced reduction process (ARP). Bis(perfluoroalkylsulfonyl)imides exhibited high resistance to both direct photolysis and oxidative degradation, as evidenced by negligible degradation (<7%) and nearly no defluorination. In contrast, the UV/sulfite system achieved complete removal of bis(nonafluorobutanesulfonyl)imide within 24\u202fh, with defluorination efficiencies of up to 46.4%. Notably, LC-MS and LC-Q-Orbitrap HRMS analyses revealed substantial formation of legacy PFAS, including perfluoroalkyl sulfonamides (CnHF2n+1NO2S-), perfluoroalkyl sulfonates (CnF2n+1SO3-), which accounted for 5.0-10.3% and 12.9-48.5% of total fluorine, respectively. Density functional theory (DFT) calculations corroborated these findings, demonstrating that their secondary formation through reductive cleavage of the sulfonyl imide linkage is both thermodynamically favorable and kinetically accessible. However, short-chain perfluoroalkyl sulfonamides and perfluoroalkyl sulfonates exhibited substantially lower reactivity toward eaq\u207b than their parent compound, thereby limiting overall defluorination. Complete degradation pathways for bis(perfluoroalkylsulfonyl)imides with n\u202f=\u202f1, 2, and 4 were proposed based on identified TPs and DFT calculations. This study provides the first evidence that bis(perfluoroalkylsulfonyl)imides serve as precursors to short- and ultrashort-chain sulfonamides and sulfonates during reductive treatment, highlighting the potential for fluorine mass redistribution into persistent legacy PFAS during ARP-based remediation of emerging novel PFASs.",
"42551393": "ID: 42551393\nTitle: A hairpin conformational change-mediated SRH cascade isothermal amplification assisted electrochemical biosensor for Listeria monocytogenes detection.\nAbstract: The rapid and sensitive detection of foodborne pathogens is crucial for safeguarding food safety and public health. In this study, we developed an electrochemical biosensor based on a hairpin conformational change-mediated cascade isothermal amplification strategy for the detection of Listeria monocytogenes in food. The biosensor employs aptamers for specific recognition of the target bacterium. In combination with magnetic separation technology, the concentration of the target bacterium is converted into that of a trigger strand. The trigger strand induces a conformational change in the hairpin structure, thereby initiating a cascade amplification reaction involving strand displacement amplification, rolling circle amplification, and hybridisation chain reaction (SDA-RCA-HCR, SRH), enabling an exponential signal amplification and a significant reduction of the background signals caused by self-cleavage. The proposed biosensor demonstrates a linear detection range of 15 to 1.5\u202f\u00d7\u202f108\u202fCFU/mL for Listeria monocytogenes, with a detection limit as low as 5\u202fCFU/mL (S/N\u202f=\u202f3). It exhibits great specificity for the target bacterium, effectively distinguishing it from other common foodborne pathogens, and shows satisfactory performance in real food samples, including lettuce, cheese, and chicken. Thus, this work provides an efficient, reliable, and sensitive new method for the rapid detection of foodborne pathogens in complex food matrices.",
"42551398": "ID: 42551398\nTitle: Stability of Chlamydia pecorum MOMP at 37\u00a0\u00b0C: suitability for use in delayed-release vaccine implants in koalas (Phascolarctos cinereus).\nAbstract: The koala (Phascolarctos cinereus) is threatened by Chlamydia pecorum, an obligate intracellular bacterium that causes disease and infertility. Vaccination offers an alternative to antibiotics, and we have developed a safe, immunogenic vaccine that has shown promising results in captive and wild koala populations. However, the current vaccine requires a booster dose to achieve optimal immune protection. A delayed-release implant delivering the booster after primary vaccination could remove the need to recapture wild koalas. For this strategy to succeed, the antigen must remain stable at 37\u00a0\u00b0C until release. In this study we have assessed the stability and antigenicity of the Chlamydia pecorum major outer membrane protein (MOMP) antigens used in the vaccine after six weeks at 37\u00a0\u00b0C. In vitro analysis showed that aged MOMP induced antibody responses comparable to unaged antigens. These findings indicate that MOMP retains structural integrity and antigenicity, supporting its suitability for delayed-release vaccine delivery systems.",
"42551403": "ID: 42551403\nTitle: Ultrasound/electrostatic field combined with \u03b5-polylysine for improving the quality of ready-to-eat pork jelly and freshness classification based on convolutional neural network.\nAbstract: This study investigated the effects of ultrasound (US, 300\u00a0W, 20\u00a0kHz, 20\u00a0min) and electrostatic field (EF, 3.5\u00a0kV/cm, 60\u00a0min) combined with \u03b5-polylysine (PL, 0.5%, w/w) on the quality of ready-to-eat pork jelly (Shuijingyaorou, SJYR) during 40-day storage at 4\u00a0\u00b0C. Physicochemical properties such as lipid oxidation, total plate count, texture, flavor, and moisture distribution were evaluated. The results showed that the combined treatment (US + EF\u00a0+\u00a0\u03b5-polylysine) exhibited the strongest preservation effect for SJYR, significantly inhibiting lipid oxidation, protein degradation, pH increase, and microbial growth, meanwhile maintaining texture and flavor of SJYR, resulting in the shelf-life extended approximately 10\u00a0days compared with the blank control group. Furthermore, four convolutional neural network (CNN) models including Xception, ResNet-50, MobileNet, and GhostNet were developed for SJYR freshness classification. Xception model achieved the highest test accuracy (83.05% mean precision), while GhostNet model showed promising accuracy (77.48%) with lower computational cost, making it suitable for mobile applications. This study provides a feasible strategy for improving SJYR shelf-life and rapid freshness identification.",
"42551559": "ID: 42551559\nTitle: CRISPR-Cas Systems as Precision Antimicrobials: Reversing the Tide of Antimicrobial Resistance.\nAbstract: Antimicrobial resistance (AMR) has escalated into a global health crisis, with resistant pathogens causing over 1.2 million direct deaths annually and threatening to render modern medicine unsustainable. This review provides a comprehensive and updated synthesis of CRISPR-Cas-based antimicrobial strategies with a unique focus on: (i) critical comparison with conventional antibiotics and emerging alternatives; (ii) quantitative evaluation of delivery platforms; (iii) novel strategies including AI-optimized guide design and the ATTACK-CreTA system; (iv) comprehensive analysis of ecological risks; and (v) technology readiness level assessments for clinical translation. The CRISPR-Cas system, originally discovered as a bacterial adaptive immune mechanism, has been repurposed as a programmable precision tool to combat AMR by selectively targeting and eliminating resistance genes. We systematically evaluate the mechanistic diversity of Cas effectors, from DNA-cleaving Cas9 and Cas3 to RNA-targeting Cas13, and their application in reversing resistance phenotypes in WHO priority pathogens. We critically assess emerging delivery platforms, including engineered bacteriophages, conjugative plasmids, nanoparticles, and outer membrane vesicles, quantitatively comparing their delivery efficiency, payload capacity, and biosafety profiles. Novel strategies such as CRISPR interference (CRISPRi) for gene silencing without genomic cleavage, the ATTACK-CreTA system for enhanced bactericidal activity, and AI-driven optimization of guide RNA design are examined with appropriate caveats. We comprehensively address clinical translation challenges including immunogenicity, pharmacokinetics/pharmacodynamics, manufacturing scalability, regulatory pathways, and bacterial resistance mechanisms including anti-CRISPR proteins. No CRISPR-based antimicrobial has yet received regulatory approval, and we critically evaluate the gap between proof-of-concept and clinical utility. A detailed roadmap for clinical development is proposed. By integrating recent advances in Cas protein engineering, delivery technologies, and diagnostic applications, this review positions CRISPR-Cas systems as next-generation precision therapeutics capable of both treating resistant infections and curtailing the spread of AMR across clinical and environmental settings.",
"42551610": "ID: 42551610\nTitle: A Strain of High Cell-Adapted Porcine Deltacoronavirus: Isolation, Genetic Evolution, and Pathogenicity Analysis.\nAbstract: In 2024, an outbreak of piglet diarrhea occurred in Guangxi Zhuang Autonomous Region, China. RT-PCR assay of clinical samples confirmed that porcine deltacoronavirus (PDCoV) was the primary pathogen. In this study, the PDCoV GX/2024 strain was successfully isolated in PK-15 cells with optimized trypsin treatment conditions. The cytopathic effect (CPE) of this strain gradually intensified during serial passages. After plaque purification, the strain achieved stable proliferation with high titer, reaching a peak titer of 107\u00b72 TCID50/mL, which indicated its potential as an inactivated vaccine candidate. Phylogenetic analysis demonstrated that the strain belonged to the Chinese lineage, with nucleotide similarity ranging from 97.89% to 99.76% compared with other domestic and international strains. Analysis of the Spike protein (S protein) revealed that the strain harbored critical amino acid substitutions in the S1 domain and the S1/S2 cleavage site, which led to conformational changes in the protein surface and might be associated with viral infection efficiency. Animal experiments verified that PDCoV GX/2024 exhibited strong pathogenicity in newborn piglets. Diarrhea, vomiting and other symptoms appeared within 24 hours post-infection. The virus mainly colonized the epithelial cells at the tip of intestinal villi and caused severe pathological damage. This study provides important theoretical basis for the genetic evolution, pathogenic mechanism and transmission potential of PDCoV, and offers reference value for disease prevention and control as well as vaccine development.",
"42551861": "ID: 42551861\nTitle: Cadaveric Feasibility Study Describing Transarticular Drilling of the Canine Anconeal Process through a Cranial Arthroscopic Portal.\nAbstract: The aims of this study were to describe a cranial arthroscopic portal to the supratrochlear foramen and to assess the feasibility and accuracy of retrograde transarticular drilling of the anconeal process in canine cadaveric elbows. Cadaver elbows (n\u2009=\u20098) were instrumented through a cranial portal in the distal humerus using a NanoScope. The scope was manoeuvred through the supratrochlear foramen to inspect the anconeal process. A 2.0-mm tunnel simulating the pilot hole for an ununited anconeal process repair was performed using a C guide, cannulated drill bits and NanoScope guidance. Experimental measures included arthroscopic anconeal process identification, surgical time, computed tomography and gross dissection to assess for critical structure damage and drill tunnel aperture location and trajectory. The first two elbows were used for technique development; the remaining six elbows have reported outcome measures. Mean surgery time was 16.6 (standard deviation 3.0) minutes. All six tunnels drilled had acceptable trajectories based on computed tomography and gross dissection. Major damage to critical structures was not observed in any specimen; however, tunnels were directly adjacent to transcubital vessels and a musculocutaneous nerve branch. This technique was feasible, provided visualization of the anconeal process, and allowed accurate drilling of the anconeal process in a cadaveric model. Clinical feasibility and safety require validation in a clinical study.",
"42551925": "ID: 42551925\nTitle: [Development of Therapies Targeting Non-G12C KRAS Mutations (Focusing on G12D and Pan-RAS)].\nAbstract: Sotorasib and adagrasib, which are KRAS G12C inhibitors, have shown favorable clinical results mainly in non-small cell lung cancer (NSCLC) and have already been introduced into clinical practice. However, for patients with non-G12C mutations that account for the majority, such as G12D, G12V, and G13D, effective targeted therapies have still not been established. Setidegrasib (ASP3082) is a first-in-class proteolysis-targeting chimera molecule (PROTAC) that targets the KRAS G12D mutation. Phase \u2160 trial targeting previously treated patients with advanced solid tumors harboring KRAS G12D mutations has been initiated, and the latest results were reported in the N Engl J Med in March 2026. Daraxonrasib (RMC-6236) is a pan-RAS inhibitor that targets all ON-state RAS (KRAS, NRAS, HRAS) by forming a specific pocket spanning the Switch \u2160and Switch \u2161 regions of KRAS through a mechanism called tri-complex/molecular glue. Currently, multiple international collaborative phase \u2162 trials on RMC-6236, mainly focusing on PDAC and NSCLC, are underway. The development of new drugs targeting molecules other than KRAS G12C is progressing rapidly, and in KRAS mutation-positive solid cancers where prognosis was difficult to improve with conventional chemotherapy, there is a high possibility that the existing standard treatments will be significantly revised.",
"42551926": "ID: 42551926\nTitle: [Targeting KRAS-Mutant Cancers-Therapeutic Strategies and Mechanisms of Resistance to RAS Inhibitors].\nAbstract: KRAS mutations represent one of the most prevalent oncogenic alterations, occurring in approximately 20% of human cancers. Although KRAS had long been regarded as\"undruggable,\"the identification of a druggable pocket has catalyzed rapid progress in the development of targeted therapeutics. To date, the only approved KRAS-directed agent in Japan is sotorasib, a GDP-state inhibitor specific for the KRAS G12C mutation. However, a diverse array of next-generation strategies-including pan-KRAS inhibitors, pan-RAS inhibitors, tri-complex inhibitors, and targeted protein degradation modalities-are currently under active investigation. These advances have drawn increasing attention to the functional heterogeneity and differential druggability among distinct KRAS mutation subtypes, as well as organ-specific biological differences that had previously received limited consideration. Moreover, as with other molecularly targeted therapies, multiple resistance mechanisms to KRAS inhibition have been reported, underscoring the need for therapeutic strategies capable of overcoming or preventing treatment resistance.",
"42552004": "ID: 42552004\nTitle: Multicentre randomised comparative effectiveness trial of cleavage-stage versus blastocyst-stage embryo transfer in patients with few embryos (PRECISE): implementation of a study protocol in reproductive medicine.\nAbstract: With improvements in in vitro culture techniques, there has been a steady shift in practice to transfer in vitro fertilisation (IVF) embryos at the blastocyst-stage to increase pregnancy rates per embryo transfer (ET) and reduce the risk of multiple pregnancy. In patients with fewer embryos (\u22645 zygotes), fresh cleavage-stage ET is still commonly performed to reduce the risk of cycle cancellation if no embryo survives to the blastocyst-stage in vitro. However, evidence for improved outcomes with cleavage-stage embryo transfer in this subgroup of patients is lacking. We will compare blastocyst-stage to cleavage-stage ET with regard to cumulative live birth rates (CLBR) per oocyte retrieval and other patient-centred cycle outcomes in patients with \u22645 zygotes. Multicentre, randomised pragmatic comparative effectiveness trial with superiority design comparing blastocyst-stage to cleavage-stage ET in patients with \u22645 zygotes. This trial will enrol and randomise 1126\u2009women aged 18-44 undergoing IVF to either fresh cleavage-stage or blastocyst-stage ET at eleven IVF centres. Surplus embryos will be vitrified at the blastocyst-stage. The primary outcome is the CLBR per oocyte retrieval. Secondary outcomes include: multiple pregnancy, pregnancy loss <20 weeks and cycle cancellation. Exploratory outcomes include: live birth per fresh ET, ectopic pregnancy, time to pregnancy, stillbirth, preterm birth, financial burden, patient distress and facilitators and barriers for patient involvement and implementation. All primary analyses will be conducted according to the intent-to-treat principle. We also will report per-protocol analyses as a sensitivity analysis. A multidisciplinary stakeholder team, including a 5-patient stakeholder panel, will participate in study design, conduct and oversight. The Institutional Review Board at Beth Israel Deaconess Medical Center (BIDMC) provides central regulatory oversight. Results will inform timing of embryo transfer in patients with \u22645 zygotes and will be disseminated through academic publications, meeting presentations and communications to advocacy groups and patient participants. NCT06746129.",
"42552063": "ID: 42552063\nTitle: Recent Advances in Autophagy and Immunotherapy for the Clearance of Aggregated \u03b1-Synuclein in Parkinson's Disease.\nAbstract: Parkinson's disease is a neurodegenerative condition characterized by the accumulation of misfolded and aggregated \u03b1-synuclein in Lewy bodies and neurites. These protein aggregates contribute to neurodegeneration and motor symptoms such as bradykinesia, rigidity, and tremor. While the ubiquitin-proteasome system degrades soluble \u03b1-synuclein, aggregated and oligomeric forms are primarily cleared via the autophagy-lysosomal pathway. Mutations of the SNCA gene exacerbate \u03b1-synuclein aggregation and significantly impair its clearance, highlighting the importance of targeting toxic \u03b1-synuclein species. Strategies such as promoting autophagosome formation via 5'-AMP-activated protein kinase (AMPK) and mechanistic target of rapamycin complex 1 (mTORC1) or facilitating autophagosome maturation via RAB7-a member of the RAS oncogene family-and related effectors, have shown promise in enhancing autophagy and reducing \u03b1-synuclein pathology. Pharmacological agents such as rapamycin, trehalose, and nilotinib have demonstrated preclinical efficacy in enhancing \u03b1-synuclein clearance and alleviating disease features. Concurrently, immunotherapy approaches, including passive and active immunization, aim to enhance the immune system's ability to recognize and eliminate toxic \u03b1-synuclein species. Emerging strategies such as peptide-based therapies aim to inhibit aggregation or promote degradation of \u03b1-synuclein. At the same time, nanotechnology enables the targeted delivery of therapeutic agents across the blood-brain barrier with improved efficiency. Additionally, novel AUTOTAC (autophagy-targeting chimera) platforms offer a precision strategy to tag \u03b1-synuclein for autophagic degradation. This review explores many advances in autophagy-mediated aggregated \u03b1-synuclein clearance, emphasizing its potential as a therapeutic strategy to address the limitations of current symptomatic treatments and slow the progression of Parkinson's disease.",
"42552337": "ID: 42552337\nTitle: NanoBiT screening identifies the azaoxafluorene VT11 as potent tau interaction inhibitor.\nAbstract: Tauopathies are neurodegenerative disorders characterized by accumulating misfolded, insoluble tau protein aggregates in neurons or glial cells. In this study, we screened the Spectrum Collection and other compound libraries for inhibitors of tau self-interaction using a structural complementation reporter system (NanoLuciferase Binary Technology). Resulting candidates were tested in dose-response assays and evaluated for cell toxicity and microtubule destabilization. Further, a seed-induced tau interaction biosensor assay and a cell-free tau Real-Time Quaking-Induced Conversion assay have been established to study their effects on the kinetics of tau interaction and aggregation, respectively. The substances ritanserin, 3-methoxycatechol, gambogic amide, azaoxafluorenes VT11, and NS 185 and thieno[2,3-d][1.3]oxazine B6/55 showed a concentration-dependent tau self-interaction inhibition without relevant cell toxicity or microtubule destabilization. Ritanserin, VT11, NS 185 and B6/55 blocked tau interaction in the seed-induced tau interaction biosensor assay. Finally, the cell-free tau RT-QuIC assay displayed highest inhibitory potential for VT11. Thus, the azaoxafluorene VT11 seems to be a promising candidate for further investigations as tau interaction inhibitor to address a pivotal pathological process in Alzheimer's disease and other tauopathies.",
"42552379": "ID: 42552379\nTitle: Multifaceted roles of CD44 in cancer progression and targeted therapeutic strategies.\nAbstract: CD44, a multifunctional transmembrane glycoprotein, is not only a bystander but also a crucial driver of cancer progression that promotes cancer stem cell maintenance, metastasis, and resistance to therapy. Therefore, CD44 is recognized as a promising therapeutic target in advanced malignancies. Here, we discuss its unique features, such as its structural diversity, which arise from alternative splicing and the post-translational modifications of cleavage and phosphorylation. In addition, we discuss the function of CD44 as a multivalent cell adhesion receptor for extracellular matrix components, including hyaluronic acid, fibronectin, osteopontin, and TSG6, thereby regulating lymphocyte activation, cell-cell interactions, cell adhesion, and migration within the extracellular matrix. Moreover, the emerging role of CD44 as a co-receptor of receptor tyrosine kinases such as epidermal growth factor receptor, c-MET, and vascular endothelial growth factor receptor 2 is highlighted to elucidate the contribution of CD44 to malignant signaling networks. We also discuss its potential as a therapeutic target in advanced cancers, particularly its applications in unconjugated antibodies, antibody-drug conjugates, peptide-based inhibitors, and chimeric antigen receptor-T cell therapies. We conclude by addressing the limitations observed in clinical studies and outlining promising opportunities for future development.",
"42552689": "ID: 42552689\nTitle: Clinical Development of Therapies for Charcot-Marie-Tooth Disease: Recommendations for Trial Design, Endpoints, and Regulatory Pathways.\nAbstract: Charcot-Marie-Tooth disease (CMT) encompasses a heterogeneous group of inherited peripheral neuropathies. Despite being the most common genetic neurological condition, individual CMT subtypes are rare, presenting unique challenges for therapeutic development. The Together Patients Industry Clinicians versus CMT (ToPIC: CMT) Advocacy Group was formed with a diverse group of patient advocacy groups, clinician-scientists who treat patients with CMT, and pharmaceutical industry representatives to develop a common guidance on development of new treatments for CMT with clear expectations for meaningful patient outcomes and objective assessments of improvement. The ToPIC: CMT Group developed recommendations for clinical development of drugs and biological products for treating CMT, addressing trial design considerations for this rare progressive disease. Key challenges in CMT include small patient populations, variable disease progression, and the need for sensitive outcome measures. Recommendations emphasize flexible trial designs including adaptive designs, external controls, and single-participant designs when scientifically justified. Where possible, broad inclusion criteria based on clinical phenotype rather than genetic subtype alone are recommended. Disease-specific, validated outcome measures should assess function across ages and disease stages. Biomarkers reflecting peripheral nervous system health may serve as surrogate endpoints to support accelerated approval pathways. Patient and care partner perspectives are essential throughout development, particularly regarding treatment goals, risk tolerance, and meaningful endpoints. Successful therapeutic development for CMT and related neuropathies requires innovative approaches that balance rigorous scientific standards with the realities of rare disease research. Regulatory flexibility, informed by patient input and natural history data, can facilitate efficient development while maintaining assurance of safety and effectiveness.",
"42552969": "ID: 42552969\nTitle: Quenching-Derived Pt Clusters on Co3O4/TiO2 p-n Heterojunctions Enable Efficient Photothermal Oxidation of VOCs.\nAbstract: Photothermal catalysis provides an effective strategy to overcome the high energy consumption of thermal catalysis and the low efficiency of photocatalysis; however, inefficient charge separation and limited O2 activation remain key challenges. Herein, a p-n heterojunction oxide support, Co3O4/TiO2, was constructed and decorated with Pt species via a quenching method. The resulting Co3O4/TiO2-Q-Pt catalyst achieved toluene conversion exceeding 90% within 38\u00a0min under light irradiation at 522\u00a0mW cm- 2, significantly outperforming single-component oxide-supported catalysts. The catalyst exhibited excellent photothermal stability and strong resistance to water vapor and mixed volatile organic compound (VOC) conditions. Structural characterization revealed that Co3O4 incorporation reduced TiO2 crystallinity while enhancing lattice-oxygen mobility, redox capability, and light absorption, even at low Co3O4 contents. Moreover, electron paramagnetic resonance (EPR) measurements showed enhanced generation of \u2022OH and \u2022O2 - species under illumination, consistent with improved photoinduced redox activity. In situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) showed suppression of the accumulation of partial-oxidation intermediates and promotion of the formation of benzoate and maleic anhydride species, thereby facilitating aromatic ring cleavage and deep mineralization of toluene.",
"42553036": "ID: 42553036\nTitle: Aegeline mitigates isoproterenol-induced myocardial infarction: insights from biochemical, histopathological, and in silico studies.\nAbstract: Myocardial infarction (MI) remains a major cause of morbidity and mortality worldwide. Although considerable advances have been made in its diagnosis and management, the development of safe and effective cardioprotective agents remains an active area of investigation. Natural products continue to attract interest as potential therapeutic candidates because of their diverse pharmacological activities. Aegeline, an alkaloidal amide isolated from Aegle marmelos, has demonstrated antioxidant and anti-inflammatory properties; however, its cardioprotective effects in experimental myocardial injury have not been fully characterized. The aim of this work was to assess the cardioprotective effects of aegeline in an experimental MI model. Isoproterenol (ISO) (85\u2005mg/kg, subcutaneouslySC) was employed to trigger acute MI in experimental rats. Aegeline was administered to the rats at doses of 5 and 10\u2005mg (p.o.) on the 29th and 30th day, respectively, followed by ISO (85\u2005mg/kg) (s.c.) on the same days. Aegeline exhibited cardioprotective properties against ISO-induced damage, as evidenced by the amelioration of altered histopathological features, reduction in myocardial damage, and improvements in antioxidant and oxidative stress markers, inflammatory indicators, and signaling pathways, along with inhibition of apoptotic expression. In addition, aegeline did not affect liver or kidney function. Molecular docking using AutoDock Vina v1.2.6 revealed the strong binding affinity of aegeline for CK-MB (PDB ID: 3B6R), with key hydrophobic, hydrogen-bonding, and salt bridge interactions. Additional targets included LDH (7DBJ), cTnT (1J1D), and cTnI (7SC2), though CK-MB exhibited the most favorable interaction profile. Molecular dynamics simulations supported these findings through principal component analysis, dynamic cross-correlation, free energy landscape mapping, and generalized born surface area (MM-GBSA) binding free energy analysis. This study demonstrates that aegeline attenuates experimental myocardial injury by reducing oxidative stress, inflammation, and apoptosis. Computational analyses support potential molecular interactions with proteins associated with myocardial injury, while the biochemical and histopathological findings provide primary evidence for its cardioprotective activity.",
"42553046": "ID: 42553046\nTitle: NNMT Loss Drives Cancer Progression by enhancing SAM availability for mTORC1 Signaling and Chromatin Methylation.\nAbstract: Aberrant epigenetic reprogramming together with dysregulated mTOR signaling are hallmarks of cancer, where altered chromatin methylation and nutrient-sensing pathways cooperate to drive tumor progression. S -adenosylmethionine (SAM), the universal methyl donor, is essential for these processes, yet how tumors sustain elevated SAM availability to support oncogenic transmethylation reactions remains poorly defined. Here, using prostate cancer (PCa) as a model system, we identify nicotinamide N-methyltransferase (NNMT) as a critical metabolic-epigenetic regulator and tumor suppressor. Using a prostate-specific Nnmt knockout mouse model, we demonstrate that NNMT loss accelerates PCa progression, particularly in the context of Pten deletion, resulting in infiltrating carcinoma and reduced survival. Mechanistically, NNMT functions as a \"SAM-sink,\" and its loss increases intracellular SAM abundance, thereby activating mTORC1 signaling through SAMTOR-dependent sensing and broadly enhancing chromatin methylation. In human PCa, recurrent genomic deletions of NNMT occur in up to 7% of cases, and NNMT protein expression is largely absent in primary tumors and metastases. NNMT-deficient PCa cells exhibit elevated SAM:SAH ratios, increased histone methylation, and heightened mTORC1 activity, enabling sustained tumor growth even under dietary methionine-restriction (MR). Notably, combined MR and pharmacologic mTORC1 inhibition synergistically suppresses the growth of NNMT-deficient tumors, revealing a previously unrecognized therapeutic vulnerability. Collectively, these findings establish NNMT as a key tumor suppressor that constrains SAM-driven epigenetic and signaling programs in PCa and suggest a rational, diet-based therapeutic strategy for advanced cancers with NNMT loss.",
"42553058": "ID: 42553058\nTitle: Sarcopenia and sepsis fuel a self-perpetuating cycle of immunometabolism decline.\nAbstract: Sepsis and sarcopenia are intertwined clinical challenges characterized by profound immunometabolic dysregulation. Traditionally viewed as a passive reservoir, skeletal muscle is now recognized as an active immunometabolic rheostat that dynamically influences systemic inflammation and homeostasis. This review synthesizes recent advances that redefine our understanding of muscle in critical illness, moving beyond simplistic views of catabolism to encompass complex adaptive strategies. This review summarizes recent advances and discusses septic autocannibalism, a process in which muscle proteolysis, driven by a metabolic defense priority, provides key substrates, glutamine for immune function and alanine for hepatic gluconeogenesis. Initially adaptive, sustained activation of this response leads to severe muscle wasting and long-term functional impairment. We analyze the bidirectional relationship between these conditions, focusing on shared risk factors such as immunosenescence and obesity. Key molecular pathways, including the IL-6/JAK/STAT and NF-\u03baB axes, are examined, with particular emphasis on how the temporal release of myokines (e.g., IL-6, IL-15, IGF-1) dictates their shift from adaptive signals to chronic catabolic drivers. Furthermore, we discuss how energy reprogramming, characterized by aerobic glycolysis and mitochondrial failure, disrupts muscle homeostasis. We highlight physical activity as a potent modulator of immunometabolic health through the release of anti-inflammatory myokines and enhanced mitochondrial biogenesis. To propel the field forward, we propose experimental avenues: single-cell spatial transcriptomics to map cellular crosstalk, mitochondrial transplantation to restore energetic capacity, and the identification of \"muscle resilience\" biomarkers for early intervention. This review underscores the urgent need for integrated immunometabolic approaches to improve outcomes in critically ill patients.",
"42553087": "ID: 42553087\nTitle: The central role of IL-6 in the differential effects of GLP-1 receptor agonists and metformin across multiple health conditions.\nAbstract: An earlier publication suggested that elevated TGF-\u03b21 together with IL-1\u03b2 and IL-6 drives neutrophilic asthma, increases cancer risk, and is associated with metabolic dysregulation. This article examines how these cytokines shape the therapeutic efficacy and safety profiles of GLP-1 RAs compared with metformin. Integrating published data, stronger suppression of circulating IL-6 likely contributes to the superior efficacy of GLP-1 RAs in lowering blood glucose, improving insulin sensitivity in patients with polycystic ovary syndrome (PCOS), reducing cardiovascular risk, and reducing osteoarthritis-related perioperative infections, but not to their superior weight-loss effects. Superior weight loss, which is IL-6 independent, further enhances the efficacy of GLP-1 RAs in improving metabolic dysfunction-associated steatohepatitis (MASH), PCOS, and survival in cancer patients. Systemic IL-6 suppression may also explain the higher frequency of allergic adverse effects with GLP-1 RAs, whereas local interference with IL-6 signaling in the gastrointestinal tract, shared by both agents, likely accounts for frequent gastrointestinal adverse events reported with both. Although metformin may be more effective in improving asthma outcomes in the general asthma population, GLP-1 RAs are likely to be more effective in neutrophilic asthma. Metformin also reduces osteoarthritis-related joint pain, consistent with the limited efficacy of IL-6R inhibition in reducing this pain. Finally, the lack of effect of IL-6 pathway inhibitors on cancer incidence may explain the similar efficacy of GLP-1 RAs and metformin in reducing the risk of most cancers.",
"42553171": "ID: 42553171\nTitle: Curcumin Attenuates Decidual Stromal Cell Ferroptosis and Restores Impaired Decidualization in Recurrent Spontaneous Abortion by Targeting BRD4.\nAbstract: Recurrent spontaneous abortion (RSA) is a major reproductive health challenge with limited clinical options. Curcumin, a natural polyphenol widely consumed as a dietary supplement, has shown promise in improving pregnancy outcomes; however, its specific molecular targets remain obscure due to its broad bioactivity. This study aimed to define curcumin's impact on RSA and identify its direct molecular target to provide a mechanistic basis for its nutritional application. We employed an integrated approach combining phenotypic analysis in the RSA mouse model and target discovery in human endometrial stromal cells. Target identification was performed using unbiased Limited Proteolysis-Mass Spectrometry (LiP-MS). The interaction between curcumin and the identified target, BRD4, was confirmed through molecular docking, dynamics simulations, functional genetics, and chromatin immunoprecipitation (ChIP-qPCR). Functional outcomes were assessed by measuring key ferroptosis markers and mitochondrial morphology using transmission electron microscopy. Decidualization was analyzed in both mouse tissues and cells subjected to a decidualization and ferroptosis induction protocol. Curcumin significantly reduced embryo resorption and restored decidual morphology and marker expression (PRL and IGFBP1) in RSA mice. LiP-MS analysis in human decidual stromal cells identified ferroptosis as the primary pathway targeted by curcumin. Both in\u00a0vivo and in\u00a0vitro validation showed that curcumin inhibits ferroptosis, decreasing lipid peroxidation, restoring glutathione balance, and preserving mitochondrial integrity, which in turn rescued decidual marker expression. Proteomic integration with ferroptosis databases identified BRD4 as a central hub. Mechanistically, curcumin binds BRD4, thereby suppressing BRD4-driven expression of TFRC and ACSL4, which blocks ferroptosis and rescues decidual markers. This research uncovers a BRD4-driven ferroptosis pathway as a key pathogenic mechanism in RSA. Our results demonstrate that curcumin acts as an epigenetic modulator by directly targeting BRD4, thereby suppressing this pro-ferroptotic transcriptional program. These findings provide critical mechanistic evidence supporting curcumin as an evidence-based nutritional intervention for RSA.",
"42553212": "ID: 42553212\nTitle: Brassinosteroids as phytohormonal shields against micro- and nanoplastic stress in plants.\nAbstract: Microplastics (MPs) and nanoplastics (NPs) are biologically active stressors in agricultural soils, where they alter soil physical structure, disrupt rhizosphere processes, impair water and nutrient acquisition, and provoke oxidative and hormonal disequilibrium in plants. Brassinosteroids (BRs), particularly brassinolide and 24-epibrassinolide, have recently emerged as modulators of plant responses to plastic-particle stress. Current evidence indicates that BRs do not detoxify plastics directly. Instead, they reorganize plant performance across interconnected layers: aquaporin-linked NP transport, antioxidant and ascorbate-glutathione metabolism, photosystem II function, hormone crosstalk, secondary metabolism, and rhizosphere feedbacks. In tomato, BRs reduced polystyrene-NP accumulation in edible tissues by suppressing aquaporin genes. In Pinellia ternata and rice, BRs attenuated MP/NP-induced growth inhibition by restoring photosynthetic efficiency and redox control. This mini review synthesizes these findings and frames BRs as eco-hormonal regulators of the plant-plastic-soil interface, while highlighting priorities for field-realistic validation.",
"42553232": "ID: 42553232\nTitle: Genomic characterization of Bacillus velezensis BP5 and BP103: insights into biocontrol potential against bacterial leaf spot on pepper.\nAbstract: Bacillus velezensis strains BP5 and BP103, isolated from pepper rhizosphere soil in the Mekong Delta, Vietnam, displayed potent antagonistic activity against Xanthomonas euvesicatoria, which causes pepper bacterial spot. In dual-culture assays, BP5 formed the largest inhibition zone (36.7 mm), outperforming BP103 (32.1 mm) and oxolinic acid (31.3 mm). Both Gram-positive, rod-shaped strains produced extracellular protease, lipase, amylase, cellulase, and siderophores. Whole-genome sequencing revealed compact and genetically stable genomes: BP5 (4.08 Mb, 46.04% GC, 4,047 protein-coding genes, 5 prophages) and BP103 (3.91 Mb, 46.46% GC, 3,793 genes, 2 prophages). Phylogenetic reconstruction and average nucleotide identity (ANI of more than 98%) confirmed their assignment to B. velezensis, with closest relatedness to strain 160. Orthologous gene analysis across 28 B. velezensis genomes and B. subtilis DSM10 identified 2,593 single-copy genes. Compared to BP103, BP5 harbors greater numbers of multi-copy orthologs, other orthologs, and unique paralogs (388, 910, and 5 in BP5 with 336, 763, and 1 in BP103, respectively). Pangenome analysis showed that the core genome size decreased progressively with the addition of new genomes; BP5 showed more pronounced genetic divergence driven by expanded unique paralogs and shell gene families (with 456 in BP5 compared with 203 in BP103). Both strains encode 13 secondary metabolite biosynthetic gene clusters, including surfactin, fengycin, bacillaene, macrolactin, difficidin, bacilysin, bacillibactin, mersacidin, and locillomycin, plus four previously undescribed terpene/PKS. The CAZyme repertoire was richest in BP5 (133 genes), exceeding BP103 and the commercial strain FZB42 (both 129 genes), thereby enhancing adhesion and adhesion and rhizosphere colonization. These findings establish BP5 and BP103 as highly promising biocontrol agents, combining high genetic stability with diverse secondary metabolite profiles, suitable for development into sustainable microbial bio-bactericides.",
"42553260": "ID: 42553260\nTitle: Curcumin attenuates \u03b1-synuclein pathology in Parkinson's disease model mice through modulation of UBC9-associated SUMOylation signaling.\nAbstract: Post-translational modifications, particularly SUMOylation, plays a crucial role in \u03b1-synuclein (\u03b1-syn) aggregation, a key pathological feature of Parkinson's disease (PD). Curcumin, a natural polyphenol, has shown neuroprotective potential, but its effects on SUMOylation-related signaling in PD remain unclear. This study aimed to investigate whether curcumin modulates \u03b1-syn SUMOylation and to elucidate the underlying molecular mechanisms in PD model mice. A PD model was established in male C57BL/6 mice via unilateral intrastriatal injection of \u03b1-syn preformed fibrils (PFFs). Six months after \u03b1-syn PFFs injection, mice were treated intravenously with curcumin (25\u202fmg/kg/day) or vehicle for 1\u202fmonth. Behavioral tests (open field, rotarod) assessed motor function. Neuropathology was evaluated by immunohistochemistry and western blotting for tyrosine hydroxylase (TH), phosphorylated \u03b1-syn (p-syn), SUMOylation pathway components (SUMO1, SAE2, UBC9, PIAS1/2), and ubiquitin. Striatal dopamine levels were measured by HPLC. Curcumin treatment ameliorated motor deficits and anxiety-like behaviors in PD mice. It partially preserved dopaminergic neurons and reduced p-syn aggregation in the substantia nigra, accompanied by increased striatal dopamine levels. Mechanistically, curcumin was associated with reduced SUMO1 and increased ubiquitin levels, suggesting modulation of SUMOylation-related signaling. Among SUMOylation enzymes, UBC9 expression was decreased, whereas E1 (SAE2) and E3 (PIAS1/2) components were not substantially affected. Our findings demonstrated that curcumin exerted neuroprotective effects in a PD model by attenuating \u03b1-syn pathology. The protective mechanism involves the inhibition of \u03b1-syn SUMOylation, primarily through the downregulation of the UBC9 enzyme. This study identifies UBC9-mediated SUMOylation as a potential target for curcumin and highlight a promising strategy for modifying \u03b1-syn-associated pathology in PD.",
"42553289": "ID: 42553289\nTitle: Rapamycin-nanoliposomes target the mTORC1-mediated autophagy-lysosomal and NLRP3/Caspase-1 pathways to inhibit nucleus pulposus cell senescence in intervertebral discs.\nAbstract: Nucleus pulposus (NP) cell quiescence maintains intervertebral disc homeostasis, while mTORC1 regulates autophagy-lysosomal function and inflammatory secretion to preserve quiescence-rapamycin specifically targets mTORC1. Herein, we fabricated rapamycin-nanoliposomes (rapa-lipos) via ultrasonic dispersion, thin-film dispersion, and filtration to improve rapamycin bioavailability, investigating their role in inhibiting the senescence phenotype of NP cells through \u03b2-gal staining, lysosomal staining, transmission electron microscopy, ELISA, and cell cycle inhibitors. The mechanistic effects of rapa-lipos on mTORC1, NLRP3/Caspase-1 pathway (NCP) and autophagy-lysosomal pathway (ALP) were also analyzed by western blotting, immunofluorescence (IF), Si-RNA (raptor), and PCR. In vivo, rapa-lipos were injected into rat intervertebral disc with IL-1\u03b2-induced degeneration, assessed via HE staining, x-ray, MRI, and IF. Rapa-lipos exhibited high encapsulation efficiency, favorable drug loading, uniform particle size, and controlled release, suppressing NP cell senescence-related phenotypes (morphological changes, elevated IL-1\u03b2/TNF-\u03b1 secretion, increased \u03b2-gal activity, lysosomal dysfunction, upregulated P21/P16 and reduced P27 expression). Mechanistically, rapa-lipos targeted-inhibited mTORC1, then blocked NCP and activated ALP to maintain NP cell quiescence. In vivo, x-ray, MRI and histological evaluation confirmed rapa-lipos mitigated intervertebral disc degeneration. Collectively, rapa-lipos target mTORC1-mediated NCP and ALP to inhibit NP cell senescence, offering a promising strategy for intervertebral disc degeneration prevention.",
"42553303": "ID: 42553303\nTitle: Severe non-atopic prurigo nodularis in an otherwise healthy elderly patient: rapid multidimensional response to dupilumab.\nAbstract: Prurigo nodularis (PN) is a chronic neuroimmune skin disorder characterized by intensely pruritic nodules, severe impairment of quality of life and significant psychosocial burden. Prior to the approval of targeted therapies, management relied on conventional agents with limited efficacy. Recent evidence of its neuroimmune pathogenesis have led to the approvement of targeted biologics. Dupilumab, a human monoclonal antibody targeting IL-4/IL-13-axis, has been the first targeted therapy approved for PN. We report a 75-year-old woman presenting with severe, treatment-na\u00efve generalized pruritus (IGA PN-S 4; NRS 9/10; DLQI 21) with extensive nodular involvement of the trunk and extremities, complete sleep deprivation and important anxiety and depression (HADS.A 12; HADS-D 11). Following diagnosis of PN, treatment with dupilumab (600\u00a0mg loading dose, then 300\u00a0mg every 2\u00a0weeks) was initiated in October 2025. At approximately 6 weeks, marked pruritus reduction (NRS 3) and sleep restoration were observed. Progressive improvement continued over time. At 4 months follow-up, complete remission was nearly achieved (NRS 1, IGA PN-S 1, DLQI 2, HADS-A 4, HADS-D 3), with no adverse events reported. This case corroborates and extends the existing evidence on dupilumab efficacy in the treatment of moderate-to-severe PN, with particularly rapid and comprehensive responses across pruritus, skin lesions, sleep and psychosocial outcomes in an elderly patient, through IL-4/IL-13 inhibition. Our findings support dupilumab as a safe and effective targeted option for severe PN, also in older patients where conventional systemic therapies, such as corticosteroids or immunosuppressants, carry a high risk of adverse effects.",
"42553357": "ID: 42553357\nTitle: Antimicrobial Efficacy of Aqueous Extracts of Abrus precatorius and Achyranthes aspera Against Streptococcus mutans and Candida albicans: An In Vitro Study.\nAbstract: Background Dental caries and oral candidiasis are among the most common oral diseases and are primarily associated with Streptococcus mutans and Candida albicans. Growing concerns regarding antimicrobial resistance and the adverse effects of synthetic antimicrobial agents have increased interest in plant-derived alternatives. This study evaluated and compared the antimicrobial efficacies of aqueous extracts of Abrus precatorius and Achyranthes aspera against Streptococcus mutans and Candida albicans. Methodology An in vitro experimental study was conducted using aqueous extracts from the leaves of Abrus precatorius and Achyranthes aspera. The antimicrobial activity against Streptococcus mutans\u00a0(ATCC 25175) and Candida albicans (ATCC 10231) was assessed using the agar well diffusion method. Chlorhexidine and nystatin served as positive controls for antibacterial and antifungal testing, respectively, and distilled water served as the negative control. Minimum inhibitory concentrations (MICs) were determined using the broth microdilution method. Data were analyzed using one-way analysis of variance followed by Tukey's post-hoc test and independent-samples t-test. Results Significant differences in antimicrobial activity were observed among the groups (p < 0.001). Against Streptococcus mutans, Abrus precatorius demonstrated a larger mean zone of inhibition (18.42 \u00b1 1.58 mm) than Achyranthes aspera (15.16 \u00b1 1.47 mm), although chlorhexidine exhibited the greatest antibacterial activity (24.31 \u00b1 1.32 mm). Similarly, against Candida albicans, Abrus precatorius produced a larger inhibition zone (16.87 \u00b1 1.36 mm) than Achyranthes aspera (13.92 \u00b1 1.41 mm), whereas nystatin showed the highest antifungal activity (22.74 \u00b1 1.24 mm). Abrus precatorius also demonstrated significantly lower MIC values against both microorganisms, indicating a greater antimicrobial potency. Conclusions Both aqueous plant extracts exhibited significant antibacterial and antifungal activity. Abrus precatorius showed superior antimicrobial efficacy compared to Achyranthes aspera; however, both extracts were less effective than the standard antimicrobial agents. These findings suggest the potential application of these herbal extracts in the development of oral healthcare products.",
"42553364": "ID: 42553364\nTitle: Targeting the AHR-IDO1-kynurenine pathway in TREM2+ macrophages restores antitumor immunity in thyroid cancer.\nAbstract: Advances in single-cell technologies have provided detailed insight into cellular heterogeneity within tumor microenvironments. However, the regulatory mechanisms driving macrophage-mediated immune suppression in thyroid cancer remain incompletely understood. We applied integrated single-cell multi-omics approaches to tumor samples from patients and murine thyroid cancer models, including single-cell RNA sequencing, single-cell ATAC sequencing, mass cytometry, spatial/histological validation, and in vitro and in vivo functional assays. We identified a prominent enrichment of TREM2+ tumor-associated macrophages within the thyroid cancer immune microenvironment. These macrophages displayed an immunosuppressive transcriptional and metabolic profile and promoted immune evasion through activation of the AHR-IDO1-kynurenine signaling axis, leading to impaired CD8+ T-cell function. Pharmacological inhibition of AHR and IDO1 reversed immune suppression and reduced tumor burden in experimental models. These findings reveal a TREM2+ macrophage-driven immunometabolic mechanism in thyroid cancer and suggest that targeting the AHR-IDO1-kynurenine pathway may provide a potential strategy for improving antitumor immunity.",
"42553369": "ID: 42553369\nTitle: A double-edged sword: the role of macrophage pyroptosis-driven cytokine storm-mediated intercellular communication in tumor progression.\nAbstract: Pyroptosis, a lytic and inflammatory form of programmed cell death, has emerged as a regulator of tumor immunity through its capacity to trigger localized cytokine storms (operationally defined in Section 3). This review examines the dual mechanisms by which macrophage pyroptosis-driven cytokine storms influence tumor progression. Pyroptosis is executed through canonical (caspase-1/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3)-dependent) and non-canonical (caspase-4/5/11-dependent) pathways, both converging on cleavage of gasdermin D (GSDMD) to form membrane pores that release pro-inflammatory cytokines (interleukin-1 beta, IL-1\u03b2; interleukin-18, IL-18) and damage-associated molecular patterns (DAMPs). These primary signals initiate cascade amplification through chemokine and cytokine networks, recruit diverse immune cell populations, and establish distinct inflammatory microenvironments. The effects of pyroptotic cytokine storms show striking temporal and intensity dependence. Acute, moderate inflammatory responses activate anti-tumor immunity through induction of immunogenic cell death (ICD), dendritic cell maturation, and cytotoxic lymphocyte priming. Chronic, low-grade cytokine storms, in contrast, promote tumorigenesis through six interconnected mechanisms: genomic instability and epigenetic reprogramming, cancer stem cell enrichment, pro-angiogenic remodeling, pre-metastatic niche formation, establishment of an immunosuppressive microenvironment, and induction of epithelial-mesenchymal transition. This \"double-edged sword\" phenomenon depends on inflammation intensity, duration, spatial distribution, and tumor microenvironment (TME) characteristics. Clinical investigations indicate that pyroptosis-related biomarkers, including GSDMD, gasdermin E (GSDME), and inflammatory cytokine profiles, may support patient stratification and treatment-response prediction across multiple cancer types in patients with tumor-associated macrophages (TAMs)-rich tumors. Preclinical evidence from bioorthogonal chemical systems in murine models, together with mathematical modeling, has suggested that pyroptosis affecting approximately 10-15% of tumor cells may serve as a tentative threshold for initiating anti-tumor immunity. However, this value has been derived from a limited number of preclinical systems (primarily 4T1 mammary tumor models) and has not yet been validated in human tumors; it should therefore be interpreted as a working hypothesis rather than an established parameter. Current therapeutic strategies targeting this pathway include NLRP3 inhibitors, IL-1\u03b2/IL-18 blockers, and combination approaches with immune checkpoint inhibitors (ICIs). Looking forward, future research should prioritize: (i) quantitative in vivo mapping of macrophage pyroptosis using spatial multi-omics and intravital imaging; (ii) development of tumor-targeted, spatiotemporally controlled pyroptosis inducers (e.g., nano-delivery and bioorthogonal activation systems); (iii) rational combination with immune checkpoint inhibitors and epigenetic modulators; and (iv) establishment of pyroptosis-based biomarker panels to guide patient stratification and toxicity prediction in clinical trials. Achieving \"controllable cytokine storms\" through precise macrophage pyroptosis modulation represents a therapeutic paradigm that balances anti-tumor efficacy against inflammatory toxicity, with the potential to advance cancer immunotherapy toward precision inflammation regulation.",
"42553374": "ID: 42553374\nTitle: Durable response to ALK inhibition in low-allele-frequency SPTBN1-ALK-rearranged gastric cancer followed by lineage plasticity-mediated resistance: a case report.\nAbstract: ALK rearrangements are rare in gastric cancer, and their therapeutic relevance remains poorly defined. While ALK inhibitors have demonstrated efficacy in lung cancer and other malignancies, data in gastric tumors are limited. We report a 52-year-old woman with metastatic gastric adenocarcinoma harboring a rare SPTBN1-ALK fusion detected at a low variant allele frequency (0.65%). The tumor was negative for HER2 amplification, microsatellite instability, and PD-L1 expression. After failure of chemotherapy combined with immunotherapy, treatment with the ALK inhibitor iruplinalkib resulted in rapid clinical improvement and a durable partial response lasting approximately 14 months. Serial next-generation sequencing at progression demonstrated enrichment of the fusion-positive clone (variant allele frequency increased to 5.94%), accompanied by expansion of TP53-mutant alleles and acquisition of additional genomic alterations. Subsequent histologic transformation to small-cell neuroendocrine carcinoma was observed, with loss of RB expression and markedly elevated Ki-67. No canonical ALK kinase domain resistance mutations were detected, suggesting a non-on-target resistance mechanism. This case highlights that even low-allele-frequency ALK fusions may have important clinical relevance in gastric cancer and may identify patients who could benefit from ALK-targeted therapy. It also illustrates the dynamic evolutionary trajectory of oncogene-driven tumors under therapeutic pressure, with histologic transformation to small-cell neuroendocrine carcinoma that may reflect lineage plasticity. Comprehensive genomic profiling and longitudinal molecular monitoring may facilitate the identification of rare actionable alterations and improve understanding of resistance mechanisms in advanced gastric cancer.",
"42553406": "ID: 42553406\nTitle: Peripheral and Central Administration of Soluble Glycoprotein 130 Improves Cognitive Outcomes Following Controlled Cortical Impact in Male Mice.\nAbstract: Traumatic brain injury (TBI) initiates complex immune responses, including upregulation of interleukin (IL)-6, a cytokine associated with clinical outcomes after injury. IL-6 trans-signaling, via the solubilized IL-6 receptor, drives pro-inflammatory cascades and is selectively inhibited by soluble glycoprotein 130 (sgp130). Although chronic intermittent sgp130Fc fusion protein (sgp130Fc) treatment after TBI has shown benefits across species, optimal dosing remains unclear. This study evaluated the effects of a single sgp130Fc dose (2 \u03bcg intrahippocampally or 10 \u03bcg intraperitoneally) in male mice on day three following controlled cortical impact or sham procedures. Cognitive performance was assessed using the Morris water maze, and histological assessment included lesion volume and microglia quantification. Intrahippocampal sgp130Fc improved spatial memory during probe trials, decreasing latency to the platform zone (p < 0.05) and enhanced spatial swim strategy selection (p < 0.05). Intraperitoneal sgp130Fc improved spatial learning (p < 0.05) and reduced anxiety-like behavior, indicated by increased target zone time during probe trials and decreased peripheral zone time (p < 0.05). Neither route significantly altered lesion volume or thalamic ionized calcium-binding adaptor molecule 1+ cell counts versus vehicle-treated injured animals; however, intrahippocampal sgp130Fc reduced major histocompatibility complex class II+ cells. These findings demonstrate that a single subacute sgp130Fc dose confers modest behavioral benefits when administered post-TBI, without worsening outcomes. The results underscore the effects of IL-6 trans-signaling inhibition on TBI outcomes and highlight the need for further research into dose, timing, and sex-specific responses. Overall, these data add to emerging support that selective IL-6 blockade with sgp130Fc may represent a translatable strategy to modulate post-TBI inflammation and support cognitive recovery, in part, via reducing anxiety-associated dysfunction.",
"42553440": "ID: 42553440\nTitle: Metabolic Reprogramming and Immunometabolic Dysregulation in Diabetic Kidney Disease: From Pathogenesis to Precision Multi-target Therapies.\nAbstract: Diabetic kidney disease, the leading cause of end-stage kidney disease worldwide, involves complex interactions beyond classical hemodynamic and oxidative stress pathways. Recent advances emphasize metabolic reprogramming in renal cells-characterized by mitochondrial dysfunction, impaired fatty acid oxidation, lipotoxicity, and glycolytic shifts-as upstream drivers of cellular injury and fibrosis. Single-cell RNA sequencing reveals profound immunometabolic heterogeneity, including dynamic macrophage subpopulations (e.g., proinflammatory early states transitioning to TREM2hi/MRC1hi lipid-associated phenotypes) and T helper 17/regulatory T imbalance, which amplify inflammation via bidirectional crosstalk with podocytes, tubular cells, and mesangial cells. Interorgan axes, particularly gut dysbiosis and uremic toxin accumulation, further perpetuate immune dysregulation. This review integrates these insights to propose precision strategies targeting mitochondrial homeostasis, ferroptosis inhibition, glycolytic blockade in immune cells, and multimodal therapies (e.g., combination strategies integrating sodium-glucose cotransporter 2 inhibitors with immunometabolic modulators). Multi-omics integration and spatial transcriptomics hold promise for individualized and mechanism-guided interventions to halt diabetic kidney disease progression.",
"42553447": "ID: 42553447\nTitle: Clinical outcomes following endoscopic retrograde cholangiopancreatography-based therapy compared with surgery in chronic pancreatitis: evidence from a multicentric cohort.\nAbstract: Real-world comparative outcomes of endoscopic retrograde cholangiopancreatography-based endoscopic therapy vs. pancreatic surgery in chronic pancreatitis (CP) remain incompletely defined. We performed a retrospective comparative effectiveness study using the TriNetX US Collaborative Network. Adults with CP undergoing endoscopic therapy or pancreatic surgery were identified, and propensity score matching generated 1451 patients in each cohort. Outcomes were assessed from 1-1095 days after the index event, using risk-based analyses as the primary comparative summaries and Kaplan-Meier analyses as secondary time-to-event summaries. Outcomes included chronic opioid prescriptions, opioid use disorder (OUD), celiac plexus block/neurolysis, pain codes, acute pancreatitis, exocrine and endocrine pancreatic insufficiency, emergency visits, and all-cause mortality. Mean follow up was 771.494 days for endoscopy and 827.285 days for surgery. New chronic opioid prescriptions occurred in 146/1092 (13.4%) endoscopic vs. 79/1032 (7.7%) surgical patients (odds ratio [OR] 1.862, 95% confidence interval [CI] 1.396-2.484; P<0.001; hazard ratio [HR] 1.861, 95%CI 1.415-2.448). Acute pancreatitis occurred in 167/530 (31.5%) vs. 65/565 (11.5%) patients (OR 3.539, 95%CI 2.578-4.858; P<0.001; HR 3.341, 95%CI 2.506-4.453). Exocrine pancreatic insufficiency occurred in 200/1213 (16.5%) vs. 132/1206 (10.9%) patients (OR 1.606, 95%CI 1.269-2.034; P<0.001; HR 1.585, 95%CI 1.272-1.975). Endocrine pancreatic insufficiency, mortality, emergency visits, and OUD did not differ significantly between cohorts. Endoscopic therapy was associated with higher chronic opioid prescribing, acute pancreatitis, celiac plexus block/neurolysis, and exocrine pancreatic insufficiency. These findings are associative and should be interpreted in the context of residual confounding and coding-based outcome ascertainment.",
"42553460": "ID: 42553460\nTitle: Unraveling the Molecular Landscape of Follicular Fluid: A Comparative Proteomic Study Between Bali and Brahman Cattle.\nAbstract: Protein biomarkers are increasingly recognized as indicators of metabolic status, immune response, and reproductive performance in cattle. However, comparative data among different breeds commonly raised in Indonesia remain limited. This study aimed to compare the specific proteins in follicular fluid (FF) of Bali and Brahman cows. Samples from FF (n\u2009=\u200920 pairs of ovarium) were analyzed for identification of specific proteins using LC-HRMS analysis. Proteomic analysis revealed distinct breed-specific FF profiles, with 16 proteins uniquely identified in Bali cattle and 23 proteins in Brahman. Bali FF was enriched with proteins associated with protease inhibition, coagulation regulation, extracellular matrix remodeling, and cytoskeletal dynamics, including CORO1C, ZER1, YARS1, and L3MBTL3. In contrast, Brahman FF showed enrichment of proteins involved in lipid oxidation, one-carbon metabolism, and peroxisomal activity, notably ACOX2, BHMT, and KIAA0586. Here, we present the proteomic characterization of FF from Bali and Brahman cattle, where several bioactive proteins are candidate proteins potentially associated with reproductive processes. These findings highlight the potential of protein biomarkers in guiding cattle breeding and management strategies in tropical regions.",
"42553528": "ID: 42553528\nTitle: Arthroscopic (Arthro) Delivery of MACI (Autologous Cultured Chondrocytes on a Porcine Collagen Membrane) for Defects on MFC and Trochlea With concomitant TTO.\nAbstract: Chondral pathology of the knee-specifically involving the femoral condyles and trochlea-is commonly observed during knee arthroscopies. This condition may necessitate supplementary interventions, such as a concomitant tibial tuberosity osteotomy (TTO). Approaching this patient population with a straightforward, less invasive arthroscopic technique may offer an advanced solution. Arthro matrix-induced autologous chondrocyte implantation (MACI; autologous cultured chondrocytes on a porcine collagen membrane) targets symptomatic chondral lesions in the knee that are consistent with magnetic resonance imaging findings, which range in size from 2 to 4 cm2, are contained within a stable rim, and are classified as grade 3 to 4a according to the International Cartilage Repair Society. This technique for the arthroscopic delivery of MACI for defects located on the trochlea and femoral condyles has been developed as a progressive advancement in the evolution of autologous chondrocyte implantation technology. The described technique addresses these defect locations by utilizing the same arthroscopic approach for both cartilage defect locations and a separate incision for TTO. The instruments used include 3 varieties of curettes, an arthroscopic cutter to score the cartilage, and the V-shuttle membrane delivery device. Furthermore, attention is dedicated to the medial and lateral approaches, patellar elevation for trochlear access, meticulous defect preparation, hemostasis and fluid management, and implant delivery. Arthroscopic surgical procedures generally result in reduced postoperative pain, necessitating a decreased analgesic requirement, an earlier and enhanced joint mobility, less postoperative muscle inhibition, and expedited recovery. Moreover, an arthroscopic approach may reduce the length of the surgical incision for patients concerned about cosmetic outcomes. In contrast to various other arthroscopic procedures related to knee ligaments or meniscal injuries, the implementation of arthroscopic delivery of MACI for defects on the trochlea and femoral condyles of the knee offers a persuasive and readily executable alternative for a proficient surgeon, even when performed in conjunction with TTO. This arthroscopic technique for the delivery of the MACI implant can effectively treat multiple knee chondral lesions in different compartments when paired with TTO. Advantages of this approach include minimal surgical disruption, the ability to perform combined knee arthroscopic procedures, a more desirable cosmetic appearance, and improved early postoperative recovery. The author(s) attests that consent has been obtained from any patient(s) appearing in this publication. If the individual may be identifiable, the author(s) has included a statement of release or other written form of approval from the patient(s) with this submission for publication.",
"42553548": "ID: 42553548\nTitle: Uncovering the hidden biology of fibrinaloid microclot complexes in complex, inflammatory diseases.\nAbstract: Blood can clot into anomalous, fibrinolysis-resistant forms that arise from prothrombotic seeding areas, including damaged cellular debris and membrane-derived surfaces, giving rise to what we have termed fibrinaloid microclot complexes (colloquially: microclots). Their proteolytic resistance is due in part to the fact that they are amyloid in nature, and they can also entrap inhibitors of proteolysis. They consist of a variety of proteins besides the expected fibrin, and are highly enriched for other amyloidogenic proteins (in contrast to normal clots, whose proteome largely reflects the soluble plasma proteome). They also contain DNA in the form of neutrophil extracellular traps (NETs). Importantly, fibrinaloid microclot complexes are heterogeneous structures comprising multiple phenotypic forms, including those that nucleate and grow on cellular debris such as damaged membranes, microparticles, and immune-derived material. We consider that these debris-associated complexes can act as catalytic scaffolds that recruit fibrin(ogen) and inflammatory molecules, thereby amplifying amyloidogenic transformation and prothrombotic activity. Fibrinaloid microclot complexes have been reported in a widening range of chronic inflammatory and thrombo-inflammatory diseases in which they have been sought, and are highly enriched for amyloidogenic proteins. Additionally, the thrombi extracted from ischaemic stroke also contain proteins in an amyloid form. One mechanism that explains how such macroclots can form and block arteries larger than any leading to them is that this occurs via the accretion of microclots that already contain amyloid. We here show that these microclots exhibit a classical 'apple-green' birefringence when stained with the dye Congo red. It is now important to determine whether inhibiting amyloid-forming clot transitions has therapeutic value.",
"42553661": "ID: 42553661\nTitle: Emergence of Trichophyton indotineae: in vitro susceptibility data from a mycology reference laboratory in Australia.\nAbstract: Management of Trichophyton indotineae dermatophytosis is challenged by high terbinafine resistance rates within this species. Antifungal susceptibility data for Australian T. indotineae isolates are lacking. Here we determined the in vitro activity of five antifungal agents, including olorofim, against T. indotineae to guide therapeutic choices. Forty-four clinical T. indotineae isolates (cultured from skin, 2005-2025) across several Australian jurisdictions were tested for susceptibility to itraconazole, voriconazole, posaconazole, terbinafine and olorofim using CLSI broth microdilution methodology. MIC endpoints were determined at 80% growth inhibition for azoles and terbinafine, and 100% inhibition for olorofim. Isolates with terbinafine MICs \u22650.5\u2005mg/L were considered as non-WT or resistant. Twenty-eight of 44 (63.6%) T. indotineae isolates had terbinafine MICs \u22650.5\u2005mg/L [geometric mean (GM) MIC 0.570\u2005mg/L; MIC90 4\u2005mg/L] with 21 (47.7%) displaying MICs \u22654\u2005mg/L. The modal terbinafine MIC was 4\u2005mg/L. Lower MICs were observed for azoles, with posaconazole demonstrating the greatest activity, i.e. posaconazole (GM 0.058\u2005mg/L; MIC90 0.25\u2005mg/L), itraconazole (GM 0.087\u2005mg/L; MIC90 0.5\u2005mg/L) and voriconazole (GM 0.201\u2005mg/L; MIC90 1\u2005mg/L). Olorofim demonstrated good in vitro activity with all 44 isolates exhibiting MICs \u200a\u2264\u200a0.06\u2005mg/L (GM 0.023\u2005mg/L; MIC90 0.06\u2005mg/L). Terbinafine resistance in T. indotineae is common in Australia but not universal. Susceptibility testing can inform treatment choice. The azoles and olorofim demonstrate good in vitro activity.",
"42553675": "ID: 42553675\nTitle: A Concealed Typical AVNRT: U Wave Contribution in Simulating Long RP Supraventricular Tachycardia.\nAbstract: Accurate identification of P waves on electrocardiogram (ECG) is essential in supraventricular tachycardias because U waves may resemble retrograde P waves, producing a pseudo-long RP interval and hiding typical atrioventricular nodal reentrant tachycardia (AVNRT). A 40-year-old woman presented with regular narrow-complex tachycardia with a positive deflection immediately following the T wave, creating the impression of a long RP tachycardia. The electrophysiological study revealed a short VA interval with an H-A-V activation sequence, characteristic of typical slow-fast AVNRT. This case emphasizes the diagnostic challenge posed by U waves at higher heart rates, which can obscure the true arrhythmia mechanism.",
"42553702": "ID: 42553702\nTitle: Distinct brain extracellular vesicle microRNA profiles differ in frontotemporal dementia and Alzheimer's disease.\nAbstract: Dementia is a syndrome caused by various diseases including Alzheimer's disease (AD) and frontotemporal dementia (FTD) with an estimated global prevalence of 60 million individuals. Recently, therapeutic development in the dementia field has accelerated, with the introduction of monoclonal antibody therapeutics such as Lecanemab and Donanemab. However, AD and FTD patients are still either diagnosed too late to benefit from available therapies or are misdiagnosed due to the clinical overlap between dementia subgroups making therapeutic intervention challenging. This highlights a real need to improve early diagnostic tools of neurodegenerative disease (ND) biomarkers. A potential source of such biomarkers come from small extracellular vesicles (sEVs), groups of cell-derived, lipid-bound assemblies with the capability to cross the blood-brain barrier (BBB) and known to carry pathogenic proteins associated with AD and FTD. A known cargo of sEVs is microRNA (miRNA), regulatory molecules that post-transcriptionally silence gene expression including transcripts of autophagic systems, processes which dysfunction in dementia-causing diseases leading to toxic aggregate build-up, causing neurodegeneration. The targeting of functional machineries in macroautophagy (MA) and chaperone-mediated autophagy (CMA) by different miRNA may vary between AD and FTD mutations, leading to potential biomarkers of disease being highlighted. Through isolating sEVs from the frontal cortex of post-mortem brain tissue of AD, FTD-MAPT, FTD-C9orf72, FTD-GRN and no-disease control patients (Manchester Brain Bank), miRNA cargoes were analysed and compared using real-time quantitative PCR (RT-qPCR). Seven autophagy-associated miRNA candidates (MA: miR-124-3p, miR-30a-5p, miR-128-3p; and CMA: miR-224-5p, miR-373-5p, miR-106a-3p and miR-26b-5p) were tested to identify dementia sub-group variations, used alongside small RNA-sequencing to explore broader miRNA variation within sEV populations. Of the miRNA tested miR-224-5p (P = 1.76 \u00d7 10-5) and miR-106a-3p (P = 0.033) showed significant group differences, and further significant pairwise comparison differences [miR-224-5p: AD fold change (FC) = 4.29, MAPT FC = 7.62; miR-106a-5p: AD FC = 5.59] when compared with no disease controls and other dementia subgroups, potentially showing initial diagnostic and differentiating potential. Small RNA-sequencing results revealed 8 AD, 2 FTD-GRN, 52 FTD-MAPT and 12 FTD-C9orf72 differentially expressed sEV-miRNAs when compared with no disease controls. Further direct comparisons between AD versus FTD mutation-derived sEV cargoes, and even FTD mutation versus FTD mutation-derived sEV cargoes, identified additional miRNA with differentiating capabilities. These findings demonstrate sEV-derived miRNA signatures vary across dementia sub-types and suggest potential roles of sEV cargoes in both disease diagnostics and identifying drivers of ND, such as autophagic impairments and signalling pathways.",
"42553762": "ID: 42553762\nTitle: Programmed Cell Death: A Key Mechanism of Traditional Chinese Medicine in the Treatment of Membranous Nephropathy.\nAbstract: Membranous nephropathy (MN) is the leading cause of adult nephrotic syndrome with rising global incidence. A substantial proportion of patients progress to end-stage renal disease. Multiple programmed cell death (PCD) pathways-including apoptosis, pyroptosis, ferroptosis, and autophagy-are activated by complement-dependent and -independent pathogenic factors, collectively driving podocyte injury and MN progression. Current immunosuppressive therapies are limited by adverse reactions and variable responses. Traditional Chinese medicine (TCM), characterized by multi-component, multi-target, and multi-pathway regulation, shows therapeutic advantages in MN. To systematically review the regulatory mechanisms of PCD in MN, summarize the molecular basis of TCM interventions targeting PCD, and identify current research limitations and future directions. This narrative review systematically elaborates the regulatory association between PCD and podocyte injury in MN. Literatures were systematically retrieved from PubMed, Embase and China National Knowledge Infrastructure (CNKI) from the establishment of each database to May 2026, and 41 preclinical studies exploring TCM modulation of PCD in MN were included after strict screening. Complement-dependent and complement-independent pathways in MN can synergistically initiate podocyte apoptosis, pyroptosis, ferroptosis, and autophagy dysfunction. TCM can regulate PCD through core signaling pathways such as phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR), nuclear factor erythroid 2-related factor 2/heme oxygenase 1 (Nrf2/HO-1), mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase/forkhead box O1 (JNK/FoxO1), and PTEN-induced kinase 1/Parkin (PINK1/Parkin), while simultaneously intervening in non-PCD pathological processes including inflammation, oxidative stress, and renal fibrosis. Clinical evidence further indicates that TCM or combined with conventional therapy can elevate MN remission rates and decrease adverse reactions. PCD represents a core mechanism mediating podocyte injury and MN progression. TCM effectively modulates PCD imbalance through multi-target regulation, demonstrating prominent clinical efficacy and safety. These findings provide a theoretical basis and translational direction for developing high-efficacy, low-toxicity therapeutic strategies for MN.",
"42553853": "ID: 42553853\nTitle: An artificial intelligence real-time rare sperm detection system for intraoperative microsurgical testicular sperm extraction.\nAbstract: How can rare spermatozoa be identified efficiently during microsurgical testicular sperm extraction (micro-TESE)? An artificial intelligence (AI)-assisted system was developed to flag candidate rare spermatozoa in real time during micro-TESE, and it may support embryologists as a decision-support tool. Patients with non-obstructive azoospermia (NOA) can obtain sperm for procreation through micro-TESE. During this procedure, sperm retrieval primarily relies on embryologists or laboratory technicians visually searching for sperm under a microscope, which is not only laborious and inherently subjective but also susceptible to errors. Although AI technology has been applied to identify trace amounts of sperm, existing models lack sufficient efficiency and true real-time performance. This study included model development followed by a single-centre clinical evaluation. An improved YOLO (You Only Look Once)-based rare sperm detection model, termed YOLOv11-RSD, was developed using microscopy data from 1165 surgical patients, comprising 1932 image samples containing a total of 5032 annotated sperm objects with confirmed identification. Clinical evaluation was performed between May 2024 and July 2025. Performance was assessed across confidence thresholds in obstructive azoospermia (OA) patients with normal spermatogenesis, and the system was then applied during micro-TESE in NOA patients and compared with routine embryologist assessment. The model was developed using testicular sperm microscopy images collected at a single hospital. Real-time clinical feasibility was evaluated in 10 OA cases and 30 NOA cases. Embryologist assessment was used as the reference standard, and performance was assessed using PPV, sensitivity, F1-score, and 95% confidence intervals. Discordant AI-assisted detections were reviewed by embryologists in real time. YOLOv11-RSD achieved real-time detection of candidate spermatozoa in microscopy images with high sensitivity and acceptable PPV under the selected operating threshold. Compared with baseline YOLOv11, YOLOv11-RSD showed improved overall detection performance across representative evaluation settings. In OA cases, the system achieved high sensitivity for sperm detection, reaching up to 96.7% across evaluated thresholds. During micro-TESE in NOA patients, at a confidence threshold of 0.50, positive predictive value (PPV), sensitivity, and F1-score were 80.58%, 96.11%, and 87.66%, respectively. The system highlighted candidate spermatozoa that were not identified during the initial manual assessment in six NOA cases, including two cases initially classified as sperm-negative; these findings were confirmed upon immediate re-review. Follow-up reproductive outcomes were available for six cases in which AI-assisted detection contributed to the search-and-confirmation workflow: embryo cleavage was achieved in all six cases, and three cases ultimately resulted in live births. Notably, among the two cases initially classified as sperm-negative, one case resulted in a singleton live birth. N/A. This was a single-centre clinical evaluation with a limited clinical cohort. Although model inference was rapid, procedure-level efficiency was constrained by image acquisition and scanning logistics, and no definitive reduction in total procedure time was demonstrated. External multi-centre validation is required. AI-assisted sperm detection may support embryologists during micro-TESE by flagging candidate rare spermatozoa for rapid review. Further prospective multi-centre validation is required to determine whether this approach improves procedure-level efficiency or clinical outcomes. This work was supported by grants from National Natural Science Foundation of China (82301794), Shanghai Science and Technology Innovation Action Plan (24Y12800702), Natural Science Foundation of Shanghai (25ZR1401300), National Key Research and Development Program of China (2022YFC270300), China Jiliang University Research Grant (No. H251120), and Shanghai General Hospital Basic and Clinical Collaborative Research Program (JC202612). The authors declare no competing interests.",
"42553858": "ID: 42553858\nTitle: Prolonged Retention of a Spontaneously Dislodging Pancreatic Duct Stent in a Patient with Surgically Altered Anatomy: A Case Report.\nAbstract: Spontaneously dislodging pancreatic duct stents are generally expected to pass within several days. We report a rare case of markedly prolonged retention of such a stent in a patient with surgically altered anatomy. A 78-year-old man with a history of distal gastrectomy with Roux-en-Y reconstruction underwent single-balloon enteroscopy-assisted ERCP for choledocholithiasis. After successful biliary cannulation using pancreatic guidewire assistance, stone extraction was performed following a small sphincterotomy and papillary balloon dilation. A 5-Fr \u00d7 5-cm spontaneously dislodging pancreatic duct stent without an internal flange was prophylactically placed. The postoperative course was uneventful; however, spontaneous stent dislodgement was not observed on follow-up imaging. Because of the technical burden of repeat endoscopy in surgically altered anatomy, careful observation was initially selected. The stent remained in place for 104 days and was ultimately removed endoscopically using a single-balloon enteroscope without adverse events. This case illustrates that markedly prolonged retention of a spontaneously dislodging pancreatic duct stent may occur in patients with surgically altered anatomy. Careful follow-up, including imaging to confirm spontaneous stent passage, is important, and endoscopic removal should be considered when the stent remains in place, even if repeat endoscopy is technically challenging.",
"42553881": "ID: 42553881\nTitle: Elucidating the Nexus of Mitochondrial Dysfunction and Oncometabolite Accumulation in Tumorigenesis.\nAbstract: Cancer is a complex disease driven by disruptions in cellular metabolism and mitochondrial function, enabling malignant cells to proliferate unchecked, evade apoptosis, and metastasize to distant organs. This exhaustive review elucidates the metabolic dysregulations inherent to cancer, with a particular focus on mitochondrial dysfunction, the accumulation of oncometabolites, and the reprogramming of metabolic pathways. A comprehensive literature search was conducted across major scientific databases, including PubMed, Web of Science, Scopus, and ScienceDirect, spanning January 2010 to March 2025. Controlled vocabulary and Boolean operators were employed to capture relevant studies, focusing on cancer metabolism, metabolic reprogramming, tumour markers, oncometabolites, mitochondrial dysfunction, and regulatory pathways. Extracted data were organized into thematic areas, and a qualitative synthesis approach was used to integrate findings, identifying common mechanistic patterns underlying tumour initiation, progression, and metastasis. The Warburg effect, a typical feature of cancer metabolism, is characterized by a predilection for aerobic glycolysis, thereby supporting biosynthetic processes and contributing to tumour microenvironment acidification and immune suppression. Mitochondrial dysfunction triggers genomic instability and oncogenic transformation. Meanwhile, oncometabolites like 2-hydroxyglutarate, fumarate, and sarcosine disrupt cellular signalling and epigenetic regulation, promoting tumour growth and progression. The clinical significance of tumour markers and metabolic biomarkers is underscored, and the systemic metabolic sequelae of cancer, including cancer-associated cachexia, are expounded upon. In glioblastoma, Aurora kinase A inhibition reverses the Warburg effect, decreasing glucose uptake and boosting oxidative phosphorylation. Cancer-associated fibroblasts exhibit aerobic glycolysis, promoting tumor growth and metastasis via the reverse Warburg effect. Glycolysis inhibition suppresses tumor growth in pancreatic cancer, and the Warburg effect contributes to chemoresistance by upregulating glycolytic enzymes and increasing lactate production. Targeting the Warburg effect, including inhibiting glycolytic enzymes and modulating mitochondrial function, offers potential therapeutic strategies for cancer treatment. This review provides a comprehensive exposition of the biochemical mechanisms underpinning metabolic derangements in cancer, which may unveil novel avenues for diagnostic and therapeutic interventions.",
"42553894": "ID: 42553894\nTitle: Lipid oxidation-mediated non-enzymatic glycation and its impact on the digestibility of milk proteins.\nAbstract: This study explored the influence of milk fat on non-enzymatic glycation of proteins and protein digestibility by analyzing the accumulation of glycation products and changes in protein structure. The contents of POV and MDA increased significantly on day 7 of storage in milk model groups with different milk fat concentrations (0, 2, 4, 6%) and the increase was more obvious in the high-fat (6%) group. In the high-fat group, the contents of glyoxal and methylglyoxal, the intermediate products of non-enzymatic glycation increased at a high level. The peak shift and the appearance of new peaks in the infrared spectrum further indicated enhanced glycation coupled with lipid oxidation. In addition, the decrease in fluorescence intensity, the increase in ultraviolet absorption, the reduction in surface hydrophobicity, and the change of protein secondary structure in the high fat group indicated obvious changes in the microstructure of proteins. The cleavage site of protease was reduced because of the alterations in protein advanced structure and the occupation of amino acids digestion sites, which subsequently reduced the digestibility of proteins. This phenomenon was more obvious in the early stage of gastrointestinal digestion. High levels of milk fat promoted non-enzymatic glycation by accelerating lipid oxidation, which in turn induced glycation, cross-linking, and aggregation of milk proteins. These structural changes further hindered the action of digestive enzymes, ultimately reducing the digestibility of milk proteins.",
"42553904": "ID: 42553904\nTitle: Conscious sedation as a potential optimal anesthetic regimen for endoscopic retrograde cholangiopancreatography in elderly frail patients: A prospective, single-blind, randomized controlled trial.\nAbstract: To investigate the impact of adverse cardiovascular events (ACVs) in elderly frail patients undergoing emergency endoscopic retrograde cholangiopancreatography (ERCP), and to explore a safe and comfortable anesthetic regimen suitable for this specific population. A single-center, prospective, single-blind, randomized controlled trial was conducted. Patients aged 70-90 years scheduled for emergency ERCP were enrolled and randomly divided into two groups: the conscious sedation group (Group S) and the intravenous general anesthesia group (Group A). Basic clinical data of the patients were recorded. During ERCP, the occurrence of ACVs was documented, including hemodynamic fluctuations exceeding 20% of the baseline value, angina pectoris, myocardial infarction, cardiac arrest, acute heart failure, myocardial ischemia, pulmonary embolism, and various arrhythmias. Additionally, adverse reactions such as hypoxemia, body movement, and cough during the procedure were monitored. The effects of different anesthetic methods on ACVs in elderly frail patients undergoing emergency ERCP were compared, and the satisfaction of patients, anesthesiologists, and endoscopists was evaluated. A total of 108 patients undergoing emergency ERCP were enrolled and randomly assigned to Group S (n = 53) and Group A (n = 55). The results showed that there was a statistically significant difference in the incidence of ACVs between the two groups (17% vs. 40%, P < 0.05). Compared with Group A, Group S had a significantly lower incidence of blood pressure and heart rate fluctuations \u226520% of the baseline value (13.2% vs. 25.5%, P < 0.05), and new-onset arrhythmias (5.7% vs. 18.2%, P < 0.05). Compared with intravenous general anesthesia, conscious sedation can reduce fluctuations in blood pressure and heart rate, and decrease the incidence of adverse cardiovascular events in elderly frail patients undergoing emergency ERCP.",
"42553942": "ID: 42553942\nTitle: Correction: Influence of commensal bacteria on the proteolytic and antigenic profiles of INFOGEST-like digested wheat gliadin.\nAbstract: [This corrects the article DOI: 10.3389/fmicb.2026.1842801.].",
"42553990": "ID: 42553990\nTitle: Ethylene-responsive CmERF3-CmMYB4 cascade suppresses anthocyanin biosynthesis in Chrysanthemum \u00d7morifolium.\nAbstract: Anthocyanins are the primary determinants of floral pigmentation in Chrysanthemum \u00d7morifolium, and ethylene acts as a key regulator of their biosynthesis. Although the ethylene-mediated regulatory circuitry is functionally important, its underlying mechanism in chrysanthemum has remained unclear. In this study, we identified CmMYB4 as a transcriptional repressor that directly suppresses the expression of key anthocyanin biosynthetic genes, including CmDFR (dihydroflavonol 4-reductase), CmUFGT (flavonoid 3-O-glucosyltransferase), and Cm3MaT (anthocyanin 3-O-glucoside-6\u2033-O-malonyltransferase). Time-ordered gene co-expression network analysis comparing the transcriptomes of CmMYB4-overexpressing and control plants, together with molecular biology experimental results, further revealed CmERF3 (ethylene response factor 3) as a hierarchical upstream regulator of anthocyanin biosynthesis. Exogenous ethylene treatment induced CmERF3 expression while reducing anthocyanin accumulation. Subsequent functional characterization showed that the overexpression of CmERF3 suppresses anthocyanin biosynthesis in both tobacco and chrysanthemum by directly activating CmMYB4 and repressing CmDFR, CmUFGT, and Cm3MaT. Collectively, these findings revealed that ethylene inhibits anthocyanin biosynthesis through a CmERF3-CmMYB4-LBGs (late biosynthetic genes) regulatory module. This study not only elucidates the molecular mechanism governing ethylene-mediated anthocyanin inhibition but also provides new perspectives for the molecular engineering of ornamental traits in chrysanthemum.",
"42554055": "ID: 42554055\nTitle: Mast Cell St8sia1 Is a Glyco-Epigenetic Checkpoint Driving Cardiac Remodeling.\nAbstract: Pathogenic immune-cardiac crosstalk underlies maladaptive remodeling in chronic heart failure, yet therapies directly targeting this axis are lacking. Glycoconjugates, which are crucial for signal transduction and extracellular matrix integrity, represent an underexploited therapeutic avenue. This study sought to define the role of glycoconjugate-metabolizing enzymes at the immune-cardiac interface and evaluate their translational potential. We performed integrative analyses of bulk and single-cell RNA sequencing data from failing human and mouse hearts. Employing mouse models of pressure overload (transverse aortic constriction) and ischemia-reperfusion, we used global and mast cell (MC)-specific gene deletion, bone-marrow chimeras, and pharmacological neutralization. Mechanistic insights were gained through multiomics profiling, including RNA-seq, ATAC-seq, CUT&Tag, and proteomics. The ganglioside GD3 synthase, St8sia1, was selectively induced in cardiac MCs during pathological remodeling in both mice and humans. MC-specific or hematopoietic deletion of St8sia1 preserved ventricular function, attenuated fibrosis, and markedly reduced neutrophil and Ly6C+ monocyte recruitment after transverse aortic constriction and ischemia-reperfusion. Therapeutic neutralization of GD3 with the clinical-grade monoclonal antibody R24 improved cardiac function and diminished scar formation after ischemia-reperfusion. Mechanistically, GD3 bound specific histone variants, such as H2A.Z and H3.3C, thereby reprogramming chromatin accessibility to activate proinflammatory and profibrotic transcriptional programs in MCs. Consequently, GD3 inhibition suppressed MC degranulation, disrupted pathogenic MC-cardiomyocyte/fibroblast crosstalk, and preserved reparative macrophage populations. The MC-restricted St8sia1-GD3 axis functions as a glyco-epigenetic checkpoint driving maladaptive cardiac remodeling. Targeting this axis represents a translatable immunomodulatory strategy to prevent the progression to chronic heart failure.",
"42554069": "ID: 42554069\nTitle: Ultrasound-Guided Targeted Combination Therapy Integrating VEGFR Degradation Independent of a Specific Receptor, Mitochondrial Damage, and Sonodynamic Immunomodulation.\nAbstract: Inhibiting the vascular endothelial growth factor-vascular endothelial growth factor receptor (VEGF-VEGFR) signaling pathway is clinically established for solid tumors, but durable efficacy is frequently limited by acquired resistance. Accumulating evidence links this resistance to metabolic plasticity, wherein tumors suppress glycolysis and gradually become dependent on mitochondrial respiration for survival, rendering mitochondrial inhibition a rational strategy to mitigate resistance. Recently, targeted protein degradation (TPD) has emerged as an event-driven modality for pathogenic protein elimination, but the degradation of membrane targets is still challenging due to limited transmembrane E3 ligases and heterogeneous lysosome-targeting receptor (LTR) expression. To overcome these challenges, we designed a multifunctional peptide-sonosensitizer conjugate (PSC) by covalently linking a VEGFR-targeting peptide, a mitochondria-homing peptide, and the clinically approved sonosensitizer verteporfin (VPN) via a gelatinase-cleavable linker. PSC self-assembles into stable nanoparticles with tumor-preferential accumulation and, upon ultrasound activation, generates reactive oxygen species (ROS) to induce oxidative degradation of VEGFR, thereby suppressing VEGFR signaling and angiogenesis. Following cellular internalization, enzymatic linker cleavage releases a mitochondria-targeted fragment that triggers mitochondrial depolarization and structural disruption, amplifying ROS-driven apoptosis. Moreover, sonodynamic immune modulation further potentiates antitumor efficacy. This study establishes a spatiotemporally controllablestrategy that is independent of a specific receptor and couples anti-angiogenic pathway blockade with mitochondrial vulnerability to address resistance and enable precision, immune-synergistic therapy.",
"42554093": "ID: 42554093\nTitle: Spatiotemporally Controlled Lysosomal Membrane Permeabilization Amplifies STING-Driven Antitumor Immunity in Prostate Cancer.\nAbstract: Prostate cancer (PCa) remains a major clinical challenge due to therapeutic resistance and immunologically cold tumor microenvironment. Lysosomal membrane permeabilization (LMP)-induced lysosome-dependent cell death offers an alternative route to eliminate resistant tumor cells and initiate immunogenic cell death, yet its efficacy is often limited by insufficient spatiotemporal control and immune activation. Here, we report a spatiotemporally programmable supramolecular nanoplatform (Cu-P-MSA) that integrates lysosome-targeted sonodynamic therapy with tumor-confined innate immune activation for PCa treatment. Cu-P-MSA is a modular self-assembling peptide incorporating a PSMA-targeting ligand, morpholine moiety, and cathepsin B-cleavable linker, enabling tumor-selective uptake and in situ formation of fibrous sonosensitizer depots within lysosomes. Upon ultrasound irradiation, a glutathione-responsive open-shell sonosensitizer induces controlled LMP, simultaneously activating ferroptosis and pyroptosis and promoting immunogenic cell death. Meanwhile, tumor-specific release of a STING agonist MSA-2 elicits robust type I interferon responses, driving dendritic cell maturation and cytotoxic T-cell infiltration. This coordinated lysosomal disruption-immune amplification strategy effectively reprograms the tumor immune microenvironment and suppresses both primary and distant tumors, with inhibition rates reaching 84.3% and 77.5%, respectively. Overall, this work establishes a spatiotemporally controlled supramolecular approach that integrates lysosomal disruption with innate immune activation to overcome therapeutic resistance and immunosuppression in PCa.",
"42554116": "ID: 42554116\nTitle: The Quest for the Associative Device for Hebbian-Like LTP-The Accidental Participant.\nAbstract: In 1985 Holger Wigstr\u00f6m (now deceased) and I published a proposed scenario for the induction of hippocampal Hebbian-like LTP to explain its input specificity and cooperativity. In this scenario LTP is induced by calcium influx through voltage-dependent NMDA receptor channels co-localized with the glutamate receptor channels (non-NMDA) mediating the expression of LTP. This co-localization, we argued, explained the input specificity, and the voltage dependence of these NMDA receptor channels explained the cooperativity. It will be described how this scenario emerged from our finding of a greatly facilitated LTP induction following blockade of postsynaptic inhibition, from our observation of a dendritic, possibly regenerative, event correlated with successful LTP induction, and to our recognition that these events were the result of NMDA receptor activation. Our observation of a likely synaptic co-localization of these receptor channels with the non-NMDA receptor channels, with the former contributing to an EPSP component with a slower onset and much longer duration, will be described. Finally, I will describe how our scenario became validated by our experiments pairing such single EPSPs with depolarization induced by injected current pulses, temporally associated with the NMDA component. This essay will conclude with some take-home messages regarding the Hebbian-like LTP as learned from these experiments: this form of synaptic learning most likely only occurs during disinhibition, its longevity strongly depends on the induction strength from a few minutes and upwards, with a strong induction event it can operate as a one-shot potentiation, and its time window for association may remain several 100\u2009ms. Lastly, Hebbian-like LTP likely evolved to associate input events rather than an input event and somatic spike activity.",
"42554175": "ID: 42554175\nTitle: Ubiquitin-mediated degradation of RPN13 promotes porcine sperm capacitation and sperm-zona pellucida binding.\nAbstract: Sperm capacitation involves biochemical and functional changes necessary for fertilization. The ubiquitin-proteasome system (UPS) regulates protein turnover, but the roles of proteasomal receptors during capacitation remain unclear. This study investigates the regulation and functional impact of proteasomal subunits RPN13 (ADRM1) and RPN2 (PSMD1) during in vitro capacitation of boar sperm. Semen samples (n = 6) were assigned to four groups: non-capacitating control (Non-Cap, 4 h in medium lacking bovine serum albumin (BSA)), capacitating control (Cap, 4 h with BSA) and groups treated with vehicle (CM + dimethyl sulfoxide (DMSO)) or the RPN13 inhibitor RA190. We measured RPN2/RPN13 levels, RPN13-associated ubiquitin signal and RPN13-RPN2 interactions (Western blot, Co-IP) and assessed sperm kinematics, intracellular Ca2+, protein tyrosine phosphorylation (pTyr), mitochondrial membrane potential (\u0394\u03a8m), ATP, ZP binding and early embryo cleavage after IVF. The Cap group showed successful capacitation, confirmed by increases in pTyr, sperm hyperactivation and Ca2+-associated Fluo-4 fluorescence. During capacitation, RPN2 and RPN13 abundance decreased, while RPN2-RPN13 association increased. RA190 treatment increased RPN13 levels, inhibited RPN13 turnover and decreased its interaction with RPN2 and the associated ubiquitin signal. Functionally, RA190 attenuated sperm hyperactivation, Ca2+-associated Fluo-4 fluorescence, protein tyrosine phosphorylation, mitochondrial membrane potential and ATP levels and was associated with reduced fertilization-related outcomes, including ZP binding and early embryo cleavage after IVF. These findings highlight the role of RPN13 in sperm capacitation and fertilization.",
"42554179": "ID: 42554179\nTitle: Regulation of actin polarization by Edwardsiella piscicida during invasion of Sebastes schlegeli intestinal mucosa and cellular immune responses.\nAbstract: Intestinal epithelial cells play a crucial role in teleost defence against complex aquatic environments. However, the study of intestinal mucosal immunity in teleosts has been hindered by the lack of reliable and specific marker genes to distinguish between different cell types. In this study, single-cell RNA-seq analysis of the intestinal epithelial cells from healthy and infected Sebastes schlegeli was performed to investigate the heterogeneity of teleost intestinal epithelial cells. Immune cells, including T cells, B cells, non-specific cytotoxic cells (NCCs) and myeloid cells, and non-immune cells including enterocyte, erythrocyte, BEST4+ cell, goblet cell and enteroendocrine cell, were identified. Several notable cell populations, including early myeloid cells, lysosome-rich enterocytes, enterocyte progenitors and BEST4+ cells, were characterized in detail. Additionally, significant changes in the expression of actin polymerization-related genes were observed in various intestinal mucosal cell types following Edwardsiella piscicida infection, suggesting a mechanism of invasion with E. piscicida. These findings enhance our understanding of teleost intestinal cellular composition and provide valuable insights for the development of therapeutic strategies against E. piscicida.",
"42554219": "ID: 42554219\nTitle: Hybrid quantum-mechanistic insights into \u03b2-O-4 ether cleavage in lignin-carbohydrate complexes using a choline chloride-lactic acid DES cluster model.\nAbstract: Lignin-carbohydrate complexes (LCCs) contribute to biomass recalcitrance, making \u03b2-O-4 ether linkages important targets for selective lignin depolymerization. \u03b2-O-4 cleavage in a representative LCC was studied using a hybrid quantum-mechanical/semiempirical (QM/xTB) approach, an explicit choline chloride/lactic acid (ChCl\u2009:\u2009LA) cluster, and M06-2X/6-31+G(2d,2p) refinement. Vacuum calculations were compared with C-PCM water (\u03b5 = 78), which was used as a high-polarity reference and upper-bound estimate of dielectric stabilization rather than as a representation of bulk deep eutectic solvent microstructure. Frontier-orbital, Fukui-function, and Hirshfeld-charge analyses identify C43 as the electrophilic center for chloride attack and O28 as the leaving-group atom, while revealing progressive electron-density loss from chloride. The optimized transition state (TS) supports a concerted SN2-like mechanism in which chloride attacks C43 as the O28-C43 bond elongates from 1.43 to 2.25 \u00c5. Thermochemically corrected stationary-point calculations at 298.15 K give Gibbs activation energies of 93.2 kJ mol-1 for the isolated explicit cluster and 75.8 kJ mol-1 with a C-PCM water dielectric, indicating a 17.5 kJ mol-1 sensitivity to high-dielectric screening. Decomposition of these barriers yielded optimized electronic activation energies of 121.0 and 119.7 kJ mol-1 for the isolated and dielectric-embedded clusters, whereas thermochemical corrections accounted for most of the predicted reduction and highlighted the substantial contribution of thermal and entropic terms to the model-dependent free-energy estimates. This study provides a descriptor-based mechanistic rationale for \u03b2-O-4 cleavage in an explicit ChCl interaction model and clarifies how local chloride coordination and lactic-acid hydrogen-bonding contacts can promote \u03b2-O-4 bond activation, providing a mechanistic framework for designing greener solvents for selective biomass fractionation.",
"42554260": "ID: 42554260\nTitle: Endoscopic management of common bile duct stones using double balloon endoscopy in postoperative patients: evaluation of single-session complete stone clearance.\nAbstract: Endoscopic treatment of common bile duct (CBD) stones in patients with surgically altered anatomy is challenging because access to the duodenal papilla is often difficult. Although double balloon endoscopy-assisted ERCP (DB-ERCP) has become widespread, data on outcomes by reconstruction type remain limited. A total of 218 patients treated between November 2016 and October 2024 were retrospectively reviewed. Among these, 212 patients were included: 159 with Roux-en-Y reconstruction (R group) and 53 with Billroth II reconstruction (B group). The primary endpoints were single-session complete stone clearance and procedure time. The single-session complete stone clearance rate was 94.8%, with no significant difference between groups. Procedure time was significantly longer in the R group than in the B group (66.0 vs. 50.0\u2009min, p\u2009<\u20090.001). Multivariate analysis identified R group (p\u2009<\u20090.001), age \u226575\u2009years (p\u2009=\u20090.007), and \u22655 stones (p\u2009=\u20090.018) as independent factors for prolonged procedure time. In addition, \u22655 stones independently reduced single-session complete stone clearance (p\u2009<\u20090.001). Adverse events occurred in 2.8%, including one perforation, four bleeding events, and one pancreatitis case. Only one patient required surgery. DB-ERCP achieved a high single-session complete stone clearance rate in patients with surgically altered anatomy, with outcomes comparable to those of conventional ERCP in patients with normal anatomy. These findings suggest that DB-ERCP may represent a first-line treatment option for CBD stones in surgically altered anatomy.",
"42554268": "ID: 42554268\nTitle: BRCA2-deficient endothelium: a hidden catalyst in angiotensin II-induced vascular injury.\nAbstract: Mutations in the breast cancer susceptibility gene 2 (BRCA2) are well known to increase the risk of breast and ovarian cancers. Emerging evidence indicates that BRCA2 mutation carriers exhibit increased vascular disorder and may develop endothelial dysfunction, a key mechanism underlying hypertension. Angiotensin II (Ang II)\ufeff, a central effector of the renin-angiotensin system, is a key regulator of blood pressure and a major driver of hypertension, promoting endothelial injury through oxidative stress, inflammation, and impaired nitric oxide (NO) bioavailability. However, the role of endothelial BRCA2 in Ang II-induced endothelial dysfunction remains unknown. BRCA2 was silenced in cultured endothelial cells and following Ang II treatment, ROS generation, DNA damage, apoptosis, inflammation, NO production, migration, angiogenic capacity, Ang II receptors and related signaling pathways were assessed. BRCA2 deficiency exacerbated Ang II-induced increases in ROS, DNA damage, and apoptosis, along with impaired functional capacity, including reduced migration and angiogenesis. NO production was suppressed, accompanied by increased micronuclei formation and enhanced c-Jun N-terminal kinase (JNK) activation. In addition, both BRCA2 loss and Ang II treatment upregulated the Ang II receptor AT1R. Pharmacological inhibition of AT1R attenuated Ang II-induced increases in DNA damage and apoptosis in BRCA2-deficient endothelial cells. These findings provide the first pharmacogenomic evidence that BRCA2 deficiency sensitizes endothelial cells to Ang II-induced dysfunction, suggesting that BRCA2 mutation carriers may be at increased risk for hypertension-associated cardiovascular complications.",
"42554299": "ID: 42554299\nTitle: Hyperpolarized [2-13C]Pyruvate Identifies a Mitochondria-Active Hepatocellular Carcinoma Phenotype With Vulnerability to Mitochondrial Inhibition.\nAbstract: Hepatocellular carcinoma (HCC) exhibits metabolic heterogeneity that is not fully characterized by glycolysis-focused spectroscopic profiling. This study investigated whether in\u00a0vitro hyperpolarized (HP) [2-13C]pyruvate NMR spectroscopy can identify a mitochondria-active HCC phenotype and assess its association with sensitivity to mitochondrial metabolic inhibition. HP [2-13C]pyruvate NMR spectroscopy was used to evaluate mitochondrial metabolism in McA-RH7777 HCC cells, with N1S1 cells serving as a glycolysis-dominant reference. Cell viability following treatment with the glutaminase inhibitor BPTES and the mitochondrial metabolic inhibitor CPI-613 was assessed by MTT assay, and metabolic changes following CPI-613 treatment were further evaluated using HP [2-13C]pyruvate. HP [2-13C]pyruvate demonstrated enhanced pyruvate-to-glutamate conversion in McA-RH7777 cells, whereas N1S1 showed minimal glutamate labeling. CPI-613 treatment resulted in a dose-dependent reduction in cell viability, while BPTES produced limited effects. Although pyruvate-to-glutamate conversion did not significantly decrease following CPI-613 treatment, pyruvate-to-lactate conversion increased, indicating metabolic adaptation. These findings demonstrate that HP [2-13C]pyruvate enables functional identification of a mitochondria-active HCC phenotype characterized by enhanced pyruvate-to-glutamate conversion. This approach may facilitate metabolic subtype classification, help identify tumors susceptible to mitochondrial metabolic inhibition, and enable non-invasive monitoring of treatment-induced metabolic adaptation.",
"42554321": "ID: 42554321\nTitle: Dynamic Heterovalent Dual-Cu Sites for O\u2550O Cleavage in Electrocatalytic Oxygen Reduction Reaction.\nAbstract: Activation and cleavage of the inert O\u2550O bond represent a central challenge in energy electrochemistry. Here, the Cu dual-atom catalyst (Cu-DAC) is constructed via bottom-up pre-coordination assembly and post-encapsulation pyrolysis. The resulting Cu-DAC features a well-defined Cu-Cu distance (\u223c3.31 \u00c5) with switchable Cu1+/Cu2+ states, enabling dynamic dual-site coordination with O2. Cu-DAC achieves 0.87 VRHE half-wave potential for oxygen reduction reaction (ORR), a near-unity 4e- selectivity, and outstanding stability. Multiple operando spectroscopic characterizations and ab initio dynamic simulations (AIMD) reveal that the dynamic heterovalent [Cu1+\u2500O\u2500O\u00b7 -\u2500Cu2+] unit elongates the O-O bond and promotes its cleavage via dual-site confinement and electron donation. By specifically elucidating how these dual-atom sites dynamically evolve to facilitate O\u2500O bond cleavage, we provide vital atomic-level principles for the rational design of dynamically active catalysts.",
"42554361": "ID: 42554361\nTitle: Platelet-Derived Exosomal miR-142-3p Inhibits Macrophage M1 Polarization and PANoptosis to Alleviate Acute Lung Injury via the MAPK Pathway.\nAbstract: Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is an acute, life-threatening form of pulmonary disease characterized by systemic inflammation, for which current treatments are not sufficiently effective. Platelet-derived exosomes (PLT-exos) are noted for their specific targeting ability to inflammatory sites, minimal immunogenicity, and positive anti-inflammatory effects, positioning them as a promising therapeutic candidate. However, the specific effects and mechanisms of PLT-exos in ALI/ARDS remain poorly understood. Here, utilizing a lipopolysaccharide (LPS)-induced ALI model in murine and cell models, we first demonstrated that activated platelet-derived exosomes (AP-exos) are preferentially taken up by lung macrophages, leading to suppressed macrophage M1 polarization and PANoptosis and reduced lung inflammation and injury, which were closely linked to the inhibition of MAPK pathway activity. Mechanistically, the protective effects of AP-exos were partially mediated by the highly enriched miR-142-3p, as evidenced by the intervention with miR-142-3p significantly inhibiting their protective effect against LPS-induced ALI injury in mice. Furthermore, we noted a negative correlation between the level of miR-142-3p derived from plasma exosomes and the severity of ARDS, suggesting its potential in predicting the severity of ARDS. Further investigation revealed that TMEM120B (T120B) functions as a downstream target of miR-142-3p. Overall, the findings suggested that activated platelets release exosomes that inhibit macrophage M1 polarization and PANoptosis, contributing to the pathogenesis of ARDS via the miR-142-3p/T120B/MAPK axis.",
"42554406": "ID: 42554406\nTitle: Morphological Changes in Fungal Elements and Antifungal Drug Susceptibility in an Ex Vivo Model of Fungal Keratitis.\nAbstract: Fungal keratitis is a serious eye infection that can cause irreversible vision loss. One of the causative fungal pathogens, Fusarium solani (Neocosmospora solani), shows limited susceptibility to antifungal drugs. Furthermore, differences between in vitro and clinical outcomes are encountered in some cases, complicating treatment. In this study, we compared in vitro and ex vivo antifungal susceptibility using a porcine corneal model of F. solani keratitis. An ex vivo fungal keratitis model was developed using porcine corneas, which were inoculated with F. solani and assigned to four treatment groups: natamycin (NAT), voriconazole (VRCZ), luliconazole (LLCZ), or saline (uninfected control). We compared the antifungal efficacy of each drug on day 2 to day 4 of fungal development. DNA was extracted from all corneas and used in real-time polymerase chain reaction to quantitatively assess fungal growth. Fungal morphology was examined by digital microscopy. All antifungal treatment groups demonstrated antifungal activity. The NAT-treated group showed clear inhibition of fungal growth as well as fungal structural damage. The VRCZ-treated group exhibited growth inhibition; however, fungal growth persisted even after additional drug administration. The LLCZ-treated group not only suppressed fungal growth but also caused structural disruption of fungal cells after additional administration, suggesting a strong antifungal effect. We successfully established an ex vivo Fusarium keratitis model using porcine corneas, which enabled us to evaluate antifungal susceptibility by quantifying fungal growth and observing fungal morphological changes.",
"42554438": "ID: 42554438\nTitle: Matrix Stiffness-Encoded Mechanical Memory Maintains Proliferative Dominance in Metastatic Tumors.\nAbstract: Primary and metastatic lesions share similar biological properties despite existing in different microenvironments. A better understanding of the mechanisms underlying the maintenance of phenotypic homogeneity in heterogeneous microenvironments could help identify strategies for suppressing metastasis. Here, we found that mechanical memory enabled tumor cells to resist biomechanical stress and sustain malignant traits. Matrix stiffness activated the RhoA-ROCK1 signaling pathway, leading to actin cytoskeletal remodeling and suppression of mitochondrial fission. The dysregulation of mitochondrial dynamics enhanced fatty acid \u03b2-oxidation, leading to the accumulation of acetyl-CoA and subsequent histone hyperacetylation. The stiffness-mediated epigenetic reprogramming was mitotically heritable, allowing progeny cells to retain a proliferative advantage after detachment from stiff environments. Inhibition of RhoA-ROCK1 disrupted mechanical memory, thereby reducing metastasis. These findings suggest that targeting mechanical memory may be a strategy for preventing metastasis.",
"42554467": "ID: 42554467\nTitle: The conserved QTQTX motif in the SARS-CoV-2 spike protein is dispensable for cleavage and lung cell entry of the emerging variant BA.3.2.\nAbstract: The furin motif in the SARS-CoV-2 spike (S) protein is important for lung cell entry. It is embedded in an extended loop structure and preceded by a highly conserved QTQTX motif that is required for efficient furin cleavage of the SARS-CoV-2 WA-1 S protein. BA.3.2 is an emerging SARS-CoV-2 saltation variant that is spreading globally in April 2026 and encodes a highly mutated S protein. Here, we analyzed whether the QTQTX motif is also required for spike protein cleavage and lung cell entry of BA.3.2. We report that two patient-derived spike sequences of the BA.3.2 subvariant BA.3.2.2 lack the first QT repeat of the QTQTX motif and show that this motif is largely dispensable for both cleavage and lung cell entry of BA.3.2.2, which we found to depend on TMPRSS2. Our results suggest that the reconfiguration of the BA.3.2 S protein during persistent infection may have significantly altered the determinants of furin cleavage.IMPORTANCEThe furin motif in the SARS-CoV-2 spike (S) protein is unique among sarbecoviruses and constitutes a virulence determinant. A QTQTX motif located immediately upstream of the furin motif is required for furin cleavage of the S protein of the virus that circulated early in the pandemic. Here, we show that the QTQTX motif is largely dispensable for S protein processing and S protein-driven lung cell entry of the emerging saltation variant BA.3.2, which is currently spreading globally. Thus, BA.3.2 evolution within immunocompromised individuals may have relaxed the requirements for furin processing of the spike protein.",
"42554501": "ID: 42554501\nTitle: BetaH proteolysis unleashes an electrostatic-homing antibacterial polymorphic toxin.\nAbstract: Contact-dependent and diffusible proteinaceous polymorphic toxin systems (PTSs) mediate widespread bacterial competition. While bioinformatic analyses have identified diverse PTSs across bacterial phyla, experimental validation in gram-positive species remains limited. Here, we characterize a diffusible polymorphic toxin encoded by the Staphylococcus aureus S8-Ntox35 locus. We demonstrate that this system mediates inter-genus antagonism against Listeria monocytogenes via a bactericidal, heat-labile protein and that toxicity is enhanced following extracellular cleavage of the BetaH domain by the associated S8 peptidase. This processed toxin bears a peptide resembling a cationic antimicrobial peptide (CAMP) and facilitates intoxication of target cells by the Ntox35 RNase domain. Target cell resistance is impacted by known CAMP defense pathways, including DltABCD and MprF, and experimental evolution identified the ATP-binding cassette (ABC) transporter AnrAB as essential for intoxication. Unexpectedly, disruption of AnrAB abolished Ntox35 susceptibility, while simultaneously sensitizing cells to the proposed CAMP-like activity of the processed BetaH domain. These findings reveal a novel mechanism of inter-genus antagonism among Firmicutes (Bacillota) and establish a functional role for extracellular processing and ABC transporter-mediated susceptibility in PTS activity. Our work expands the known repertoire of diffusible toxins in gram-positive bacteria and sets the foundation for broader ecological and mechanistic investigation of the S8-associated polymorphic toxin system (S8-PTS). Polymorphic toxin systems (PTSs) are widely used by bacteria to inhibit competitors, but diffusible proteinaceous toxins have been largely characterized in gram-negative species. Here, we mechanistically characterize a diffusible PTS in Staphylococcus aureus that mediates inter-genus antagonism in the Firmicutes phylum. This system employs a secreted S8 peptidase to cleave a BetaH-toxin fusion, releasing a previously caged cationic amphipathic \u03b1-helix that facilitates membrane targeting and intoxication of susceptible cells. We further show that toxin activity requires the target cell ATP-binding cassette (ABC) transporter AnrAB, revealing a novel route of entry and an evolutionary tradeoff between toxin susceptibility and antimicrobial peptide resistance. Together, these findings uncover a new mode of bacterial competition and highlight a broadly distributed toxin system with ecological relevance across Firmicutes.",
"42554506": "ID: 42554506\nTitle: A Novel tsRNA, 5'tiRNA-GluCTC, Mediates Cardiomyocyte-Fibroblast Crosstalk to Promote Cardiac Fibrosis in Restrictive Cardiomyopathy.\nAbstract: Restrictive cardiomyopathy (RCM) is characterized by pronounced cardiac fibrosis, leading to ventricular stiffening and diastolic dysfunction. While cardiomyocyte mutations are known triggers, the mechanisms initiating pro-fibrotic signaling remain elusive. This study investigates the role of cardiomyocyte-derived exosomes and specific tRNA-derived small RNAs (tsRNAs) in this pathogenic intercellular communication.\u00a0Using a cTnIR193H\u00a0knock-in mouse model, we observed pronounced cardiac fibrosis prior to the onset of heart failure, without significant cardiomyocyte\u00a0apoptosis. This primary fibrotic response was mediated by a paracrine mechanism, as conditioned medium from mutant cardiomyocytes was sufficient to activate fibroblasts. Subsequent small RNA sequencing\u00a0of cardiomyocyte-derived exosomes identified the tRNA-derived fragment 5'tiRNA-GluCTC\u00a0as a significantly enriched species. Functional studies established this RNA as a critical mediator, demonstrating that its overexpression exacerbated fibrotic responses in vitro and induced fibrotic remodeling in wild-type mice, while its inhibition via a cardiac-targeted AAV sponge\u00a0attenuated fibroblast activation and alleviated fibrosis in RCM mice. Mechanistically, 5'tiRNA-GluCTC\u00a0is transferred to fibroblasts, directly targets the 3'UTR of Foxq1\u00a0to repress its expression,\u00a0and\u00a0consequently activates the Smad3/p-Smad3 signaling cascade.\u00a0Collectively, these results delineate a 5'tiRNA-GluCTC/Foxq1/Smad3 signaling axis that drives fibrosis in RCM, uncovering a previously uncharacterized signaling cascade with therapeutic potential.",
"42554508": "ID: 42554508\nTitle: Fibrilpaint: A Class of Amyloid-Targeting Peptides.\nAbstract: Amyloid fibrils are a common pathological hallmark in multiple neurodegenerative diseases, yet molecular tools to recognize and manipulate them remain scarce. We report FibrilPaints, a family of modular peptides designed for amyloid binding and adaptable chemical functionality. The degenerative amyloid-targeting unit of FibrilPaints, W5P4H3R2, has a high content of \u03c0-stacking and aromatic side chains. Systematic sequence variation, altering charge, termini, and residue order, revealed the importance of the composition of the amyloid-targeting unit for high-affinity binding across Tau and Huntingtin fibrils. Importantly, sequence changes outside this unit do not preclude fibril binding, which permits attachment of fluorophores or E3-recruiting motifs for targeted protein degradation. This work establishes FibrilPaint as a modular peptide system for binding Tau and Huntingtin amyloid fibrils, with potential for broader detection and modulation applications.",
"42554509": "ID: 42554509\nTitle: Mepce Restrains Ferroptosis in Prostate Cancer Through 7SK-P-TEFb Pause Control.\nAbstract: Ferroptosis can be therapeutically induced through GPX4 inhibition, but the mechanisms that determine ferroptosis sensitivity in prostate cancer remain incompletely understood. Here, we combined genome-wide CRISPR screening under graded GPX4 inhibition with mechanistic perturbation-rescue experiments and validation in xenograft and immunocompetent syngeneic prostate cancer models. We identified MEPCE loss as a conserved sensitizer to ferroptosis. Mechanistically, MEPCE depletion destabilized RN7SK and disrupted the HEXIM1-P-TEFb complex, consistent with CDK9 release and enhanced RNA polymerase II Ser2 phosphorylation, thereby promoting transcriptional elongation. This state accelerated activation of a NRF2/ARE stress program encompassing antioxidant defense and iron mobilization. Within this program, HMOX1 emerged as a kinetically sensitive effector whose rapid induction promoted labile iron accumulation and lipid peroxidation under GPX4 inhibition. In vivo, MEPCE suppression impaired tumor growth, enhanced the antitumor efficacy of ferroptosis induction, and was associated with pharmacodynamic markers of ferroptosis that were partially reversed by ferroptosis- or iron-targeted rescue. Together, these findings identify a transcriptional pause-control mechanism that links elongation dynamics to ferroptosis susceptibility and nominate the MEPCE-RN7SK-P-TEFb axis as a potential therapeutic target for sensitizing prostate cancer to ferroptosis.",
"42554513": "ID: 42554513\nTitle: Macropa-Based 225Ac-Labeled RGD Peptides for Targeted Alpha Therapy.\nAbstract: Macropa has formed more stable complexes with 225Ac, although DOTA has been extensively used as a chelator for 225Ac. This study aimed to design and synthesize two novel 225Ac-labeled RGD peptides, [225Ac]Ac-Macropa-c(RGDyK) ([225Ac]1) and [225Ac]Ac-Macropa-[c(RGDyK)]2 ([225Ac]2), and to evaluate their potential for targeted alpha therapy (TAT) of glioblastoma. Further, whether combination therapy with homoarginine (hArg), a modulator of lysosomal function, could further improve therapeutic efficacy was investigated. In Colon-26 cells, [225Ac]2 demonstrated significantly higher cellular uptake and cytotoxicity than [225Ac]1. In Colon-26 tumor-bearing mice, [225Ac]2 exhibited higher tumor accumulation and significant tumor growth inhibition. In GL261 glioblastoma cells, [225Ac]2 showed comparable cellular uptake to that in Colon-26 cells, and combination with hArg in vitro improved its cytotoxicity. However, no significant difference in therapeutic efficacy was observed between the hArg-treated and control groups in the orthotopic GL261 glioblastoma mouse model. The results of this study indicate the potential of 225Ac-labeled RGD peptides incorporating Macropa as a chelator as promising agents for TAT. However, the further optimization of combination strategies and pharmacokinetics is required.",
"42554519": "ID: 42554519\nTitle: Assembly of Bioactive Superstructures via Metal-Phenolic Complexation for Blood Purification.\nAbstract: Hyperbilirubinemia, which stems from bilirubin accumulation caused by liver failure, can lead to jaundice, multi-organ damage, and mortality. Hemoperfusion is the most effective detoxification approach, and its efficacy critically depends on the adsorbent system. However, most developed adsorbents exhibit poor hemocompatibility, posing risks of coagulation and bacterial infection. Herein, we report a facile and highly biocompatible strategy to fabricate bioactive superstructures for blood purification, enabling toxin removal. The formation of the bioactive superstructures is mediated by the assembly of polydopamine/ZnII networks on natural polymer templates, followed by surface modification with human serum albumin. The obtained superstructures enable rapid bilirubin clearance owing to the synergistic effects of metal coordination and polyphenol-mediated noncovalent interactions (hydrogen bonding and hydrophobic and electrostatic interactions). Furthermore, these bioactive superstructures exhibit excellent antibacterial efficacy, achieving > 99.9% killing efficiency of both Escherichia coli and Staphylococcus aureus, while enhancing anticoagulant performance by 2.6-fold (compared to the control) through inhibition of platelet adhesion/activation and modulation of the intrinsic coagulation pathway. This work expands our understanding of metal-phenolic-mediated superstructure assembly. Furthermore, the efficient toxin removal, antibacterial protection, and anticoagulant activity of the developed bioactive superstructures are expected to underpin the rational design of blood-contacting materials based on metal-phenolic complexation.",
"42554584": "ID: 42554584\nTitle: Magneto-NIR-II-Programmed Cascade Nanozymes Unlocking Blood-Brain Barrier Translocation and Autophagic Resistance in Glioblastoma.\nAbstract: Glioblastoma (GBM) remains a highly aggressive central nervous system malignancy, and its treatment is hindered by poor drug accumulation across the blood-brain barrier (BBB) and autophagy-mediated repair. To address these barriers, rare-earth-doped Nd0.02Fe2.98S4@HA nanozymes (NFSH) are constructed as magneto-NIR-II-programmed cascade nanozymes for trans-BBB delivery, multimodal imaging, and ferroptosis amplification. Hyaluronic acid (HA)-mediated CD44 targeting and oriented magnetic field-enhanced BBB permeability promote tumor enrichment, while Nd3+ doping endows NFSH with strengthened superparamagnetism, near-infrared second window (NIR-II) photodynamic activity, and NIR-II fluorescence capability. Under alternating magnetic field (AMF) and NIR-II laser stimulation, NFSH activates catalase-, peroxidase-, glutathione oxidase-, and nicotinamide adenine dinucleotide (NADH) oxidase-like cascade catalysis, which amplifies reactive oxygen species (ROS) production, consumes glutathione, and induces ferroptosis. In the acidic tumor microenvironment, AMF further promotes H2S release, disrupts lysosomal autophagic degradation, and aggravates mitophagy inhibition through NADH depletion-mediated ATP deficiency. This cascade mechanism enhances ferroptosis and reshapes the tumor immune microenvironment by relieving hypoxia and promoting M2-to-M1 macrophage polarization. In addition, NFSH enables NIR-II fluorescence and T2-weighted magnetic resonance imaging for real-time visualization of treatment. This strategy provides an integrated trans-BBB theranostic platform for autophagy-suppressed ferroptosis therapy against GBM.",
"42554589": "ID: 42554589\nTitle: Why do bisphosphonates work in the treatment of CRPS? A translational pharmacological perspective to solve a conundrum.\nAbstract: Bisphosphonates (BPs) have consistently yielded positive results in the treatment of Complex Regional Pain Syndrome (CRPS). However, biochemical and histopathological evidence suggests that osteoclasts, the primary pharmacological target of BPs, is not a key driver in the early pathogenetic steps of CRPS. To critically review and integrate current evidence on pharmacological mechanisms of action of BPs in CRPS and to propose unifying hypotheses explaining their clinical efficacy beyond antiosteoclastic activity. Literature from in vitro studies, animal models, and randomized clinical trials was integrated. Approximately 80 primary and review articles were critically reviewed. BPs efficacy in CRPS is best explained by a bone-driven pharmacological model by which local drug accumulation is achieved in the site of disease. This enables modulation of inflammatory and nociceptive processes on adjacent non-bone cells, including macrophages, neutrophils, keratinocytes, and nociceptive fibers. Inhibition of the mevalonate pathway in these cells may reduce inflammatory cytokines, nerve growth factor (NGF), and reactive oxygen species (ROS), while modulating nociceptive signaling pathways. Additional mechanisms, including modulation of vascular tone and neuropeptide signaling, may further contribute to therapeutic effects. This hypothesis may inform optimal treatment strategies and together improve understanding of CRPS pathophysiology.",
"42554595": "ID: 42554595\nTitle: Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy.\nAbstract: During glioma progression, monocytes abundantly infiltrate but primarily differentiate into immunosuppressive macrophages to promote tumor growth. Redirecting monocyte differentiation offers a compelling yet underexplored therapeutic opportunity. In this work, we found M1-polarized macrophage-derived extracellular vesicles (M1-EVs) efficiently induced monocytes to differentiate into anti-tumor macrophages via tumor necrosis factor alpha (TNF-\u03b1)-mediated signaling. Despite promising, the therapeutic efficacy of M1-EVs was constrained by insufficient glioma accumulation and CD47-mediated phagocytic inhibition. To address this challenge, we further engineered M1-EVs with dual-targeting specificity by genetically incorporating a tumor-directed chimeric antigen receptor (CAR) against IL13R\u03b12 or EGFRvIII together with CD47-blocking SIRP\u03b1 variants. The resulting dual-targeting EVs (M1-CS-EVs) exhibited enhanced blood-brain barrier (BBB) penetration and glioma accumulation while locally disrupting CD47-SIRP\u03b1 interactions. In three orthotopic glioma models, M1-CS-EVs elicited a potent anti-tumor immune response and enhanced tumor phagocytosis, significantly suppressing tumor growth while prolonging animal survival. Our findings establish a platform technology for directing monocyte differentiation toward anti-tumor phenotypes, offering a broadly applicable strategy for glioma treatment."
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"42554509": "Zhang T, Zhan X, Zhang J, Tan M, Chu C et al. (2026). Mepce Restrains Ferroptosis in Prostate Cancer Through 7SK-P-TEFb Pause Control.. Advanced science (Weinheim, Baden-Wurttemberg, Germany). ID: 42554509.",
"42554513": "Echigo H, Munekane M, Fuchigami T, Mishiro K, Kikunaga H et al. (2026). Macropa-Based 225Ac-Labeled RGD Peptides for Targeted Alpha Therapy.. Molecular pharmaceutics. ID: 42554513.",
"42554519": "Wang P, Chen J, Chen S, Peng B, Zhou Z et al. (2026). Assembly of Bioactive Superstructures via Metal-Phenolic Complexation for Blood Purification.. Angewandte Chemie (International ed. in English). ID: 42554519.",
"42554584": "Liu R, Wu Y, Zhang S, Zhao H, Zheng J et al. (2026). Magneto-NIR-II-Programmed Cascade Nanozymes Unlocking Blood-Brain Barrier Translocation and Autophagic Resistance in Glioblastoma.. Advanced science (Weinheim, Baden-Wurttemberg, Germany). ID: 42554584.",
"42554589": "Varenna M, Crotti C, Di Taranto R, Orsini F, Zucchi F (2026). Why do bisphosphonates work in the treatment of CRPS? A translational pharmacological perspective to solve a conundrum.. Expert review of clinical pharmacology. ID: 42554589.",
"42554595": "Pan Y, Liu X, Meng QF, Cao L, Li R et al. (2026). Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy.. Advanced science (Weinheim, Baden-Wurttemberg, Germany). ID: 42554595."
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"mvcReports": [
{
"id": "mvc_dp_suggested_experiments_1785956283059",
"title": "Suggested Experiments Report",
"plan": {
"title": "SUGGESTED EXPERIMENTS : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Experimental Protocol Metrics"
},
{
"type": "synthesis",
"title": "Executive Analysis of Suggested Experiments",
"content": "The proposed research focuses on the intersection of BoNT-A (botulinum neurotoxin type A) and extracellular vesicle (EV) trafficking. Evaluated protocols emphasize two primary research vectors: 1) Systemic tracking of fluorescently tagged EVs post-sciatic nerve injection, specifically examining accumulation in the deep cervical lymph nodes [ID: Run1_Eval1_synthesis], and 2) Mechanistic cellular analysis, including the quantification of retrograde-to-exocytic flux using live-cell confocal imaging [ID: Run2_Eval1_synthesis]. Further studies are suggested to test the influence of SNARE inhibition on EV efflux in spinal dorsal horn models and the accumulation of pathological proteins via pulse-chase methodology [ID: Run1_Eval1_synthesis; ID: Run2_Eval1_synthesis]."
},
{
"type": "logic_network",
"title": "Proposed Experimental Logic Pathways"
},
{
"type": "comparison_matrix",
"title": "Methodological Comparison Matrix",
"headers": [
"Focus Area",
"Key Technique",
"Targeted Model"
],
"rows": [
[
"Systemic Flux",
"Fluorescent Tagging",
"Sciatic Nerve / Cervical Lymph Nodes"
],
[
"Cellular Mechanics",
"Live-cell Confocal",
"Spinal Dorsal Horn / Soma"
],
[
"Protein Trafficking",
"Pulse-Chase",
"Compartmentalized Neuronal Cells"
]
]
},
{
"type": "bottlenecks",
"title": "Literature Synthesis Gaps",
"content": "The current dataset provides experimental recommendations but lacks: 1) Specific longitudinal data regarding the time-course of EV accumulation; 2) Direct quantification of baseline vs. experimental BoNT-A dosage efficacy; and 3) Established control parameters for distinguishing between active axonal transport and passive diffusion of labeled EVs."
}
]
}
},
{
"id": "mvc_dp_suggested_studies_1785956298934",
"title": "Suggested Studies Report",
"plan": {
"title": "SUGGESTED STUDIES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Study Distribution Metrics"
},
{
"type": "synthesis",
"title": "Executive Analysis of Research Trajectories",
"content": "The identified studies focus on the intersection of BoNT-A (Botulinum neurotoxin serotype A) and cellular transport/secretion mechanisms. Current literature [Run1_Eval1_synthesis] emphasizes retrograde axonal transport rates and EV (extracellular vesicle) secretion profiles following SNAP25 cleavage. A secondary evaluative focus [Run2_Eval1_synthesis] shifts to metabolic impacts, specifically somatic lysosomal enzyme activity. These data suggest a dual-mechanism investigation path: tracking physical protein spread versus intracellular organelle function in treated neurons."
},
{
"type": "comparison_matrix",
"title": "Methodological Comparison Matrix",
"headers": [
"Focus Area",
"Mechanism",
"Primary Target"
],
"rows": [
[
"Axonal Transport",
"Retrograde Kinetics",
"BoNT-A Derivatives"
],
[
"Secretory Profile",
"SNAP25 Cleavage",
"Spinal Interneurons"
],
[
"Metabolic Function",
"Lysosomal Enzyme Activity",
"Somatic Compartment"
]
]
},
{
"type": "logic_network",
"title": "Systemic Research Logic Map"
},
{
"type": "bottlenecks",
"title": "Literature Gap Assessment"
}
]
}
},
{
"id": "mvc_dp_swansons_literature_based_discovery_candidates_1785956313064",
"title": "Swansons Literature Based Discovery Candidates Report",
"plan": {
"title": "SWANSONS LITERATURE BASED DISCOVERY CANDIDATES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity and Discovery Metrics"
},
{
"type": "synthesis",
"title": "Executive Summary of BoNT-A Therapeutic Candidates",
"content": "The analysis of Swanons Discovery candidates reveals two primary clinical hypotheses for Botulinum neurotoxin type A (BoNT-A). The first identifies a mechanism where SNAP25 cleavage via BoNT-A inhibits SNARE-dependent exocytosis, potentially curtailing the trans-synaptic propagation of pathogenic EVs [ID: 37037273, 37394036]. The second hypothesis suggests that BoNT-A retrograde transport routes cargo into the somatic lysosomal degradation pathway, offering a potential mechanism to clear intracellular protein aggregates associated with tauopathy [ID: 25878289, 42553548]. Both models emphasize the repurposing of existing BoNT-A biological pathways to modulate neurodegenerative disease progression."
},
{
"type": "logic_network",
"title": "Conceptual Bridge Mapping"
},
{
"type": "comparison_matrix",
"title": "Discovery Hypothesis Comparison",
"headers": [
"Hypothesis Type",
"Primary Mechanism",
"Clinical Target"
],
"rows": [
[
"Propagation Inhibition",
"SNARE Cleavage",
"ALS/Pathogenic EVs"
],
[
"Aggregate Clearance",
"Endolysosomal Upregulation",
"Tauopathy/Microclots"
]
]
},
{
"type": "bibliography",
"title": "Source Literature Index"
}
]
}
},
{
"id": "mvc_dp_contradictions_between_evidences_1785956326474",
"title": "Contradictions Between Evidences Report",
"plan": {
"title": "CONTRADICTIONS BETWEEN EVIDENCES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Global Data Metrics"
},
{
"type": "synthesis",
"title": "Executive Analysis",
"content": "The clinical evaluation of current literature indicates no direct contradictions regarding primary biological mechanisms [Run1_Eval1_synthesis]. While the existence of retrograde transport remains supported by existing data, evidence suggests variability in the spatial distribution of cleavage within central nervous system processes [Run2_Eval1_synthesis]. Current research gaps involve a lack of explicit resolution regarding the clinical implications of this spatial variance [Run2_Eval1_synthesis]. Overall, primary pathways are consistent, though localized downstream effects require further empirical investigation."
},
{
"type": "contradiction_topology",
"title": "Conflict Node Mapping",
"content": "None identified in current literature set regarding primary mechanisms or retrograde transport existence."
},
{
"type": "bottlenecks",
"title": "Literature Gap Assessment"
},
{
"type": "data_bar_chart",
"title": "Literature Gap Distribution",
"xAxisLabel": "Gap Category",
"data": [
{
"label": "Primary Mechanisms",
"value": 0
},
{
"label": "Retrograde Transport",
"value": 0
},
{
"label": "Spatial Cleavage Distribution",
"value": 1
}
]
}
]
}
},
{
"id": "mvc_dp_repurposed_solutions_1785956339585",
"title": "Repurposed Solutions Report",
"plan": {
"title": "REPURPOSED SOLUTIONS : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity and Scope Metrics"
},
{
"type": "synthesis",
"title": "Executive Analysis: BoNT-A Repurposing",
"content": "The analyzed data highlights two distinct clinical repurposing strategies for Botulinum Neurotoxin type A (BoNT-A). The first strategy shifts the application from conventional neuromuscular paralysis to the spatiotemporal regulation of pathological extracellular vesicle (EV) secretion [ID: Run1_Eval1_synthesis]. The second strategy leverages the toxin\u2019s inherent retrograde transport capabilities to serve as a precision delivery vector for degradation-inducing motifs to the neuronal soma, specifically targeting neurodegenerative disease pathologies [ID: Run2_Eval1_synthesis]. Current evidence is limited to theoretical mechanisms; significant gaps exist regarding longitudinal toxicity, precise dosage scaling for intracellular delivery, and the engineering requirements for cargo-motif attachment."
},
{
"type": "comparison_matrix",
"title": "Comparative Repurposing Pathways",
"headers": [
"Focus Area",
"Target Mechanism",
"Clinical Goal"
],
"rows": [
[
"EV Modulation",
"Spatiotemporal inhibition",
"Pathological secretome control"
],
[
"Neuro-Therapeutic",
"Retrograde axonal transport",
"Soma-targeted protein degradation"
]
]
},
{
"type": "logic_network",
"title": "Mechanistic Logic Pathway"
},
{
"type": "bottlenecks",
"title": "Evidence Gaps and Research Constraints"
}
]
}
},
{
"id": "mvc_dp_BoNT_induced_lysosomal_flux_1785956353720",
"title": "BoNT Induced Lysosomal Flux Report",
"plan": {
"title": "BONT INDUCED LYSOSOMAL FLUX : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity Scorecard"
},
{
"type": "synthesis",
"title": "Clinical Executive Summary",
"content": "Current literature confirms that Botulinum neurotoxin A (BoNT/A) is effectively routed to lysosomal compartments [ID: Run2_Eval1_synthesis]. However, there is a critical evidence gap regarding the co-trafficking or sequestration of exogenous pathological proteins, such as Tau, within this flux mechanism. This hypothesis remains strictly an experimental inference, as current data lacks direct observational confirmation of this synergistic interaction."
},
{
"type": "bottlenecks",
"title": "Identified Evidence Gaps",
"content": "Strong: Direct evidence linking BoNT/A-induced lysosomal flux to the clearance or sequestration of extracellular pathological proteins like Tau."
},
{
"type": "gap_distribution",
"title": "Literature Gap Strength",
"data": [
{
"label": "Confirmed",
"value": 20
},
{
"label": "Inferred",
"value": 80
}
]
}
]
}
},
{
"id": "mvc_dp_EV_somatic_sequestration_rate_1785956366552",
"title": "EV Somatic Sequestration Rate Report",
"plan": {
"title": "EV SOMATIC SEQUESTRATION RATE : CUSTOM ANALYSIS",
"evidence_tier": "INSUFFICIENT",
"panels": [
{
"type": "metrics",
"title": "Evidence Integrity Scorecard"
},
{
"type": "synthesis",
"title": "Executive Analysis of Sequestration Dynamics",
"content": "The clinical evaluation of the 'EV Somatic Sequestration Rate' indicates a critical lack of empirical resolution. Based on the available evaluation, there is insufficient evidence to quantify the shift in extracellular vesicle (EV) concentration between the soma and terminal release sites [ID: Run2_Eval1_synthesis]. Consequently, no comparative baseline can be established, resulting in a data gap that precludes predictive modeling or therapeutic targeting at this time."
},
{
"type": "gap_distribution",
"title": "Literature Gap Assessment"
},
{
"type": "bottlenecks",
"title": "Identified Evidence Bottlenecks"
},
{
"type": "logic_network",
"title": "Process Logic Map"
}
]
}
},
{
"id": "mvc_dp_synaptic_vs_somatic_clearance_ratio_1785956379317",
"title": "Synaptic Vs Somatic Clearance Ratio Report",
"plan": {
"title": "SYNAPTIC VS SOMATIC CLEARANCE RATIO : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity Metrics"
},
{
"type": "synthesis",
"title": "Clinical Synthesis of Clearance Dynamics",
"content": "Current literature provides evidence of a clear physiological shift toward somatic processing of the toxin [ID: Run2_Eval1_synthesis]. However, a quantitative determination of the Synaptic Vs Somatic Clearance Ratio remains unavailable, representing a critical knowledge gap in current evidentiary records."
},
{
"type": "gap_distribution",
"title": "Literature Gap Assessment"
},
{
"type": "bottlenecks",
"title": "Missing Analytical Parameters"
}
]
}
}
],
"aggregatedDatapoints": {
"suggested_experiments": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": [
"Assess if BoNT-A injection at sciatic nerve termini influences the rate of fluorescently tagged EV accumulation in the deep cervical lymph nodes.",
"Test whether selective SNARE inhibition via BoNT-A in the spinal dorsal horn affects the efflux of brain-derived EVs in a mouse model."
]
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Quantify the ratio of retrograde-to-exocytic vesicle flux in the presence vs. absence of BoNT/A using live-cell confocal imaging. Analyze the accumulation of tagged pathological proteins in the soma using pulse-chase methods in compartments."
}
],
"suggested_studies": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": [
"A systematic analysis of retrograde axonal transport rates of BoNT-A derivatives in the context of neurodegenerative protein spread.",
"Investigation into the impact of BoNT-A-induced SNAP25 cleavage on the secretion profile of EVs in spinal interneurons."
]
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Compare somatic lysosomal enzyme activity levels in BoNT/A-treated vs. control neurons."
}
],
"swansons_literature_based_discovery_candidates": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": {
"Discovered Hypothesis (A to C)": "BoNT-A mediated inhibition of synaptic exocytosis can reduce the pool of neuronally-derived EVs available for trans-synaptic propagation, thereby limiting disease seeding.",
"Literature A (Origin)": "BoNT-A synaptic silencing (Source 37037273)",
"Literature C (Target)": "Intercellular propagation of pathogenic proteins/EVs in ALS (Source 37394036)",
"The Intersecting Bridge B": "SNARE-dependent exocytosis (SNAP25)",
"Biological Rationale": "Since SNARE complex integrity is required for both neurotransmitter release and the unconventional secretion of EVs, BoNT-A induced cleavage of SNAP25 effectively halts the exocytic machinery used by pathogenic EVs to spread along connected neural circuits."
}
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "- Discovered Hypothesis (A to C): BoNT/A-induced somatic endolysosomal upregulation could serve as a compensatory clearing mechanism for tau-containing microclots in tauopathy-prone neurons. - Literature A (Origin): BoNT/A retrograde transport to the soma and fusion with lysosomes (ID: 25878289) - Literature C (Target): Amyloidogenic protein clearing in neurodegeneration (ID: 42553548) - The Intersecting Bridge B: LC3-positive autophagosomes and lysosomal fusion. - Biological Rationale: BoNT/A routes cargo into the somatic lysosomal degradation pathway; thus, stimulating this specific retrograde pathway may mechanically facilitate the clearance of other non-toxin-related intracellular protein aggregates."
}
],
"contradictions_between_evidences": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": "None identified regarding the primary mechanisms, though the therapeutic outcome of BoNT in central nervous system processes remains a point of clinical study."
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "None identified in the current literature set regarding retrograde transport existence, though spatial distribution of cleavage varies."
}
],
"repurposed_solutions": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": "Repurpose BoNT-A as a tool for spatiotemporally controlled cessation of pathological EV secretion, rather than just neuromuscular paralysis."
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Utilization of BoNT/A-induced retrograde transport as a targeted vehicle to deliver degradation-inducing motifs to the neuronal soma for neurodegenerative diseases."
}
],
"BoNT_induced_lysosomal_flux": [
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "The evidence indicates that BoNT/A is routed to lysosomes, but whether this specifically draws in other exogenous pathological proteins like Tau remains an experimental inference."
}
],
"EV_somatic_sequestration_rate": [
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Insufficient data to quantify the specific change in EV reaching the soma compared to terminal release."
}
],
"synaptic_vs_somatic_clearance_ratio": [
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Evidence confirms a shift toward somatic processing of the toxin, but a definitive clearance ratio of general synaptic-to-somatic cargo is not provided."
}
]
},
"stats": {
"promptTokens": 374326,
"completionTokens": 27781,
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},
"zenodo_doi": "10.5281/zenodo.21812604"
}