{
"claim": "anti-lamin b1 (phospho t575) antibody - nuclear envelope marker",
"timestamp": "2026-08-05T18:33:39.511Z",
"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- \"LBR_phosphorylation_sites\": Identify specific phosphorylation sites on LBR that increase in occupancy following lamin A overexpression in TKO cells.\n- \"LBR_A_type_lamin_interaction\": Determine if lamin A directly phosphorylates LBR or recruits specific kinases to the nuclear envelope to induce displacement.\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 ... tags at the very beginning of your response.\n\n=======================================================\nCONTEXT LITERATURE (STATIC CACHE):\nID: 42554576\nTitle: Clinical utility of adrenal venous sampling for functional lateralization in ACTH-independent autonomous cortisol secretion: relevance in non-decisive imaging.\nAbstract: Determining functional laterality in adrenocorticotropic hormone (ACTH)-independent autonomous cortisol secretion remains challenging when imaging demonstrates bilateral or non-decisive adrenal findings. We evaluated whether adrenal venous sampling (AVS) improves lateralization in this clinically challenging subgroup. In this single-center retrospective cohort study, we identified 66 patients with ACTH-independent autonomous cortisol secretion assessed between April 2016 and October 2025. Among them, 21 patients with bilateral or non-decisive imaging findings underwent successful AVS and constituted the study population. All patients underwent contrast-enhanced computed tomography (CT), adosterol scintigraphy, and AVS under low-dose dexamethasone suppression without exogenous ACTH stimulation. Postoperative remission was used as outcome-based evidence of true unilateral disease. Lateralization indices were calculated using cortisol-based and metanephrine-corrected ratios. Among patients with bilateral or non-decisive CT findings (n\u2009=\u200921), AVS identified unilateral cortisol secretion in 11 (52%). Postoperative remission was confirmed in 9 of these 11 patients (82%), supporting true unilateral disease. The median follow-up period was 4.0 years (range, 1.6-7.0 years), and remission status remained unchanged throughout follow-up. Neither adrenal size difference (2.0\u2005mm vs 1.5\u2005mm; p\u2009=\u20090.59) nor adrenal size ratio (1.11 vs 1.10; p\u2009=\u20090.48) on CT predicted AVS-defined lateralization. In patients with bilateral or non-decisive CT findings, AVS frequently identified surgically remediable unilateral cortisol secretion that would not have been recognized by imaging alone. These findings support the incremental diagnostic value of AVS and suggest that functional assessment should be considered when imaging alone is insufficient to determine laterality in ACTH-independent autonomous cortisol secretion.\n\nID: 42554561\nTitle: Ultrasound Cavitation of Microbubbles Augments Limb Perfusion and Promotes Ulcer Healing in Chronic Limb-Threatening Ischemia.\nAbstract: In patients with peripheral artery disease and chronic limb-threatening ischemia (CLTI), there is a lack of effective treatment options when arterial revascularization is not feasible. In preclinical models, ultrasound (US) cavitation of microbubble contrast agents can augment limb perfusion through multiple shear-mediated vasoactive pathways. This study aims to evaluate whether US cavitation increases limb tissue perfusion and accelerates healing of ischemic ulcers in patients with CLTI. Patients (n = 24) with CLTI and chronic nonhealing ulcers were randomized to standard care or cavitation therapy, which was performed every 3 days \u00d7 6. Definity was infused, and US (1.3 MHz, mechanical index: 1.3) was performed for 15 minutes at the wound site and simultaneously over the ipsilateral femoral artery to leverage conducted vasodilation. The primary endpoints at day 18 were change in ulcer area and ulcer microvascular perfusion. Complete or partial ulcer healing occurred in almost all cavitation-treated patients, with a significant reduction in ulcer size only in the cavitation group and not the control group (median: -36.6% [Q1-Q3: -15.4% to -65.0%] vs +3.4% [Q1-Q3: -14.7% to +20.0%]; P = 0.039). In cavitation-treated patients, there was a 2.5-fold increase in ulcer margin perfusion, a 1.6-fold increase in functional microvascular density on super-resolution US microangiography, and a 1.8-fold increase in femoral artery flow (P < 0.05 for all parameters). These flow parameters did not increase in control patients. In patients with CLTI and nonhealing ulcers, periodic therapy with cavitation of microbubbles using US parameters approved for diagnostic imaging promotes healing of ischemic ulcers, likely through improvement in tissue perfusion and microvascular vascularity.\n\nID: 42554556\nTitle: Dyspnea as an Anginal Equivalent: Multimodality Diagnosis of Dynamic, Severe Ischemic Mitral Regurgitation.\nAbstract: Dynamic ischemic mitral regurgitation (IMR), provoked by ischemia or exercise, may escape detection at rest and is associated with adverse outcomes. A 65-year-old man with ischemic cardiomyopathy (left ventricular ejection fraction, 20%) and recurrent left circumflex in-stent restenosis refractory to multiple interventions presented with progressive dyspnea and chest pain. Resting filling pressures were mildly abnormal, though symptoms remained out of proportion to findings. During balloon angioplasty of the left circumflex, transient ischemia provoked large V waves on the pulmonary catheter wedge pressure tracing; simultaneous echocardiography confirmed severe MR that resolved on deflation of the intracoronary balloon. Stress echocardiography corroborated exercise-induced severe MR. Despite revascularization and medical therapy optimization, symptoms persisted, and mitral transcatheter edge-to-edge repair (M-TEER) was performed with successful resolution of symptoms. Dynamic IMR may be underdiagnosed when diagnostic studies are performed at rest. This case demonstrates concordant invasive and noninvasive provocative strategies for diagnosis, with M-TEER for refractory symptoms. Dynamic IMR requires a high index of suspicion and provocative multimodality testing. M-TEER may benefit patients with severe IMR refractory to revascularization and optimized medical therapy.\n\nID: 42554555\nTitle: Microaxial Flow Pump Thrombosis Presenting as Predominant Direct Hyperbilirubinemia.\nAbstract: Clinically significant hemolysis and device thrombosis are uncommon with the Impella 5.5, and presentation with predominant direct hyperbilirubinemia has not been described. A 57-year-old man with refractory cardiogenic shock underwent Impella 5.5 placement as a bridge to transplant. He developed severe direct-predominant hyperbilirubinemia (total bilirubin 18.4 mg/dL; direct 11.7 mg/dL) with rising lactate dehydrogenase (1,395 U/L) and undetectable haptoglobin, despite normal purge pressures and no device alarms. Duplex ultrasonography excluded biliary obstruction. Device exchange revealed a large, organized thrombus in the motor/outflow housing, after which hemolysis was resolved, and biochemical improvement followed, allowing successful bridging to heart transplantation. Occult Impella thrombosis may present as direct-predominant hyperbilirubinemia, an atypical pattern that can mimic hepatobiliary disease and confound the diagnostic evaluation. Direct hyperbilirubinemia during Impella support should prompt evaluation for subclinical device thrombosis. Timely device exchange can reverse hemolysis and preserve transplant candidacy.\n\nID: 42554518\nTitle: LC-MS/MS Determination of Hair GHB: Is Segmental Analysis Useful for Exogenous Discrimination?\nAbstract: Discrimination between endogenous and exogenous Gamma-hydroxybutyrate (GHB) in hair matrix is a major challenge in the forensic context. According to the UNODC, to verify a possible intake, GHB concentration should be compared with the subject's basal level. Most published studies investigated endogenous GHB on a single hair segment, without considering possible intra-individual variation. The present study aimed to detect endogenous GHB in hair using a newly validated method, with a focus on the relevance of segmental analysis. Two hundred subjects were tested. After the removal of the first 0.5\u2009cm from the proximal end, each hair strand was divided into three segments of 1\u2009cm. A water extraction was performed during the pulverization process, and the extract was injected into a LC-MS/MS system. Data have been statistically analyzed. Overall, endogenous GHB concentration ranged from < LOD (0.1\u2009ng/mg) to 6.6\u2009ng/mg, with 27% of samples having a value < LOD in at least two segments. Within the remaining subjects, neither gender nor age significantly influenced the concentration. On the other hand, results confirmed that hair treatment can modify endogenous GHB concentrations. Intra-individual variability was limited, with the highest percentage of cases showing minimal changes in concentration (0-0.1\u2009ng/mg) between contiguous segments. However, in up to 4% of cases, the variation resulted to be higher than 1\u2009ng/mg between contiguous segments. The present study then confirms the need to perform in forensic cases the segmental analysis, to be sure that any potential increase of GHB depends exclusively on external administration.\n\nID: 42554514\nTitle: Computed Tomography-Based Anatomical and Morphometric Analysis of the Left and Right Gastric Arteries in a Vietnamese Population.\nAbstract: Anatomical variations of the left gastric artery (LGA) and right gastric artery (RGA) are clinically relevant for gastric surgery and endovascular interventions. This study aimed to characterize the origin, anatomical distribution and morphometric features of the LGA and RGA using 64-slice computed tomography (CT). A cross-sectional descriptive study was conducted on 385 individuals who underwent contrast-enhanced abdominal CT at Binh Dan Hospital between September 2024 and April 2025. The LGA originated from the celiac trunk in 91.9% of cases and was most frequently located between the lower one-third of T12 and the upper one-third of L1 (>\u200961%). Its mean diameter and length were 2.7\u2009\u00b1\u20090.3\u2009mm and 30.3\u2009\u00b1\u20099.3\u2009mm, respectively, with the Type I branching pattern predominating, characterized by branches supplying the oesophagus, gastroesophageal junction and the lesser curvature. The mean distance from the abdominal aorta to the LGA was 19.6\u2009\u00b1\u20096.1\u2009mm. The RGA was identified in 88.3% of cases and most commonly arose from the proper hepatic artery (72.9%), with a mean diameter of 1.6\u2009\u00b1\u20090.2\u2009mm. An arterial arcade along the lesser curvature was observed in 81.3% of cases. These findings provide quantitative anatomical data that may support accurate identification of vascular landmarks during gastric surgery and facilitate appropriate catheter selection for endovascular procedures involving the LGA and RGA.\n\nID: 42554504\nTitle: Amino-Rich Microporous Organic Network Composite Membranes for Efficient Extraction of Domoic Acid From Complex Matrices.\nAbstract: Microporous organic networks (MONs) have attracted considerable attention because of their high porosity, modifiable structure and excellent stability. However, their rigid structure makes it difficult to form membranes, leading to limited separation applications. MONs' powder suffers from issues of dusting and severe aggregation, leading to difficulties in separation and recycling. Herein, MONs' powder was converted into flexible and reusable membranes via phase inversion. The membranes were applied to enrich and extract domoic acid (DA, an algal toxin) from complex matrices. The adsorption performance and mechanism of three MONs membranes with different amino content were investigated and compared. The results indicate that the adsorption energy of the membranes for DA is positively correlated with the number of amino groups. Under optimized conditions, by combining syringe filter-based membrane micro-solid-phase extraction (SF-M-\u00b5SPE), with high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), the developed method showed a low detection limit (0.5 pg mL-1), a wide linear range (1.5\u223c1000.0 pg mL-1, R \u2265 0.9999) and good repeatability (RSD \u2264 3.8%, n = 6). The membranes were successfully used to extract DA from seafood. There was trace DA (9.4 pg mL-1) found in oyster samples. This work demonstrates the great potential of MONs membranes in sample pretreatment.\n\nID: 42554489\nTitle: Rapid, low-cost, and portable molecular detection of HIV-1 subtype B and HCV 1a directly from blood.\nAbstract: Accessible molecular diagnostics for human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV) remain constrained by the cost and infrastructure requirements of existing molecular point-of-care platforms. In this study, we describe a portable, proof-of-concept molecular workflow with a total runtime of 32 min for the simultaneous qualitative detection of HIV-1 subtype B and HCV genotype 1a directly from contrived whole blood. The workflow employs a battery-powered heating device, ambient-stable reagents, and lateral-flow dipstick readout. Contrived whole blood samples were diluted twofold with nuclease-free water and added directly to rehydrated reverse-transcription loop-mediated amplification (RT-LAMP) reactions without nucleic acid extraction, and reaction parameters were systematically varied to determine lower limits of detection. Both viral targets were consistently detected at clinically relevant input levels following a 30 min isothermal amplification step, with visual lateral flow results available within 2 min. In addition, CRISPR-Cas12a detection was used as an orthogonal method to verify RT-LAMP target amplicons during assay development. Together, this work establishes the analytical feasibility of a rapid, low-cost, portable molecular workflow and provides a foundation for future development of decentralized HIV-1 and HCV molecular diagnostics.IMPORTANCEOngoing changes in human immunodeficiency virus (HIV) and hepatitis C virus (HCV) epidemiology highlight the need for more adaptable and accessible diagnostic tools. In this study, we developed a fast, low-cost test that detects HIV-1 and HCV from a small blood sample. The test is designed to work outside traditional laboratories, using minimal equipment. It does not require complex sample preparation, refrigeration, or expensive machines, and produces results in about 30 min. The workflow combines a rapid genetic amplification method with an easy-to-read test strip and a small, battery-powered heater, making it portable and suitable for use in low-resource or remote settings. An additional molecular method was used during development to confirm that the test was accurately detecting the correct viral targets. By focusing on simplicity, speed, and affordability, this work aims to expand access to reliable HIV and HCV testing in settings where these infections often occur together and laboratory access is limited.\n\nID: 42554475\nTitle: Monolithic integration of SiC diode detectors into scanning X-ray microscopy optical elements for in situ beam diagnostics.\nAbstract: Reliable monitoring of the incident X-ray beam intensity and position is essential in soft X-ray microspectroscopy, where beam instabilities directly affect data normalization and achievable spatial resolution. Conventional beam diagnostics are often implemented using offline detectors located far from the sample or by measuring downstream on empty regions of the sample support membrane, not fully reflecting the experimental conditions experienced during measurements. In this work, the development of functionalized soft X-ray optical elements based on monolithic silicon carbide (SiC) diode detectors, enabling real-time in situ beam diagnostics directly within a scanning transmission X-ray microscopy (STXM) setup, is presented. Two complementary device geometries are investigated: order-sorting aperture (OSA)-based detectors and center-stop-based detectors. The devices are fabricated using plasma-focused-ion-beam milling and substrate thinning, allowing the integration of optical and sensing functionalities with minimal changes to the beamline and endstation setup. The performance of the functionalized optics is experimentally validated at the PolLux STXM beamline of the Swiss Light Source. The devices provide simultaneous access to the transverse beam position and the incident beam intensity I0 through the unfocused 0th-order component. Using this component, the OSA detector enabled simultaneous measurements of the intensity and transverse displacement of the beam from the focused beam illuminating the sample. Between 700 and 1000\u2005eV, the SiC measured current exhibited linear correlation to the avalanche photodiode (APD) signal, with coefficient of determination R2 = 0.997. Position-sensitive detection was possible with a four-sector diode, where all sectors were electrically continuous and beam responsive, while gaps between the quadrants were insensitive. These results demonstrate the feasibility of SiC-based functionalized X-ray optics as beam diagnostic tools for soft X-ray microscopy and spectroscopy.\n\nID: 42554452\nTitle: Seeing Beyond the Surgeon's Eye: Artificial Intelligence for Postoperative Monitoring of Free Vascularized Flaps: A Narrative Review.\nAbstract: Postoperative monitoring of free vascularized flaps (FVFs) is critical for early detection of ischemia and timely salvage. Although traditional monitoring relies heavily on clinical examination and adjunctive devices, these approaches are labor\u2011intensive, subjective, and resource\u2011dependent. Recent advances in artificial intelligence (AI), particularly machine learning and deep learning, have enabled automated, data\u2011driven approaches to postoperative free\u2011flap monitoring. However, the clinical readiness, methodological robustness, and translational challenges of these AI\u2011based systems remain incompletely understood. This review examines recent studies on AI\u2011assisted postoperative free\u2011flap monitoring, focusing on technical methodologies, clinical applications, translational relevance, current limitations, and future research directions. A systematic-narrative hybrid review was conducted by searching PubMed, Embase, Web of Science, and Google Scholar from January 2000 to February 2, 2026. Eligible studies reported clinically relevant outcomes, including flap viability, ischemia detection, venous congestion, thrombosis prevention, and salvage rates. Studies focused on preoperative prediction models, non\u2011AI monitoring techniques, editorials, and opinion articles were excluded. Two independent reviewers performed study selection and data extraction, with disagreements resolved by consensus. Nine studies met the inclusion criteria. Most studies used supervised machine\u2011learning models, primarily using visible\u2011light images, infrared imaging, photoplethysmography, hyperspectral imaging, or multimodal sensor data for flap surveillance. All but 1 study were retrospective, and no prospective human clinical trials were identified. AI\u2011based systems demonstrated promising performance in detecting arterial ischemia and venous congestion, often achieving diagnostic accuracies comparable to or exceeding those of traditional monitoring methods. Diagnostic accuracy across studies ranged from 82% to 98.4%. Key challenges included limited data sets, lack of external validation, heterogeneous methodologies, absence of standardized outcome metrics, and concerns about generalizability across diverse skin tones and clinical settings. AI\u2011assisted postoperative free\u2011flap monitoring is a rapidly evolving and promising adjunct to conventional surveillance. Although current evidence suggests benefits for early ischemia detection and workload reduction, significant barriers-particularly methodological heterogeneity, data scarcity, and limited prospective validation-must be addressed before widespread clinical implementation. Future progress will depend on standardized data sets, transparent model development, multicenter prospective studies, and ethically sound, interoperable AI frameworks that enable equitable and scalable precision microsurgical care.\n\nID: 42554414\nTitle: Impact of Point-of-Care Ultrasound on Clinical Decision-Making During Common Critical Care Scenarios: Linking Educational Gaps to Clinical Practice.\nAbstract: As point-of-care ultrasound (POCUS) becomes mainstream, educational efforts to teach image acquisition, interpretation, and incorporation of findings into clinical decision-making become more relevant to ensure quality and competency. Despite expanding POCUS use, limited data exist on how novice learners translate newly acquired POCUS skills into diagnostic and management decisions. In this educational study, we investigated the impact of incorporating POCUS into patient assessment on diagnostic and management decisions of novice POCUS learners. Learners participated in a one-day POCUS course, consisting of high-yield lectures, case discussions, and hands-on training. Diagnostic and management skills and confidence in those skills were assessed by administering clinical vignettes prior to the course without the use of POCUS, prior to the course with access to POCUS images, and at the conclusion of the course with access to POCUS images. A one-day POCUS course improved learners' overall diagnostic accuracy, management decisions, and confidence level in their clinical decision-making across clinical scenarios where POCUS is indicated. Despite overall improvement, access to POCUS images had varied effects on changes in diagnostic accuracy and management decisions. With training, access to POCUS images can augment diagnostic accuracy and enhance management where POCUS is indicated. Furthermore, learners display greater confidence in their decisions when they have access to POCUS images. These findings should be interpreted considering important methodological limitations, including the absence of a control group and potential testing effects from repeated case exposure. Clinical evaluation of this phenomenon in real patient encounters is required.\n\nID: 42554395\nTitle: Cardiac Amyloidosis in Asia: What We Know and Don't Know.\nAbstract: \n\nID: 42554351\nTitle: The Selective Synthesis and Photothermal Conversion Properties of U- and S-Shaped Flexible Double-Metallacycle Assemblies.\nAbstract: Molecular double-metallacycles have drawn considerable interest for their unique architectures and broad application prospects, yet selective synthesis remains a significant challenge. This work reports a novel triimidazolyl flexible ligand precursor L1 containing a phenyl unit with three methyl groups and three methylene spacers that enable flexibility for the coordination of imidazole units. Together with three half-sandwich rhodium building blocks (B1-B3) of varying conjugation sizes, L1 was assembled via coordination-driven self-assembly and silver-ion bridging to afford one U-shaped and two S-shaped double-metallacycles in high yields. The greater steric demand of B2 and B3 favored the S-shaped topology. The formation of all target structures was clearly confirmed by SC-XRD (Single-crystal x-ray diffraction analysis), NMR (Nuclear magnetic resonance) spectroscopy, IR (Infrared) spectroscopy, and ESI-TOF-MS (Electrospray Ionization-Time of Flight Mass) spectrometry. TGA (Thermogravimetric analysis) indicated good thermal stability for compounds 1-3. Photothermal conversion experiments revealed that S-shaped double-metallacycle 3 achieves notable photothermal conversion efficiency (21.46%-22.95%) in the NlR (near-infrared) region. EPR (Electron paramagnetic resonance) studies further corroborated that radical generation from the building blocks enhances photothermal performance.\n\nID: 42554333\nTitle: A Platform for High-Throughput Chemical Analysis of Foods: Characterization of Extra Virgin Olive by Direct Mass Spectrometry and Machine Learning.\nAbstract: Poor diet is now the leading cause of early death globally. In part, this is because our complex food supply chains are increasingly at risk of overprocessing, contamination, low nutrient content, and economically motivated fraud. Chemical testing can offer insights into these concerns, but testing methods are frequently impractical. Extra virgin olive oil (EVOO) is a premium food of high nutritional value, but because of its growing popularity and high price, it can be a target for mislabeling, substitution, dilution, and/or false claims of origin. Rapid and accurate testing methods for its characterization are therefore increasingly important. We used two direct forms of mass spectrometry (MS)-laser desorption/ionization (LDI) MS and direct analysis in real time (DART) MS-to obtain complex chemical signatures of edible oils. The data generated on a set of reference samples were then used to develop and train three independent machine learning (ML) models that assess key characteristics of a test oil. We also developed a proof-of-concept DART-MS/MS assay add-on for the quantification of bioactive phenols in EVOO. Our approach accurately predicts several attributes of an edible oil based on novel markers and intricate patterns within the acquired data. Further, pure reference standards and an isotopically-labeled internal standard allow accurate quantification of the constituent phenols. Because there is no chromatography, both the mass fingerprints and quantification can be performed in seconds-minutes. The method uses low (milliliter) volumes of sample and green solvents, and when combined with ML, it offers rapid data analysis and comprehensive result interpretation.\n\nID: 42554328\nTitle: Evaluation of medial plantar, dorsal sural nerve conduction and sural/radial amplitude ratio in prediabetic patients.\nAbstract: Distal sensory polyneuropathy (DSPN) is a common complication of Diabetes Mellitus (DM). Patients often experience disturbing neuropathic symptoms such as numbness and burning. These symptoms can also be seen in prediabetic patients who do not meet the diagnostic criteria for DM. The sural nerve responses are evaluated as standard in electrophysiologic technique. However, the sural nerve may not always show abnormalities in those with mild neuropathy. Therefore, different nerves can be studied to detect mild neuropathy. Our study aimed to investigate both the superiority of electrophysiological tests evaluating mild neuropathy over each other and the sensitivity of these tests in patients with preDM. To do that, it focused on studying medial plantar, dorsal sural and sural/ radial amplitude ratio (SRAR) in prediabetic or DM patients. The study employed patients with prediabetes and DM. Our study included 92 (65 female, 27 male) patients. The neuropathic symptoms of the patients were evaluated using the Self-Leeds Assessment of Neuropathic Symptoms and Sign (LANSS) pain scale. In addition to standard electrophysiological studies, nerve conduction studies were conducted on dorsal sural, medial plantar and radial nerve. SNAP (sensory nerve action potential) amplitudes of prediabetic patients without neuropathic symptoms are compared with healthy controls. Although the mean amplitude of dorsal sural and medial plantar responses in the preDM patient group with high LANSS scores is at the lower limit of normal, it was significantly lower than those with low LANSS score. There was no significant difference in the SRAR rates between the two groups. Among the prediabetic patients with neuropathic symptoms, 13 (56%) showed below normal dorsal sural responses and 16 (69%) exhibited below normal medial plantar responses. The findings of our study show that dorsal sural and medial plantar responses may be affected in prediabetic patients with neuropathic complaints, even if sural nerve conduction is normal. Medial plantar nerve SNAP found to be more sensitive. In contrast, SRAR was evaluated as having a low contribution compared to dorsal sural and medial plantar responses in detecting a mild polyneuropathy. It is important to look for polyneuropathy in prediabetic patients with sensitive electrophysiological methods. A distalis szenzoros polyneuropathia (DSPN) a diabetes mellitus (DM) gyakori sz\u00f6v\u0151dm\u00e9nye. A betegek gyakran zavar\u00f3 neuropathi\u00e1s t\u00fcneteket, p\u00e9ld\u00e1ul zsibbad\u00e1st \u00e9s \u00e9g\u0151 \u00e9rz\u00e9st tapasztalnak. Ezek a t\u00fcnetek praediabeteses betegekben is el\u0151fordulhatnak, akik m\u00e9g nem teljes\u00edtik a DM diagnosztikai krit\u00e9riumait. Az elektrofiziol\u00f3giai vizsg\u00e1latok sor\u00e1n rutinszer\u0171en a n. suralis v\u00e1laszait \u00e9rt\u00e9kelik. Enyhe neuropathia eset\u00e9n azonban a suralis ideg vizsg\u00e1lata nem mindig mutat elt\u00e9r\u00e9st. Ez\u00e9rt az enyhe neuropathia kimutat\u00e1s\u00e1ra m\u00e1s idegek vizsg\u00e1lata is sz\u00fcks\u00e9ges lehet. Vizsg\u00e1latunk c\u00e9lja annak meghat\u00e1roz\u00e1sa volt, hogy az enyhe neuropathia kimutat\u00e1s\u00e1ra alkalmazott elektrofiziol\u00f3giai vizsg\u00e1latok k\u00f6z\u00fcl melyek \u00e9rz\u00e9kenyebbek, valamint ezen m\u00f3dszerek \u00e9rz\u00e9kenys\u00e9g\u00e9nek felm\u00e9r\u00e9se praediabeteses betegekben. Ennek \u00e9rdek\u00e9ben a n. plantaris medialis \u00e9s a n. suralis dorsalis, valamint a suralis/radialis amplit\u00fad\u00f3ar\u00e1ny (SRAR) vizsg\u00e1lat\u00e1t v\u00e9gezt\u00fck praediabeteses \u00e9s diabeteses betegekben. A vizsg\u00e1latba praediabeteses \u00e9s DM-betegek ker\u00fcltek bevon\u00e1sra. \u00d6sszesen 92 f\u0151 (65 n\u0151, 27 f\u00e9rfi) vett r\u00e9szt a vizsg\u00e1latban. A neuropathi\u00e1s t\u00fcneteket a Self-Leeds Assessment of Neuropathic Symptoms and Signs (LANSS) f\u00e1jdalomsk\u00e1la seg\u00edts\u00e9g\u00e9vel \u00e9rt\u00e9kelt\u00fck. A standard elektrofiziol\u00f3giai vizsg\u00e1latok mellett idegvezet\u00e9si vizsg\u00e1latokat v\u00e9gezt\u00fcnk a suralis dorsalis, a plantaris medialis \u00e9s a radialis idegen is. A neuropathi\u00e1s t\u00fcneteket nem mutat\u00f3 praediabeteses betegek SNAP- (szenzoros idegi akci\u00f3s potenci\u00e1l) amplit\u00fad\u00f3it eg\u00e9szs\u00e9ges kontrollszem\u00e9lyek \u00e9rt\u00e9keivel hasonl\u00edtottuk \u00f6ssze. B\u00e1r a magas LANSSpontsz\u00e1m\u00fa praediabeteses betegekben a suralis dorsalis \u00e9s a plantaris medialis v\u00e1laszok \u00e1tlagos amplit\u00fad\u00f3ja m\u00e9g a norm\u00e1ltartom\u00e1ny als\u00f3 hat\u00e1r\u00e1n bel\u00fcl volt, szignifik\u00e1nsan alacsonyabbnak bizonyult az alacsony LANSS-pontsz\u00e1m\u00fa betegekhez k\u00e9pest. A SRAR-\u00e9rt\u00e9kek tekintet\u00e9ben nem volt szignifik\u00e1ns k\u00fcl\u00f6nbs\u00e9g a k\u00e9t csoport k\u00f6z\u00f6tt. A neuropathi\u00e1s t\u00fcneteket mutat\u00f3 praediabeteses betegek k\u00f6z\u00fcl 13 f\u0151n\u00e9l (56%) a suralis dorsalis v\u00e1lasz amplit\u00fad\u00f3ja a norm\u00e1l\u00e9rt\u00e9k alatt volt, m\u00edg 16 f\u0151n\u00e9l (69%) a plantaris medialis v\u00e1lasz mutatott k\u00f3rosan alacsony amplit\u00fad\u00f3t. Eredm\u00e9nyeink azt mutatj\u00e1k, hogy neuropathi\u00e1s panaszokkal rendelkez\u0151 praediabeteses betegekben a suralis dorsalis \u00e9s a plantaris medialis idegv\u00e1laszok m\u00e1r akkor is k\u00e1rosodhatnak, amikor a n. suralis vezet\u00e9se m\u00e9g norm\u00e1lis. A plantaris medialis ideg SNAP-amplit\u00fad\u00f3ja \u00e9rz\u00e9kenyebb vizsg\u00e1lati param\u00e9ternek bizonyult. Ezzel szemben a SRAR csak csek\u00e9ly t\u00f6bblet\u00e9rt\u00e9ket ny\u00fajtott az enyhe polyneuropathia kimutat\u00e1s\u00e1ban a suralis dorsalis \u00e9s a plantaris medialis ideg vizsg\u00e1lat\u00e1hoz k\u00e9pest. Praediabeteses, neuropathi\u00e1s panaszokkal jelentkez\u0151 betegek eset\u00e9ben fontos a polyneuropathia keres\u00e9se nagy \u00e9rz\u00e9kenys\u00e9g\u0171 elektrofiziol\u00f3giai m\u00f3dszerekkel.\n\nID: 42554326\nTitle: Carotid in-stent restenosis and re-restenosis treated with drug-eluting balloon dilation - case report and literature review.\nAbstract: According to relevant multicenter studies, significant in-stent stenosis develops in 5-6% of cases after internal carotid artery stenting. We present the risks, pathomechanism, diagnostics and treatment options of in-stent stenosis, the evidence, and the discrepancy between conservative and endovascular treatment. If the in-stent stenosis reaches a significant level, repeated endovascular treatment may be considered. The results of drug-eluting balloon dilation are better than balloon dilation. We present the case of a 72-year-old female patient. In an asymptomatic state, 80% stenosis of both internal carotid arteries was confirmed. Both sides were stented without complications. The patient did not have drug resistance, and the patient's compliance was very good. After 4 months, the patient was asymptomatic, and significant bilateral in-stent restenosis was confirmed. We performed drug-eluting balloon dilation in two sessions. In 3 months, we confirmed 85% in-stent restenosis again on the left side, due to which we performed repeated drug-eluting balloon dilation. The patient was asymptomatic and complaintfree. According to the last controls, there is currently no stenosis within the stents. Restenosis in the carotid stent can be effectively managed using drug-eluting balloon dilation, ensuring a high degree of safety. Our case, when a restenosis develops within a short time after drug-eluting ballon dilation, is very rare. Nagy tanulm\u00e1nyok szerint arteria carotis interna stentel\u00e9s ut\u00e1n 5-6%-ban alakul ki stenten bel\u00fcli jelent\u0151s sz\u0171k\u00fclet. Cikk\u00fcnkben bemutatjuk a carotisstenten bel\u00fcl bek\u00f6vetkez\u0151 sz\u0171k\u00fclet rizik\u00f3it, patomechanizmus\u00e1t, diagnosztik\u00e1j\u00e1t \u00e9s kezel\u00e9si lehet\u0151s\u00e9geit, a konzervat\u00edv \u00e9s endovascularis kezel\u00e9s evidenci\u00e1j\u00e1t \u00e9s diszkrepanci\u00e1j\u00e1t. Ha a stenten bel\u00fcli sz\u0171k\u00fclet szignifik\u00e1ns szintet \u00e9r el, ism\u00e9telt endovascularis ell\u00e1t\u00e1s j\u00f6het sz\u00f3ba. A gy\u00f3gyszerrel bevont ballonos t\u00e1g\u00edt\u00e1s eredm\u00e9nye jobb, mint a sima ballonos t\u00e1g\u00edt\u00e1s\u00e9. Egy 72 \u00e9ves n\u0151beteg\u00fcnk eset\u00e9t mutatjuk be. T\u00fcnetmentes \u00e1llapotban mindk\u00e9t arteria carotis interna 80%-os sz\u0171k\u00fclete igazol\u00f3dott. Mindk\u00e9t oldalt sz\u00f6v\u0151dm\u00e9nymentesen stentelt\u00fck. A betegn\u00e9l gy\u00f3gyszer-rezisztencia nem igazol\u00f3dott, a beteg a javasolt gy\u00f3gyszeres kezel\u00e9seket alkalmazta. N\u00e9gy h\u00f3nap m\u00falva a beteg t\u00fcnetmentes \u00e1llapot\u00e1ban mindk\u00e9t oldalon a stenteken bel\u00fcl szignifik\u00e1ns restenosis igazol\u00f3dott, ami miatt k\u00e9t \u00fcl\u00e9sben gy\u00f3gyszerrel bevont ballonos t\u00e1g\u00edt\u00e1st v\u00e9gezt\u00fcnk. \u00dajabb 3 h\u00f3nap m\u00falva bal oldalon ism\u00e9telten 85%-os stenten bel\u00fcli sz\u0171k\u00fcletet igazoltunk, ami miatt ism\u00e9telt gy\u00f3gyszerrel bevont t\u00e1g\u00edt\u00e1st v\u00e9gezt\u00fcnk. A beteg v\u00e9gig t\u00fcnet- \u00e9s panaszmentes volt, utols\u00f3 kontrollok alapj\u00e1n jelenleg a stenteken bel\u00fcl sz\u0171k\u00fclet nincs. A carotisstenten bel\u00fcli restenosis nagy biztons\u00e1ggal kezelhet\u0151 gy\u00f3gyszerrel bevont ballonos t\u00e1g\u00edt\u00e1ssal. Nagyon ritka, hogy t\u00e1g\u00edt\u00e1st k\u00f6vet\u0151en kialakul \u00fajabb sz\u0171k\u00fclet r\u00f6vid id\u0151n bel\u00fcl.\n\nID: 42554324\nTitle: Globoid cell leukodystrophy (Krabbe disease).\nAbstract: Background - Globoid cell leukodystrophy or Krabbe disease is a rare condition caused by biallelic pathogenic variants in the GALC gene. Accumulation of the metabolic intermedier galactosyl ceramide and psychosine leads to the death of myelin-producing cells. Demyelination results in large amounts of myelin debries that exceed the clearence capacity of microglial cells and macrophages and cause their transformation into globoid cells.Case presentation - We present the case of a currently 17 months old girl whose motor and cognitive development was noted to be delayed at 4 month of age. She had episodes of febrile enteritis followed by focal seizures and spasms with increasing frequency. Her neurological exam was significant for irritability, lack of eye contact, lethargy, increased limb tone, axial hypotonia, few spontaneous movements, sluggish deep reflexes, frequent myoclonus, and automatisms (chewing motions). The seizures were difficult to control. Her cranial MRI revealed confluent white matter abnormalities in the hemispheral white matter, cerebellum and brainstem suggesting an underlying metabolic abnormality. Whole exome sequencing identified two likely pathogenic variants in trans within the GALC gene, establishing the diagnosis of globoid cell leukodystrophy. At the time of diagnosis, hematopoietic stem cell transplantation (HSCT) was not recommended due to the advanced disease. Antiseizure medications and supportive therapy have been the means of controlling her symptoms.Conclusion -\u00a0Infantile Krabbe disease is a severe progressive disorder leading to death in 2-3 years. The only currently approved disease modifying therapy is HSCT that is effective only in early stages and is not without risks itself. There are numerous preclinical and clinical studies involving various forms of corrective gene therapy that may halt or reverse the course of this dismal condition. Bevezet\u00e9s \u2013 A globoidsejtes leukodystrophia vagy Krabbe-k\u00f3r egy olyan ritka betegs\u00e9g, amelyet a GALC g\u00e9n biall\u00e9likus patog\u00e9n vari\u00e1nsai okoznak. A galaktocerebrozid \u00e9s pszichozin nev\u0171 metabolikus intermedierek felhalmoz\u00f3d\u00e1sa a myelintermel\u0151 sejtek pusztul\u00e1s\u00e1hoz vezet. A demyelinisatio sor\u00e1n keletkez\u0151, nagy mennyis\u00e9g\u0171 myelint\u00f6rmel\u00e9k lebont\u00e1sa meghaladja a microglia \u00e9s makrof\u00e1g sejtek kapacit\u00e1s\u00e1t, aminek k\u00f6vetkezt\u00e9ben globoid sejtekk\u00e9 alakulnak \u00e1t.Esetbemutat\u00e1s \u2013 A tanulm\u00e1nyban egy jelenleg 17 h\u00f3napos kisl\u00e1ny eset\u00e9t mutatjuk be, akin\u00e9l 4 h\u00f3napos korban a motoros \u00e9s kognit\u00edv fejl\u0151d\u00e9s elmarad\u00e1s\u00e1t \u00e9szlelt\u00e9k. Majd l\u00e1zas enteritisepiz\u00f3dok jelentkeztek, amelyeket egyre gyakoribb\u00e1 v\u00e1l\u00f3 fok\u00e1lis rohamok \u00e9s g\u00f6rcs\u00f6k k\u00f6vettek. Neurol\u00f3giai st\u00e1tusz\u00e1b\u00f3l kiemelend\u0151 a nagyfok\u00fa irritabilit\u00e1s, szemkontaktus hi\u00e1nya, letargia, fokozott v\u00e9gtagt\u00f3nus, axi\u00e1lis hipot\u00f3nia, kev\u00e9s spont\u00e1n mozg\u00e1s, renyhe m\u00e9lyreflexek, gyakori myoclonusok \u00e9s automatizmusok (cs\u00e1mcsog\u00e1s). Rohamai nehezen voltak kontroll\u00e1lhat\u00f3ak. Koponya-MRIvizsg\u00e1lata az agyf\u00e9ltek\u00e9k feh\u00e9r\u00e1llom\u00e1ny\u00e1ban, a kisagyban \u00e9s az agyt\u00f6rzsben konfluens jelzavart mutatott, ami metabolikus rendelleness\u00e9gre utalt. Teljes exomszekven\u00e1l\u00e1s sor\u00e1n a GALC g\u00e9nben k\u00e9t, transz poz\u00edci\u00f3ban elhelyezked\u0151, val\u00f3sz\u00edn\u0171leg patog\u00e9n vari\u00e1nst azonos\u00edtottunk, ami meger\u0151s\u00edtette a globoid-sejtes leukodystrophia diagn\u00f3zis\u00e1t. A diagn\u00f3zis id\u0151pontj\u00e1ban a haematopoeticus \u0151ssejt-transzplant\u00e1ci\u00f3 (HSCT) a betegs\u00e9g el\u0151rehaladott st\u00e1diuma miatt nem volt javasolhat\u00f3. Ez\u00e9rt a t\u00fcnetek kezel\u00e9s\u00e9re antiepilepszi\u00e1s szereket \u00e9s sokr\u00e9t\u0171 t\u00e1mogat\u00f3 ter\u00e1pi\u00e1t alkalmaztunk.K\u00f6vetkeztet\u00e9s \u2013 A csecsem\u0151kori Krabbek\u00f3r s\u00falyos, progressz\u00edv betegs\u00e9g, amely 2-3 \u00e9ven bel\u00fcl hal\u00e1lhoz vezet. Jelenleg az egyetlen j\u00f3v\u00e1hagyott betegs\u00e9gm\u00f3dos\u00edt\u00f3 ter\u00e1pia a HSCT, ami csak korai f\u00e1zisban hat\u00e9kony, \u00e9s nem kock\u00e1zatmentes. Sz\u00e1mos preklinikai \u00e9s klinikai tanulm\u00e1ny van folyamatban k\u00fcl\u00f6nb\u00f6z\u0151 g\u00e9nter\u00e1pi\u00e1s megk\u00f6zel\u00edt\u00e9sekkel, amelyek meg\u00e1ll\u00edthatj\u00e1k vagy visszaford\u00edthatj\u00e1k ennek a s\u00falyos \u00e1llapotnak a lefoly\u00e1s\u00e1t.\n\nID: 42554319\nTitle: Scrub Typhus Presenting as Fulminant Myocarditis with Cardiogenic Shock: The Diagnostic Value of an Eschar.\nAbstract: \n\nID: 42554318\nTitle: Integrated Metabolomic and Metagenomic Profiling Reveals Distinct Microbial-Metabolic Signatures in the Adenoma-Carcinoma Sequence of Colorectal Cancer.\nAbstract: Colorectal cancer (CRC) arises via the stepwise adenoma-carcinoma sequence (ACS). Gut microbial dysbiosis and host metabolic reprogramming jointly correlate with CRC onset and advancement, yet their stage-specific crosstalk across ACS remains largely unclear. Limited multi-omics research on microbial-metabolic interactions throughout ACS hinders the development of early diagnostic biomarkers and preventive strategies. Here, we combined untargeted mucosal metabolomics and fecal shotgun metagenomic sequencing in 36 participants, covering healthy controls, ACS, and CRC patients. We systematically analyzed microbial composition, functions, differential metabolites, and enriched pathways and integrated multi-omics data to screen stage-specific signatures. Distinct gut microbial profiles and progressive functional shifts toward pathogenicity and abnormal carbohydrate metabolism were observed along ACS. Mucosal metabolism was continuously disrupted, with prominent alterations in taurine-hypotaurine, sphingolipid, and bile acid pathways. Core differential metabolites showed excellent diagnostic performance. Microbe-metabolite interactions were progressively enhanced to form a concerted pro-tumor axis. This study characterizes unique ACS-stage microbial-metabolic features. Dysregulated metabolic pathways and key microbe-metabolite crosstalk are closely associated with CRC progression, offering novel non-invasive biomarkers and premalignant intervention targets.\n\nID: 42554316\nTitle: Magnetic resonance imaging markers predict disease progression in early-stage multiple system atrophy: a 2-year prospective cohort study.\nAbstract: Magnetic resonance imaging (MRI) markers are identified as important indicators for the diagnosis of multiple system atrophy (MSA). However, whether these MRI markers can predict the disease progression of MSA remain undefined. We aimed to investigate the relationship between MRI markers and disease progression in patients with early MSA. The patients were divided into MRI-positive and MRI-negative groups based on MSA-specific MRI markers. Disease progression was evaluated using the Unified MSA Rating Scale (UMSARS), Montreal Cognitive Assessment and Frontal Assessment Battery. A repeated measures ANCOVA was used to compare the rate of disease progression between the two groups. A multiple linear regression model was used to assess the association between MRI subtype and disease progression. A total of 144 patients with early MSA were enrolled and 73 patients completed the 2-year follow-up. Patients with MSA and MSA of the parkinsonian subtype (MSA-P) in the MRI-positive group exhibited significantly faster disease progression on the total UMSARS score over a 2-year follow-up compared to those in the MRI-negative group (p\u2009=\u20090.002 and p\u2009=\u20090.020, respectively). Multiple linear regression analysis revealed that MRI-positive status was a significant predictor of more severe disease progression on the UMSARS total score in patients with MSA and MSA-P at the 1- and 2-year follow-up after adjusting for age, sex, and baseline disease duration (all p\u2009<\u20090.05). This study highlights that MRI markers are valuable imaging predictors of disease progression in early MSA, in addition to their diagnostic role.\n\nID: 42554307\nTitle: The 21-gene recurrence score assay as a tool for predicting recurrence risk and guiding adjuvant treatment selection in early breast cancer.\nAbstract: Estrogen receptor-positive (ER+), HER2-negative breast cancer is the most common breast cancer subtype. While adjuvant endocrine therapy reduces recurrence risk, identifying which patients benefit from the addition of chemotherapy remains a key clinical challenge. The Oncotype DX\u00ae 21-gene Recurrence Score assay (Exact Sciences, via Genomic Health, Inc.) was developed to address this by quantifying distant recurrence risk and informing chemotherapy decisions in early-stage ER+/HER2- disease. This diagnostic profile reviews the development, validation, and clinical evidence for Oncotype DX, including findings from the TAILORx and RxPONDER prospective trials and the subsequent development of hybrid tools integrating genomic and clinicopathological data. Alternative multiparameter molecular tests (MammaPrint, Prosigna, EndoPredict, Breast Cancer Index) are summarized and compared. We review international guideline recommendations, decision impact studies, cost-effectiveness evidence, and ongoing trials. Oncotype DX has strong prognostic evidence and has meaningfully reduced chemotherapy use, though its case as a biomarker predictive of therapeutic effect from chemotherapy rests on trial designs with important limitations. Its independent prognostic contribution beyond comprehensive clinicopathological assessment requires further clarification, and cost-effectiveness varies substantially by indication and healthcare setting.\n\nID: 42554301\nTitle: BOLD fMRI activations and LC contrast associated with successful episodic memory in healthy older adults and MCI.\nAbstract: We examined whether locus coeruleus (LC) contrast, an indirect proxy for neuronal density and structural health, was related to delayed episodic memory in healthy older adults (OAs) and those with mild cognitive impairment (MCI) and its association with encoding-related brain activity, controlling for emotional salience. Participants (27 younger adults, 26 OAs, and 24 MCI) memorized emotional and neutral images during functional magnetic resonance imaging (MRI). Delayed memory was tested after 4 h and compared to immediate memory. LC contrast was measured via LC-sensitive MRI. Memory was lower, independent of delay or emotional valence, and LC contrast was more strongly reduced in those with MCI compared to OAs. In OAs, LC contrast correlated with delayed memory and was mildly associated with LC activity during successful encoding. All groups remembered emotional images better than neutral ones, though memory was greatly impaired in those with MCI. LC contrast was associated with lower brain activity during encoding and emotional salience processing. Its relationship to delayed memory was evident in those with MCI.\n\nID: 42554285\nTitle: Temporal order of clinical, imaging, and biomarker changes in frontotemporal lobar degeneration-associated syndromes.\nAbstract: The temporal sequence of clinical, imaging, and biological changes in sporadic frontotemporal lobar degeneration (FTLD)-associated syndromes remains poorly characterized, and a comprehensive biomarker cascade model is lacking. We developed a data-driven biomarker cascade model in 489 patients across the FTLD spectrum (211 behaviorial variant frontotemporal dementia [bvFTD], 129 primary progressive aphasia [PPA], 71 corticobasal syndrome [CBS], 66 progressive supranuclear palsy [PSP], and 12 FTD associated with amyotrophic lateral sclerosis [FTD-ALS]; 1904 patient-visit observations). Plasma, magnetic resonance imaging (MRI), and clinical biomarkers were modeled using sigmoid trajectories fitted to covariate-adjusted longitudinal data. Plasma glial fibrillary acidic protein departed from normality earliest, followed by Trail Making Test Part B (TMT-B), white matter lesion volume, and neurofilament light chain. Insular atrophy showed the steepest transition among MRI measures; clinical dementia rating dementia staging instrument plus National Alzheimer's Coordinating Center behavior and language domains sum of boxes declined most steeply overall. TMT-B inflected earliest in bvFTD, whereas insula atrophy dominated in PPA. This first data-driven temporal cascade of multimodal biomarkers in sporadic FTLD-associated syndromes offers a framework for disease staging and stage-specific clinical trial design.\n\nID: 42554244\nTitle: Electrocatalytically Amplified Immunosensing Enabled by Titanium Oxynitride-Supported Ru Nanodots on Graphene Oxide Hybrid Architecture.\nAbstract: Electrocatalytically amplified electrochemical immunosensing is a powerful strategy for sensitive and interference-free detection of biomarkers. Herein, we report a high-performance electrochemical immunosensor enabled by a hierarchical hybrid architecture comprising Ru nanodots anchored on titanium oxynitride nanoflakes dispersed on graphene oxide (Ru/TiON-GO), linked via APTES to biorecognition building blocks. The engineered heterostructure exhibits advanced electrocatalytic activity, arising from synergistic electronic coupling between Ru nanodots and the highly conductive TiON-GO support, effectively promoting interfacial electron transport between the electrocatalytic surface and the [Fe(CN)6]3-/4- redox probe. Leveraging this electrocatalytic platform, a prostate-specific antigen (PSA) impedimetric immunosensor was constructed, achieving a limit of detection of 0.06\u00a0ng mL-1 (2.3 pM) and a wide linear response covering clinically relevant concentration range. The immunosensor demonstrates excellent selectivity in human serum, operating in interference-free mode even in the presence of common coexisting biomolecules, highlighting suitability for medical diagnostics. Density functional theory calculations further elucidate the origin of the enhanced electrocatalytic performance, with charge density difference plots and density of states analysis revealing Ru-driven electron redistribution and increased density of states near the Fermi level. This work establishes the Ru/TiON-GO nanocomposite as a robust electrocatalytic platform for advanced immunosensing applications, paving the way toward next-generation electrochemical diagnostic devices.\n\nID: 42554243\nTitle: Processionary Moth Exposure in Europe: Toxic-Irritant and Immunoglobulin E-Mediated Endotypes, Diagnosis and Management.\nAbstract: Processionary moths (Thaumetopoea spp.), particularly the oak processionary moth (Thaumetopoea processionea), are an increasingly relevant cause of allergic and toxic disease in Europe, occurring in the context of reported range expansion, changing environmental conditions and persistent contamination with airborne urticating hairs (setae). Clinical manifestations range from toxic-irritant dermatitis and ocular injury to respiratory symptoms, while a subset of exposed individuals develops genuine IgE-mediated disease, including rhinoconjunctivitis, asthma, and rare cases of anaphylaxis. These presentations reflect two mechanistically distinct endotypes: (i) toxin-driven epithelial penetration by the urticating hairs with neuro-inflammatory activation and (ii) allergen-specific sensitisation to defined molecular components such as Tha p 1 and Tha p 2. Accurate endotype differentiation is clinically relevant, as symptom latency, severity, and risk of systemic progression differ substantially. Diagnostic evaluation relies on exposure history and phenotype recognition, complemented in selected cases and experienced centres by skin testing using research-grade extracts and emerging component-resolved approaches. Basophil activation testing may provide additional mechanistic insights in selected cases. Management comprises prompt decontamination, anti-inflammatory therapy, urgent ophthalmologic assessment when eye involvement is suspected, and guideline-based management of anaphylaxis. The environmental persistence of setae and recurrent seasonal exposure highlight the need for improved surveillance, validated molecular diagnostics, and translational research defining immune mechanisms and biomarkers.\n\nID: 42554217\nTitle: Advances in Nucleic Acid-Based Amplification Strategies for Dual-Mode Biosensing of Pathogenic Bacteria: From Molecular Mechanisms to Integrated Analytical Systems.\nAbstract: Rapid, accurate, and reliable detection of pathogenic bacteria remains a critical need in clinical, food, and environmental monitoring. In today's context, nucleic acid amplification-mediated biosensing have emerged as a prominent approach to improve detection sensitivity, whereas dual-mode signal readout approaches have more enhanced analytical robustness and reliability. This review outlines recent advances in nucleic acid signal amplification strategies, including enzyme-based methods like LAMP, RPA, and RCA, as well as enzyme-free approaches like HCR, CHA, and EDR. Special attention is given to incorporating these amplification methods into dual-mode biosensing systems that combine both optical and electrochemical transduction mechanisms. This integration enables complementary signal generation and improves detection accuracy by reducing false-positive and false-negative results. This study critically examines the advancement of nucleic acid signal amplification strategies (NASAS)-mediated dual-mode sensing systems for detecting major pathogenic bacteria, including Escherichia coli, Salmonella, Listeria monocytogenes, Staphylococcus aureus, and Vibrio species, focusing on selectivity, sensitivity, assay design, and real-sample applicability. Finally, the review highlights present challenges related to system integration, standardization, and point-of-care applications. Additionally, it outlines potential future directions for rendering nucleic acid amplification-based dual-mode probes into practical diagnostic devices. Overall, this study affords a comprehensive synthesis of emerging approaches and design mechanisms for next-generation diagnostic scaffold for pathogen analysis.\n\nID: 42554216\nTitle: Phosphonic Acid-Enriched Nanogels as High Relaxivity Platforms for Enhanced Magnetic Resonance Imaging and Drug Delivery.\nAbstract: Clinically small-molecule magnetic resonance imaging (MRI) contrast agents face limitations due to safety concerns, low relaxivity, narrow imaging windows, and restricted targeting capabilities. This study synthesized biocompatible nanogels via the polymerization of N-vinyl caprolactam and vinyl phosphonic acid. The nanogels containing phosphonic acid groups could tightly bind with Mn2+, which could improve the biosafety by preventing the release of free Mn2+. Meanwhile, the strong ligand effect improved the longitudinal relaxivity of encapsulated Mn2+ to 29.8 mM- 1s- 1, which was roughly 3.5 times, 10.6 times, and 6.6 times higher than that of manganese chloride (MnCl2), mangafodipir (MnDPDP) and gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA), respectively. These nanogels exhibited the capability of targeting bone metastases and bone tissues in vivo owing to the phosphonic acid groups. Moreover, these nanogels-based contrast agents prolonged the effective imaging window due to the increased circulation time. They also exhibited a loading capacity for clinical hydrophobic drugs, such as atovaquone (ATO), thereby potentially improving the pharmacokinetics. This work highlights the prospect of poly(vinyl phosphonic acid) based nanogels as multifunctional nanoplatforms for diagnostic and therapeutic applications.\n\nID: 42554207\nTitle: Manganese Ion-Based NanoPlatforms for Tumor Theranostics.\nAbstract: Manganese-ion-based nanoparticles have emerged as highly versatile platforms that seamlessly integrate diagnostic and therapeutic functions, enabling precision cancer theranostics. In this review, we discuss manganese-based nanomaterials through a design-oriented framework that connects synthesis, structure, function, application, and limitation. Mainstream preparation strategies, including thermal decomposition, permanganate reduction, hydrothermal/solvothermal synthesis, template-assisted construction, coprecipitation, and mild biomimetic routes, are compared with respect to physicochemical control, Mn activation behavior, scalability, and biomedical suitability. The diagnostic applications of manganese-based nanoplatforms are discussed, including activatable T1-weighted magnetic resonance imaging (MRI) and multimodal imaging systems, as well as Mn-activated DNAzyme platforms for gene regulation. We further review therapeutic modalities enabled by Mn2 +, encompassing chemodynamic therapy (CDT), nanozyme-mediated redox regulation within the tumor microenvironment, stimulus-responsive drug and prodrug delivery, magnetic hyperthermia, and their synergistic combinations. Finally, we critically analyze key biosafety considerations along with current translational challenges and future perspectives for developing clinically viable, image-guided combination cancer therapies based on manganese-ion nanoplatforms.\n\nID: 42554201\nTitle: Plasma Metabolomes Identify Distinct Asthma Metabotypes: Findings From the COREA Cohort.\nAbstract: Phenotype-based asthma classification has limitations that have motivated endotype-based approaches grounded in pathophysiology. However, existing biomarkers offer limited specificity and clinical accessibility. Plasma metabolomics, which reflects disease-specific metabolic states, offers a promising strategy for asthma classification. This study aimed to identify metabolic subgroups (metabotypes) of adult asthma using plasma metabolomics, with potential implications for personalized treatment. Plasma samples from 407 patients with asthma in the Cohort for Reality and Evolution of Adult Asthma in Korea (COREA) were analyzed using Biocrates AbsoluteIDQ p400 HR kit with liquid chromatography-mass spectrometry. After preprocessing, 281 metabolites were natural-log-transformed, standardized, and partitioned by k-means clustering. The number of clusters was determined by a multi-criteria assessment combining cluster validity indices, consensus clustering, stability analysis, and cross-algorithm agreement. Four metabotypes with distinct lipid-class signatures were identified. Group 1 (n\u2009=\u2009123) was characterized by elevated ether-linked phosphatidylcholines, comprising younger patients with the earliest symptom onset and female predominance. Group 2 (n\u2009=\u200953) showed elevated lyso-phosphatidylcholines and altered amino acid metabolism (elevated glutamate, reduced glutamine), representing a metabolically intermediate, non-T2-high subgroup. Group 3 (n\u2009=\u2009158) exhibited globally reduced sphingomyelins and broadly lower phosphatidylcholines in middle-aged, non-obese patients. Group 4 (n\u2009=\u200973) showed markedly elevated triacylglycerols (TG) and diacylglycerols (DG) with the highest body mass index (BMI), consistent with a non-T2, obesity-related metabotype. The TG/DG signature of Group 4 remained robust after adjustment for BMI, age, and sex. Plasma metabolomics identifies clinically meaningful asthma metabotypes, supporting integration of metabolomic profiling into personalized asthma management.\n\nID: 42554183\nTitle: Wnt signalling pathway dysfunction in non-obstructive azoospermia: identifying key genes and potential therapeutic targets.\nAbstract: This study aimed to investigate the involvement of the Wnt signalling pathway in the pathogenesis of non-obstructive azoospermia (NOA) and to identify potential diagnostic and therapeutic targets. We obtained the gene expression profiles of NOA patients from the GEO database, screened for differentially expressed genes (DEGs) and NOA-associated co-expressing gene modules, then intersected the DEGs with related genes in the Wnt signalling pathway and identified key genes. LASSO regression analysis was used to find hub genes, and qRT-PCR and WB methods were used to verify the expression of key genes in NOA patients. The immune infiltration and GSEA were conducted in order to investigate the relationship between hub genes and immune cells infiltration and NOA-associated pathways; furthermore, we built the miRNA-mRNA-TF regulatory network and predicted possible small molecules drug targets of the hub gene. Four candidate genes were related to NOA: CSNK1G2, GNG3, H2AFB1 and PARD6A. The key hub gene was CSNK1G2 based on the results of the LASSO regression analysis, and its expression level was significantly down-regulated in NOA patients. The immune infiltration analysis and GSEA results showed that the expression level of CSNK1G2 had a close relationship with immune cells infiltration and NOA-related pathways. Furthermore, we built a miRNA-mRNA-TF regulatory network and predicted the possible small molecule drugs of CSNK1G2. Our findings suggest that the Wnt signalling pathway, particularly CSNK1G2, plays a crucial role in the pathogenesis of NOA. CSNK1G2 may be a novel target for the diagnosis and treatment of NOA.\n\nID: 42554173\nTitle: Plasma Concentrations of Amyloid-\u03b2 Peptides, BACE-1, and Tau Proteins in Alzheimer's Disease.\nAbstract: The aim of this study was to compare, using ELISA, plasma levels of amyloid beta (A\u03b2)40, A\u03b242, beta-site amyloid precursor protein cleaving enzyme 1 (BACE-1), total tau (t-tau), and phosphorylated tau (p-tau), as well as the A\u03b242/A\u03b240 ratio, between patients with Alzheimer's disease (AD) and healthy controls, and to evaluate their diagnostic performance in relation to demographic and lifestyle factors. This study is a single-center, cross-sectional, case-control study. Twenty-four individuals diagnosed with AD and 37 healthy volunteers included in the study. Alongside the analysis of plasma samples obtained from the participants, demographic data were analyzed to assess the potential influence of lifestyle and environmental factors on disease development. Analysis of the case data showed that increasing age was a risk factor for AD, higher education level was associated with an increased risk of AD, and tea consumption was inversely associated with AD. While age is a well-known risk factor, both the increased risk of AD associated with higher education and the relatively protective effect of tea consumption against AD are supported by the literature. Evaluation of the levels of A\u03b240, A\u03b242, BACE-1, t-tau, and p-tau and of the A\u03b242/A\u03b240 ratio revealed no significant differences between the patient and control groups. Additionally, A\u03b240, A\u03b242, BACE-1 showed correlations in both the control and Alzheimer's groups, whereas t-tau did not. None of the investigated plasma biomarkers (A\u03b240, A\u03b242, BACE-1, t-tau, p-tau, and the A\u03b242/A\u03b240 ratio) discriminated Alzheimer's patients from healthy controls, with all receiver operating characteristic area under the curve values below 0.62. These findings indicate that in this cohort, plasma levels of these individual markers did not provide diagnostic value; larger longitudinal studies including cerebrospinal fluid comparisons and multi-marker panels are needed.\n\nID: 42554166\nTitle: Predictive Value of Endoscopy in the Identification of Complete Response After Neoadjuvant Therapy for Rectal Adenocarcinoma.\nAbstract: Large-scale evidence on the role of endoscopy alone as the assessment tool to identify complete response after neoadjuvant therapy in locally advanced rectal cancer, particularly across different neoadjuvant therapy regimens, is scarce. This study aims to evaluate the diagnostic accuracy of endoscopy for predicting luminal complete response after neoadjuvant therapy and compare its performance between patients receiving neoadjuvant chemotherapy and neoadjuvant chemoradiotherapy. Retrospective cohort study. The study was conducted at a teaching hospital. A total of 349 locally advanced rectal cancer patients (223 neoadjuvant chemotherapy, 126 neoadjuvant chemoradiotherapy) who underwent neoadjuvant therapy and restaging endoscopy were included. Three reviewers assessed endoscopic images independently to determine the occurrence of endoscopic complete response. The diagnostic accuracy for predicting pathological luminal complete response (ypT0) was calculated. The overall accuracy of endoscopy for predicting complete response was 92.0% (321/349), with a positive predictive value of 69.6%, sensitivity of 69.6% and specificity of 95.4%. A flat scar was the most predictive feature (positive predictive value = 70.3%). The sensitivity of endoscopic performance was significantly higher in the neoadjuvant chemoradiotherapy group compared to the neoadjuvant chemotherapy group (90.0% vs. 53.8%, p = 0.008), despite comparable overall accuracy and area under the curve. The addition of biopsy did not significantly improve diagnostic ability. Inter-reviewer agreement was substantial to excellent (\u03ba=0.754-0.883). No video or artificial intelligence was involved. Endoscopy is an optional tool for identifying luminal complete response after neoadjuvant therapy. Its sensitivity is lower after neoadjuvant chemotherapy than after neoadjuvant chemoradiotherapy, warranting caution against missed diagnoses. Beyond the flat scar, other features like small flat ulcers also have a non-negligible complete response rate. The role of routine biopsy still needs further exploration. See Video Abstract.\n\nID: 42554132\nTitle: Characterizing the Milk Microbiome in Subclinical Mastitis: A Pilot 16S rRNA-Based Study in Cattle and Water Buffalo.\nAbstract: In the dairy sector of Bangladesh, subclinical mastitis (SCM) is a substantial and frequently undiagnosed challenge, with reported prevalence rates of 60%-77% in cattle and approximately 52% in buffaloes. Due to its complex characteristics and progressive development, efficient diagnosis and management are essential for enhancing dairy productivity. This pilot study employed 16S rRNA amplicon sequencing using Oxford Nanopore's MinION to investigate the milk microbiota of healthy and mastitic cattle and buffalo. A total of 423 clustered nucleotide sequences were identified in the samples, indicating significant taxonomic diversity: 11 phyla, 26 classes, 58 orders, 120 families and 272 genera. Distinct phylum-level patterns were observed, with Firmicutes predominating in healthy milk and a relative increase in Proteobacteria and Actinobacteriota in mastitic samples. At the genus level, Streptococcus and Lactococcus were predominant in mastitic samples, whereas Staphylococcus and Lactococcus were more prevalent in healthy milk. The results indicate that although overall microbial diversity was relatively consistent across groups, mastitis correlated with alterations in bacterial community composition, with notable differences between cattle and buffalo. This study suggests a potential association between SCM and microbial shifts; however, microbiome profiling cannot yet be recommended for diagnostic application. Clinical applicability requires validation in large-scale studies with individual-level sampling.\n\nID: 42554121\nTitle: Clinical value of intraoperative frozen section in the surgical management of ovarian tumors: diagnostic accuracy and correlation with final histopathology.\nAbstract: Intraoperative frozen section (FS) analysis is widely used during surgery for ovarian tumors to guide the extent of the operative procedure. It plays a particularly significant role in young women, in whom fertility-sparing surgery is considered. Despite its high clinical utility, FS has technical limitations that may result in discrepancies compared with final histopathological diagnosis. To assess the concordance between intraoperative frozen section diagnosis and final histopathological examination of ovarian tumors, and to evaluate the diagnostic accuracy, sensitivity, specificity, and predictive values of FS. The analysis included finally 100 women aged 11-78 years from among 114 who were hospitalized and underwent surgery for ovarian tumors conducted at a tertiary Gynecology and Obstetrics Clinical Hospital from January to June 2025. Frozen section diagnoses were compared with final histopathological results. Concordance rates, patterns of diagnostic discrepancies, and diagnostic performance parameters (sensitivity, specificity, positive predictive value [PPV], and negative predictive value [NPV]) were calculated using final histopathology as the reference standard. Overall concordance between frozen section and final histopathology was 85%. Diagnostic discrepancies were observed in 15% of cases, with only 2% considered clinically significant. Frozen section showed excellent diagnostic performance for benign tumors (sensitivity 100%, specificity 94.1%) and malignant tumors (specificity and PPV 100%). The lowest diagnostic accuracy was observed for borderline tumors, which accounted for most clinically relevant discrepancies. The most frequent source of discordance involved hemorrhagic cysts, which were ultimately diagnosed as endometriotic cysts on final histopathology. Intraoperative frozen section analysis is a reliable and safe diagnostic tool that effectively supports surgical decision-making in ovarian tumors. Its high diagnostic accuracy and predictive values confirm its clinical value, particularly in tailoring the extent of surgery. Special caution is warranted in the interpretation of borderline tumors, for which final histopathological examination remains essential.\n\nID: 42554111\nTitle: Diagnostic Accuracy and Optimal Thresholds of Salivary Pepsin (Peptest) in Laryngopharyngeal Reflux: A Systematic Review and Meta-Analysis.\nAbstract: Salivary pepsin has been proposed as a non-invasive biomarker for diagnosing laryngopharyngeal reflux (LPR); however, its diagnostic utility remains uncertain due to inconsistent findings across studies. This meta-analysis evaluated the diagnostic accuracy of a rapid lateral flow test (Peptest) compared to the reference standard of multichannel intraluminal impedance-pH (MII-pH) monitoring. Systematic review and meta-analysis of cohort studies. Patients with LPR. The primary outcomes were diagnostic odds ratio (DOR), summary receiver operating characteristic curve and area under the curve (AUC). In total, 11 studies comprising 928 samples were included. The diagnostic performance of salivary pepsin for LPR was analysed at cut-off values of 16, 45-50 and 100\u2009ng/mL. The pooled DOR and AUC were 3.49 and 0.684 at 16\u2009ng/mL, 6.08 and 0.772 at 45-50\u2009ng/mL and 4.78 and 0.717 at 100\u2009ng/mL, respectively. Multiple-time sampling significantly improved sensitivity compared to single-time sampling across all cut-off values (0.82 vs. 0.33-0.63 at 16\u2009ng/mL; p\u2009<\u20090.0001), although specificity was slightly lower. The Peptest demonstrates potential as a simple, non-invasive diagnostic alternative to 24-h-MII-pH monitoring for LPR. Among the evaluated thresholds, a cut-off of 45-50\u2009ng/mL provided the highest diagnostic accuracy. Collecting multiple saliva samples may further enhance detection sensitivity. However, in patients with a high clinical suspicion of LPR but negative Peptest results, 24-h-MII-pH monitoring remains necessary to confirm the diagnosis due to the test's limited negative predictive value.\n\nID: 42554097\nTitle: Operando Diagnosis of Acidic CO2 Electroreduction Degradation Mechanism in a Zero-Gap Electrolyzer via the Distribution of Relaxation Times.\nAbstract: Acidic CO2 electroreduction (CO2R) in membrane electrode assemblies (MEAs) offers a promising route for high-performance electrolysis but remains limited by poor stability. Here, we integrate operando electrochemical impedance spectroscopy (EIS) with distribution of relaxation times (DRT) analysis to diagnose the key processes governing acidic CO2R in MEAs. Comparative evaluation of Nafion and phosphoric-acid-doped polybenzimidazole (PA-PBI) membranes shows that PA-PBI effectively suppresses salt precipitation, providing a reliable basis for interpreting DRT spectra. By systematically adjusting electrolysis parameters, distinct features associated with interfacial contact resistance, anodic/cathodic ionic conduction, charge transfer, and CO2 transport are resolved in DRT spectra. Furthermore, stability tests with DRT analysis reveal rapid failure of Nafion-based MEA (\u223c0.5\u00a0h) due to salt accumulation, whereas the PA-PBI-based MEA operates for 103\u00a0h before CO2 mass-transport limitations arise. These findings demonstrate operando EIS-DRT as a powerful diagnostic tool and provide mechanistic guidance for designing long-lasting acidic CO2R MEAs.\n\nID: 42554095\nTitle: Silent Surveillance: Neonatal Nurses' Recognition, Interpretation and Escalation of Early Clinical Deterioration-An Interpretive Description Study.\nAbstract: To explore how neonatal nurses in Level III (high-acuity) neonatal intensive care units recognise, interpret, and escalate early signs of clinical deterioration. A qualitative interpretive description study underpinned by critical realism. Twenty-two registered nurses were purposively recruited from four Level III units in the Jouf Region of northern Saudi Arabia using criterion-based maximum-variation sampling. Semi-structured interviews were conducted in Arabic or English and analysed in their original language. Situation awareness and invisible work scholarship served as sensitising lenses. Rigour was supported through member reflection, negative-case analysis, peer debriefing and an audit trail. Four interrelated themes were identified: (1) 'the infant tells you before the monitor does', reflecting recognition of subtle deviation from the infant's baseline; (2) 'building the case', in which nurses tested and assembled evidence to make concern clinically credible before escalation; (3) 'the escalation calculus', describing how anticipated physician responses, hierarchy and previous dismissal shaped escalation and (4) 'the unit works against you', showing how alarm burden, documentation demands and staffing variability constrained vigilance. Recognition and escalation formed a cyclical, organisationally situated process rather than a single response to abnormal observations. Escalation involved interpretive, relational and legitimising work before early concern became actionable. Participants' accounts suggest that timely response depends on clinical expertise and environments that protect bedside attention and legitimise early escalation. Neonatal services should protect surveillance capacity through staffing and workload design, alarm reduction, simulation of ambiguous deterioration and constructive escalation feedback. The study identifies case-building as a distinct stage between noticing deterioration and escalating concern and indicates how organisational conditions may constrain early nursing surveillance. Consolidated Criteria for Reporting Qualitative Research. Patients and the public were not involved.\n\nID: 42554082\nTitle: Liquid Biopsy in Head and Neck Squamous Cell Carcinoma: A Systematic Review of the Scientific Literature.\nAbstract: Head and neck squamous cell carcinoma (HNSCC) presents a significant global health challenge, characterized by late-stage diagnosis and high rates of locoregional recurrence. Traditional tissue biopsy, while considered the gold standard, is invasive and often fails to capture the spatial and temporal heterogeneity of the tumor. Liquid biopsy has emerged as a transformative, noninvasive paradigm for molecular diagnostics, leveraging circulating biomarkers found in blood and saliva. This systematic review critically evaluates current evidence regarding the diagnostic accuracy, prognostic value, and molecular validity of circulating tumor cells, circulating tumor DNA (ctDNA), and the transcriptomic landscape of coding and noncoding RNAs (miRNAs, lncRNAs, and circRNAs) in HNSCC. Our review highlights the specificity of ctDNA for monitoring minimal residual disease, particularly in human papillomavirus- and Epstein-Barr virus-driven malignancies, where viral kinetics serve as sensitive biomarkers for recurrence. Furthermore, we explore the utility of \"Saliva-Exosomics,\" suggesting that salivary exosomes may offer high sensitivity for early-stage oral cavity lesions due to anatomical proximity. The review also synthesizes data on novel RNA species, such as tumor-educated platelets and circular RNAs, which act as stable systemic biosensors. Despite promising results, significant heterogeneity in isolation protocols and detection platforms (e.g., next-generation sequencing vs. digital PCR) remains a barrier to widespread clinical implementation. We conclude that integrating multianalyte liquid biopsies into clinical practice holds the potential to personalize therapeutic strategies, improve survival outcomes, and redefine the standard of care in head and neck oncology.\n\nID: 42554080\nTitle: Utility of Amniotic Fluid Biomarkers to Predict Preterm Delivery for Clinical Chorioamnionitis in Women With Intact Membranes.\nAbstract: Clinical chorioamnionitis is a major contributor to preterm birth and neonatal morbidity. Diagnosis in women with intact membranes is challenging, particularly in the subclinical phase where maternal signs are subtle and conventional serum markers are limited in predictive value. To evaluate the diagnostic performance of selected novel amniotic fluid biomarkers (interleukin-6, interleukin-10, tumour necrosis factor-\u0251, matrix metalloproteinase-8), and conventional amniotic and serum biomarkers in predicting delivery for clinical chorioamnionitis within 14\u2009days of amniocentesis in women with suspected subclinical infection and intact membranes. A prospective cohort study was conducted at a single Australian tertiary hospital. Thirty-two women with singleton pregnancies and suspected subclinical chorioamnionitis underwent amniocentesis. Amniotic fluid and serum were analysed for selected biomarkers. The primary outcome was delivery for clinical chorioamnionitis within 14\u2009days of amniocentesis. ROC curves were constructed to assess biomarker performance. Delivery for clinical chorioamnionitis occurred in 11 of 31 evaluable cases (35%). Matrix metalloproteinase-8, lactate dehydrogenase, and glucose were the strongest predictors (area under the curve (AUC) of 0.93 (95% CI 0.85-1.0), 0.93 (95% CI 0.85-1.0), and 0.92 (95% CI 0.8-1.0) respectively). Amniotic fluid biomarkers outperformed serum markers (C-reactive protein AUC 0.70 (95% CI 0.49-0.91) and white cell count AUC 0.67 (95% CI 0.46-0.86)). Novel amniotic fluid biomarkers did not demonstrate superior diagnostic performance over conventional amniotic fluid biomarkers including lactate dehydrogenase and glucose. Amniotic fluid biomarkers show promise in identifying women with subclinical chorioamnionitis. These findings support further validation of scalable biomarker-based testing to guide early intervention and improve perinatal outcomes.\n\nID: 42153377\nTitle: Antagonistic contributions of A-type and B-type lamins to LBR localization and dynamics.\nAbstract: Lamin B receptor (LBR) is an inner nuclear membrane protein that organizes peripheral heterochromatin and cooperates with lamins to tether chromatin at the nuclear periphery. However, how individual lamin isoforms regulate LBR localization and anchorage remains unclear. Using mouse embryonic fibroblasts lacking all endogenous lamins, we examined how individual lamin isoforms regulate LBR behavior. Expression of lamin B1 or B2 was sufficient to anchor LBR at the nuclear envelope, whereas lamin A expression increased LBR mobility and led to its displacement from the nuclear envelope. This effect was mediated by phosphorylation of LBR and was recapitulated by lamin A overexpression in wild-type cells. Collectively, these findings define isoform-specific and antagonistic roles for A-type and B-type lamins in regulating LBR anchorage at the nuclear envelope. In addition, they indicate a lamin A-dependent mechanism that may reflect a broader developmental process, since LBR and lamin A sequentially tether peripheral heterochromatin during development.\n\nID: 41757046\nTitle: Antagonistic contributions of A-type and B-type lamins to LBR localization and dynamics.\nAbstract: Lamin B receptor (LBR) is an inner nuclear membrane (INM) protein that plays crucial roles in maintaining nuclear architecture and organization of peripheral heterochromatin. Lamins and LBR both contribute to chromatin tethering at the nuclear periphery, and the expression of LBR and A-type lamins is tightly regulated during development to ensure a faithful transition between different chromatin tethering modalities. Despite its well-established association with B-type lamins, the contributions of individual lamin isoforms to LBR localization and anchorage have not been systematically examined. Here, we used mouse embryonic fibroblasts (MEFs) lacking all endogenous lamins (triple lamin knockout: TKO) to assess how specific lamin isoforms and domains regulate LBR subcellular localization and mobility. Whereas ectopic expression of either lamin B1 or lamin B2 was sufficient to tether LBR to the nuclear envelope in TKO cells, expression of lamin A increased the lateral mobility of LBR at the nuclear membrane, resulting in its displacement from the nuclear envelope to the ER. The lamin A-induced displacement of LBR was mediated by phosphorylation of LBR. Overexpression of lamin A in wild-type MEFs similarly increased LBR phosphorylation and promoted its displacement from the nuclear envelope. Collectively, these findings define isoform-specific and antagonistic roles for A-type and B-type lamins in regulating LBR anchorage at the nuclear envelope. In addition, they indicate a lamin A-dependent mechanism that may reflect a broader developmental process, since LBR and lamin A sequentially tether peripheral heterochromatin during development.\n\nID: 32013726\nTitle: Cholesterol derivatives induce dephosphorylation of the histone deacetylases Rpd3/HDAC1 to upregulate autophagy.\nAbstract: Histone deacetylases (HDACs) are important for global gene expression and contribute to numerous physiological events. Deacetylase Rpd3 in yeast and its conserved homolog HDAC1 in mammals oppositely regulate autophagy; however, how Rpd3/HDAC1 is regulated to mediate autophagy remains unclear. Here, we showed autophagy occurrence in silkworm (Bombyx mori) required BmRpd3, wherein steroid hormone 20-hydroxyecdysone (20E) signaling regulated its protein level and nuclear localization negatively. Inhibition of MTOR led to dephosphorylation and nucleo-cytoplasmic translocation of BmRpd3/HsHDAC1. Besides, cholesterol, 20E, and 27-hydroxycholesterol could all induce massive dephosphorylation and cytoplasmic localization of BmRpd3/HsHDAC1, and thus autophagy by affecting MTORC1 activity. In addition, three phosphorylation sites (Ser392, Ser421, and Ser423) identified in BmRpd3 were conserved in HsHDAC1. Single or triple phosphorylation-site mutation attenuated the phosphorylation levels of BmRpd3/HsHDAC1, leading to their cytoplasmic localization and autophagy activation. In general, cholesterol derivatives, especially hydroxylated cholesterol, caused dephosphorylation and nucleo-cytoplasmic shuttling of BmRpd3/HsHDAC1 through inhibition of MTOR signaling to facilitate autophagy in B. mori and mammals. These findings improve our understandings of BmRpd3/HsHDAC1-mediated autophagy induced by cholesterol derivatives and shed light on their potential as a therapeutic target for neurodegenerative diseases and autophagy-related studies.Abbreviations: 20E: 20-hydroxyecdysone; 27-OH: 27-hydroxycholesterol; ACTB: actin beta; AMPK: AMP-activated protein kinase; Atg: autophagy-related; BmSqstm1: Bombyx sequestosome 1; CQ: chloroquine; HDAC: histone deacetylase; LMNB: Lamin B1; MTOR: mechanistic target of rapamycin kinase; PE: phosphatidylethanolamine; SQSTM1/p62: sequestosome 1; TUBA1A: tubulin alpha 1a.\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 ... tags at the very beginning of your response.\n\n=======================================================\nCONTEXT LITERATURE (STATIC CACHE):\nID: 42465301\nTitle: The role of electrostatic interactions in the phase separation of HP1\u03b1 and its protein binding partners.\nAbstract: Heterochromatin protein 1\u03b1 (HP1\u03b1) is an intrinsic component of heterochromatin domains where it is involved in a diverse set of functions including heterochromatin spreading and organization, chromatin compaction and transcriptional silencing. It has been suggested that HP1\u03b1 functions through a phase separation mechanism, a process that has been observed in vitro in the presence of N-terminal phosphorylation, nucleic acids and nucleosome arrays. HP1\u03b1 can also interact with numerous binding partners that contain a specific motif called an HP1 access code (HAC). HACs recognize and bind to an interface formed by the chromoshadow (CSD) domains in the HP1\u03b1 homodimer, the functional form of the protein. It has been shown that some HP1\u03b1 binding partners can enhance its phase separation ability while others disrupt the process. Here, we focus on the interactions between HP1\u03b1 and three binding partners, namely the p150 subunit of the chromatin assembly factor 1 (CAF-1), the N-terminal domain of the lamin B receptor (LBR), and the mitotic protein Shugoshin 1 (Sgo1). Using phase separation assays, we show that CAF-1 prevents HP1\u03b1 phase separation while LBR and Sgo1 enhance it. Binding assays, mutational studies, NMR spectroscopy and computational analysis allow us to dissect the contributions of the HAC motifs, the charge patterns of the binding partner sequences and the role of N-terminal phosphorylation on HP1\u03b1 in condensate formation. Our results demonstrate that each binding partner uniquely balances these contributions to modulate the properties of HP1\u03b1, while electrostatic interactions dominate the regulation of phosphorylated HP1\u03b1. These results suggest that HP1\u03b1's binding partners play an important role in the modulation of its properties and the regulation of its functions in distinct biological contexts.\n\nID: 42361996\nTitle: Lespedeza bicolor root extract ameliorates LPS-induced acute lung injury through p38 MAPK/NF-\u03baB pathway.\nAbstract: Acute lung injury (ALI) is a severe inflammatory disorder lacking effective therapeutic options. Lespedeza bicolor, a traditional medicinal plant, and its root extract (LBR) contain bioactive constituents with potential anti-inflammatory properties. This study aimed to evaluate the protective effects and underlying mechanisms of LBR against ALI. Network pharmacology combined with transcriptomic and enrichment analyses was used to identify potential targets and signaling pathways of LBR-derived active compounds. The anti-ALI efficacy of LBR was assessed in LPS-induced RAW264.7\u202fcells and mouse models through histopathological examination, measurement of inflammatory cytokines, and immunohistochemistry. RNA sequencing, western blotting, and immunofluorescence assays were further performed to validate key signaling pathways. A total of 157 overlapping targets between LBR and ALI-associated genes were identified, mainly enriched in inflammatory and immune-related pathways, particularly the MAPK and NF-\u03baB cascades. LBR significantly inhibited LPS-induced pro-inflammatory cytokine expression, phosphorylation of p38 MAPK and p65, and nuclear translocation of NF-\u03baB p65 in vitro. In vivo, LBR improved lung histopathology, reduced inflammatory infiltration, and alleviated pulmonary edema. Mechanistically, LBR suppressed activation of the p38 MAPK/NF-\u03baB axis with comparatively weaker effects on ERK and JNK pathways. In conclusion, LBR protects against LPS-induced ALI by modulating inflammatory responses via inhibition of the p38 MAPK/NF-\u03baB signaling pathway, supporting its potential as a promising multi-target natural therapeutic for ALI.\n\nID: 42252781\nTitle: Weakening the nuclear envelope: Lamin B receptor in melanoma metastasis.\nAbstract: Nuclear envelope fragility has been identified as a hallmark of the metastatic process. Indeed, cancer cells undertake a challenging journey through diverse environments during metastasis, notably squeezing through narrow spaces and dense collagen matrices. In their report, Baird et\u00a0al. identify a key player in nuclear fragility in confined cancer cell migration: the Lamin B receptor. Its overexpression and clustering in specific regions of the nuclear envelope increase nuclear distortion and fragility, rendering nuclei more susceptible to rupture with consequent chromatin extrusion and genomic instability. Elucidating the mechanisms underlying this process, particularly the receptor's sterol reductase activity, represents a promising step towards the identification of novel therapeutic targets.\n\nID: 42153377\nTitle: Antagonistic contributions of A-type and B-type lamins to LBR localization and dynamics.\nAbstract: Lamin B receptor (LBR) is an inner nuclear membrane protein that organizes peripheral heterochromatin and cooperates with lamins to tether chromatin at the nuclear periphery. However, how individual lamin isoforms regulate LBR localization and anchorage remains unclear. Using mouse embryonic fibroblasts lacking all endogenous lamins, we examined how individual lamin isoforms regulate LBR behavior. Expression of lamin B1 or B2 was sufficient to anchor LBR at the nuclear envelope, whereas lamin A expression increased LBR mobility and led to its displacement from the nuclear envelope. This effect was mediated by phosphorylation of LBR and was recapitulated by lamin A overexpression in wild-type cells. Collectively, these findings define isoform-specific and antagonistic roles for A-type and B-type lamins in regulating LBR anchorage at the nuclear envelope. In addition, they indicate a lamin A-dependent mechanism that may reflect a broader developmental process, since LBR and lamin A sequentially tether peripheral heterochromatin during development.\n\nID: 42124592\nTitle: Envelope-Limited Chromatin Sheets (ELCS) Formation in The Nuclear Envelope of HL-60/S4 Cells.\nAbstract: Envelope-Limited Chromatin Sheets (ELCS) can be induced in human promyelocytic HL-60/S4 cells by treatment with retinoic acid (RA). After 4 days, the differentiated granulocytes exhibit multilobed nuclei with outgrowths of the nuclear envelope (NE) and associated heterochromatin extending into the surrounding cytoplasm (ELCS). These fascinating structures reveal a periodic meshwork of 30 nm chromatin fibers, when viewed by Cryo-electron microscopy. Genetic and biochemical evidence indicates that RA increases the synthesis of Lamin B Receptor (LBR), which is a key enzyme for Cholesterol biosynthesis and is an essential bridge between the NE and peripheral heterochromatin. This article is in part a review of our microscopic data on the structure of ELCS, and in part a description of related transcription changes that result in the formation of ELCS. In addition, this article contains a structural and biochemical comparison of RA-induced granulocytes with phorbol ester (TPA) induced HL-60/S4 macrophages, which lack nuclear lobulation, do not form ELCS, and exhibit a reduction in LBR and Cholesterol biosynthesis. From our perspective, ELCS can be viewed as \"fabric\" outgrowths of the nuclear envelope, frequently connecting nuclear lobes and capable of sustaining the twisting and squeezing distortions imposed upon nuclear shape, as the granulocytes traverse narrow tissue channels.\n\nID: 42017968\nTitle: Tau oligomerization induces nuclear lamina invagination and chromatin remodeling in Alzheimer's disease.\nAbstract: The aggregation of the microtubule-associated protein tau into oligomeric complexes is strongly correlated with the onset and progression of neurodegeneration in Alzheimer's disease (AD). Increasing evidence implicates nuclear membrane disruption in AD and related tauopathies; however, whether this is a cause or consequence of neurodegeneration remains unresolved. Here, we show that nuclear lamina disruption emerges at the early Braak stages, coinciding with the initial formation of pathological tau aggregates in post-mortem AD brain tissue. Using the tauopathy mouse model (P301S PS19), we demonstrate that oligomeric tau (oTau) directly binds to the Lamin B Receptor (LBR), inducing nuclear envelope invaginations as revealed by electron microscopy. These structural alterations are accompanied by chromatin remodeling and gene expression dysregulation. To dissect the underlying mechanism, we employed a light-inducible OptoTau system (4R1N Tau::mCherry::Cry2Olig) in human iPSC-derived neurons, enabling real-time visualization of tau aggregation dynamics. This system revealed selective recruitment of oTau to the nuclear envelope and direct interactions with LBR and Lamin B2, leading to nuclear deformation and activation of the protein translational stress response. Together, these findings identify nuclear membrane disruption as an early and potentially causative event in tau-mediated neurodegeneration, establishing a mechanistic link between tau oligomerization, nuclear stress, and chromatin remodeling. Targeting nuclear destabilization may offer new therapeutic avenues for mitigating AD pathogenesis.\n\nID: 41757046\nTitle: Antagonistic contributions of A-type and B-type lamins to LBR localization and dynamics.\nAbstract: Lamin B receptor (LBR) is an inner nuclear membrane (INM) protein that plays crucial roles in maintaining nuclear architecture and organization of peripheral heterochromatin. Lamins and LBR both contribute to chromatin tethering at the nuclear periphery, and the expression of LBR and A-type lamins is tightly regulated during development to ensure a faithful transition between different chromatin tethering modalities. Despite its well-established association with B-type lamins, the contributions of individual lamin isoforms to LBR localization and anchorage have not been systematically examined. Here, we used mouse embryonic fibroblasts (MEFs) lacking all endogenous lamins (triple lamin knockout: TKO) to assess how specific lamin isoforms and domains regulate LBR subcellular localization and mobility. Whereas ectopic expression of either lamin B1 or lamin B2 was sufficient to tether LBR to the nuclear envelope in TKO cells, expression of lamin A increased the lateral mobility of LBR at the nuclear membrane, resulting in its displacement from the nuclear envelope to the ER. The lamin A-induced displacement of LBR was mediated by phosphorylation of LBR. Overexpression of lamin A in wild-type MEFs similarly increased LBR phosphorylation and promoted its displacement from the nuclear envelope. Collectively, these findings define isoform-specific and antagonistic roles for A-type and B-type lamins in regulating LBR anchorage at the nuclear envelope. In addition, they indicate a lamin A-dependent mechanism that may reflect a broader developmental process, since LBR and lamin A sequentially tether peripheral heterochromatin during development.\n\nID: 41735607\nTitle: LBR and LAP2 mediate heterochromatin tethering to the nuclear periphery to preserve genome homeostasis.\nAbstract: In most eukaryotic cells, euchromatin is localized in the nuclear interior, whereas heterochromatin is enriched at the nuclear envelope (NE). This conventional chromatin organization is established by heterochromatin tethering to the NE; however, its importance for cellular homeostasis is largely unexplored. One tether is constituted by the lamin B receptor (LBR) in mammals, but the enigmatic nature of other redundant tethers has hampered functional analyses. Here we demonstrate that downregulation of abundant, ubiquitous NE proteins can induce the global detachment of heterochromatin from the NE and its repositioning to the nuclear interior. We identify LBR and lamina-associated polypeptide 2 (LAP2) as key factors for peripheral heterochromatin positioning in differentiated and pluripotent mammalian cells. Their long-term loss leads to changes in three-dimensional chromatin organization and a reduction in repressive epigenetic marks, especially H3K27me3. These changes are associated with massive deregulation of gene expression, activation of antiviral innate immunity, and defects in cell fate determination.\n\nID: 41706894\nTitle: Lamin B receptor upregulation in metastatic melanoma causes nuclear envelope fragility in confined migration during cancer invasion.\nAbstract: Cancer cells migrate through regions of tissue confinement during metastasis, causing nuclear envelope (NE) rupture, which generates heritable DNA damage and disease aggression. We sought to determine if NE rupture was intrinsic to malignant transformation and seek a mechanistic cause. We found that metastatic cells from multiple cancer subtypes have increased NE fragility in confinement compared to their benign counterparts. Meta-analysis of transcriptomic data from clinical samples of melanoma progression together with an siRNA-based live-cell confinement screen revealed that lamin B receptor (LBR) transcriptional upregulation correlates with melanoma progression and NE fragility. LBR is an inner nuclear membrane protein (INM) that scaffolds lamins and chromatin and has sterol reductase activity in the cholesterol biosynthesis pathway. Applying superresolution and atomic force microscopies to characterize the cellular and biophysical events leading to NE rupture revealed that upregulated LBR increased nuclear deformability and can generate local ruptures of the INM, promoting blebs in the nuclear membrane that burst to release nuclear contents into the cytoplasm. Structure-function analysis showed that LBR's sterol reductase activity is required for its promotion of NE fragility in cells confined in vitro. Use of tumor organoids and an in vivo melanoma model revealed that upregulation of LBR was associated with increased NE fragility, metastatic invasion, and decreased patient survival. Thus, upregulation of LBR in melanoma promotes nuclear deformability, while LBR's sterol reductase activity causes fragility and instability of the nuclear membrane, and these changes in the nucleus provide a possible mechanism for increased genetic heterogeneity in melanomas.\n\nID: 41561380\nTitle: Calcium waves and nuclear tension changes coordinate mechanical stress dissipation in locally folded epithelia.\nAbstract: Epithelia are continuously exposed to biomechanical forces such as compression, stretch, and shear stress arising from their dynamic microenvironments. Changes in tension, including stretch, trigger cell rearrangements, divisions, and transcriptional responses until mechanical stress is dissipated. Here, we focus on epithelial folding, a fundamental process by which flat monolayers transform into 3D functional tissues. Using an innovative method for fold generation combined with live imaging, mechanobiology tools, and chemical screening, we uncover the role of calcium waves in the mechanical adaptation of folded epithelia at both tissue and nuclear levels. Folding-associated tension induces nuclear flattening that is recovered within minutes through calcium waves spreading outward across the epithelium. By creating an LBR-overexpressing mutant that relaxes the nuclear envelope, we show that, despite calcium waves, nuclear tension is required for shape recovery via cPLA2-dependent contractility. Our results identify the mechanism of nuclear shape recovery and reveal nuclei as internal tension sensors.\n\nID: 41059452\nTitle: A Family of LBR Biallelic Pathogenic Variants Resulting in Rhizomelic Skeletal Dysplasia with Pelger-Hu\u00ebt Anomaly.\nAbstract: The lamin-B receptor (LBR) gene has two primary functions: maintaining the structural integrity of the nuclear envelope and playing a role in cholesterol biosynthesis. Heterozygous variants in the LBR gene have been associated with Pelger-Hu\u00ebt anomaly (PHA, OMIM #169400), while homozygous or compound heterozygous mutations have been associated with rhizomelic skeletal dysplasia, with or without PHA (OMIM #618019) and Greenberg dysplasia (OMIM #215140). We report a 4-year-old boy presenting with short stature and short limbs and his mother exhibiting milder findings. Genetic analysis revealed a heterozygous c.1640A>G (p.Asn547Ser) and c.43C>T (p.Arg15*) variants in the LBR gene in both the boy and his mother. The father was identified as a heterozygous carrier of the c.43C>T (p.Arg15*) variant. Peripheral blood smears confirmed the PHA in the patient and his parents. The phenotypic differences observed between the mother and male child in our study highlight the genetic variability and regressive nature of LBR-related skeletal dysplasias. This report shows the complexity of LBR-related phenotypes and expands the clinical spectrum of LBR mutations in rhizomelic skeletal dysplasia with PHA.\n\nID: 41002030\nTitle: Coordinated control of genome-nuclear lamina interactions by topoisomerase 2B and lamin B receptor.\nAbstract: Lamina-associated domains (LADs) are megabase-sized genomic regions anchored to the nuclear lamina (NL). Factors controlling the interactions of the genome with the NL have largely remained elusive. Here, we identified DNA topoisomerase 2 beta (TOP2B) as a regulator of these interactions. TOP2B binds predominantly to inter-LAD (iLAD) chromatin and its depletion results in a partial loss of genomic partitioning between LADs and iLADs, suggesting that this enzyme might protect specific iLADs from interacting with the NL. TOP2B depletion affects LAD interactions with lamin B receptor (LBR) more than with lamins. LBR depletion phenocopies the effects of TOP2B depletion, despite the different positioning of the two proteins in the genome. This suggests a complementary mechanism for organizing the genome at the NL. Indeed, co-depletion of TOP2B and LBR causes partial LAD/iLAD inversion, reflecting changes typical of oncogene-induced senescence. We propose that a coordinated axis controlled by TOP2B in iLADs and LBR in LADs maintains the partitioning of the genome between the NL and the nuclear interior.\n\nID: 40739853\nTitle: cTAGE5/MEA6 Regulates LBR Localization to Maintain Nuclear Envelope Integrity and Safeguard Against Aging.\nAbstract: cTAGE5/MEA6 plays a pivotal role in COPII complex assembly, ER-to-Golgi trafficking, and secretion. However, whether cTAGE5/MEA6 is involved in other cellular functions remains unclear. Here, we show that conditional cTAGE5 knockout results in embryonic lethality during development and premature aging in adult mice. cTAGE5 deficiency leads to abnormal nuclear structure and disturbed cell proliferation in MEF cells. Further mechanistic studies reveal that cTAGE5 localizes not only to the ER exit sites but also to other ER structures, where it interacts with the lamin B receptor (LBR). Loss of cTAGE5 disrupts LBR's localization to the inner nuclear membrane, leading to its retention in the ER and instability. This results in abnormal nuclear (envelope) morphology and cellular senescence, likely driven by activation of the P53/P21 senescence pathway. Thus, our study uncovers cTAGE5's role in maintaining nuclear envelope integrity and highlights its function and potential mechanism in preventing cellular senescence and animal aging.\n\nID: 40369063\nTitle: STAT5 and STAT3 balance shapes dendritic cell function and tumour immunity.\nAbstract: Immune checkpoint blockade (ICB) has transformed cancer therapy1,2. The efficacy of immunotherapy depends on dendritic cell-mediated tumour antigen presentation, T cell priming and activation3,4. However, the relationship between the key transcription factors in dendritic cells and ICB efficacy remains unknown. Here we found that ICB reprograms the interplay between the STAT3 and STAT5 transcriptional pathways in dendritic cells, thereby activating T cell immunity and enabling ICB efficacy. Mechanistically, STAT3 restrained the JAK2 and STAT5 transcriptional pathway, determining the fate of dendritic cell function. As STAT3 is often activated in the tumour microenvironment5, we developed two distinct PROTAC (proteolysis-targeting chimera) degraders of STAT3, SD-36 and SD-2301. STAT3 degraders effectively degraded STAT3 in dendritic cells and reprogrammed the dendritic cell-transcriptional network towards immunogenicity. Furthermore, STAT3 degrader monotherapy was efficacious in treatment of advanced tumours and ICB-resistant tumours without toxicity in mice. Thus, the crosstalk between STAT3 and STAT5 transcriptional pathways determines the dendritic cell phenotype in the tumour microenvironment and STAT3 degraders hold\u00a0promise for cancer immunotherapy.\n\nID: 40355051\nTitle: Novel LBR pathogenic variants with loss of sterol reductase activity participate in the pathogenesis of skeletal dysplasia via dysregulating canonical Wnt pathway.\nAbstract: Biallelic pathogenic variants in the lamin B receptor (LBR) with impaired sterol reductase function are associated with the development of perinatal lethal Greenberg dysplasia (GRBGD) and mild nonfatal skeletal dysplasia with or without Pelger-Huet anomaly (PHASK), as well as other related hereditary skeletal dysplasia. However, the underlying molecular mechanism remains unclear. In this study, we found two novel pathogenic variants of LBR, namely missense mutation (c.1011\u00a0T\u00a0>\u00a0G, NM_002296.4; p.Cys337Trp, NP_002287.2) and LBR gene deletion (Chr1q42.12 (225,515,082-225,633,464), NC_000001.10). LBR is a novel substrate of FBW7, which is degraded by GSK3\u03b2/FBW7-mediated proteasome pathway and whose C337W mutation promotes its degradation through enhanced interaction with FBW7. Wild-type but not C337W mutant LBR is upregulated by WNT3A-mediated inactivation of GSK3\u03b2/FBW7 axis and then participated in WNT3A-activated Wnt pathway through its mediated cholesterol synthesis. MC3T3-E1 cells with Lbr knockdown or cholesterol removal exhibited reduced mineralized nodules in the presence of WNT3A, but addition of cholesterol in the culture medium reversed this phenotype. Collectively, we detected two novel variants in LBR and our study revealed for the first time that disruption of cholesterol synthesis by LBR impairs Wnt pathway and thus disrupts the cell osteogenic differentiation, providing new insights into the pathogenesis of skeletal dysplasia caused by LBR variation.\n\nID: 40279158\nTitle: Major nuclear locales define nuclear genome organization and function beyond A and B compartments.\nAbstract: Models of nuclear genome organization often propose a binary division into active versus inactive compartments yet typically overlook nuclear bodies. Here, we integrated analysis of sequencing and image-based data to compare genome organization in four human cell types relative to three different nuclear locales: the nuclear lamina, nuclear speckles, and nucleoli. Although gene expression correlates mostly with nuclear speckle proximity, DNA replication timing correlates with proximity to multiple nuclear locales. Speckle attachment regions emerge as DNA replication initiation zones whose replication timing and gene composition vary with their attachment frequency. Most facultative LADs retain a partially repressed state as iLADs, despite their positioning in the nuclear interior. Knock out of two lamina proteins, Lamin A and LBR, causes a shift of H3K9me3-enriched LADs from lamina to nucleolus, and a reciprocal relocation of H3K27me3-enriched partially repressed iLADs from nucleolus to lamina. Thus, these partially repressed iLADs appear to compete with LADs for nuclear lamina attachment with consequences for replication timing. The nuclear organization in adherent cells is polarized with nuclear bodies and genomic regions segregating both radially and relative to the equatorial plane. Together, our results underscore the importance of considering genome organization relative to nuclear locales for a more complete understanding of the spatial and functional organization of the human genome.\n\nID: 39417178\nTitle: Lespedeza bicolor root extract exerts anti-TNBC potential by regulating FAK-related signalling pathways.\nAbstract: Lespedeza bicolor is a shrub plant that has been widely distributed in East Asia. The methanol extract from its LBR has been shown to exhibit anticancer and anti-bacterial effects. However, its anticancer efficacy in TNBC remains uncertain. This work aimed to study the anti-TNBC effect of LBR ethanol extract and its underlying mechanism. LBR triggered the cell death in TNBC through inhibiting cell proliferation, S-phase cell arrest, and induction of apoptosis. RNA-seq analysis revealed that the genes altered by LBR treatment were predominantly enriched in the cell adhesion. Notably, LBR inhibited phosphorylation and distribution of FAK. Furthermore, LBR demonstrated significant anticancer activity in xenograft tumors in mice through inhibiting cancer cell growth and inducing apoptosis. This work demonstrated the anticancer efficiency of LBR in TNBC without causing significant adverse effect, which providing a foundation for developing LBR based chemotherapeutic agents for breast cancer therapy.\n\nID: 39252008\nTitle: Calpain-1 weakens the nuclear envelope and promotes the release of neutrophil extracellular traps.\nAbstract: The inducers of neutrophil extracellular trap (NET) formation are heterogeneous and consequently, there is no specific pathway or signature molecule indispensable for NET formation. But certain events such as histone modification, chromatin decondensation, nuclear envelope breakdown, and NET release are ubiquitous. During NET formation, neutrophils drastically rearrange their cytoplasmic, granular and nuclear content. Yet, the exact mechanism for decoding each step during NET formation still remains elusive. Here, we investigated the mechanism of nuclear envelope breakdown during NET formation. Immunofluorescence microscopic evaluation revealed a gradual disintegration of outer nuclear membrane protein nesprin-1 and alterations in nuclear morphology during NET formation. MALDI-TOF analysis of NETs that had been generated by various inducers detected the accumulation of nesprin-1 fragments. This suggests that nesprin-1 degradation occurs before NET release. In the presence of a calpain-1, inhibitor nesprin-1 degradation was decreased in calcium driven NET formation. Microscopic evaluation confirmed that the disintegration of the lamin B receptor (LBR) and the collapse of the actin cytoskeleton occurs in early and later phases of NET release, respectively. We conclude that the calpain-1 degrades nesprin-1, orchestrates the weakening of the nuclear membrane, contributes to LBR disintegration, and promoting DNA release and finally, NETs formation.\n\nID: 38839376\nTitle: Reconstitution of nuclear envelope subdomain formation on mitotic chromosomes in semi-intact cells.\nAbstract: In metazoans, the nuclear envelope (NE) disassembles during the prophase and reassembles around segregated chromatids during the telophase. The process of NE formation has been extensively studied using live-cell imaging. At the early step of NE reassembly in human cells, specific pattern-like localization of inner nuclear membrane (INM) proteins, connected to the nuclear pore complex (NPC), was observed in the so-called \"core\" region and \"noncore\" region on telophase chromosomes, which corresponded to the \"pore-free\" region and the \"pore-rich\" region, respectively, in the early G1 interphase nucleus. We refer to these phenomena as NE subdomain formation. To biochemically investigate this process, we aimed to develop an in vitro NE reconstitution system using digitonin-permeabilized semi-intact mitotic human cells coexpressing two INM proteins, emerin and lamin B receptor, which were labeled with fluorescent proteins. The targeting and accumulation of INM proteins to chromosomes before and after anaphase onset in semi-intact cells were observed using time-lapse imaging. Our in vitro NE reconstitution system recapitulated the formation of the NE subdomain, as in living cells, although chromosome segregation and cytokinesis were not observed. This in vitro NE reconstitution required the addition of a mitotic cytosolic fraction supplemented with a cyclin-dependent kinase inhibitor and energy sources. The cytoplasmic soluble factor(s) dependency of INM protein targeting differed among the segregation states of chromosomes. Furthermore, the NE reconstituted on segregated chromosomes exhibited active nucleocytoplasmic transport competency. These results indicate that the chromosome status changes after anaphase onset for recruiting NPC components.\n\nID: 38712240\nTitle: Neutrophils secrete exosome-associated DNA to resolve sterile acute inflammation.\nAbstract: Acute inflammation, characterized by a rapid influx of neutrophils, is a protective response that can lead to chronic inflammatory diseases when left unresolved. Secretion of LTB 4 -containing exosomes is required for effective neutrophil infiltration during inflammation. In this study, we show that neutrophils release nuclear DNA in a non-lytic, rapid, and repetitive manner, via a mechanism distinct from suicidal NET release and cell death. The packaging of nuclear DNA occurs in the lumen of nuclear envelope (NE)-derived multivesicular bodies (MVBs) that harbor the LTB 4 synthesizing machinery and is mediated by the lamin B receptor (LBR) and chromatin decondensation. Disruption of secreted exosome-associated DNA (SEAD) in a model of sterile inflammation in mouse skin amplifies and prolongs the presence of neutrophils, impeding the onset of resolution. Together, these findings advance our understanding of neutrophil functions during inflammation and the physiological significance of NETs, with implications for novel treatments for inflammatory disorders.\n\nID: 38185834\nTitle: Fly clock, my clock, and lamin B receptor.\nAbstract: In the fruit fly Drosophila melanogaster, circadian rhythm was disrupted when the inner nuclear membrane protein lamin B receptor (LBR) was depleted from its clock neurons (Proc. Natl. Acad. Sci. USA 118, e2019756118. 2021; https://doi.org/10. 1073/pnas.2019756118 and Research 6, 0139, 2023; https://doi.org/10.34133/research.0139). Ordinarily, the clock proteinPERIOD (PER) forms foci close to the inner nuclear membrane in the circadian clock's repression phase. The size, number, and location of foci near the nuclear membrane oscillate with a 24-h rhythm. When LBR was absent the foci did not form. The PER foci bring per and other clock genes close to the nuclear envelope, where their transcription is silenced. Then, in the circadian clock's activation phase, the PER protein gradually gets degraded and the foci disappear. The clock genes, including per, relocate to the nucleus interior where they resume transcription. Rhythmic re-positioning of clock genes between nucleus periphery and interior, correlates with their repression and activation in the circadian cycle. Absence of LBR disrupted this rhythm. Phosphorylation of PER promoted the formation of foci whereas dephosphorylation by protein phosphatase 2A causedthem to disappear. LBR promoted focus formation by destabilizing the catalytic subunit of protein phosphatase 2A. The lbr gene is no stranger to this journal. The first hint that vertebrate LBR is also a sterol biosynthesis enzyme, specifically, a sterol C14 reductase, was reported here (J. Genet. 73, 33-41, 1994; https://www.ias.ac.in/article/fulltext/jgen/073/01/0033-0041). Mutations in the human Lbr gene cause a range of phenotypes--from the relatively benign Pelger-Huet anomaly to the perinatally lethal Greenberg skeletal dysplasia.Drosophila, like all insects, is a sterol auxotroph. The fly orthologue of vertebrate lbr genes encodes a protein (dLBR) that shares several properties with vertebrate LBR proteins, with one notable exception. While human LBR complemented theyeast Saccharomyces cerevisiae erg24 mutant which lacks sterol C14 reductase activity, dLBR did not (J. Cell. Sci. 117, 2015-28, 2004; https://doi.org/10.1242/jcs.01052). Despite not possessing sterol reductase activity, dLBR retains significant sequence homology with vertebrate LBRs which have this activity. An undergraduate summer trainee in my laboratory obtained early (unpublished) evidence that dLBR lost sterol reductase activity during evolution. She transferred adult drosophila flies to vials containing a medium made of agar, dextrose, and dried and powdered mycelium of the filamentous fungus Neurospora crassa. On medium made with wild-type mycelium, theflies mated, laid eggs, hatched larvae, and developed pupae which eclosed progeny adult flies. The life cycle was no different than on 'regular' fly food composed of agar, dextrose and yeast extract. However, on a medium made with mycelium from a sterol C14 reductase null mutant, the flies laid eggs which hatched and released larvae, but the larvae failed to pupate, and no adult progeny flies emerged. This was because the fly lacks a sterol C14 reductase. The wild-type sterol, ergosterol, is a precursor of the steroid hormone ecdysone needed for molting and metamorphosis. Can expression of vertebrate LBR in dLBR-depleted fly clock neurons restore circadian rhythm? Can expression of vertebrate LBR enable flies to complete their life cycle on mutant Neurospora medium? Does LBR regulate the vertebrate clock in a like manner? If yes, then is the sterol reductase activity dispensable in this role? These are some questions that came to my mind on a recent morning walk. The walk itself was a much-cherished outcome of my circadian clock.\n\nID: 37880237\nTitle: Lamin A upregulation reorganizes the genome during rod photoreceptor degeneration.\nAbstract: Neurodegenerative diseases are accompanied by dynamic changes in gene expression, including the upregulation of hallmark stress-responsive genes. While the transcriptional pathways that impart adaptive and maladaptive gene expression signatures have been the focus of intense study, the role of higher order nuclear organization in this process is less clear. Here, we examine the role of the nuclear lamina in genome organization during the degeneration of rod photoreceptors. Two proteins had previously been shown to be necessary and sufficient to tether heterochromatin at the nuclear envelope. The lamin B receptor (Lbr) is expressed during development, but downregulates upon rod differentiation. A second tether is the intermediate filament lamin A (LA), which is not normally expressed in murine rods. Here, we show that in the rd1 model of retinitis pigmentosa, LA ectopically upregulates in rod photoreceptors at the onset of degeneration. LA upregulation correlated with increased heterochromatin tethering at the nuclear periphery in rd1 rods, suggesting that LA reorganizes the nucleus. To determine how heterochromatin tethering affects the genome, we used in vivo electroporation to misexpress LA or Lbr in mature rods in the absence of degeneration, resulting in the restoration of conventional nuclear architecture. Using scRNA-seq, we show that reorganizing the nucleus via LA/Lbr misexpression has relatively minor effects on rod gene expression. Next, using ATAC-seq, we show that LA and Lbr both lead to marked increases in genome accessibility. Novel ATAC-seq peaks tended to be associated with stress-responsive genes. Together, our data reveal that heterochromatin tethers have a global effect on genome accessibility, and suggest that heterochromatin tethering primes the photoreceptor genome to respond to stress.\n\nID: 37223461\nTitle: Phosphorylation Promotes the Accumulation of PERIOD Protein Foci.\nAbstract: Circadian clock drives the 24-h rhythm in our behavior and physiology. The molecular clock consists of a series of transcriptional/translational feedback loops operated by a number of clock genes. A very recent study reported that the clock protein PERIOD (PER) is organized into discrete foci at the nuclear envelope in fly circadian neurons, which is believed to be important for controlling the subcellular localization of clock genes. Loss of inner nuclear membrane protein lamin B receptor (LBR) leads to disruption of these foci, but how they are regulated is yet unknown. Here, we found that PER foci are likely phase-separated condensates, the formation of which is mediated by intrinsically disordered region in PER. Phosphorylation promotes the accumulation of these foci. Protein phosphatase 2A, which is known to dephosphorylate PER, hampers the accumulation of the foci. On the other hand, the circadian kinase DOUBLETIME (DBT) which phosphorylates PER enhances the accumulation of the foci. LBR likely facilitates PER foci accumulation by destabilizing the catalytic subunit of protein phosphatase 2A, MICROTUBULE STAR (MTS). In conclusion, here, we demonstrate a key role for phosphorylation in promoting the accumulation of PER foci, while LBR modulates this process by impinging on the circadian phosphatase MTS.\n\nID: 36611919\nTitle: SR Protein Kinase 1 Inhibition by TAF15.\nAbstract: Although SRPKs were discovered nearly 30 years ago, our understanding of their mode of regulation is still limited. Regarded as constitutively active enzymes known to participate in diverse biological processes, their prominent mode of regulation mainly depends on their intracellular localization. Molecular chaperones associate with a large internal spacer sequence that separates the bipartite kinase catalytic core and modulates the kinases' partitioning between the cytoplasm and nucleus. Besides molecular chaperones that function as anchoring proteins, a few other proteins were shown to interact directly with SRPK1, the most-studied member of SRPKs, and alter its activity. In this study, we identified TAF15, which has been involved in transcription initiation, splicing, DNA repair, and RNA maturation, as a novel SRPK1-interacting protein. The C-terminal RGG domain of TAF15 was able to associate with SRPK1 and downregulate its activity. Furthermore, overexpression of this domain partially relocalized SRPK1 to the nucleus and resulted in hypophosphorylation of SR proteins, inhibition of splicing of a reporter minigene, and inhibition of Lamin B receptor phosphorylation. We further demonstrated that peptides comprising the RGG repeats of nucleolin, HNRPU, and HNRNPA2B1, were also able to inhibit SRPK1 activity, suggesting that negative regulation of SRPK1 activity might be a key biochemical property of RGG motif-containing proteins.\n\nID: 36598800\nTitle: Differentiation-dependent changes in lamin B1 dynamics and lamin B receptor localization.\nAbstract: The nuclear lamina serves important roles in chromatin organization and structural support, and lamina mutations can result in laminopathies. Less is known about how nuclear lamina structure changes during cellular differentiation-changes that may influence gene regulation. We examined the structure and dynamics of the nuclear lamina in human-induced pluripotent stem cells (iPSCs) and differentiated germ layer cells, focusing on lamin B1. We report that lamin B1 dynamics generally increase as iPSCs differentiate, especially in mesoderm and ectoderm, and that lamin B receptor (LBR) partially redistributes from the nucleus to cytoplasm in mesoderm. Knocking down LBR in iPSCs led to an increase in lamin B1 dynamics, a change that was not observed for ELYS, emerin, or lamin B2 knockdown. LBR knockdown also affected expression of differentiation markers. These data suggest that differentiation-dependent tethering of lamin B1 either directly by LBR or indirectly via LBR-chromatin associations impacts gene expression.\n\nID: 36454702\nTitle: The injury-induced transcription factor SOX9 alters the expression of LBR, HMGA2, and HIPK3 in the human kidney.\nAbstract: Induction of SRY box transcription factor 9 (SOX9) has been shown to occur in response to kidney injury in rodents, where SOX9-positive cells proliferate and regenerate the proximal tubules of injured kidneys. Additionally, SOX9-positive cells demonstrate a capacity to differentiate toward other nephron segments. Here, we characterized the role of SOX9 in normal and injured human kidneys. SOX9 expression was found to colocalize with a proportion of so-called scattered tubular cells in the uninjured kidney, a cell population previously shown to be involved in kidney injury and regeneration. Following injury and in areas adjacent to inflammatory cell infiltrates, SOX9-positive cells were increased in number. With the use of primary tubular epithelial cells (PTECs) obtained from human kidney tissue, SOX9 expression was spontaneously induced in culture and further increased by transforming growth factor-\u03b21, whereas it was suppressed by interferon-\u03b3. siRNA-mediated knockdown of SOX9 in PTECs followed by analysis of differential gene expression, immunohistochemical expression, and luciferase promoter assays suggested lamin B receptor (LBR), high mobility group AT-hook 2 (HMGA2), and homeodomain interacting protein kinase 3 (HIPK3) as possible target genes of SOX9. Moreover, a kidney explant model was used to demonstrate that only SOX9-positive cells survive the massive injury associated with kidney ischemia and that the surviving SOX9-positive cells spread and repopulate the tubules. Using a wound healing assay, we also showed that SOX9 positively regulated the migratory capacity of PTECs. These findings shed light on the functional and regulatory aspects of SOX9 activation in the human kidney during injury and regeneration.NEW & NOTEWORTHY Recent studies using murine models have shown that SRY box transcription factor 9 (SOX9) is activated during repair of renal tubular cells. In this study, we showed that SOX9-positive cells represent a proportion of scattered tubular cells found in the uninjured human kidney. Furthermore, we suggest that expression of LBR, HMGA2, and HIPK3 is altered by SOX9 in the kidney tubular epithelium, suggesting the involvement of these gene products in kidney injury and regeneration.\n\nID: 36124691\nTitle: Nuclear envelope protein lamin B receptor protects the genome from chromosomal instability and tumorigenesis.\nAbstract: Lamin B Receptor (LBR) is an inner nuclear membrane protein that assembles the nuclear envelope post mitosis. Here we show that LBR depletion induces mitotic defects accompanied by recurrent chromosomal losses. In addition, LBR knockdown results in nuclear aberrations such as nuclear blebs and micronuclei, with chromosomes showing higher frequency of losses, being enriched within the micronucleus. Furthermore, doxycycline-induced conditional depletion of LBR significantly increased tumor volumes that form within the subcutaneous xenografts of mice. Of note, the tumor-derived primary cells recapitulated chromosomal losses and gains, revealing a novel role for LBR as a tumor suppressor. Co-immunoprecipitation of LBR uncovered an association of LBR with telomere-associated factors. Interestingly, qPCR array-based gene expression profiling showed a significant upregulation of telomere repeat-binding factor 1 (TRF1) upon LBR depletion. Remarkably, TRF1 knockdown in the background of LBR depletion maintains chromosomal stability, unraveling a novel mechanism involving LBR and TRF in the maintenance of chromosomal stability in colorectal cancer cells.\n\nID: 36044892\nTitle: Bi-allelic loss-of-function variants in TMEM147 cause moderate to profound intellectual disability with facial dysmorphism and pseudo-Pelger-Hu\u00ebt anomaly.\nAbstract: The transmembrane protein TMEM147 has a dual function: first at the nuclear envelope, where it anchors lamin B receptor (LBR) to the inner membrane, and second at the endoplasmic reticulum (ER), where it facilitates the translation of nascent polypeptides within the ribosome-bound TMCO1 translocon complex. Through international data sharing, we identified 23 individuals from 15 unrelated families with bi-allelic TMEM147 loss-of-function variants, including splice-site, nonsense, frameshift, and missense variants. These affected children displayed congruent clinical features including coarse facies, developmental delay, intellectual disability, and behavioral problems. In silico structural analyses predicted disruptive consequences of the identified amino acid substitutions on translocon complex assembly and/or function, and in\u00a0vitro analyses documented accelerated protein degradation via the autophagy-lysosomal-mediated pathway. Furthermore, TMEM147-deficient cells showed CKAP4 (CLIMP-63) and RTN4 (NOGO) upregulation with a concomitant reorientation of the ER, which was also witnessed in primary fibroblast cell culture. LBR mislocalization and nuclear segmentation was observed in primary fibroblast cells. Abnormal nuclear segmentation and chromatin compaction were also observed in approximately 20% of neutrophils, indicating the presence of a pseudo-Pelger-Hu\u00ebt anomaly. Finally, co-expression analysis revealed significant correlation with neurodevelopmental genes in the brain, further supporting a role of TMEM147 in neurodevelopment. Our findings provide clinical, genetic, and functional evidence that bi-allelic loss-of-function variants in TMEM147 cause syndromic intellectual disability due to ER-translocon and nuclear organization dysfunction.\n\nID: 35972904\nTitle: Shared and Distinctive Neighborhoods of Emerin and Lamin B Receptor Revealed by Proximity Labeling and Quantitative Proteomics.\nAbstract: Emerin and lamin B receptor (LBR) are abundant transmembrane proteins of the nuclear envelope that are concentrated at the inner nuclear membrane (INM). Although both proteins interact with chromatin and nuclear lamins, they have distinctive biochemical and functional properties. Here, we have deployed proximity labeling using the engineered biotin ligase TurboID (TbID) and quantitative proteomics to compare the neighborhoods of emerin and LBR in cultured mouse embryonic fibroblasts. Our analysis revealed 232 high confidence proximity partners that interact selectively with emerin and/or LBR, 49 of which are shared by both. These included previously characterized NE-concentrated proteins, as well as a host of additional proteins not previously linked to emerin or LBR functions. Many of these are TM proteins of the ER, including two E3 ubiquitin ligases. Supporting these results, we found that 11/12 representative proximity relationships identified by TbID also were detected at the NE with the proximity ligation assay. Overall, this work presents methodology that may be used for large-scale mapping of the landscape of the INM and reveals a group of new proteins with potential functional connections to emerin and LBR.\n\nID: 35650273\nTitle: Genetic architecture of band neutrophil fraction in Iceland.\nAbstract: The characteristic lobulated nuclear morphology of granulocytes is partially determined by composition of nuclear envelope proteins. Abnormal nuclear morphology is primarily observed as an increased number of hypolobulated immature neutrophils, called band cells, during infection or in rare envelopathies like Pelger-Hu\u00ebt anomaly. To search for sequence variants affecting nuclear morphology of granulocytes, we performed a genome-wide association study using band neutrophil fraction from 88,101 Icelanders. We describe 13 sequence variants affecting band neutrophil fraction at nine loci. Five of the variants are at the Lamin B receptor (LBR) locus, encoding an inner nuclear membrane protein. Mutations in LBR are linked to Pelger-Hu\u00ebt anomaly. In addition, we identify cosegregation of a rare stop-gain sequence variant in LBR and Pelger Hu\u00ebt anomaly in an Icelandic eight generation pedigree, initially reported in 1963. Two of the other loci include genes which, like LBR, play a role in the nuclear membrane function and integrity. These GWAS results highlight the role proteins of the inner nuclear membrane have as important for neutrophil nuclear morphology.\n\nID: 35453551\nTitle: Diagnostic and Clinical Value of Specific Autoantibodies against Kelch-like 12 Peptide and Nuclear Envelope Proteins in Patients with Primary Biliary Cholangitis.\nAbstract: Primary biliary cholangitis (PBC) is a chronic autoimmune liver disease characterized by the presence of antimitochondrial and antinuclear antibodies in patients\u2019 serum. Here, we analyzed the reactivity of autoantibodies against a novel autoantigen, kelch-like 12 (KLHL12) protein, in a cohort of 138 PBC and 90 non-PBC patients. Additionally, we compared the reactivity of KLHL12 with antinuclear envelope antibodies: anti-gp210, anti-p62, and anti-LBR. Commercially available kits and an \u2018in-house\u2019 ELISA were used in the studies. Antinuclear envelope antibodies were detected in 65% of PBC patients and the presence of these antibodies was observed more frequently in patients diagnosed with later stages (III/IV) of PBC, according to Ludwig\u2019s classification (p < 0.05) and were found to correlate with a higher concentration of bilirubin. Overall, anti-KLHL12 antibodies were found more frequently in PBC patients than in non-PBC controls (p < 0.001). Anti-KLHL12 antibodies were detected in 36% of the tested PBC cohort, including PBC patients negative for antimitochondrial antibodies. Presence of anti-KLHL12 was also associated with a higher concentration of bilirubin and correlated with fibrosis (p < 0.05). Anti-KLHL12 antibodies were detected in 30% of PBC individuals positive for antinuclear envelope antibodies, while anti-KLHL12 and antinuclear envelope antibodies were found in 17% of all PBC cases. Concluding, our data confirm that antibodies against the KLHL12 protein are highly specific for PBC and when used in combination with other markers, may significantly increase the diagnosis of PBC.\n\nID: 35239049\nTitle: CTCF supports preferentially short lamina-associated domains.\nAbstract: More than one third of the mammalian genome is in a close association with the nuclear lamina, thus these genomic regions were termed lamina-associated domains (LADs). This association is fundamental for many aspects of chromatin biology including transcription, replication, and DNA damage repair. LADs association with the nuclear envelope is thought to be dependent on two major mechanisms: The first mechanism is the interaction between nuclear membrane proteins such as LBR with heterochromatin modifications that are enriched in LADs chromatin. The second mechanism is based on proteins that bind the borders of the LADs and support the association of the LADs with the nuclear envelope. Two factors were suggested to support the second mechanism: CCCTC-binding factor (CTCF) and YY1 based on their enriched binding to LADs borders. However, this mechanism has not been proven yet at a whole genome level. Here, to test if CTCF supports the LADs landscape, we generated melanoma cells with a partial loss of function (pLoF) of CTCF by the CRISPR-Cas9 system and determined the LADs landscape by lamin B ChIP-seq analysis. We found that under regular growth conditions, CTCF pLoF led to modest changes in the LADs landscape that included an increase in the signal of 2% of the LADs and a decrease in the signal of 8% of the LADs. However, CTCF importance for the LADs landscape was much higher upon induction of a chromatin stress. We induced chromatin stress by inhibiting RNA polymerase II, an intervention that is known to alter chromatin compaction and supercoiling. Notably, only in CTCF pLoF cells, the chromatin stress led to the dissociation of 7% of the LADs from the lamina. The CTCF-dependent LADs had almost three times shorter median length than the non-affected LADs, were enriched in CTCF binding at their borders, and were higher in their facultative-status (cell-type specific). Thus, it appears that CTCF is a key factor in facilitating the association of short facultative LADs with the nuclear lamina upon chromatin stress.\n\nID: 34638576\nTitle: Regulation of ER Composition and Extent, and Putative Action in Protein Networks by ER/NE Protein TMEM147.\nAbstract: Nuclear envelope (NE) and endoplasmic reticulum (ER) collaborate to control a multitude of nuclear and cytoplasmic actions. In this context, the transmembrane protein TMEM147 localizes to both NE and ER, and through direct and indirect interactions regulates processes as varied as production and transport of multipass membrane proteins, neuronal signaling, nuclear-shape, lamina and chromatin dynamics and cholesterol synthesis. Aiming to delineate the emerging multifunctionality of TMEM147 more comprehensively, we set as objectives, first, to assess potentially more fundamental effects of TMEM147 on the ER and, second, to identify significantly TMEM147-associated cell-wide protein networks and pathways. Quantifying curved and flat ER markers RTN4 and CLIMP63/CKAP4, respectively, we found that TMEM147 silencing causes area and intensity increases for both RTN4 and CLIMP63, and the ER in general, with a profound shift toward flat areas, concurrent with reduction in DNA condensation. Protein network and pathway analyses based on comprehensive compilation of TMEM147 interactors, targets and co-factors then served to manifest novel and established roles for TMEM147. Thus, algorithmically simplified significant pathways reflect TMEM147 function in ribosome binding, oxidoreductase activity, G protein-coupled receptor activity and transmembrane transport, while analysis of protein factors and networks identifies hub proteins and corresponding pathways as potential targets of TMEM147 action and of future functional studies.\n\nID: 34234015\nTitle: Clock proteins regulate spatiotemporal organization of clock genes to control circadian rhythms.\nAbstract: Circadian clocks regulate \u223c24-h oscillations in gene expression, behavior, and physiology. While the genetic and molecular mechanisms of circadian rhythms are well characterized, what remains poorly understood are the intracellular dynamics of circadian clock components and how they affect circadian rhythms. Here, we elucidate how spatiotemporal organization and dynamics of core clock proteins and genes affect circadian rhythms in Drosophila clock neurons. Using high-resolution imaging and DNA-fluorescence in situ hybridization techniques, we demonstrate that Drosophila clock proteins (PERIOD and CLOCK) are organized into a few discrete foci at the nuclear envelope during the circadian repression phase and play an important role in the subnuclear localization of core clock genes to control circadian rhythms. Specifically, we show that core clock genes, period and timeless, are positioned close to the nuclear periphery by the PERIOD protein specifically during the repression phase, suggesting that subnuclear localization of core clock genes might play a key role in their rhythmic gene expression. Finally, we show that loss of Lamin B receptor, a nuclear envelope protein, leads to disruption of PER foci and per gene peripheral localization and results in circadian rhythm defects. These results demonstrate that clock proteins play a hitherto unexpected role in the subnuclear reorganization of core clock genes to control circadian rhythms, revealing how clocks function at the subcellular level. Our results further suggest that clock protein foci might regulate dynamic clustering and spatial reorganization of clock-regulated genes over the repression phase to control circadian rhythms in behavior and physiology.\n\nID: 33958580\nTitle: Small heat-shock protein HSPB3 promotes myogenesis by regulating the lamin B receptor.\nAbstract: One of the critical events that regulates muscle cell differentiation is the replacement of the lamin B receptor (LBR)-tether with the lamin A/C (LMNA)-tether to remodel transcription and induce differentiation-specific genes. Here, we report that localization and activity of the LBR-tether are crucially dependent on the muscle-specific chaperone HSPB3 and that depletion of HSPB3 prevents muscle cell differentiation. We further show that HSPB3 binds to LBR in the nucleoplasm and maintains it in a dynamic state, thus promoting the transcription of myogenic genes, including the genes to remodel the extracellular matrix. Remarkably, HSPB3 overexpression alone is sufficient to induce the differentiation of two human muscle cell lines, LHCNM2 cells, and rhabdomyosarcoma cells. We also show that mutant R116P-HSPB3 from a myopathy patient with chromatin alterations and muscle fiber disorganization, forms nuclear aggregates that immobilize LBR. We find that R116P-HSPB3 is unable to induce myoblast differentiation and instead activates the unfolded protein response. We propose that HSPB3 is a specialized chaperone engaged in muscle cell differentiation and that dysfunctional HSPB3 causes neuromuscular disease by deregulating LBR.\n\nID: 33753404\nTitle: The Diverse Cellular Functions of Inner Nuclear Membrane Proteins.\nAbstract: The nuclear compartment is delimited by a specialized expanded sheet of the endoplasmic reticulum (ER) known as the nuclear envelope (NE). Compared to the outer nuclear membrane and the contiguous peripheral ER, the inner nuclear membrane (INM) houses a unique set of transmembrane proteins that serve a staggering range of functions. Many of these functions reflect the exceptional position of INM proteins at the membrane-chromatin interface. Recent research revealed that numerous INM proteins perform crucial roles in chromatin organization, regulation of gene expression, genome stability, and mediation of signaling pathways into the nucleus. Other INM proteins establish mechanical links between chromatin and the cytoskeleton, help NE remodeling, or contribute to the surveillance of NE integrity and homeostasis. As INM proteins continue to gain prominence, we review these advancements and give an overview on the functional versatility of the INM proteome.\n\nID: 42554579\nTitle: Being a university teacher in a landscape of practice-A qualitative study of critical incidents.\nAbstract: Acceptance of teachers' need for pedagogical training to maintain quality in health professions education has grown. However, to enhance interest in becoming a professional teacher, a better understanding of experiences related to initial contact with teaching is needed. The framework of landscape of practice (LoP) offers a perspective highlighting identification processes as academics and clinicians participate in and navigate across familiar and new communities. The aim of this study was to explore how boundary crossing into teaching practices is expressed in educators' narrative constructions of critical incidents from teaching situations at a medical university. Teachers' written critical incident analyses (CIA) of teaching situations (N\u00a0=\u00a069), submitted during two iterations of a faculty development course at a medical university, were collected. The assignments were analysed using qualitative content analysis. One overarching theme, Teaching appears as an individual venture, and four subthemes, Internalised self-blame and external attribution of responsibility; Encountering teaching situations marked by limited control and psychological safety; Relational dynamics and communicative challenges; Pedagogical commitment and professional agency, were identified. The findings mirrored strong reactions, abandonment and isolation, but also engagement and responsibility for high-quality education. The findings show that engaging in educational practice, developing a professional identity as a teacher is a complex, often isolated endeavour, sometimes disconnected from primary professional practice. Relating these findings to identification processes described within the framework of LoP, demonstrates a need to support individual processes as well as the connections and collaboration between different communities in a LoP. To address this, three interrelated actions are needed: support from the teachers' primary communities of practice and the wider landscapes of practice; strengthening structural and cultural conditions that recognise education as a scholarly activity and an integral part of the university's LoP; and increased educational awareness and competence at management level.\n\nID: 42554540\nTitle: Body Composition in Heart Failure: A Magnetic Resonance Imaging and Dual X-Ray Absorptiometry Assessment in the UK Biobank Study.\nAbstract: Metabolic and musculoskeletal abnormalities in patients with heart failure (HF) may not be fully captured by body mass index. Body composition was studied in participants with HF and matched control subjects in the UK Biobank Imaging Study, and findings were compared among those with HF with reduced ejection fraction, HF with mildly reduced ejection fraction, and HF with preserved ejection fraction. Participants with HF and available ejection fraction (n = 185) were matched 1:5 to age-, sex-, and body mass index-matched control subjects (n = 925). Whole-body magnetic resonance-quantified visceral adipose tissue (VAT), abdominal subcutaneous adipose tissue, liver fat, thigh fat-free muscle volume (MV), and muscle fat infiltration (MFI) were assessed. Personalized z-scores adjusted for sex and body size were derived. Adverse muscle composition (AMC) was defined as low MV z-score and high MFI. Dual-energy x-ray absorptiometry was used to measure total fat and appendicular lean mass. Compared with control subjects, HF participants had higher VAT (z-score = 0.46 \u00b1 1.1 vs 0.07 \u00b1 1.0; P < 0.001), MFI (8.4 \u00b1 2.3% vs 7.6 \u00b1 2.0%; P < 0.001), and AMC (36% vs 17%; P < 0.001) and lower MV (z-score = -0.7 \u00b1 1.0 vs -0.1 \u00b1 0.9; P < 0.001). Despite the muscle derangements identified by magnetic resonance imaging assessment, the prevalence of sarcopenia diagnosed using combined grip strength and dual-energy x-ray absorptiometry-derived lean mass parameters varied considerably depending on the criteria applied. Among HF subtypes, HF with mildly reduced ejection fraction showed the highest VAT and lowest MV, HF with preserved ejection fraction had greater fat and MFI, and HF with reduced ejection fraction had the greatest prevalence of AMC and the weakest grip strength. Participants with HF showed higher VAT and adverse muscle changes, with weaker grip strength. Sarcopenia prevalence varied substantially across definitions, reflecting substantial variability and discrepancies among current sarcopenia definitions and criteria. Traditional anthropometric indexes underestimate the burden of body composition derangements in HF.\n\nID: 42554525\nTitle: TagR, a newly identified member of the MarR family of transcriptional regulators, represses the NRPS operon in Klebsiella oxytoca.\nAbstract: Toxigenic Klebsiella oxytoca strains produce the pyrrolobenzodiazepine enterotoxins tilimycin (TM) and tilivalline (TV), which contribute to the development of antibiotic-associated hemorrhagic colitis. The biosynthesis of these toxins depends on the nonribosomal peptide synthetase (NRPS) operon located within the til pathogenicity island. Although several global and signal-responsive regulators of NRPS operon expression have been identified, the regulatory network governing enterotoxin biosynthesis remains incompletely characterized. In this study, we identified a previously unrecognized transcriptional regulator encoded within the til pathogenicity island of K. oxytoca. This protein, designated TagR (Tilivalline-associated genes repressor), is a member of the MarR family and acts as a negative regulator of NRPS operon expression. Structural prediction, molecular dynamics simulations, and biochemical analyses demonstrated that TagR exhibits the characteristic architecture of MarR family regulators and forms a stable homodimer. Deletion of tagR led to significant upregulation of the NRPS-associated genes npsA, thdA, and npsB, while complementation restored transcriptional repression. Electrophoretic mobility shift assays confirmed that TagR binds directly and specifically to the regulatory region upstream of the NRPS operon, supporting a mechanism of direct transcriptional repression. Consistent with these findings, loss of TagR significantly increased the cytotoxicity of K. oxytoca culture supernatants toward HeLa cells. Collectively, these results identify TagR as a direct repressor of the NRPS operon and expand the regulatory framework governing enterotoxin biosynthesis in toxigenic K. oxytoca. This study provides new insight into the transcriptional control of virulence-associated genes and establishes TagR as a previously unrecognized component of the regulatory network controlling TM and TV production.IMPORTANCEElucidating the mechanisms by which toxigenic Klebsiella oxytoca regulates enterotoxin production is critical for understanding the pathogenesis of antibiotic-associated hemorrhagic colitis. TagR is identified as a previously unrecognized MarR family regulator that directly represses the nonribosomal peptide synthetase (NRPS) operon responsible for tilimycin (TM) and tilivalline (TV) biosynthesis. This discovery uncovers a novel regulatory mechanism governing toxin production and offers new perspectives on virulence regulation in this emerging intestinal pathogen.\n\nID: 42554520\nTitle: Total synthesis and structural characterization of a novel protein scaffold from the snail Biomphalaria glabrata.\nAbstract: Disulfide-rich miniproteins constitute compact and highly stable scaffolds of growing interest for molecular and structural engineering. Schistosomins are ~80-residue proteins conserved across gastropods that form a long-standing orphan family whose structure and biological roles have remained unknown. Here, we report the total chemical synthesis and structural characterization of a schistosomin isoform from Biomphalaria glabrata, a medically relevant intermediate host of the parasite Schistosoma mansoni. Using state-of-the-art solid-phase peptide synthesis, chemoselective peptide ligation, and controlled oxidative folding, we obtained homogeneous, well-folded schistosomin suitable for biophysical and structural studies. High-resolution X-ray crystallography reveals a previously undescribed disulfide-rich fold defining a new class of miniprotein scaffold. Nano differential scanning fluorimetry and circular dichroism experiments demonstrate the remarkable thermal stability of this scaffold. Complementary in silico analyses suggest that the two naturally occurring isoforms, which differ by a single residue, exhibit highly similar structural and dynamic properties. Finally, transcript and protein analyses across snail tissues provide the first spatial expression map of schistosomin in a medically relevant mollusk. Together, this work establishes schistosomin as a novel and robust miniprotein scaffold and provides a structural and biological framework for exploring its function and potential applications.\n\nID: 42554515\nTitle: Allosteric effects of CDR-H3 mutations modulate binding and neutralization of a conserved SARS-CoV-2 RBD-targeting antibody.\nAbstract: SARS-CoV-2 antigenic evolution continues to erode the activity of first-generation monoclonal antibodies, underscoring the value of antibodies that recognize conserved features within the spike receptor-binding domain (RBD). As a model for breadth-oriented engineering, we assessed the RBD-directed antibody XG83, which has a CDRH3-dominated paratope. Utilizing the XG83-Wuhan RBD crystal structure as a reference, we integrated MOE alanine scanning and residue scanning with CDRH3 (A116-Y132) to identify chemically reasonable replacements and test interaction to an Omicron BA.1 RBD. The parental CDRH3-centric pose in comparative docking had a better score (-280.18) than the mutant MuXG83 (-254.2), along with 100-ns molecular dynamics showed that MuXG83 was less stable (with higher RMSD/RMSF with less favorable interaction energy). The ELISA results against Omicron BA.1 RBD demonstrated that XG83 had 40% stronger binding than MuXG83; convergence happened only at the highest concentration, concordant with SPR studies and the fact that mutation increased dissociation. According to this binding gap, BA.1 pseudovirus neutralization demonstrated that XG83 was much more powerful than MuXG83, showing that the E118L/F130H CDR-H3 alterations decreased functional activity against Omicron BA.1. These results imply that allosteric influences on interface stability and conformational dynamics by non-epitope CDR-H3 residues can affect antibody performance. Functional testing was confined to Omicron BA.1; therefore, larger variant-panel studies are needed to ascertain if such mutations affect antibody breadth. Our findings highlight a structure-guided approach for optimizing paratopes and indicate that non-epitope (potentially allosteric) changes to CDRH3 are also important while investigating potential development and neutralization before advancement. This study shows how modest allosteric characteristics in CDR-H3 control the delicate balance between neutralizing potency and breadth, making a timely and significant addition to SARS-CoV-2 antibody engineering. Using crystallography, alanine scanning, residue scanning, docking, molecular dynamics, and experimental ELISA and neutralization assays, this study offers a structure-guided framework for rational paratope optimization. The discovery of CDR-H3 residues that regulate long-range stability rather than just direct epitope contacts reveals an unappreciated aspect of antibody design and explains why some alterations improve anticipated interactions but degrade functional performance. Importantly, the comparison of wild-type XG83 and modified MuXG83 shows how allosteric tuning affects antibody-antigen compatibility in developing variations like Omicron. These findings illuminate conserved RBD recognition and offer strategies for building next-generation therapeutic antibodies that are more resistant to viral evolution.\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: 42554497\nTitle: Dynamic Fluorescence Visualization of Nonequilibrium Supramolecular Assembly Regulated by a Proton Reservoir.\nAbstract: Living systems sustain nonequilibrium supramolecular assemblies through continuous energy input and concomitant dissipation, with adenosine triphosphate (ATP) serving as a universal chemical fuel that regulates dynamic processes by coupling hydrolysis to downstream chemical or conformational changes. Biomimetic recreation of this behavior remains challenging because fuel-mediated regulation in artificial assemblies is typically coupled to building-block structure, limiting independent control over assembly dynamics and real-time monitoring. Here we report amphiphile-based fluorescent assemblies regulated by a time-programmable proton reservoir, where the fuel-consuming esterification-hydrolysis cycle is decoupled from the building blocks and controls assembly through programmed acid availability. The amphiphilic building blocks undergo stimulus-responsive assembly with fluorescence color tunable from blue to green, yellow, and white, and quantum yields spanning 1.7%-71.9%. The proton reservoir temporarily holds acid in hydrolysable esterified forms and subsequently regenerates acid over prescribed timescales; by molecular design, the acid-masking period is tuned from 0 to 250\u00a0min and acid regeneration from 10 to 120\u00a0min. Coupling this cycle to the fluorescent assembly regulates nonequilibrium organization, enables catalytic nucleophilic reactions, and provides an intrinsic optical signal for visualizing fuel consumption and assembly transformation in real time.\n\nID: 42554484\nTitle: Miro1 Protects Against Acute Kidney Injury Through Modulating Mitochondrial Homeostasis via Interaction With Sirt6.\nAbstract: Acute kidney injury (AKI) is highly prevalent worldwide but lacks specific treatments. Mitochondrial dysfunction plays a central role in its pathogenesis, making mitochondrial protection a promising therapeutic strategy. Miro1 is a key regulator of mitochondrial dynamics, yet its involvement in AKI remains unexplored. In this study, we found that Miro1 expression was significantly downregulated in renal tubular cells from AKI patients and in multiple murine AKI models. Functional studies demonstrated that Miro1 loss aggravated tubular cell injury and mitochondrial dysfunction, whereas Miro1 overexpression exerted protective effects. Mechanistically, we identified Sirt6 as a novel interacting partner of Miro1. Sirt6 directly deacetylated Miro1 at lysine residue K182, which inhibited its ubiquitin-proteasome degradation and maintained Miro1 protein stability. Disruption of Sirt6 activity led to enhanced Miro1 downregulation and increased tubular cell apoptosis, while Sirt6 activation reversed these detrimental effects in a deacetylase-dependent manner. Reciprocal rescue experiments further confirmed that Miro1 serves as a critical downstream effector of Sirt6. Collectively, our findings uncover the Sirt6-Miro1 axis as a previously unrecognized protective mechanism in AKI, wherein Sirt6 stabilizes Miro1 via deacetylation to preserve mitochondrial homeostasis and alleviate kidney injury. This axis may represent a novel therapeutic target for AKI intervention.\n\nID: 42554476\nTitle: Structure design and implementation of a water cooling sub-20\u2005nm multilayer Kirkpatrick-Baez mirror system in the Structural Dynamics beamline ID23 at HEPS.\nAbstract: The first water-cooling nano multilayer Kirkpatrick-Baez mirror system in the Structural Dynamics beamline (ID23) at High Energy Photon Source (HEPS) has been implemented. An Invar gantry is engineered to achieve a balance between light weight and stability. The cooling system, including eutectic gallium-indium (eGaIn), copper braids and a cooling water circuit with a multi-bend copper pipe in a compact space, decouples movement and mitigates vibration. The mirror cooling holders introduce less than 0.5\u2005nm RMS height error and 0.1\u2005\u00b5rad RMS slope error in the mirror surface shapes. A series of stability tests is applied to verify the mechanism structure. In a 1\u2005h test assessment using a water-cooling flow of 4.5\u2005L\u2005min-1, the system demonstrated a positional stability of 5.96\u2005nm and an angular stability of 86.76\u2005nrad from 1\u2005Hz to 500\u2005Hz. The focal spot size of 13.39\u2005nm \u00d7 15.15\u2005nm (H \u00d7 V) at a photon energy of 21.8\u2005keV demonstrates the system's performance.\n\nID: 42554465\nTitle: Pseudomonas aeruginosa mgtC gene is under the control of PhoP and CbrAB regulators, and its expression can be visualized in macrophages.\nAbstract: The MgtC virulence factor is important during the intramacrophage stage in both classical intracellular pathogens, such as Salmonella Typhimurium, and in extracellular bacteria that transiently encounter intracellular environments during infection, such as Pseudomonas aeruginosa. In these different pathogens, mgtC expression is induced in vitro by magnesium ion depletion, a condition reported to mimic the macrophage environment. Here, we developed an unstable GFP reporter system to monitor in real time the transcriptional activation of the P. aeruginosa mgtC promoter. After in vitro validation in magnesium-defined media, this reporter system allowed visualization of the mgtC promoter induction in a subset of bacteria when P. aeruginosa localized inside cultured macrophages. In addition, although rare under our experimental conditions, in vivo activation of the mgtC promoter was observed for the first time within macrophages of live, infected zebrafish larvae, a cutting-edge vertebrate model for real-time imaging. While MgtC regulation in Salmonella is mediated by the magnesium-responsive PhoPQ two-component system, its regulation in P. aeruginosa remained unknown. The use of mutant strains for two-component regulatory systems revealed that the PhoP regulator, but not by its cognate sensor PhoQ, was required to activate P. aeruginosa MgtC expression in vitro. Unexpectedly, CbrAB, a two-component system specific to Pseudomonas species, was also involved in P. aeruginosa MgtC regulation. Both PhoP and CbrB regulatory proteins were found to directly bind the mgtC promoter, supporting a dual transcriptional control. These findings reveal substantial differences in mgtC gene regulation in different bacterial pathogens, reflecting distinct strategies to drive appropriate expression of a shared virulence factor involved in macrophage adaptation.IMPORTANCEThe adaptation of bacterial pathogens to the host intracellular microenvironment requires tight and rapid regulation of specific genes, and investigating the in vivo transcriptional dynamics of such genes is a major challenge. Here, we focused on the expression of mgtC, a gene important for adaptation to the intramacrophage environment in classical intracellular pathogens, such as Salmonella Typhimurium, and bacteria with a transient intracellular lifestyle, such as Pseudomonas aeruginosa. An unstable GFP reporter system was designed to monitor the transcriptional dynamics of P. aeruginosa mgtC. The use of this reporter system in a state-of-the-art vertebrate model for live imaging, the zebrafish embryo, allowed in vivo tracking of P. aeruginosa mgtC promoter activation inside macrophages in a living host. Furthermore, the expression of P. aeruginosa mgtC was found to be regulated through a mechanism distinct from that of Salmonella MgtC, since it involves the PhoP regulatory protein, but not the PhoQ sensor, and the Pseudomonas-specific CbrAB two-component system, reflecting diverse, finely tuned strategies to control a virulence factor shared by several major human pathogens.\n\nID: 42554464\nTitle: The FleQ-FleN circuit balances flagellar number against fitness in Pseudomonas aeruginosa.\nAbstract: Evolutionary constraints governing flagellar number in bacterial pathogens remain poorly understood. While related Pseudomonas species are hyperflagellated, Pseudomonas aeruginosa maintains strict monoflagellation through the FleQ-FleN regulatory circuit. Here, we demonstrate that FleN dosage is essential for maintaining monoflagellation and fitness. Confirming earlier reports, wild-type P. aeruginosa displayed unipolar monoflagellation, whereas \u2206fleN mutants developed multiple flagella, frequently more than four, in unipolar or bipolar arrangements. These hyperflagellated cells showed marked fitness defects, including reduced growth, attenuated virulence in a nematode infection model, and a competitive disadvantage in co-culture. The virulence defect reflected hyperflagellation rather than the loss of motility, since a non-flagellated \u2206fliC mutant retained full pathogenicity. Notably, \u2206fleN cells rapidly evolved suppressor mutations in fleQ that partially restored growth and motility without always restoring monoflagellation. Five independent suppressor alleles mapped to key FleQ domains, four in the AAA+ ATPase domain, and one in the DNA-binding domain, consistent with reduced FleQ activity that rebalances the circuit. Single-cell holographic tracking showed that suppressor strains swim with heterogeneous dynamics, including subpopulations that reach wild-type (WT) speeds, despite carrying multiple flagella. Quantitative proteomics indicated that the fitness burden extends beyond flagellar components, with protein-abundance changes across metabolism, stress responses, and signaling. Conversely, selection under high-viscosity conditions drove wild-type cells to acquire enhanced spreading through a recurrent fleN mutation, fleNV178G; multiflagellation is therefore accessible. Together, these findings indicate that the FleQ-FleN circuit balances the flagellar number against fitness, holding P. aeruginosa at a single flagellum while retaining the latent capacity to reach multiflagellated states through single mutations.IMPORTANCEBacterial flagella are extracellular appendages that rotate to propel the cell and enable swimming motility. While some bacteria have multiple flagella, many pathogenic species like Pseudomonas aeruginosa have just one. Surprisingly, mutants of P. aeruginosa with multiple flagella performed worse, that is, they grew more slowly, were less infectious in laboratory animals, and were outcompeted by wild-type bacteria. Even when some mutant bacteria evolved compensatory changes, they still struggled compared to single-flagellum bacteria. This reveals an important evolutionary trade-off: while multiple flagella might seem advantageous for movement, having just one flagellum allows the bacteria to grow faster and cause more severe infections. This plasticity likely explains why P. aeruginosa is so successful both in the environment and as a human pathogen.\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: 42554420\nTitle: Straightforward Synthesis, Spectroscopic Characterizations, Molecular Docking, Molecular Dynamics Simulations, and Comprehensive DFT Calculations of Novel Tetrazole 1,5 and 2,5-Disubstituted Tetrazole Scaffolds.\nAbstract: New 1,5- and 2,5-disubstituted tetrazole derivatives were successfully synthesized under phase-transfer catalysis conditions in good to excellent yields (21%-97%). The structures of the obtained compounds were confirmed by 1H and 13C NMR spectroscopy, as well as single-crystal X-ray diffraction analysis. Density functional theory calculations performed at the B3LYP/DNP 3.5 level showed that compound F possesses the smallest HOMO-LUMO energy gap (3.390\u2009eV), suggesting enhanced chemical reactivity. Monte Carlo/SAA simulations demonstrated strong adsorption tendencies on metallic surfaces, particularly for compound 1j on the Fe(110) surface, with an adsorption energy of -229.944 kcal mol-1. Molecular docking investigations against Abl kinase targets demonstrated favorable binding affinities, with docking scores reaching -11.7 and -8.3 kcal mol-1 for the most active derivatives. In addition, 100\u2009ns molecular dynamics simulations confirmed the stability of the 2HZI-3d and 4TWP-2j complexes throughout the simulation period. These findings highlight the potential of the synthesized tetrazole derivatives as promising candidates for further biological and corrosion-related applications.\n\nID: 42554407\nTitle: Annexin A1 as a Key Modulator of Inflammatory, Glial, and Angiogenic Signaling Pathways in Diabetic Retinopathy.\nAbstract: Annexin A1 (AnxA1) regulates inflammation and neovascularization in ocular diseases, influencing growth factor signaling and cytoskeletal dynamics. Accordingly, this study aimed to investigate the role of the AnxA1 protein in diabetic retinopathy (DR) and to understand how its absence affects inflammation and vascular dysfunction in DR. DR model was induced 12 weeks after streptozotocin injection in wild-type (AnxA1+/+) and AnxA1 knockout (AnxA1-/-) mice. We evaluated the consequences of AnxA1 deficiency on retinal morphology, M\u00fcller cell activation, inflammation, and angiogenesis associated with DR. We also performed an in silico analysis in wild-type animals with DR induced for 6 weeks and in a dataset of patients with DR. Furthermore, a choroidal neovascularization model was used to evaluate the role of AnxA1 in angiogenesis signaling. AnxA1 deficiency exacerbated selective systemic inflammatory alterations and promoted retinal neurodegeneration associated with DR. AnxA1-/- diabetic animals exhibited increased circulating IL-6, IL-17, chemokine (C-X-C motif) ligand 1, and monocyte chemoattractant protein-1 levels, retinal monocyte chemoattractant protein-1 and IL-4 upregulation, dysregulated MAPK/STAT3 signaling, and altered gliosis responses. In addition, AnxA1 deficiency was associated with a proangiogenic signaling profile characterized by increased retinal levels of EGF, VEGF-A, and endothelin-1 in DR, whereas the upregulation of VEGF-C was predominantly observed in the choroidal neovascularization model. Endogenous AnxA1 modulates inflammatory-glial and proangiogenic signaling pathways in the diabetic retina. Its deficiency is associated with selective inflammatory dysregulation and enhanced angiogenesis-related signaling during DR progression.\n\nID: 42554395\nTitle: Cardiac Amyloidosis in Asia: What We Know and Don't Know.\nAbstract: \n\nID: 42554385\nTitle: Liver for the Cardiologist: Why the Liver Belongs in the Cardiometabolic Workflow (CKM to CKML).\nAbstract: \n\nID: 42554384\nTitle: Cardiologists as Champions of Kidney Health in the CKM Era.\nAbstract: \n\nID: 42554359\nTitle: Tuning Water Transport in Subnanometer-Sized Artificial Water Channels.\nAbstract: Water transport through nanochannels is not only essential for biological functions but also holds promise for the development of advanced flow sensors and membranes. In this study, a series of artificial water channels with subnanometer-sized pores\u00a0and tunable hydrophilicity were designed by mimicking the architecture of natural water channels. Experimental measurements and molecular dynamics (MD) simulations reveal that water transport can be modulated by pore hydrophilicity and electric field. Increasing pore hydrophilicity is associated with enhanced water permeability, consistent with continuous wetting and more uniform dipole alignment within the single-file water wires. In contrast, a hydrophobic backbone impedes water transport due to dewetting effects and broader dipole orientation distributions. In addition, a static electric field significantly facilitates water transport and leads to a membrane polarization-dependent water transport behavior by promoting more ordered water dipole configurations. These findings provide molecular-level and external-field-based strategies to regulate water transport in subnanometer channels and may guide the design of energy-efficient desalination membranes.\n\nID: 42554338\nTitle: Exploring Trade-offs Between Carbon Storage and Timber Production with Alternative Rotation and Harvest Policies: Insights from a Real-World Forest Management Area.\nAbstract: Forests provide essential ecosystem services, particularly timber production and carbon sequestration, both of which play critical roles in climate change mitigation and sustainable forest management. However, the long-term consequences of alternative management strategies on carbon-timber trade-offs remain insufficiently understood. This study addresses this gap by analysing the combined effects of rotation length, harvest regulation strategies, and tree-size dynamics on forest carbon storage and timber production using the ET\u00c7AP decision support system (DSS). Four management scenarios were developed by combining short and long rotation periods with area-control and volume-control harvesting policies, while explicitly accounting for tree-size-dependent carbon dynamics. Simulations were conducted for a representative forest planning unit in T\u00fcrkiye over a 100-year planning horizon. Results showed that longer rotations and volume-control harvesting consistently projected the largest increase in carbon storage by promoting the development and retention of larger trees, whereas shorter rotations and area-control harvesting enhanced timber production but intensified trade-offs with carbon sequestration. While tree-size dynamics influenced the magnitude of carbon accumulation, they did not substantially alter the overall trade-off patterns among management strategies. These results indicate that management decisions affecting age structure and harvesting regimes are important determinants of long-term carbon-timber outcomes. As the analysis is based on deterministic simulations that exclude major disturbance processes such as wildfire, pest outbreaks, and climate extremes, the results need to be interpreted as scenario-based projections. Nevertheless, the study highlights the value of integrating ecosystem-based forest management with decision support systems to systematically evaluate long-term management alternatives and support climate-informed forest planning.\n\nID: 42554278\nTitle: Conformational asymmetry of human immunodeficiency virus-1 protease flaps increases along the darunavir drug resistance pathway.\nAbstract: Under selective drug pressure, the structure and dynamics of HIV-1 protease (PR) evolve to confer resistance to the drug. Here, we report on the conformational changes in PR flaps for drug-resistance mutations using 19F nuclear magnetic resonance (NMR) spectroscopy. We prepared wild-type (wt) PR and its three mutant variants containing 4, 10, or 11 mutations, namely 4Mut, 10Mut, and 11Mut, that were previously found in a viral passage study against darunavir (DRV). The proteins were uniformly labeled with 15N and with 19F amino-acid specifically in the 5-indole positions of two conserved tryptophan residues, W6 and W42, which are located near the dimer interface and at the edge of the flap region of PR, respectively. In solution, 19F NMR spectra of these proteins in the inhibitor-free forms showed a downfield shift of the W42 resonance as the number of mutations increased. In the DRV-bound form, the W42 resonance again shifted with increasing number of mutations and, importantly, split into two peaks, reflecting different local electronic environments of the two flaps of the dimer. Solid-state 19F magic angle spinning (MAS) NMR spectra of wt-PR and 10Mut also exhibited two W42 resonances in DRV-bound crystalline forms. These observations indicate that drug-resistance mutations increase the asymmetry between the two flaps in the PR-inhibitor interaction in the inhibitor-bound forms.\n\nID: 42554274\nTitle: Molecular Architecture of Temporal Variability Network Dysfunction in Bipolar Disorder.\nAbstract: Characterized by recurrent fluctuations in mood states, bipolar disorder (BD) is widely conceptualized as a disconnection syndrome associated with dysregulated brain dynamics. Nevertheless, the molecular mechanisms underlying this aberrant connectivity dynamics in BD remain elusive. Using resting-state electroencephalography (EEG) data from BD patients and healthy controls, this study first delineated the characteristic alterations in temporal variability of functional connectivity in BD and further elucidated their underlying molecular mechanisms and clinical relevance. Current findings revealed significantly reduced temporal variability within large-scale brain subnetworks, most notably in the dorsal attention, somatomotor, and visual networks. Importantly, these neurodynamic signatures effectively predicted the symptom severity in individuals with BD.\u00a0Moreover, the spatial patterns of these dynamic alterations are associated with the expression of BD risk genes enriched in synaptic function and metabolic pathways, as well as with the spatial organizations of various neurotransmitter receptors, including CB1, mGluR5, H3, and MOR.\u00a0Collectively, these results provide evidence for a multiscale pathophysiological framework that links genetic susceptibility and chemoarchitectural alterations to dynamic brain network instability, ultimately underpinning the core clinical manifestations in BD.\n\nID: 42554263\nTitle: Epigenetic aging in Alzheimer's disease: Relation to proteome.\nAbstract: Alzheimer's disease (AD) arises from heterogeneous biological processes, and long-term environmental exposures may become biologically embedded, as reflected in epigenetic clocks. We calculated blood DNA methylation-based epigenetic clocks and constructed protein co-abundance networks of cerebrospinal fluid (CSF) proteomics data. We performed association analysis of epigenetic age acceleration with network modules, followed by functional and cell-type enrichment analyses, association analyses of hub proteins with AD endophenotypes, and pseudotime trajectory analysis. Six network modules were significantly associated with epigenetic age acceleration and were enriched in pathways related to neuronal connectivity, proteostasis, immune activation and remodeling, immune signaling, and immunoepigenetic regulation. Hub proteins demonstrated significant associations with baseline amyloid/tau/neurodegeneration biomarkers and longitudinal cognitive changes. Pseudotime analysis revealed continuous, non-linear variation in epigenetic age acceleration along the inferred trajectory. Our CSF proteomics study identified neuronal, proteostatic, and immune-related molecular signatures associated with epigenetic age acceleration in AD.\n\nID: 42554262\nTitle: The association of cardiovascular health with new-onset pulmonary hypertension and the mediating role of proteomic signatures.\nAbstract: The cardiovascular health (CVH) metrics have been reported to play an important role in the development of noncommunicable chronic diseases, yet its link to pulmonary hypertension (PH) risk and the underlying biological mechanisms remain unclear. This study aimed to investigate the association of CVH with PH risk and elucidate the mediating role of plasma proteomic signatures. A total of 279 220 participants without PH at enrollment of the UK Biobank were included. Cox regression was used to quantify the association between CVH and incident PH. Proteome-wide association analysis, mediation analysis, and functional enrichment analysis were conducted to identify protein mediators. Key hub proteins were further validated at the transcriptional level through quantitative polymerase chain reaction (qPCR) in an animal model of PH, as well as at the protein level, and by macrophage-specific knockdown of interleukin (IL)-6 and CCL4 to evaluate its impact on rat pulmonary artery smooth muscle cell (PASMC) migration and proliferation. Over a median 13.2-year follow-up, 1325 PH cases occurred. Compared to the lowest CVH, participants with moderate and high CVH had 59% [hazard ratio (HR): 0.41; 95% confidence interval (CI): 0.33-0.49] and 82% (HR: 0.18; 95% CI: 0.14-0.23) lower risk, respectively. Proteomic analyses revealed that this association was significantly mediated by a distinct plasma protein signature. Pathway enrichment analysis indicates that proteins are significantly enriched in inflammatory/immune pathways, and key hub proteins were identified as participating in the central mechanism pathway. In the lung tissue of PH rat models, the mRNA and protein expression levels of IL-6 and C-C motif chemokine ligand 4 (CCL4) were significantly elevated. Furthermore, functional assays demonstrated that knockdown of IL-6 or CCL4 in macrophages significantly attenuated the migration and proliferation of rat PASMCs in vitro. High CVH level, defined by Life's Essential 8 (LE8), is significantly linked to a reduced risk of developing PH. This protective effect is primarily mediated by a proteomic signature, revealing the role of signaling pathways such as cytokine-cytokine receptor interaction in the prevention of PH.\n\nID: 42554236\nTitle: MODY Mutations in Transcription Factors HNF1A and HNF1B Affect the Production of Interferon Signaling Proteins: Evidence at the Proteome Level.\nAbstract: This discovery study investigated the impact of MODY-associated mutations in hepatocyte nuclear factors HNF1A and HNF1B on the cellular proteome, aiming to identify affected pathways and advance understanding of diabetes pathogenesis. Human induced pluripotent stem cells (hiPSCs) carrying the HNF1A frameshift mutation (p.Pro291fsinsC) were differentiated into pancreatic progenitors, and renal proximal tubule epithelial cells (RPTECs) were engineered to overexpress HNF1B with the S148L point mutation. Label-free quantitative proteomics was performed using data-dependent and data-independent acquisition on Orbitrap and timsTOF mass spectrometers. Pathway enrichment was analyzed using Qiagen IPA, Hallmark gene sets, and STRING networks. Comprehensive proteome coverage (over 7000 proteins) revealed consistent downregulation of oxidative phosphorylation, mitochondrial function, and interferon signaling pathways. Both models exhibited suppression of innate immune responses, with overlapping downregulated proteins, including members of the OAS, IFIT, and MX1 families. Using label-free proteomics, we show that MODY-associated mutations in HNF1A and HNF1B suppress mitochondrial function and interferon signaling and are additionally associated with reduced abundance of predicted targets such as A1CF as well as diabetes-related proteins including SCGN, supporting their role in diabetes pathogenesis.\n\nID: 42554217\nTitle: Advances in Nucleic Acid-Based Amplification Strategies for Dual-Mode Biosensing of Pathogenic Bacteria: From Molecular Mechanisms to Integrated Analytical Systems.\nAbstract: Rapid, accurate, and reliable detection of pathogenic bacteria remains a critical need in clinical, food, and environmental monitoring. In today's context, nucleic acid amplification-mediated biosensing have emerged as a prominent approach to improve detection sensitivity, whereas dual-mode signal readout approaches have more enhanced analytical robustness and reliability. This review outlines recent advances in nucleic acid signal amplification strategies, including enzyme-based methods like LAMP, RPA, and RCA, as well as enzyme-free approaches like HCR, CHA, and EDR. Special attention is given to incorporating these amplification methods into dual-mode biosensing systems that combine both optical and electrochemical transduction mechanisms. This integration enables complementary signal generation and improves detection accuracy by reducing false-positive and false-negative results. This study critically examines the advancement of nucleic acid signal amplification strategies (NASAS)-mediated dual-mode sensing systems for detecting major pathogenic bacteria, including Escherichia coli, Salmonella, Listeria monocytogenes, Staphylococcus aureus, and Vibrio species, focusing on selectivity, sensitivity, assay design, and real-sample applicability. Finally, the review highlights present challenges related to system integration, standardization, and point-of-care applications. Additionally, it outlines potential future directions for rendering nucleic acid amplification-based dual-mode probes into practical diagnostic devices. Overall, this study affords a comprehensive synthesis of emerging approaches and design mechanisms for next-generation diagnostic scaffold for pathogen analysis.\n\nID: 42554209\nTitle: Mutation-driven alterations in the AcrB efflux pump modulate antibiotic resistance in Klebsiella pneumoniae.\nAbstract: To investigate the global prevalence of the acrB efflux transporter among antibiotic resistance genes (ARGs) in Klebsiella pneumoniae and to determine how mutations within AcrB drug-binding pockets influence structural stability and antibiotic interactions. A global genomic analysis of 42 ARGs reported in K. pneumoniae isolates worldwide was performed to identify the prominence of acrB. A total of 92 AcrB protein sequences were assessed for drug-binding pocket mutations relative to the wild-type strain NCTC9637. De novo protein structures were generated using AlphaFold2 and evaluated through molecular dynamics (MD) simulations. MD analyses were conducted with multiple antibiotics to assess the impact of pocket mutations on drug-binding affinities. acrB emerged as the most widely distributed ARG, conferring resistance to at least 12 antibiotic classes. Three isolates, NCTC9617, NCTC9645, and NCTC13635, displayed multiple drug-binding pocket mutations when compared with the wild type. MD simulations revealed marked structural instability in NCTC9645, while NCTC13635 showed convergence towards a wild-type-like conformation. Docking analyses demonstrated that binding pocket mutations altered antibiotic-binding affinities, suggesting possible alterations in substrate recognition and transport. AcrB is a key driver of multidrug resistance in K. pneumoniae, and mutations within its drug-binding pockets can significantly impact protein stability and antibiotic-binding behaviour. Monitoring these structural variations may help predict emerging resistance patterns and guide therapeutic decision-making. Multidrug-resistant Klebsiella pneumoniae infections are a growing clinical threat with limited therapeutic options. By integrating global resistance gene surveillance with structural modelling of AcrB efflux pump variants, this study identifies drug-binding pocket mutations that modulate antibiotic affinity and efflux pump stability. These insights provide a molecular basis for predicting treatment failure, guiding precision antibiotic therapy, and advancing the development of efflux pump inhibitors as adjunctive treatments for drug-resistant Gram-negative infections.\n\nID: 42554193\nTitle: Emerging bismuth-based perovskite single crystals for advanced optoelectronics.\nAbstract: Bismuth-based perovskite single crystals have emerged as a promising lead-free alternative to solve the toxicity and stability challenges of lead-based halide perovskites. Owing to their unique structural diversity, ranging from low-dimensional halide perovskites and double perovskites to layered perovskite oxyhalides, these materials exhibit exceptional optoelectronic properties, including tunable bandgap, high environmental stability, higher ion migration activation energy, and long carrier lifetime. This review provides a systematic overview of recent advances in bismuth-based perovskite single crystals. We first discuss their classification and crystal structure characteristics, followed by a detailed analysis of their electronic band structure, self-trapped exciton luminescence, charge carrier dynamics, and optical properties. Subsequently, various growth strategies are introduced for bismuth-based single crystals, including the slow evaporation method, temperature reduction crystallization method, anti-solvent diffusion method, hydrothermal method, melt crystallization method, and vapor transport method. The applications of these single crystals in high-performance photodetectors and X-ray detectors are comprehensively summarized. Finally, the key challenges and future directions toward scalable synthesis, surface passivation interface engineering, multifunctional integration, and emerging applications are discussed.\n\nID: 42554189\nTitle: Positional isomerism-driven solvatochromism and excited-state behavior in ortho- and para-hydroxy bromobenzylidene Schiff bases: a combined experimental and DFT investigation.\nAbstract: This study presents a comprehensive investigation of how hydroxyl positional isomerism governs the solvatochromic response and excited-state behavior of bromobenzylidene Schiff base derivatives. Two structurally related isomers, SB-oOH and SB-pOH, were synthesized and systematically analyzed using combined spectroscopic and quantum chemical approaches. The results reveal a striking contrast in photophysical behavior arising solely from the substituent position. SB-oOH exhibits weak solvent-dependent spectral shifts, attributed to intramolecular O-H\u22efN hydrogen bonding that restricts \u03c0-electron delocalization and stabilizes a localized excitation (LE) state. In contrast, SB-pOH displays pronounced positive solvatochromism, large Stokes shifts, and strong solvent sensitivity, consistent with increased polarizability and stronger light-matter interaction, whereas SB-oOH maintains a more rigid and less responsive electronic structure. Quantitative solvatochromic analysis using linear solvation energy relationship (LSER) models demonstrates that SB-oOH is primarily influenced by nonspecific dielectric interactions, while SB-pOH is strongly governed by both solvent polarity and hydrogen-bonding effects. Theoretical calculations based on density functional theory (DFT) and time-dependent DFT (TD-DFT) support these findings, revealing enhanced HOMO-LUMO separation, a reduced energy gap, and increased electronic delocalization in the para-substituted system. Furthermore, optical band gap and refractive index analyses confirm that the enhanced ICT characteristic of SB-pOH, as supported by the combined experimental and theoretical results, is consistent with increased polarizability and stronger light-matter interactions, whereas SB-oOH maintains a more rigid and less responsive electronic structure. Thus, this study demonstrates that subtle structural variation through hydroxyl positional isomerism can induce profound changes in electronic structure and excited-state dynamics. These findings provide a clear structure-property relationship and offer a rational strategy for designing Schiff base-based functional materials with tunable optoelectronic properties. This study highlights positional isomerism as an effective molecular design tool for controlling excited-state processes.\n\nID: 42554186\nTitle: From Quantification to Translation: Advanced Bioanalytical Strategies for Evaluating Targeted Drug Delivery Systems Across Molecular, Cellular, and In\u00a0Vivo Scales.\nAbstract: Targeted Drug Delivery Systems (TDDS) have been proposed as an innovative approach for enhancing therapeutic efficiency through selective targeting of drugs. Although there has been substantial progress in the area of targeted therapy, the application of TDDS into clinical practice has been impeded by difficulties in their bioanalysis. Existing bioanalytical methods designed for small-molecule drugs are not sufficient for proper characterization of the complex structure, dynamics, and functionality of the advanced delivery systems. This article reviews various methods of bioanalysis used in the assessment of TDDS at the molecular,cellular, tissue,and in vivo levels. Special emphasis is placed on problems such as separation of encapsulated and released drugs, the formation of the protein corona, matrix effects, biodistribution analysis, and detection of very low drug concentrations. Additionally, the review highlights important translational bottlenecks, such as non-standardization of protocols, regulatory challenges, method validation difficulties, and scaling problems.Emerging approaches like artificial intelligence-enabled bioanalysis, multi-omics, smart biosensing, personalized assessment, and digital twin technology are analyzed as possible remedies to tackle the existing challenges.Collectively, these advancements underscore the importance of integrated and standardized bioanalytical frameworks for bridging preclinical evaluation with clinical translation, thereby enabling the successful development and implementation of next-generation targeted drug delivery systems.\n\nID: 42554185\nTitle: The dynamics of the reaction between Co3O4/SiO2 and synthesis gas in a magnetic field.\nAbstract: The reaction of a Co3O4/SiO2 catalyst with synthesis gas (CO\u2009:\u2009H2 = 1\u2009:\u20091) was studied using in situ magnetometry in programmable temperature and isothermal modes. Changes in magnetic induction and the formation rate of CO2 were used to monitor the dynamics of the reaction. Then, a simple kinetic model was used to estimate the rate ratios of the following reactions: Co3O4 + CO \u2192 CoO + CO2 and CoO + CO \u2192 Co + CO2. The effect is observed both in the synthesis gas and in pure CO. It was demonstrated that an external magnetic field exerts a multidirectional influence on the rates of these reactions. As the magnetic field strength increases, the rate of the first reaction decreases, while that of the second increases. This suggests that the process is spin-dependent.\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: 42554171\nTitle: The rise of carnivoran mammals in Europe through the lens of body mass.\nAbstract: Carnivoran mammals (cats, dogs, bears, seals, etc.) and their kin radiated during the Palaeogene, among other carnivorous clades (hyaenodonts, mesonychians, and oxyaenodonts). This radiation has often been framed as a competition ultimately won by carnivorans, but the data supporting this hypothesis largely originate from North America. We derive body mass (BM) from dental measurements in European hyaenodonts, mesonychians, oxyaenodonts, and carnivoramorphans ranging from the latest Palaeocene to the end of the Oligocene (MP 6-30; 57.2-23.3 Ma) and compare those with North American data. We show that European assemblages tell a different story, with a marked increase of the disparity of carnivoramorphan BMs at and after the Middle Eocene Climatic Optimum (MECO; 40 Ma), with no clear effect on hyaenodonts. The 'Grande Coupure' (Eocene-Oligocene transition, approx. 34-33.5 Ma) marks the onset of a progressive decrease in the mean BM of hyaenodonts, while carnivoramorphans continue their rise initiated around the MECO. The observed BM patterns therefore do not follow the expected double-wedge pattern of competitive replacement and are possibly best explained by climate change-induced biodiversity dynamics.\n\nID: 42554170\nTitle: Guidelines for Studying Mutagenesis in Quiescent Schizosaccharomyces pombe.\nAbstract: All living organisms can enter a non-dividing state known as quiescence, which often functions as a survival strategy. In many cases, quiescent cells retain the ability to re-enter the cell cycle when conditions become favorable. Therefore, preserving the integrity of non-dividing cells is essential not only to prevent their deterioration, but also to ensure proper population re-establishment once growth resumes. This also applies to the genome, whose stability must be maintained during both proliferation and quiescence. For several years, we have investigated this aspect of quiescence in fission yeast, which enters a G0 state in response to nitrogen starvation. We have shown that wild-type G0 cells accumulate mutations over time, with a mutational spectrum distinct from that observed during proliferation. We are now extending this work to mutants defective in various DNA repair pathways. In doing so, we developed a robust protocol ensuring highly reproducible results when assessing mutagenesis in quiescent cells. Here, we describe this protocol in detail.\n\nID: 42554148\nTitle: A computational framework for K\u00e1rm\u00e1n gaiting in robotic fish: spatio-temporal perception and CPG-based reinforcement learning.\nAbstract: Navigating in unsteady wake flows, such as K\u00e1rm\u00e1n vortex streets, presents a formidable challenge for biomimetic autonomous underwater vehicles. Biological fish achieve this by utilizing their lateral line sensory systems to perceive local flow gradients and adopting an energy-efficient swimming pattern known as the K\u00e1rm\u00e1n gait. To translate this biological phenomenon into a practical robotics engineering solution, this paper proposes a fully computational framework focusing on the modeling and simulation of a spatio-temporal sensory system to autonomously generate the K\u00e1rm\u00e1n gait. To overcome the unrealistic assumption of full-state observability common in existing reinforcement learning studies, we model a multi-point lateral line array coupled with a frame-stacking mechanism. This allows the simulated agent to reconstruct the spatio-temporal topology of the surrounding unsteady flow relying exclusively on local pressure and velocity gradients. The sensory model is integrated with a spatio-temporal perceptual twin delayed deep deterministic policy gradient (STP-TD3) algorithm, which drives a Hopf-oscillator-based central pattern generator. Through rigorous high-fidelity computational fluid dynamics simulations, we quantitatively evaluate the autonomous emergence of the K\u00e1rm\u00e1n gait by assessing the agent's kinematic energy proxy-mapped from joint actuation effort. Results reveal that the agent expends significantly less mechanical effort navigating through the turbulent vortex street compared to swimming in steady water, suggesting the active exploitation of the local wake dynamics. The results theoretically underscore the necessity of distributed STP for biomimetic robots, providing a robust algorithmic blueprint for future physical deployments in complex aquatic environments.\n\nID: 42554138\nTitle: Precisely Timed On-Demand Droplet Ejection and Collision in Free Space for Controlled Reaction Initiation.\nAbstract: Precise initiation and synchronization of chemical reactions in free space remain major challenges for time-resolved studies in structural biology, chemistry, and materials science. Although modern probes provide femtosecond resolution, experimental access to early reaction dynamics is limited by the ability to trigger, mix, and deliver nano-to-picoliter liquid samples with deterministic spatial and temporal control. Existing liquid jets and drop-on-demand systems cannot reliably define reaction start times in flight or synchronize them to external pulse patterns. We present an acousto-pneumatic droplet ejection platform that delivers single sub-nanoliter droplets on-demand with microsecond timing accuracy and reproducible trajectories, enabling deterministic one-drop-per-trigger operation synchronized to external signals at repetition rates up to 100\u00a0Hz. Deterministic mid-air droplet collisions define reaction initiation directly at the probe and are compatible with demanding pulse structures such as X-ray free-electron lasers. The programmable platform offers high-resolution, time-resolved experimentation, spanning ultrafast X-ray studies and precision nanoliter dispensing.\n\nID: 42554126\nTitle: Vibrio Community Structure Shapes the Diversity of Carbenicillin-Hydrolysing Class A \u03b2-Lactamase Circulating in European Coastal Environments.\nAbstract: Coastal environments are increasingly recognised as reservoirs of known antibiotic resistance genes (ARGs), but are less frequently identified as sources of novel ARGs. Here, we investigated class A \u03b2-lactamases circulating in European coastal environments used for oyster farming. We examined their diversity, function, and the ecological factors associated with their geographic distribution and environmental dynamics. A high diversity of carbenicillinases was detected in the culturable microbiome of European oysters. The Harveyi and Splendidus clades were key Vibrio lineages structuring the geography of carbenicillinase diversity. The Harveyi clade was primarily associated with the circulation of known carbenicillinases in Mediterranean samples, whereas the Splendidus clade contributed previously uncharacterized carbenicillinase sequences across all Europe. A one-year seasonal monitoring revealed that Vibrio alginolyticus drives the circulation of blaCARB-42 in the Mediterranean Thau lagoon, with dynamics strongly associated with seawater temperature. blaCARB-42 conferred intrinsic resistance to both carboxypenicillins and aminopenicillins in V. alginolyticus, which was found in most other species of the Harveyi clade with additional resistances to aztreonam, third-generation cephalosporins and aminoglycosides. Since the Harveyi clade includes major human pathogens, these findings have direct implication for environmental and One Health surveillance, as rising seawater temperatures may increase coastal exposure to antibiotic-resistant Harveyi clade Vibrio.\n\nID: 42554071\nTitle: Molecular Backbone Regulation for Enhanced Ion Retention in Nonvolatile Organic Electrochemical Synaptic Transistors.\nAbstract: Organic electrochemical synaptic transistors (OESTs) provide a promising platform for high-performance neuromorphic devices by enabling ion-driven synaptic weight modulation. Most studies have primarily adopted interface-dominant organic strategies to maintain ion doping, while relatively overlooking the intrinsic molecular-level effects on ion implantation. From a molecular structure control perspective, the role of the polymer backbone remains poorly understood, resulting in an unclear correlation between ion dynamics and thin-film microstructure. In this study, we present a molecular design strategy that enhances ion doping stability through polymer backbone regulation. This strategy induces a favorable thin-film microstructure that promotes dense packing and enhanced crystallinity, enabling efficient ion implantation and transport. These structural characteristics have effectively emulated enhanced nonvolatile memory properties and biological synaptic operations, including long-term potentiation and depression. Furthermore, high accuracy was achieved in artificial neural network (ANN) simulations using the MNIST dataset. These results suggest that molecular-level control of thin-film microstructure governs the synaptic performance of OESTs, providing valuable molecular design guidelines for high-performance neuromorphic devices.\n\nID: 42554056\nTitle: EpiCure (Epithelial Curation): a versatile and handy tool for curation of epithelial segmentation.\nAbstract: Despite advances in deep-learning and bioimage analysis, manual curation of segmented and tracking data is still required to extract accurate quantitative single cell temporal information from large tissue/embryo movies. However, very few tools address specifically this challenge. We present here EpiCure (Epithelial Curation), a versatile tool designed to streamline and accelerate manual curation of segmentation and tracking in 2D movies of large epithelial tissues. EpiCure uses temporal information and morphometric parameters to automatically identify segmentation and tracking errors and provides user-friendly tools to correct them. It focuses on ergonomics and offers visualization options to help navigating in movies covering a large number of cells, speeding up the detection and curation of errors. EpiCure is highly interoperable, supports input from diverse segmentation tools and includes multiple export filters enabling seamless integration with downstream analysis pipelines. Using movies from several animal models, we highlight the importance of curating cell segmentation and tracking for accurate downstream analysis, and how EpiCure helps extracting single cell dynamics and detecting cellular events on large dataset.\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: 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: 42553425\nTitle: Sweet apple e-cigarette vapor differentially modulates the mTOR pathway in oral squamous cell carcinoma cell lines.\nAbstract: Oral squamous cell carcinoma (OSCC) is the most common head and neck cancer and is associated with high recurrence and poor prognosis. This study investigated the effects of Sweet Apple e-cigarette vapor extract (Apple EVE), with and without nicotine, on mTOR pathway activation in OSCC cell lines Ca9-22 and Cal 27. Cells were exposed for 6 hours to 10% Apple EVE generated from \"Reds Apple Juice\" in the presence or absence of nicotine (6 mg), with untreated cells as controls. Phosphorylation levels of mTOR pathway components (p-mTOR, p-p70S6K, p-4EBP1, and p-AKT) were assessed by Western blot and quantified by densitometry normalized to \u03b2-actin. Cell invasion was evaluated using a Matrigel-coated real-time xCELLigence assay. Data were analyzed using the Mann-Whitney U test (p < 0.05). Nicotine-containing Apple EVE significantly increased p-mTOR in both cell lines. In Ca9-22 cells, it decreased p-p70S6K and p-4EBP1, whereas in Cal 27 cells it increased p-AKT. Apple EVE without nicotine induced more modest and variable effects. No significant changes in cell invasion were observed in either cell line. Apple EVE, particularly when combined with nicotine, differentially modulates mTOR signaling in a cell line-specific manner in OSCC cells. These findings highlight the complex effects of flavored e-cigarette aerosols on cancer-related pathways and warrant further investigation into the functional consequences of these early signaling changes.\n\nID: 42553297\nTitle: Targeting mitochondria for the treatment of neurodegenerative diseases.\nAbstract: Mitochondria are central regulators of cellular metabolism, redox balance, calcium signaling, and cell survival, making them essential for neuronal function. Because neurons rely heavily on mitochondrial oxidative phosphorylation to meet their high energetic demands, mitochondrial dysfunction has emerged as a key pathogenic driver in major neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. Defects in mitochondrial bioenergetics, excessive reactive oxygen species production, impaired mitochondrial dynamics, disrupted mitophagy, and dysregulated calcium handling collectively contribute to neuronal damage, synaptic dysfunction, and neuroinflammation. These insights have prompted growing interest in therapeutic strategies that directly target mitochondria to restore organelle homeostasis. Recent advances in chemical biology and nanomedicine have enabled the development of mitochondria-targeted ligands, peptide-based targeting systems, and carrier or nanotechnology-enabled delivery platforms designed to overcome biological barriers and selectively deliver therapeutic cargos to mitochondria within the central nervous system. In this Review, we summarize mitochondrial pathological mechanisms in neurodegenerative diseases and discuss emerging mitochondria-targeted therapeutic strategies, highlighting delivery technologies, therapeutic modalities, and translational challenges. Although most strategies remain at the preclinical or proof-of-principle stage, these advances are beginning to shape a conceptual framework for precision mitochondrial medicine, with the longer-term goal of developing disease-modifying interventions for neurodegenerative disorders.\n\nID: 42552750\nTitle: Plasma p-tau217 detects Alzheimer's disease co-pathology in cerebral amyloid angiopathy: Comparison to CSF biomarkers in the ANGMAR cohort.\nAbstract: Cerebral amyloid angiopathy (CAA) frequently co-occurs with Alzheimer's disease (AD), generating mixed vascular-neurodegenerative phenotypes. Plasma phosphorylated tau (p-tau)217 is a robust biomarker of AD, but its performance in CAA remains unclear. We studied 231 participants, including 50 patients with CAA (Boston v2.0), 154 with AD, and 27 cognitively unimpaired controls. Plasma p-tau217 was measured on an automated platform and compared to cerebrospinal fluid (CSF)-defined AD status. Associations with magnetic resonance imaging (MRI) markers of CAA burden were evaluated. Among CAA participants, 29 met CSF criteria for AD co-pathology. Plasma p-tau217 discriminated CAA patients with and without AD co-pathology (area under the curve\u00a0=\u00a00.920) and showed no association with MRI markers of CAA burden. Predefined cut-offs (\u2265 0.27 pg/mL and \u2265 0.34 pg/mL) yielded high accuracy for identifying AD co-pathology within CAA. Plasma p-tau217 shows high diagnostic accuracy for identifying AD co-pathology in CAA and is not associated with MRI markers of CAA burden, supporting its specificity for AD-related pathology.\n\nID: 42552582\nTitle: AHG1-AFP interaction as a regulatory node in the ABA response during seed germination.\nAbstract: Seed dormancy and germination are tightly regulated by complex signaling networks that integrate internal and external cues, including the endogenous phytohormone abscisic acid (ABA). ABA HYPERSENSITIVE GERMINATION 1 (AHG1), a group A type 2C protein phosphatase (PP2C), is thought to modulate the activity of transcription factors such as ABA INSENSITIVE 5 (ABI5) in seeds and during germination. AHG1 is regulated by DELAY OF GERMINATION 1 (DOG1), a key regulator of seed dormancy, through physical interaction. We previously reported that AHG1 also interacts with ABI FIVE BINDING PROTEIN 2 (AFP2), a member of the AFP family; however, the molecular basis of AHG1-AFP coordination has remained unclear. In this study, we show that AHG1 interacts with all AFP family members and that AFP3 binds AHG1 and ABI5 through adjacent but distinct amino acid residues within its C-domain, allowing simultaneous association with both proteins. In addition, AHG1 modulates the phosphorylation status of AFP3 at Ser60 in a DOG1-dependent manner, suggesting that DOG1-AHG1 regulates AFP3 post-translationally. Transcriptomic analyses of AHG1- or AFP3-overexpressing lines revealed that these factors are associated with the regulation of a shared set of ABA-responsive genes, including AFPs, and that AFP3 overexpression is predominantly associated with altered expression of genes involved in transcriptional regulation. Large-scale protein interaction analyses showed that AFPs interact with multiple classes of transcription factors, suggesting their involvement in diverse regulatory pathways, including ABA signaling. Together, these findings demonstrate that DOG1 regulates ABI5 function and modulates ABA responses, at least in part, by controlling AHG1-mediated dephosphorylation of AFPs.\n\nID: 42552554\nTitle: Interaction between p-AMPK and RIPK3 reveals a mechanism by which AdipoAI suppresses necroptosis to attenuate periodontitis.\nAbstract: Necroptosis is implicated in the pathogenesis of various inflammatory diseases, including periodontitis. This study aimed to investigate the molecular mechanisms underlying necroptosis in gingival fibroblasts (GFs) and to evaluate the therapeutic potential of AdipoAI, a novel adiponectin receptor agonist, administered locally, along with its regulatory effect on necroptosis. Analysis of single-cell RNA sequencing data using the AUCell scoring system demonstrated significant activation of necroptosis in human periodontal tissues, with GFs identified as the primary cellular target. Integrated approaches, including molecular docking, co-immunoprecipitation, and immunofluorescence, revealed a physical association between phosphorylated AMP-activated protein kinase (p-AMPK) and Receptor-Interacting Protein Kinase 3 (RIPK3). Additionally, we established a mouse model of experimental periodontitis and an LPS/AZD'5582/z-VAD-fmk (LAZ)-induced necroptosis model in hGFs. Techniques including Western blotting, flow cytometry, and transmission electron microscopy were employed to evaluate the effects of AdipoAI administration. Local administration of AdipoAI significantly alleviated gingival inflammation and alveolar bone resorption in mice. In hGFs, AdipoAI effectively suppressed the activation of p-RIPK3 and phosphorylated Mixed Lineage Kinase Domain-Like protein (p-MLKL) and reduced the production of inflammatory factors. Mechanistic investigations revealed that AdipoAI binds to Adiponectin Receptor 1 (AdipoR1) - adiponectin receptor interacting protein (APPL1) to activate the AMPK signaling pathway, facilitating the association between p-AMPK and RIPK3, thereby inhibiting RIPK3/MLKL-mediated necroptosis. This study is the first to unveil a mechanism by which AdipoAI suppresses GF necroptosis through p-AMPK-mediated RIPK3 regulation, offering new strategic insights and experimental evidence for targeted periodontitis therapy.\n\nID: 42552539\nTitle: Impaired NADH-linked mitochondrial respiration disrupts ventral midbrain neuronal programs in POLG disease.\nAbstract: POLG (DNA polymerase \u03b3 catalytic subunit)-related mitochondrial diseases are among the most severe primary mitochondrial disorders and are characterized by progressive neurodegeneration with prominent dopaminergic involvement. However, the cell type-specific mechanisms linking mitochondrial DNA instability to neuronal vulnerability remain incompletely defined. Using patient-derived midbrain organoids and single-cell RNA sequencing, we investigated how POLG mutations alter mitochondrial and neuronal programs at subtype resolution. We analyzed dopaminergic neuronal populations and ventral midbrain neurons to define disease-associated transcriptional changes. To evaluate therapeutic improvement, POLG organoids were treated chronically with nicotinamide riboside (NR), followed by single-cell transcriptomic profiling and pathway enrichment analysis. POLG mutations induced a coordinated downregulation of genes associated with oxidative phosphorylation and synaptic signaling, particularly in terminally differentiated dopaminergic neurons. This transcriptional alteration involved genes encoding respiratory chain complexes I-V, mitochondrial translation machinery, and ATP synthase components, suggesting disruption of mitochondrial bioenergetic programs at the transcriptomic level. Among dopaminergic subtypes, DA2 neurons and ventral midbrain neurons showed the most pronounced transcriptional alterations, indicating maturation-dependent vulnerability. NR treatment was associated with altered expression of genes involved in oxidative phosphorylation, NADH dehydrogenase activity, respiratory chain assembly, and synaptic pathways. Following NR exposure, dopaminergic subpopulations exhibited changes in cell-type proportions and partial normalization of mitochondrial- and synaptic-related transcriptional programs. These findings identify transcriptional alterations in pathways related to mitochondrial respiration. The data further suggests that modulation of NAD\u207a metabolism is associated with transcriptional changes in mitochondrial and neuronal pathways in this disease context.\n\nID: 42552448\nTitle: CHAF1A promotes RAD51 loading and homologous recombination to drive tumor radioresistance.\nAbstract: Aberrant activation of DNA damage repair (DDR) pathways drives therapeutic resistance in cancer. Although Chromatin Assembly Factor 1 subunit A (CHAF1A) is well characterized for its role in chromatin assembly, its specific function in the DNA damage response has remained poorly defined. Here, we demonstrate that CHAF1A functions as a key regulator of DNA repair and radioresistance. Upon DNA double-strand breaks, CHAF1A is rapidly recruited to damage sites in an ATM-associated manner. Intriguingly, CHAF1A recruitment further enhances ATM phosphorylation and amplifies the DNA damage signal. Consequently, depletion of CHAF1A compromises the efficiency of both homologous recombination (HR) and non-homologous end joining (NHEJ). Mechanistically, CHAF1A governs repair pathway utilization in a cell cycle-dependent manner: during S phase, CHAF1A interacts with RAD51 in collaboration with PCNA, promoting the loading of RAD51 at DNA damage sites; whereas in non-S-phase cells, CHAF1A preferentially promotes the recruitment of KU70 to support NHEJ. Functionally, silencing CHAF1A markedly enhances tumor radiosensitivity, as robustly validated in cell-derived xenograft models and supported by proof-of-concept evidence from a patient-derived xenograft model. Collectively, our findings establish CHAF1A as a pivotal regulator of DNA damage repair in lung cancer models and position it as a promising therapeutic target for overcoming radioresistance. Cancer cells often become resistant to radiotherapy by repairing the DNA damage that radiation causes. In this study, we discovered that a protein called CHAF1A plays a key role in helping cancer cells repair this damage and survive radiation treatment. Using cell experiments, genetic approaches, and mouse tumor models\u2014including tumors derived directly from patients (as a proof-of-concept)\u2014we found that CHAF1A is recruited to sites of DNA damage and activates two major DNA repair pathways depending on the cell cycle stage. When we blocked CHAF1A, cancer cells became much more sensitive to radiation, and tumors shrank significantly more after treatment. These findings suggest that targeting CHAF1A could be a promising new strategy to make radiotherapy more effective for cancer patients, potentially improving treatment outcomes and reducing the chance of cancer returning after radiation therapy.\n\nID: 42552421\nTitle: p75 neurotrophin receptor signaling through the RhoA/ROCK pathway contributes to Tau-mediated neurodegeneration.\nAbstract: Therapeutic development in Alzheimer's Disease (AD) has for the most part been focused on reducing \u03b2-amyloid load. Nevertheless, neurofibrillary tangles (NFTs), produced by aggregation of hyper-phosphorylated Tau protein, correlate with neurodegeneration and cognitive impairment significantly better than amyloid accumulation in AD patients. Here we report that P301S mice, a model of Tau-mediated neurodegeneration, carrying mutant variants of the p75 neurotrophin receptor (p75NTR) deficient in RhoA/ROCK signaling are protected from neurodegeneration and cognitive impairment. Both p75\u2206DD, lacking the death domain, and triple mutant p75KKEA, unable to interact with RhoGDI, decreased insoluble Tau species, reduced gliosis, neurodegeneration and synapse loss, and improved spatial learning and memory in P301S mice. Intriguingly, p75C259A, a variant unresponsive to neurotrophins but still competent for RhoA signaling induced by myelin-derived ligands, did not afford any neuroprotection. P301S neurons expressing p75\u2206DD or p75KKEA, but not p75C259A, showed reduced phospho-Tau and ROCK and GSK3\u03b2 activity, the two main kinases responsible for Tau phosphorylation. In line with this, treatment with myelin-associated glycoprotein (MAG) enhanced Tau phosphorylation and ROCK activity in P301S neurons expressing wild type p75NTR or p75C259A, but not p75\u2206DD or p75KKEA. Together, these results indicate that p75NTR contributes to AD Tauopathy by enhancing the activity of the RhoA-ROCK pathway.\n\nID: 42552398\nTitle: CDK2-mediated phosphorylation of TRF1 S11 facilitates DNA damage repair by remodeling telomeric chromatin.\nAbstract: Telomeres, coated by the shelterin complex, prevent end-to-end fusions and aberrant DNA repair, yet how telomere-binding proteins coordinate chromatin remodeling during the DNA damage response remains unclear. Here we show that the telomeric protein TRF1 is phosphorylated at serine 11 (S11) in response to DNA double-strand breaks, a modification that enhances cellular resistance to DNA damage. We found that CDK2 directly mediates this phosphorylation, which triggers recruitment of the histone methyltransferase SETD5 to telomeric chromatin. SETD5-dependent deposition of H3 trimethylation at lysine 36 (H3K36me3) promotes local chromatin decompaction and enables subsequent recruitment of the phosphatase PPP4C to dephosphorylate \u03b3H2AX. Loss of TRF1 S11 phosphorylation results in persistent \u03b3H2AX foci, delayed DNA repair, and compromised telomere integrity. Our results define a CDK2-TRF1-SETD5-PPP4C signaling axis that orchestrates phosphorylation-dependent chromatin remodeling at telomeres to ensure genome maintenance during DNA damage stress.\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: 42552377\nTitle: NF-\u03baB-active tumors with matrix CAFs and suppressive immunity as key resistance mechanisms to chemoradiation in rectal cancer.\nAbstract: Most patients with intermediate-to-high-risk locally advanced rectal cancer (LARC) undergo neoadjuvant chemoradiotherapy (NCRT), but reliable predictive biomarkers and well-defined resistance mechanisms remain lacking. We sought biomarkers and molecular mechanisms of NCRT resistance in LARC. We analyzed paired pre-NCRT and post-NCRT biopsies from 26 patients with intermediate-to-high-risk LARC using single-cell RNA sequencing and spatial transcriptomics. Suboptimal responders showed pre-NCRT activation of NF-\u03baB, Wnt, Notch, VEGF, fatty acid, and glutamate pathways, whereas optimal responders exhibited higher cell-cycle and oxidative phosphorylation activity in tumor cells. We experimentally validated that NF-\u03baB inhibition increased chemoradiation sensitivity in colorectal cancer cells. Matrix cancer-associated fibroblasts (mCAFs), characterized by transforming growth factor-\u03b2/extracellular matrix programs, preferentially interacted with NF-\u03baB-high tumor cells and immunosuppressive populations in suboptimal responders, whereas optimal responders exhibited cytotoxic CD8\u207a T cell expansion and conventional dendritic cell-2 enrichment. Spatial mapping revealed mCAF-dense, extracellular matrix-rich niches colocalized with NF-\u03baB-rich tumor-adjacent regions in suboptimal cases. Synchronous rectal and sigmoid tumors from one patient shared suboptimal response with NF-\u03baB activation and mCAF enrichment, suggesting the resistance arises from adaptive epithelial-stromal programs rather than site-specific genetics. Collectively, integrated multicompartment signatures, rather than a single molecular profile, define an NCRT-resistant ecosystem in which tumor NF-\u03baB activation, mCAF enrichment, and immune suppression converge to sustain non-response. In line with this framework, combined NF-\u03baB and oxidative phosphorylation scores stratified response status with robust performance (area under the curve\u2009=\u20090.875) and highlighted resistance-associated therapeutic targets.\n\nID: 42552044\nTitle: The Tau-mitochondria connection and its impact on cellular energy metabolism in Alzheimer's disease.\nAbstract: Alzheimer's disease (AD) is a complex multifactorial neurodegenerative disease process resulting in progressive cognitive deterioration and synaptic dysfunction. The primary research approach in AD has traditionally focused on amyloid- pathology however an increasingly evidence suggests that tau protein is a key mediator of neuronal damage via a direct action on mitochondrial bioenergetics. In this chapter we look at the nature of the tau-mitochondrial interface, and propose a paradigm of tau-induced energy failure in AD. Physiologically tau provides stability to the microtubules and is involved in transport mechanisms within cells. In AD, tau is excessively post-translationally modified hyperphosphorylated and truncated tau species form toxic oligomers that incorrectly translocate to mitochondria, interacting pathologically with critical proteins such as voltage-dependent anion channel 1 (VDAC1) and adenine nucleotide translocase (ANT), impeding the mitochondrial ATP/ADP exchange and reducing oxidative phosphorylation efficiency. Tau also further damages mitochondria by excessive fission, inhibition of axonal transport and Inhibition of mitophagy by interrupting PINK1-Parkin signaling. In turn, the build-up of dysfunctional mitochondria leads to ROS production, mtDNA damage and calcium imbalance creating a vicious cycle toward oxidative stress and tau pathology. At the cellular level they cause an energy depletion of the synapse and at the systems level cause glucose hypometabolism and activation of neuroinflammation. The chapter additionally discusses novel therapeutic approaches that target both tau and mitochondrial abnormalities, namely antisense oligonucleotides (ASO), mitochondria targeted compounds and mitophagy modifiers, stressing that it would be more effective to utilize a cocktail of these inhibitors. As a whole, in the context of decreased bioenergetics, the tau-mitochondria axis is an important factor to consider in the successful treatment of AD.\n\nID: 42552038\nTitle: Metabolic therapeutic targets in Alzheimer's disease.\nAbstract: Alzheimer's disease (AD) is increasingly recognized as a disorder marked not only by amyloid-\u03b2 and tau pathology, but also by profound disturbances in brain energy metabolism that arise early in disease progression. Accumulating evidence indicates that impairments in glucose utilization, insulin signaling, and mitochondrial function precede neurodegeneration and contribute directly to synaptic failure and cognitive decline. This chapter presents a comprehensive overview of Alzheimer's disease through the lens of metabolic dysfunction, highlighting disrupted neuronal bioenergetics as a central and unifying feature of pathogenesis. We examine key metabolic pathways implicated in AD, including cerebral glucose hypometabolism, brain insulin resistance, impaired glycolysis, mitochondrial oxidative phosphorylation deficits, oxidative stress, and altered mitochondrial dynamics. The chapter further discusses therapeutic strategies aimed at restoring metabolic homeostasis, such as insulin sensitization, enhancement of glucose transport, activation of mitochondrial biogenesis, modulation of the electron transport chain, and the use of mitochondria-targeted antioxidants. In parallel, alternative energy approaches-including ketone metabolism, fatty acid oxidation, and pentose phosphate pathway activation-are explored as promising avenues to bypass glucose-dependent energy deficits and reinforce neuronal resilience. Emerging directions in metabolic therapeutics are also highlighted, including combination treatment strategies, NAD+-sirtuin and AMPK signaling, and the expanding role of the gut microbiome-brain metabolism axis. By integrating insights from experimental models, neuroimaging studies, and clinical trials, this chapter underscores the potential of metabolic interventions to enable early, disease-modifying strategies for Alzheimer's disease.\n\nID: 42551770\nTitle: Aging preserves mTORC1 but attenuates JNK-SMAD2L signaling sensitivity to passive stretch-induced tension development in isolated mouse skeletal muscle.\nAbstract: Aging is associated with impaired skeletal muscle mass and function, often attributed to reduced sensitivity to anabolic stimuli. This study investigated whether aging influences the sensitivity of key anabolic signaling pathways to mechanical tension development in skeletal muscle. Using an ex vivo model, extensor digitorum longus (EDL) muscles from adult (16\u202fweeks) and old (24\u202fmonths) female mice were subjected to a standardized passive stretch protocol, with contralateral muscles serving as controls. During recovery, phosphorylation of proteins related to downstream mTORC1 and JNK-SMAD2-L signaling were assessed by immunoblotting. Passive stretch significantly increased phosphorylation of mTORC1-related proteins (mTOR, p70S6K, rpS6, and 4E-BP1) in both adult and old muscles, with no significant differences between age groups, indicating preserved mTORC1 signaling sensitivity to mechanical tension with aging. In contrast, the magnitude of activation of JNK and SMAD2-L signaling was attenuated in old muscles. Our findings reveal that mechanosensitive anabolic signaling is differentially affected by aging. While the intrinsic capacity for mTORC1 activation in response to mechanical tension appears to be preserved with aging, JNK-SMAD2L signaling exhibits reduced mechanosensitivity in aged muscle. This divergence suggests that aging selectively impairs tension-sensitive transcriptional pathways, potentially constraining muscle remodeling despite preserved translational signaling capacity. These findings further imply that age-related deficits observed in vivo may, at least in part, arise from systemic influences rather than intrinsic defects adhering to mTORC1 mechanotransduction.\n\nID: 42551761\nTitle: Integrated network pharmacology, molecular docking, and experimental validation reveal synergistic inhibition of EGFR and PI3K-Akt/JAK2-STAT3 pathways by Esculin and Esculetin to exert anti-colorectal cancer effects.\nAbstract: Colorectal cancer (CRC) remains a global health challenge with limited efficacy of single-target therapies. Esculin and esculetin, the main coumarins of Cortex Fraxini, have been reported to exhibit anti-tumor activities in various cancer cell lines in preclinical studies. This study systematically elucidated the synergistic anti-CRC mechanisms of esculin and esculetin by integrating network pharmacology, bioinformatics, molecular docking, molecular dynamics simulations, and in vitro validation. Network pharmacology predicted 23 overlapping targets, with 10 core proteins significantly enriched in the PI3K-Akt and JAK2-STAT3 pathways, further supported by bioinformatics analysis. Molecular docking and 100-ns molecular dynamics simulations revealed that the esculin-esculetin complex exhibited stronger binding affinity and stable interaction with EGFR compared with each monomer. In vitro, the natural 5:1 combination synergistically inhibited HCT116 and HT29 cell proliferation, suppressed colony formation, migration, and invasion, downregulated the phosphorylation of PI3K, Akt, JAK2 and STAT3, reduced EGFR, TOP1 and CDK2 mRNA, decreased the Bcl-2/Bax ratio, and increased cleaved caspase-3 levels. Quantitative validation by EdU, TUNEL, and flow cytometry confirmed concentration-dependent inhibition of proliferation and induction of apoptosis. These findings demonstrate that esculin and esculetin co-target EGFR and simultaneously block downstream PI3K-Akt and JAK2-STAT3 dual pathways, providing a multi-component, multi-target, multi-pathway synergistic model for C. fraxini-based combination therapy against CRC.\n\nID: 42551679\nTitle: Multifunctional phosphorylated cellulose fabric with enhanced antibacterial and mechanical properties based on a Tris buffering strategy.\nAbstract: Cellulose fabric, as a popular textile material, demands multifunctional modification to broaden its application landscape. Although phosphorylation is an effective approach to endow flame retardancy, it often severely impairs the mechanical properties of cellulose fabric. To address this challenge, a buffering strategy was developed to alleviate this problem in this study while concurrently achieving antibacterial performance. A multifunctional flame-retardant and antibacterial cotton fabric was prepared via a one-pot process using triphosphate acid ATMP as cross-linker, and Tris as both a buffer and an N-halamine precursor. ATMP/Tris-modified fabric exhibited excellent flame retardancy with 31.7% of LOI value and self-extinguishing performance at a low weight gain. Simultaneously, the formation of N-halamine structures conferred superior antibacterial efficacy against E. coli and S. aureus, resulting in complete inactivation within a few minutes without any leaching. The multifunctional fabric exhibited notable washing durability, retaining 83% of its original LOI value after repeated laundering. Notably, in addition to maintaining good flexibility, the incorporation of Tris significantly improved the whiteness, air permeability, and breaking force of cellulose fabric compared to the ATMP-modified counterpart. Therefore, this work offers a simple strategy to enhance the comprehensive properties of flame-retardant multifunctional cellulose fabric.\n\nID: 42551397\nTitle: A chemical toolkit for sulfofucose metabolism: Scalable access to sulfofucose and sulfofucolytic intermediates.\nAbstract: Sulfosugars are organosulfur metabolites that contribute to global sulfur cycling, yet the metabolism of the rarer sulfofucose (SFuc) remains poorly defined, in part due to limited access to authentic standards. We report a scalable five-step synthesis of SFuc from d-galactose, including a uniformly 13C6-labelled isotopologue, and the preparation of defined and putative sulfofucolytic intermediates: sulfogalactonate, sulfotagatose, and sulfotagatose-1-phosphate. Single-crystal X-ray diffraction was crucial for redefining the composition of a tagatose-uronate calcium salt, enabling its reliable use as an intermediate en route to sulfotagatose. Chemoenzymatic phosphorylation furnished sulfotagatose-1-phosphate as a diagnostic standard. Together, these compounds establish a practical chemical toolkit to support pathway assignment, enzyme annotation, and discovery of sulfofucose-degrading bacteria.\n\nID: 42551228\nTitle: Celastrol Ameliorates Systemic Lupus Erythematosus via Restoring Nt5e/ adenosine-dependent Th17/Treg Balance.\nAbstract: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by the breakdown of immune tolerance, systemic inflammation, and multi-organ damage. The imbalance between T helper 17 (Th17) cells and regulatory T (Treg) cells is a key driver in SLE pathogenesis. This study aimed to investigate whether Celastrol (Cel), a natural triterpenoid derived from Tripterygium wilfordii, exerts therapeutic effects in SLE by modulating the Th17/Treg balance, and to explore the underlying molecular mechanism. An in vivo study using the MRL/lpr lupus-prone mouse model was conducted to evaluate therapeutic efficacy, complemented by in vitro assays and molecular analysis to identify target pathways. MRL/lpr mice were orally administered Cel and disease progression was assessed via survival analysis, spleen pathology, autoantibody levels, and renal histopathology. T cell subsets were analyzed using flow cytometry and Western blot. Proteomic profiling was performed to identify potential molecular targets, followed by molecular docking, molecular dynamics simulations, cellular thermal shift assay (CETSA), MicroScale Thermophoresis (MST) and surface plasmon resonance (SPR) to validate target engagement. AB680, a pharmacological inhibitor of ecto-5'-nucleotidase (Nt5e), was used to confirm the functional role of Nt5e. Cel treatment significantly improved survival, reduced splenomegaly, suppressed anti-dsDNA/ANA autoantibody levels, and alleviated renal damage in lupus-prone mice. It promoted Treg cell differentiation and inhibited Th17 polarization, evidenced by increased Foxp3 expression and decreased STAT3 phosphorylation. Nt5e was identified as a direct binding target of Cel. Target engagement was validated through CETSA, MST and SPR, and inhibition of Nt5e by AB680 partially reversed Cel's therapeutic effects, indicating that Nt5e mediates its immunomodulatory action. Our results indicate that Cel restores the Th17/Treg balance via upregulating Nt5e, thereby ameliorating SLE-associated injury. This study thus provides novel insights into the mechanism of Cel treating SLE and underscores the therapeutic potential of targeting Nt5e in SLE.\n\nID: 42550925\nTitle: Identification and Characterization of OsSSIP, a Conserved Immune Peptide Regulating Rice Defense.\nAbstract: Rice blast, caused by Magnaporthe oryzae, poses a serious threat to crop yield; however, immune-active small secreted peptides associated with this disease remain largely uncharacterized. In this study, we identified OsSSIP, a pathogen-inducible peptide in rice. OsSSIP-related similar sequences are widely distributed across angiosperms, while canonical signal peptides appear to be predominantly enriched in Poaceae. Strikingly, the SSP-like homologs of OsSSIP share a conserved N-terminal Tyr-Cys core motif that is essential for OsSSIP activity. Exogenous application of OsSSIP triggered early reactive oxygen species burst, mitogen-activated protein kinase phosphorylation, and defense-related gene expression, while also inhibiting rice root growth. Overexpression of ProOsSSIP enhanced disease resistance accompanied by transient growth suppression, whereas knockout mutants exhibited compromised immunity. A scalable recombinant five-copy OsSSIP protein produced in Escherichia coli effectively mimicked the native peptide. Both OsSSIP and the recombinant protein elicited immune responses not only in rice and wheat but also in dicots such as tomato, soybean, and Arabidopsis. Furthermore, two chemically synthesized peptides derived from non-signal-peptide similar sequences also displayed immune activity. Collectively, this work demonstrates a previously unappreciated role of OsSSIP in rice immunity against blast disease and highlights the potential of exogenous OsSSIP as an eco-friendly immune stimulator for broad-spectrum crop disease control.\n\nID: 42550812\nTitle: Functional CCR7A-mediated cellular responses are negatively modulated by the splice variant CCR7B.\nAbstract: C-C chemokine receptor 7 (CCR7) directs immune cell homing to secondary lymphoid organs and has been implicated in cancer metastasis through its ligands CCL19 and CCL21. Human CCR7 pre-mRNA undergoes alternative splicing to generate five transcripts that encode three protein isoforms with distinct N-termini, termed CCR7A, CCR7B, and CCR7C, but their comparative properties and cross-regulation are not well defined. Here, we cloned these three isoforms and systematically characterized their expression, localization, signaling, and mutual interactions in mammalian cells under both strong (CMV) and weaker (ubiquitin C, UbiC) promoter control to reduce overexpression-related artifacts. Variant-specific RT-PCR revealed that transcripts encoding CCR7A (V1) and CCR7B (V2) predominate in diverse human cell lines, whereas CCR7C-encoding variants (V3-V5) are weakly expressed. EGFP imaging and HiBiT-based assays showed efficient plasma-membrane targeting of CCR7A, partial membrane localization and prominent perinuclear accumulation of CCR7C, and largely cytosolic retention of CCR7B. Under UbiC-driven expression, CCR7A mediated robust CCL19- and CCL21-induced Gi/o and Gq-like activation, intracellular Ca\u00b2\u2009\u207a\u2009mobilization, ERK phosphorylation, GRK3-dependent G\u03b21 recruitment, and \u03b2-arrestin1 binding, whereas CCR7C displayed weaker and mainly CCL19-biased signaling. CCR7B did not respond to either chemokine in any signaling readout and thus behaved as a non-signaling isoform. NanoBiT-based assays and co-immunoprecipitation demonstrated that all three isoforms form homo- and heterodimers, with particularly strong association between CCR7A and CCR7B. Co-expression of CCR7B reduced CCR7A surface expression and markedly attenuated chemokine-induced Ca\u00b2\u2009\u207a\u2009responses, mini-Gi interaction, and \u03b2-arrestin1 recruitment, while confocal microscopy revealed redistribution of CCR7A-EGFP from the plasma membrane to intracellular compartments. Moreover, MDA-MB-231 breast cancer cells, which express CCR7A and CCR7B transcripts, did not migrate toward CCL19 or CCL21 despite preserved motility toward low-serum medium. These findings identify CCR7A as the dominant functional isoform, CCR7C as a weak CCL19-biased receptor with inefficient membrane targeting, and CCR7B as a non-signaling dominant-negative isoform that dampens CCR7A-mediated responses, suggesting that CCR7 splicing fine-tunes chemokine responsiveness in immune and cancer cells.\n\nID: 42550765\nTitle: Functional restoration of immune defects in STAT1 gain-of-function disease following stem cell gene editing.\nAbstract: Germline gain-of-function (GOF) mutations in the signal transducer and activator of transcription 1 (STAT1) gene cause a dominantly inherited inborn error of immunity (IEI) characterized by chronic mucocutaneous candidiasis, autoimmunity, severe opportunistic infections and an increased risk of malignancy. Allogeneic hematopoietic stem cell (HSC) transplantation (HSCT) is curative but is associated with increased risk of morbidity and mortality in STAT1 GOF patients compared to other IEI. To develop a curative, autologous alternative to HSCT, we evaluated gene editing strategies in STAT1 GOF model cell lines, primary T cells, and patient-derived HSCs. Universal and mutation-specific strategies using CRISPR/Cas-mediated homology-directed repair (HDR) were limited by low efficacy (<25%), poor viability, and a lack of allele-specificity. In contrast, adenine base editing corrected the recurrent and highly pathogenic p.T385M mutation with upwards of 90% efficiency in patient T cells and HSCs without significant unintended on- or off-target genomic aberrations. Gene editing functionally restored total STAT1 expression (p<0.0217), STAT1 phosphorylation (p<0.0056), interferon-stimulated gene expression (OAS1; p=0.0005) and improved IL-17 production (p<0.0001). Edited HSCs retained multilineage differentiation capacity and sustained engraftment with persistence of the corrected allele at 16 weeks in humanized immunodeficient mice. These data demonstrate efficient and precise correction of STAT1 GOF mutations by base editing, with maintenance of the correction through long-term engraftment in vivo. This represents the first application of gene editing to correct a dominant gain-of-function mutation causing immunodeficiency, with potential applicability to other genetic disorders associated with heterozygous and gain-of-function mutations.\n\nID: 42550764\nTitle: DRP1-mediated mitochondrial fragmentation is a druggable vulnerability in multiple myeloma.\nAbstract: Mitochondrial dynamics is a key regulator of cellular homeostasis, orchestrating metabolic reprogramming that fuels tumor progression and treatment resistance. In multiple myeloma (MM), however, the functional relevance of mitochondrial remodeling has not been fully defined. Using ultrastructural analyses, we reveal that MM cells display a highly fragmented mitochondrial network, a phenotype further exacerbated in both cell lines and primary MM cells resistant to proteasome inhibitors. Transcriptomic profiling across multiple patient-derived datasets consistently demonstrated upregulation of DNM1L gene, which encodes the mitochondrial fission GTPase DRP1, particularly in relapsed and refractory MM, and revealed a significant association with inferior overall survival. Disrupting mitochondrial fission, either through genetic targeting of DNM1L or pharmacologic inhibition of DRP1 with the selective small molecule inhibitor Drpitor1a, resulted in pronounced mitochondrial dysfunction, impaired oxidative phosphorylation, and potent anti-myeloma activity in vitro, culminating in a hybrid cell death program with a predominant apoptotic component accompanied by ferroptotic features. These effects were recapitulated in vivo in a bortezomib-resistant xenograft model, where either DNM1L depletion or DRP1 inhibition produced similar outcomes. Mechanistically, the transcription factor c-MYC upregulated DNM1L expression, and DRP1-dependent mitochondrial fragmentation sustained MYC-driven oxidative metabolism and lipid synthesis. Altogether, these findings establish aberrant mitochondrial fission as a pathogenic hallmark of MM and highlight DRP1 inhibition as a promising therapeutic approach, especially for relapsed or refractory disease.\n\nID: 42550422\nTitle: Plasma Extracellular Vesicles from Glioblastoma Patients Affect Phenotype and Activate Receptor Tyrosine Kinases in Glioblastoma Cells.\nAbstract: Extracellular vesicles are important transmitters of oncogenic signals between cancer cells. Despite numerous studies of vesicles from model glioblastoma (GBM) cells, information on the effects of vesicles from GBM patients on the development of model GB cells is lacking. In this study, plasma vesicles from healthy donors (HVs) and GBM patients (GVs) were studied to affect expression of differentiation (GFAP and GLT-1) and stemness (CD9 and CD133) markers and expression and activation of the oncogenic receptor tyrosine kinases EGFR and PDGFR\u03b2 in GBM cells (primary GBM011 and model U251 MG cell lines) and rat and human normal astrocytes. GVs reduced GFAP expression in GBM011 cells and stimulated GFAP and CD9 expression in U251 MG cells. GVs increased EGFR and PDGFR\u03b2 expression in both GBM cell lines and enhanced EGFR (Y1086) and PDGFR\u03b2 (Y751) phosphorylation in U251 MG cells. HVs stimulated CD9 expression in U251 MG cells and PDGFR\u03b2 expression in GBM011 cells, but decreased EGFR and PDGFR\u03b2 expression in U251 MG cells. Thus, both GVs and HVs influence the GBM phenotype and receptor tyrosine kinase activation.\n\nID: 42550413\nTitle: Energy metabolism in the kidney and its role in chronic kidney disease.\nAbstract: Chronic kidney disease (CKD) is associated with dysregulated lipid metabolism, particularly in proximal kidney tubules, where fatty acid oxidation serves as the primary energy source. The proximal tubules' reliance on fatty acid oxidation rather than glycolysis underscores their unique metabolic profile, consistent with the absence of key glycolytic enzymes. This dysregulation contributes to maladaptive hypertrophy in CKD, where surviving nephrons exhibit compensatory hypertrophy to maintain kidney function. Here, we focus on two key regulators of lipid metabolism: peroxisome proliferator-activated receptor alpha (PPAR\u03b1) and adenosine monophosphate (AMP)-activated protein kinase (AMPK). Recent multi-omics studies have identified PPAR\u03b1 as an important determinant of proximal tubule cell size and a mediator of compensatory hypertrophy. In CKD models, AMPK activity decreases, impairing cellular responses to energy stress, as indicated by altered AMP/ATP ratios. This defective energy sensing may be exacerbated by uremic metabolites that diminish AMPK function. Unc-51-like autophagy activating kinase 1 (ULK1) has been identified as a regulator of AMPK activity through specific phosphorylation sites that enhance AMP sensitivity. Future research should assess whether targeting these pathways restores metabolic homeostasis and mitigates CKD progression by enhancing AMPK activity and lipid metabolism.\n\nID: 42550401\nTitle: Zinc sulfate modulates the C3H10T1/2 mesenchymal stem cell line secretome and NF\u03baB signaling during inflammation to influence selected immune cell responses.\nAbstract: Zinc (Zn) is essential for immune cell function, while mesenchymal stem cells (MSCs) exert immunomodulatory effects primarily through the secretion of soluble factors. Considering the ability of MSCs and Zn to modulate the immune and inflammatory systems, this study investigated, in vitro, the effects of Zn supplementation on MSC responses to inflammatory stimuli and the subsequent modulation of macrophages and lymphocytes. Using the C3H10T1/2 line as a MSC model, we determined that 1\u00a0\u00b5M ZnSO4 enhanced MSC metabolic activity without affecting viability or cell-cycle distribution, whereas higher concentrations reduced cell viability. Under lipopolysaccharide (LPS) stimulation, Zn inhibited NF\u03baB phosphorylation and increased AMPK phosphorylation, indicating anti-inflammatory and adaptive metabolic responses. Similarly, under TNF-\u03b1 stimulation, Zn also reduced NF\u03baB phosphorylation. Zn supplementation altered MSC secretory profiles, reducing IL-6, IL-10, and nitric oxide (NO) production while increasing TGF-\u03b2 and prostaglandin E2 (PGE2) levels, indicating that Zn modifies MSC-derived soluble factor production under inflammatory conditions. Conditioned media from Zn-treated MSCs attenuated IL-6 and IL-12 production in macrophages, indicating a reduced pro-inflammatory cytokine response, whereas lymphocyte responses were unaffected. Importantly, Zn modulation of cytokine production was observed under LPS stimulation but not under TNF-\u03b1 exposure, suggesting that Zn preferentially interferes with signaling pathways triggered by microbial stimuli. Overall, this study provides mechanistic insight into how Zn affects the secretory profile and inflammatory signaling pathways of C3H10T1/2 cells. These findings support further studies in primary MSCs to determine whether Zn supplementation may represent a useful strategy for modulating MSC-mediated immune regulation in therapeutic settings.\n\nID: 42550324\nTitle: Correction: Succinate and lactate produced as conserved biomarkers through chronic and transient substrate-level phosphorylation: from microorganisms to cancer.\nAbstract: \n\nID: 42550257\nTitle: Environmental enrichment modulates parvalbumin interneuron deficits and plasticity-related signaling after early postnatal NMDA receptor hypofunction in male rat visual cortex.\nAbstract: Sensory processing deficits in schizophrenia have been linked to dysfunction of cortical inhibitory interneurons, particularly parvalbumin-expressing (PV+) populations. NMDA receptor hypofunction during development is known to disrupt interneuron maturation, but its long-term impact on visual cortex circuitry and the potential for experience-dependent modulation in adulthood remain unclear. Here, we examined the effects of early postnatal N-methyl-D-aspartate (NMDA) receptor blockade with MK-801 on the number of PV+\u2009and somatostatin-expressing (SST+) interneurons in the primary visual cortex (V1) and assessed whether environmental enrichment (EE) in adulthood modulates these alterations by analyzing molecular changes using Western blot. Male Long-Evans rats received MK-801 (0.5\u00a0mg/kg) or saline from postnatal day 10-20, followed by EE exposure from P55-73. Stereological analyses revealed a marked reduction of PV-immunoreactive cells in layers II/III and IV, while SST+ populations were largely preserved. EE increased the number of PV-immunoreactive cells across groups, and modestly enhanced SST+ cells in layer IV, although no treatment \u00d7 housing interaction was detected, indicating a general enrichment-related effect. At the molecular level, MK-801 reduced expression of the NMDA receptor subunit NR1 and increased Akt phosphorylation, whereas EE enhanced PSD95 expression, ERK phosphorylation, and GABAA \u03b22/3 subunit levels, without increasing NR1 levels. These findings indicate that early NMDA receptor hypofunction induces long-lasting, subtype-specific alterations in inhibitory circuitry in V1. EE in adulthood engages molecular pathways associated with synaptic plasticity and modulates interneuron immunoreactivity, suggesting that inhibitory circuits retain some capacity for experience-dependent remodeling despite persistent receptor-level deficits.\n\nID: 42549985\nTitle: Inherent \u03b1-Helix Around Tyrosine 183 in the Intrinsically Disordered Tail of the Multi-Site Docking Platform Gab1.\nAbstract: The biological function of intrinsically disordered proteins is frequently coupled to short linear motifs, which serve as protein binding sites. This goes often hand in hand with local transient structural element formation. The intrinsically disordered C-terminal region of the multi-site docking protein Grb2-associated binder 1 (Gab1) contains several well-characterized phosphotyrosine pairs, whereas the Tyr162 and Tyr183 epitopes have remained practically unstudied. Here, we combine computational prediction, circular dichroism and high-resolution NMR spectroscopy (chemical shift and relaxation analyses) to structurally characterize a Gab1 fragment containing residues 142-203. The here determined NMR structure shows an \u03b1-helix for residues Pro180 to Ile187, with Tyr183 positioned centrally, while all other parts of the fragment, including the region around Tyr162, are disordered. This helical conformation is maintained upon changes in pH, variation of peptide lengths and phosphorylation status. The inherent helix around Tyr183 distinguishes the local structural environment of Tyr162 and Tyr183 within the intrinsically disordered Gab1 tail and provides a structural framework for future studies on interaction partners and potential functional roles of this structural motif.\n\nID: 42549799\nTitle: Hepatocyte MLKL drives obesity-driven hepatocellular carcinoma progression via mitochondrial dysfunction independent of necroptosis in MASLD.\nAbstract: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a leading cause of hepatocellular carcinoma (HCC), particularly in obesity, yet mechanisms linking hepatocyte dysfunction to tumorigenesis remain unclear. Mixed lineage kinase domain-like protein (MLKL), the effector of necroptosis, is elevated in MASLD, but its hepatocyte-intrinsic role in obesity-driven MASLD-HCC is unknown. Using a long-term Western diet (WD)-induced MASLD-HCC model in hepatocyte-specific MLKL knockout (MlklHepKO) mice, we defined MLKL's hepatocyte-intrinsic function. WD increased hepatocyte MLKL protein expression without detectable necroptosis activation, indicating a necroptosis-independent role. MLKL deficiency did not alter WD-induced inflammation, fibrosis, or liver injury but increased hepatic lipid accumulation while reducing lipotoxic lipid species and preserving mitochondrial function. WD-fed MlklHepKO mice developed fewer and smaller tumors with reduced incidence, multiplicity, proliferation, and stemness. Transcriptomic analysis revealed upregulation of mitochondrial oxidative phosphorylation pathways in MlklHepKO livers. WD suppressed the mitochondrial fusion protein and tumor suppressor MFN2, whereas MLKL deficiency restored MFN2 expression post-translationally. In HCC cells, MLKL deletion reduced proliferation, improved mitochondrial respiration, and decreased glycolysis; these effects were reversed by MFN2 deletion. MLKL localized to nuclear and mitochondrial compartments, consistent with organelle-intrinsic functions. The human MLKL inhibitor necrosulfonamide (NSA) suppressed HepG2 xenograft growth, and elevated MLKL expression in human HCC correlated with poorer overall survival. Hepatocyte MLKL promotes MASLD-associated HCC through a non-necroptotic mechanism involving MFN2 suppression, impaired mitochondrial function, and increased tumor proliferation and stemness. These findings identify MLKL as a potential therapeutic target in MASLD-associated HCC.\n\nID: 42549686\nTitle: LMTK2 Inhibits Metastasis of Colon Cancer Cells Through p38 MAPK.\nAbstract: Metastasis is the leading cause of death in patients with colon cancer. Although Lemur tyrosine kinase 2 (LMTK2) has been implicated in cell proliferation, its role in metastasis is unclear. In this study, we found that low LMTK2 expression correlates with metastatic status (M1 stage) and a higher metastasis rate in a large colon cancer cohort. Additionally, LMTK2 expression is negatively correlated with pro-metastatic genes and positively correlated with metastasis suppressors, a pattern that was validated in 50 clinical colon cancer specimens. Functional studies demonstrated that LMTK2 potently inhibits cell migration and invasion in\u00a0vitro and suppresses metastasis in a mouse model in\u00a0vivo. Mechanistically, LMTK2 disrupts the interaction between p38 MAPK and its upstream kinases MKK3/6, thereby inhibiting p38 activation and subsequent cell migration and invasion. Notably, the kinase-deficient LMTK2 (K168R) mutant also suppresses p38 phosphorylation and impedes cell migration and invasion. These results uncover a novel anti-metastatic function of LMTK2, which acts through inhibition of p38 independently of its kinase activity.\n\nID: 42549431\nTitle: Injectable Artificial Photoreceptors: STEM Cell Functional Integration and in vivo Electrophysiological Validation in Retinal Degeneration Models.\nAbstract: In this study, an injectable artificial photoreceptor (APR) is engineered to transduce visible light into cell-level electrical stimulation via localized surface plasmon resonance to restore vision in patients with vision loss. APRs comprise gold nanoparticles (AuNPs) with poly (vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) dielectric coating and are integrated onto a high-permittivity barium titanate core. This yields a stable nanocomposite with maximum absorbance in the green band at around 525\u2009nm. APRs were evaluated in vitro using human pluripotent stem cell-derived retinal ganglion cells (hRGCs) exposed to light stimulation. In vivo efficacy was assessed following intravitreal injection of APRs in the rd1 mouse model, and functional outcomes were evaluated using visual evoked potentials (VEPs) and pupillary light reflex (PLR) testing. In vitro, optical stimulation of hRGCs in the presence of APRs increased multielectrode-array (MEA) firing. This shifted cellular metabolism toward mitochondrial oxidative phosphorylation without inducing apoptosis. In rd1 mice, intravitreal APR delivery produced robust restoration of light-evoked retinal activity in explant MEAs and improved functional readouts in vivo, validated by enhanced PLR and increased N1 amplitudes on flash VEPs. This study demonstrates that a fully injectable nanophotonic prosthesis can restore light responsiveness in a retinal degeneration model without the need for implanted electronics or genetic modification.\n\nID: 42549418\nTitle: Synthesis and anti-inflammatory activity evaluation of pterostilbene-based phthalide derivatives.\nAbstract: In recent years, phthalide and pterostilbene have been widely studied as important natural products. In this project, we designed and synthesized two series (a total of 37) of pterostilbene-phthalide heterozygous derivatives. The activities of all synthesized compounds were evaluated by LPS-induced RAW264.7 macrophages. The results demonstrated that all phthalide-phthalimide hybrids effectively inhibited NO release, with F22 showing the most significant inhibitory effect. Moreover, MTT assays confirmed that none of the compounds showed significant cytotoxicity. Mechanistic studies revealed that F22 significantly inhibited inducible iNOS and COX-2 expression and effectively suppressed the phosphorylation of I\u03baB and p65 proteins in the NF-\u03baB signaling pathway as well as phosphorylation of p-JNK, p-p38, and p-ERK in the MAPK pathway. These findings indicate that the anti-inflammatory activity of F22 is mediated through inhibition of the NF-\u03baB/MAPK signaling pathways. In vivo studies demonstrated that F22 effectively alleviated dextran sulfate sodium salt-induced acute colitis in mice. In summary, F22 exhibits promising potential as an anti-inflammatory agent and holds significant value for the development of novel anti-inflammatory drugs.\n\nID: 42549075\nTitle: Genome-wide characterization of the CIPK gene family in mung bean and functional validation of VrCIPK5 in drought stress response.\nAbstract: Calcineurin B-like protein-interacting protein kinases (CIPKs) act as core regulators in plant calcium (Ca2+) signaling pathways and mediate abiotic stress adaptation. Mung bean (Vigna radiata L.) is an economically important legume that is widely grown in arid and semi-arid regions; however, drought stress significantly reduces its yield and quality. Despite this, genome-wide identification and functional analysis of the CIPK gene family have not been reported in mung bean, limiting our understanding of drought-resistance mechanisms and stress-tolerant variety breeding. In this study, 23 VrCIPK genes were identified from the mung bean genome, which were unevenly distributed across 6 chromosomes, with the remaining genes located on scaffolds. Phylogenetic analysis classified the VrCIPK family into five groups (A-E), characterized by high structural conservation of the N-terminal kinase and the C-terminal NAF/FISL regulatory domains. Gene Ontology annotation further indicated their conserved involvement in protein phosphorylation and calcium signal transduction. Collinearity analysis revealed that CIPK genes in mung bean and other species have relatively conserved evolutionary patterns. Segmental duplication contributed to VrCIPK gene family expansion, and most duplicated gene pairs underwent purifying selection during evolution. Gene structure and motif analyses showed that VrCIPK genes within the same group shared conserved structural features. Cis-acting element profiling revealed abundant hormone- and stress-responsive elements in VrCIPK promoters, indicating diverse transcriptional regulatory potential. Transcriptomics analysis and quantitative real-time PCR demonstrated that VrCIPK5 was significantly induced under drought stress. The heterologous overexpression of VrCIPK5 in tobacco conferred enhanced drought tolerance by promoting proline accumulation, increasing antioxidant enzyme (superoxide dismutase, peroxidase, and catalase) activities, accelerating stomatal closure, and upregulating downstream stress-responsive genes (NtSOD, NtCAT, NtP5CS1, NtLEA5, and NtRD29A), thus alleviating reactive oxygen species overaccumulation and membrane lipid peroxidation. Collectively, these findings fill the research gap in the genome-wide identification and functional analysis of the CIPK gene family in mung bean and provide key genetic resources and theoretical support for breeding drought-resistant mung bean varieties.\n\nID: 42549063\nTitle: Functional polysaccharide-based biosorbents as efficient green alternatives for radionuclide removal from radioactive waste: a review.\nAbstract: Radioactive waste presents persistent environmental and health challenges, creating an urgent need for remediation materials that are not only effective and selective but also sustainable. Conventional treatment methods often suffer from high cost, limited selectivity, and secondary waste generation, prompting growing interest in biopolymer-based alternatives. Polysaccharides including chitosan, alginate, cellulose, starch, carrageenan, and pectin have emerged as highly promising platforms for radioactive ion removal due to their natural abundance, biodegradability, and rich surface chemistry. Their functional groups (hydroxyl, amino, carboxyl, and sulfate) enable strong coordination with diverse radionuclides, including actinides, fission products, and activation products. This review critically examines the structure-property relationships governing radionuclide adsorption by polysaccharide-based materials, with particular attention to chemical modification strategies, including grafting, cross-linking, phosphorylation, and hybridization. These approaches significantly enhance adsorption capacity, selectivity, and stability, with reported Uranium(vi) uptake exceeding 600 mg g-1 for engineered systems. Advanced composites, including polysaccharide-polysaccharide hybrids and polysaccharide-based metal-organic frameworks, are highlighted for their potential in treating complex radioactive waste streams. Future perspectives focus on multifunctional material design, regeneration efficiency, and scalability to advance polysaccharide-based adsorbents toward practical nuclear waste management applications.\n\nID: 42548835\nTitle: HAT regimen attenuates NF-\u03bab-driven megakaryocyte apoptosis and neuronal cell death in sepsis: convergent mechanisms protecting thrombocytopenia and cognitive function.\nAbstract: Sepsis-induced NF-\u03baB hyperactivation drives two devastating cell death cascades: megakaryocyte apoptosis causing thrombocytopenia (incidence 35%-59%), and neuronal apoptosis with microglial-mediated neuroinflammation causing cognitive dysfunction in up to 70% of survivors. Mechanistically, NF-\u03baB-driven upregulation of pro-apoptotic mediators (cleaved caspase-3, cytochrome c release) impairs megakaryopoiesis while simultaneously inducing hippocampal neuronal death and synaptic loss. The HAT regimen (hydrocortisone, ascorbic acid, and thiamine) modulates complementary nodes of this NF-\u03baB/apoptosis axis, yet its cell-autonomous mechanisms of protecting megakaryocytes and neurons from sepsis-induced programmed cell death remain uncharacterized. We employed a multi-level translational approach to interrogate HAT-mediated cell survival mechanisms. A retrospective cohort of 184 propensity score-matched sepsis patients with thrombocytopenia provided clinical validation. Mechanistic studies used cecal ligation and puncture (CLP) in C57BL/6 mice, with cell death profiling (TUNEL, cleaved caspase-3, Annexin V), blood-brain barrier integrity assays, and synaptic protein quantification. In vitro apoptosis and proliferation assays used MEG-01 megakaryoblasts as a preliminary screening platform (note: MEG-01 harbors BCR-ABL, which constitutively elevates baseline NF-\u03baB activity; primary mechanistic conclusions are grounded in the CD34+ primary system) and primary CD34+ hematopoietic stem cells, BV-2 microglia, and primary hippocampal neurons, combined with NF-\u03baB pathway dissection (p65 nuclear translocation, IKK phosphorylation, I\u03baB\u03b1 dynamics), genetic validation by p65 siRNA knockdown, and transcriptomic/proteomic profiling, delineated component-specific pro-survival mechanisms. Pharmacological synergy was formally quantified by Chou-Talalay CI analysis. E-value sensitivity analyses were applied to primary clinical endpoints. HAT synergistically suppressed NF-\u03baB p65 nuclear translocation by 52% in megakaryoblasts and 54% in hippocampal tissue, reducing pro-inflammatory cytokines by 35%-42%. In megakaryocytes, HAT inhibited apoptosis by 48.6% and enhanced proplatelet formation by 52.4%, corresponding to accelerated platelet recovery (78.5% vs. 42.3% increase at day 7, P < 0.001) and reduced 28-day mortality (22.8% vs. 34.8%, P = 0.038) in patients. In the CLP model, HAT reduced hippocampal neuronal apoptosis (TUNEL+ cells -54.8%; cleaved caspase-3+ neurons -58.2%), attenuated microglial activation (Iba-1+ cells -48.6%), preserved blood-brain barrier integrity (Evans blue extravasation -62.4%), and maintained synaptic protein expression (PSD-95 + 42.6%; synaptophysin +38.4%), translating to significant cognitive and psychological benefit in survivors. Each HAT component contributed distinct anti-apoptotic mechanisms-hydrocortisone suppressed p65 translocation, ascorbic acid blocked ROS-mediated IKK activation, and thiamine restored mitochondrial membrane potential-producing formally synergistic (Chou-Talalay CI = 0.61 in megakaryocytes and CI = 0.58 in hippocampal neurons, both <1.0) pro-survival effects exceeding individual component efficacy. HAT group membership remained an independent predictor of preserved cognition on multivariable regression adjusting for ventilation duration and ICU stay (adjusted OR 0.38, 95% CI 0.18-0.79, P = 0.009). HAT therapy protects two distinct cell populations-megakaryocytes and hippocampal neurons-from sepsis-induced programmed cell death through convergent, component-specific suppression of NF-\u03baB-driven apoptotic signaling. These mechanistic findings reframe HAT as a broad-spectrum anti-apoptotic intervention, providing a cellular and molecular rationale consistent with its dual clinical benefit against thrombocytopenia and cognitive dysfunction in sepsis. Given the null findings of major HAT RCTs in unselected populations, these mechanisms particularly support biomarker-enriched trial designs.\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- \"LBR_phosphorylation_sites\": Identify specific phosphorylation sites on LBR that increase in occupancy following lamin A overexpression in TKO cells.\n- \"LBR_A_type_lamin_interaction\": Determine if lamin A directly phosphorylates LBR or recruits specific kinases to the nuclear envelope to induce displacement.\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 The proposed experimental framework centers on two primary research threads regarding nuclear envelope dynamics. Run 1 focuses on the biochemical validation of Lamin B1 (phospho T575) through immunoblotting and immunofluorescence in LBR-deficient models [ID: Run1_Eval1]. Conversely, Run 2 shifts the analytical scope toward the LBR-Lamin A interaction, utilizing mass spectrometry and phospho-mimetic mutagenesis to investigate potential displacement mechanisms [ID: Run2_Eval1].
Significant evidentiary gaps remain, as there is currently no data provided confirming the baseline kinetics of these phosphorylation events or the specific inhibitory pathways linked to T575, which are cited as necessary for verification.
"
},
{
"type": "comparison_matrix",
"title": "Experimental Strategy Comparison",
"headers": [
"Research Focus",
"Methodological Approach",
"Primary Objective"
],
"rows": [
[
"Lamin B1 (T575)",
"Western Blot / IF Microscopy",
"Nuclear Envelope Localization"
],
[
"LBR-Lamin A",
"Mass Spectrometry / Mutagenesis",
"Displacement Mechanism"
]
]
},
{
"type": "bottlenecks",
"title": "Key Evidence Gaps and Requirements",
"content": [
"Medium: Absence of established baseline kinetics for T575 phosphorylation.",
"Medium: Lack of identified specific kinase inhibitors for T575 validation.",
"Strong: Missing experimental confirmation on whether phosphorylation is sufficient for displacement."
]
},
{
"type": "data_bar_chart",
"title": "Distribution of Experimental Modalities",
"xAxisLabel": "Methodology",
"data": [
{
"label": "Proteomics",
"value": 1
},
{
"label": "Imaging/IF",
"value": 1
},
{
"label": "Blotting",
"value": 1
},
{
"label": "Mutagenesis",
"value": 1
}
]
}
]
}
},
{
"id": "mvc_dp_suggested_studies_1785955048424",
"title": "Suggested Studies Report",
"plan": {
"title": "SUGGESTED STUDIES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Evaluation Scorecard"
},
{
"type": "synthesis",
"title": "Clinical Synthesis of Proposed Research Trajectories",
"content": "The proposed research tracks coalesce around two central themes: nuclear envelope (NE) remodeling and the regulatory role of Lamin A/LBR phosphorylation. Research from Run 1 focuses on mapping Lamin B1 phosphorylation sites during envelope remodeling and comparing phosphospecific staining across cellular conditions [ID: Run1_Eval1]. Research from Run 2 extends this by investigating the kinases recruited by Lamin A that modulate LBR, specifically questioning how these phosphorylation states impact LBR binding affinities to heterochromatin [ID: Run2_Eval1]. A significant gap exists in empirical data confirming the cross-talk between Lamin B1 site-specific phosphorylation and LBR-mediated chromatin recruitment."
},
{
"type": "logic_network",
"title": "Research Dependency Pathways"
},
{
"type": "bottlenecks",
"title": "Literature Gaps and Missing Evidence"
},
{
"type": "comparison_matrix",
"title": "Comparison of Proposed Study Focus",
"headers": [
"Focus Area",
"Primary Target",
"Mechanistic Goal"
],
"rows": [
[
"Run 1",
"Lamin B1",
"Nuclear Envelope Remodeling"
],
[
"Run 2",
"LBR / Lamin A",
"Chromatin Interaction Dynamics"
]
]
},
{
"type": "data_bar_chart",
"title": "Proposed Study Distribution",
"xAxisLabel": "Research Evaluation Cycle",
"data": [
{
"label": "Run 1 (Structural Focus)",
"value": 2
},
{
"label": "Run 2 (Regulatory Focus)",
"value": 2
}
]
}
]
}
},
{
"id": "mvc_dp_swansons_literature_based_discovery_candidates_1785955061537",
"title": "Swansons Literature Based Discovery Candidates Report",
"plan": {
"title": "SWANSONS LITERATURE BASED DISCOVERY CANDIDATES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Discovery Metrics & Signal Integrity"
},
{
"type": "synthesis",
"title": "Executive Analysis of LBR Regulatory Pathways",
"content": "Analysis of literature-based discovery candidates reveals a dual-modality regulatory framework for the Lamin B receptor (LBR). Run 1 identifies a potential role for hydroxylated cholesterol derivatives in modulating HDAC1-mediated chromatin compaction via MTOR-linked phosphorylation [ID: 32013726, 41757046]. Run 2 highlights LBR phosphorylation as a mechanical response mechanism, where LBR mobility facilitates nuclear tension recovery during cellular confinement [ID: 41706894, 41561380]. Both hypotheses converge on the 'Intersecting Bridge' of LBR phosphorylation, though they diverge in biological output: chromatin compaction vs. mechanical tension adaptation."
},
{
"type": "logic_network",
"title": "Hypothesis Pathway Convergence"
},
{
"type": "comparison_matrix",
"title": "Discovery Hypothesis Comparison",
"headers": [
"Parameter",
"Hypothesis 1: Chromatin",
"Hypothesis 2: Mechanics"
],
"rows": [
[
"Originator",
"Cholesterol/HDAC1",
"LBR/Nuclear Fragility"
],
[
"Target",
"LBR/Chromatin Compaction",
"LBR/Tension Recovery"
],
[
"Mediator",
"MTOR Signaling",
"Phosphorylation State"
]
]
},
{
"type": "bibliography",
"title": "Verified Literature References"
}
]
}
},
{
"id": "mvc_dp_contradictions_between_evidences_1785955074948",
"title": "Contradictions Between Evidences Report",
"plan": {
"title": "CONTRADICTIONS BETWEEN EVIDENCES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Evidence Integrity Metrics"
},
{
"type": "synthesis",
"title": "Executive Analysis",
"content": "A comprehensive review of literature regarding Lamin B1 phosphorylation sites was conducted across multiple evaluation runs. In both Run1 and Run2, zero instances of contradictory evidence or data conflicts were identified within the provided context. The findings indicate a high degree of internal consistency regarding the current documentation state, characterized by an absence of adversarial claims. Data gaps remain regarding the specific phosphorylation mechanisms due to the null result in identified contradictions."
},
{
"type": "gap_distribution",
"title": "Literature Gap Assessment",
"data": [
{
"label": "None Identified",
"value": 100
}
]
},
{
"type": "comparison_matrix",
"title": "Run-by-Run Conflict Status",
"headers": [
"Evaluation Phase",
"Contradictions Detected"
],
"rows": [
[
"Run1",
"None"
],
[
"Run2",
"None"
]
]
}
]
}
},
{
"id": "mvc_dp_repurposed_solutions_1785955089722",
"title": "Repurposed Solutions Report",
"plan": {
"title": "REPURPOSED SOLUTIONS : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity Metrics"
},
{
"type": "synthesis",
"title": "Executive Summary: Repurposed Solutions",
"content": "The analysis of 'Repurposed Solutions' highlights two distinct therapeutic strategies targeting nuclear stability [ID: Run1_Eval1_synthesis, Run2_Eval1_synthesis]. Current data points suggest leveraging cholesterol derivatives to influence MTOR-dependent pathways, alongside the use of LBR phosphorylation inhibitors to potentially mitigate nuclear fragility in metastatic phenotypes [ID: Run1_Eval1_synthesis, Run2_Eval1_synthesis]. Significant gaps remain regarding the synergistic potential of combining these pharmacological agents and their long-term clinical efficacy in vivo."
},
{
"type": "comparison_matrix",
"title": "Repurposed Strategy Comparison",
"headers": [
"Mechanism",
"Primary Target",
"Clinical Goal"
],
"rows": [
[
"Cholesterol Derivatives",
"MTOR-dependent localization",
"Nucleo-cytoplasmic control"
],
[
"LBR Inhibitors",
"Nuclear envelope phosphorylation",
"Reversal of nuclear fragility"
]
]
},
{
"type": "bottlenecks",
"title": "Literature Evaluation Gaps"
}
]
}
},
{
"id": "mvc_dp_LBR_phosphorylation_sites_1785955103106",
"title": "LBR Phosphorylation Sites Report",
"plan": {
"title": "LBR PHOSPHORYLATION SITES : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Data Integrity Scorecard"
},
{
"type": "synthesis",
"title": "Executive Analysis of LBR Phosphorylation",
"content": "The clinical investigation into 'LBR Phosphorylation Sites' [ID: Run2_Eval1_synthesis] yields a critical data gap. While the provided evidence confirms a global increase in LBR phosphorylation, it fails to characterize specific residue-level sites. Consequently, the mapping of functional phosphorylation domains remains undefined within the current literature scope."
},
{
"type": "gap_distribution",
"title": "Literature Gap Density: Site Specificity"
},
{
"type": "bottlenecks",
"title": "Identified Evidence Bottlenecks"
},
{
"type": "verification_audit",
"title": "Evidence Verification Audit"
}
]
}
},
{
"id": "mvc_dp_LBR_A_type_lamin_interaction_1785955116625",
"title": "LBR A Type Lamin Interaction Report",
"plan": {
"title": "LBR A TYPE LAMIN INTERACTION : CUSTOM ANALYSIS",
"evidence_tier": "EVALUATED",
"panels": [
{
"type": "metrics",
"title": "Evidence Integrity Scorecard",
"data": [
{
"label": "Evidence Confidence",
"value": "Medium"
},
{
"label": "Data Points Extracted",
"value": 1
},
{
"label": "Mechanism Status",
"value": "Inconclusive"
}
]
},
{
"type": "synthesis",
"title": "Executive Summary: LBR Phosphorylation Dynamics",
"content": "Current data establishes that Lamin A facilitates LBR phosphorylation and subsequent displacement [ID: Run2_Eval1_synthesis]. However, the causal mechanism remains a significant literature gap, as the evidence does not specify whether this process relies on a direct interaction between Lamin A and LBR or the recruitment of intermediary, unidentified kinases [ID: Run2_Eval1_synthesis]."
},
{
"type": "bottlenecks",
"title": "Literature Gaps and Constraints",
"content": "The primary bottleneck is the 'Mechanistic Ambiguity' regarding the kinase source. The current literature confirms the functional outcome (displacement) but lacks the structural or molecular evidence to distinguish between direct interaction and recruitment-based signaling."
},
{
"type": "gap_distribution",
"title": "Knowledge Gap Mapping",
"data": [
{
"label": "Known Interaction Outcome",
"value": 50
},
{
"label": "Unknown Kinase Mechanism",
"value": 50
}
]
}
]
}
}
],
"aggregatedDatapoints": {
"suggested_experiments": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": [
"Assess the reactivity of anti-Lamin B1 (phospho T575) antibodies using western blot against wild-type and LBR-deficient MEF lysates.",
"Perform immunofluorescence microscopy using the anti-Lamin B1 (phospho T575) antibody in cells treated with inhibitors of kinases potentially targeting T575 to verify nuclear envelope localization."
]
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": [
"Perform mass spectrometry-based phosphoproteomic screening of LBR isolated from TKO cells expressing ectopic Lamin A to identify displacement-associated phosphorylation sites.",
"Evaluate LBR-Lamin A interaction using phospho-mimetic LBR mutants to determine if phosphorylation per se is sufficient for displacement."
]
}
],
"suggested_studies": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": [
"Systematic mapping of Lamin B1 phosphorylation sites during nuclear envelope remodeling.",
"Comparative analysis of phosphospecific antibody staining patterns at the nuclear periphery vs ER in cells with high/low lamin A expression."
]
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": [
"Comprehensive mapping of kinases recruited to the NE by Lamin A that target LBR.",
"Investigation into whether LBR phosphorylation state influences the affinity of its binding to chromatin-associated heterochromatin proteins."
]
}
],
"swansons_literature_based_discovery_candidates": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": "- Discovered Hypothesis (A to C): Hydroxylated cholesterol derivatives may act as negative regulators of Lamin B receptor (LBR) anchorage to the nuclear envelope by modulating HDAC1-mediated chromatin compaction.\n- Literature A (Origin): Cholesterol derivatives modulate HDAC1 dephosphorylation (ID: 32013726).\n- Literature C (Target): LBR anchorage is regulated by phosphorylation/mobility states (ID: 41757046).\n- The Intersecting Bridge B: MTOR signaling/Protein phosphorylation.\n- Biological Rationale: Since both LBR anchorage and HDAC1 localization are controlled by phosphorylation and MTOR signaling, steroid-derived metabolic cues likely coordinate the state of the nuclear envelope and chromatin compaction."
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": {
"Discovered Hypothesis (A to C)": "LBR phosphorylation state regulates cellular response to mechanical force in the nuclear envelope.",
"Literature A (Origin)": "LBR displacement is linked to nuclear envelope fragility in confined migration (ID: 41706894).",
"Literature C (Target)": "Nuclei function as internal tension sensors where LBR levels influence shape recovery (ID: 41561380).",
"The Intersecting Bridge B": "Phosphorylation-dependent mobilization of LBR.",
"Biological Rationale": "Since Lamin A-induced phosphorylation of LBR increases mobility (ID: 41757046) and decreased LBR levels relate to NE tension recovery (ID: 41561380), the phosphorylation state of LBR likely mediates the nuclear mechanical response to confinement."
}
}
],
"contradictions_between_evidences": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": "None identified within the provided context regarding Lamin B1 phosphorylation sites."
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "None detected in the current literature set."
}
],
"repurposed_solutions": [
{
"pentamatrix": "Run1_Eval1_synthesis",
"data": "Use of cholesterol derivatives to manipulate MTOR-dependent nuclear protein localization."
},
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Use of LBR phosphorylation inhibitors or modulators to potentially reverse nuclear fragility in metastatic cells."
}
],
"LBR_phosphorylation_sites": [
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Specific sites are not identified in the text; only the global increase in phosphorylation is noted."
}
],
"LBR_A_type_lamin_interaction": [
{
"pentamatrix": "Run2_Eval1_synthesis",
"data": "Lamin A increases LBR phosphorylation and promotes its displacement, but whether this is through direct interaction or recruitment of unidentified kinases is not explicitly detailed."
}
]
},
"stats": {
"promptTokens": 260630,
"completionTokens": 20570,
"totalTokens": 281200
},
"zenodo_doi": "10.5281/zenodo.21812420"
}