Gene Literature Dashboard

<h4>Background</h4>Transitioning from pediatric to adult HIV care is critical for adolescents and young adults living with HIV (AYLH). This study assessed the timing and factors associated with this transition in Asia.<h4>Setting</h4>Retrospective study of AYLH enrolled in Therapeutics Research, Education, and AIDS Training Asia Pediatric HIV Observational Database clinics between May 1991 and December 2020.<h4>Methods</h4>Eligible AYLH were categorized into <18-year-old transitions, and ≥18-year-old transitions or continued pediatric care.<h4>Results</h4>The analysis included 1803 AYLH from 6 countries. Of these, 92.7% (n = 1672) had perinatally acquired HIV, and 69.9% (n = 1260) were ≥18-year-old transitions or continued pediatric care. Excluding AYLH who remained in pediatric care, median age at transition was 16.1 years (interquartile range [IQR]: 14.9-17.0) for <18-year-old transitions and 20.0 years (IQR: 19.1-21.2) for ≥18-year-old transitions. At age 18, among 1368 AYLH with available viral load data, 72.9% (n = 998) had viral loads <50 copies/mL (78.9% in <18-year-old transitions [n = 310/393] vs 70.6% in ≥18-year-old transitions [n = 688/975]; P = 0.002). The median CD4 count was 635 cells/μL (IQR: 450-842). Factors independently associated with transitions at ≥18-year-old or continued pediatric care included residence in an upper-middle-income country (adjusted odds ratio [aOR] 10.68 [95% confidence interval (CI): 7.76 to 14.68], P < 0.001), living with family (aOR 2.81 [95% CI: 1.99 to 3.97], P < 0.001), and previous antiretroviral therapy class switching (aOR 1.91 [95% CI: 1.34 to 2.72], P < 0.001).<h4>Conclusion</h4>Most AYLH in this cohort transitioned at ≥18 years of age. About one-fourth had unsuppressed viral loads at age 18, irrespective of transition status, underscoring the need for personalized treatment approaches that support antiretroviral therapy adherence from pediatric care, preparing AYLH for transition to adult clinics.

Myelinogenesis is insufficient in numerous myelin-related diseases in the CNS, leading to functional impairments. Myelinogenesis couples with angiogenesis to ensure adequate need of oxygen and nutrients for oligodendrocyte (OL) differentiation. However, approaches to synchronize myelino-vascular coupling remain unavailable. We hypothesize the identification of shared signaling pathways in vascular cells and oligodendroglia may yield novel strategies to promote myelin repair through strengthening the blood vessel-myelination coupling. Here, single-cell sequencing and in situ hybridization revealed high expression of G-protein-coupled receptor 30 (Gpr30) in both vascular cells and oligodendroglia, with selective enrichment in pericytes and oligodendrocyte precursor cells (OPCs). Cell-specific deletion of GPR30 in pericytes driven by PDGFRβ^CreERT2 resulted in enhanced angiogenesis and myelination in developing brains. GPR30 deletion in OPCs or antagonizing GPR30 by G15 resulted in increased MBP-positive cell density and enhanced nanofiber wrapping capacity in vitro, thereby demonstrating an inhibiting role of GPR30 on OPC differentiation. To elucidate the coordinative role of GPR30 in both cell types, we employed NG2^CreERT to induce a conditional knockout of GPR30 in both NG2-positive pericytes and OPCs. The conditional deletion of GPR30 enhanced myelination and increased vascular density in developing brains. Further, GPR30 cKO or G15 treatment enhanced myelin repair and functional recovery in the chronic neonatal hypoxia and lysolecithin-induced demyelination model, suggesting that antagonizing GPR30 is a promising strategy to synchronize angiogenesis with myelination to promote myelinogenesis. These findings establish GPR30 antagonism as a promising approach to enhance myelin repair through synchronizing pericyte-mediated angiogenesis and OPC differentiation.

Glial cells, including radial glia, oligodendrocyte precursor cells (OPCs), oligodendrocytes, astrocytes, and microglia, are active and dynamic regulators of central nervous system (CNS) development, homeostasis, and disease. Through extensive interactions with neurons, other glial populations, and the vasculature, they form highly specialized communication networks that are essential for normal brain function. While transcriptomic approaches have revealed extensive glial heterogeneity and enabled the prediction of putative signaling networks, a critical challenge remains in validating and translating these findings at the level of distinct protein complexes existing both within and between the various glial cell types. This is largely due to the fact that traditional proteomic technologies lack spatial resolution and/or fail to capture protein interaction networks. Proximity labeling (PL) has emerged as a powerful strategy to overcome these limitations by enabling cell-type-specific mapping of protein networks and subcellular proteomes, with spatial and temporal precision. Emerging studies have applied PL enzymes, such as BioID, TurboID, and HRP, across diverse glial populations, starting to uncover protein networks supporting their interactions with neurons and vascular elements, allowing metabolic support, maintenance of microenvironment homeostasis and cell-cell communication (including synaptic modulation). In this review, we summarize the main PL enzymes, discuss key studies across different glial cell types, and examine the technical challenges and future perspectives of applying PL to investigate glial biology. By complementing transcriptomic data with spatially resolved proteomic insights, PL provides a unique opportunity to deepen our understanding of glial cell biology in health and disease.

Fluoride is widely used for caries prevention due to its effects on mineralized tissues, yet its potential role as a modifier of systemic metal homeostasis remains insufficiently explored. This systematic review synthesizes preclinical evidence on the association between fluoride exposure and changes in metal and semi-metal concentrations across biological matrices. A comprehensive search strategy was conducted across major databases without language or date restrictions, following SyRF, CAMARADES and PRISMA 2020 guidelines. Thirty-one animal studies were included, encompassing multiple species, exposure conditions and analytical approaches. Despite substantial methodological heterogeneity, consistent patterns emerged. Fluoride exposure was associated with element-specific redistribution of the metallome rather than uniform change. Essential elements were predominantly depleted, most consistently zinc, copper and manganese, whereas the toxic metals lead and cadmium tended to be retained. This contrast between homeostatically regulated essential elements that are lost and non-regulated toxic metals that accumulate supports the hypothesis that fluoride differentially modifies the distribution and retention of co-existing elements. The novelty of this review lies in integrating metallomic outcomes across experimental models, highlighting fluoride as a potential systemic modulator rather than a tissue-specific agent. Although variability in study design and risk of bias limits causal inference, the consistent directionality of findings across models reinforces their biological plausibility and translational relevance.

This study employed an exploratory qualitative methodology to examine how IVF clinics in the United States develop and implement policies regarding the transfer of embryos that test positive for genetic conditions through preimplantation genetic testing for monogenic disorders (PGT-M). The study explored who is involved in creating these policies, and the key factors influencing these decisions. Semi-structured interviews were conducted with genetic counselors working in IVF clinics across the United States. Participants were asked about clinic policies regarding PGT-M positive embryos, their role in policy creation, and how patient requests are handled. Transcripts were analyzed thematically to identify patterns in clinic practices, policy rationales, and decision-making processes. Findings showed considerable variation in how clinics approach PGT-M affected embryo transfers. While one clinic follows strict guidelines that prohibit the transfer of embryos with pathogenic variants (including likely pathogenic variants or variants of uncertain significance [VUS]), others allow case-by-case consideration, particularly when patients face barriers to further IVF cycles. Common policy frameworks prioritize transfer of unaffected embryos, with affected embryos considered only after other options are exhausted. Key factors influencing policy decisions included ethical considerations, clinic values, provider discretion, legal and insurance frameworks, and concerns about the child's future quality of life. Despite formal protocols, many clinics still assess individual cases, underscoring the complexity of balancing standardization with patient-centered care. Genetic counselors often serve as intermediaries, advocating for patients while navigating institutional limitations. This study highlights the need for clearer, yet flexible, guidelines on PGT-M positive embryo transfer to ensure consistency while preserving patient autonomy. Greater integration of genetic counselors into policy development and clinical decision-making can support more ethically grounded and informed reproductive care.

The Min pig, a representative northern Chinese indigenous breed, carries a unique ancestral background shaped by the historical phylogeography of Northeast Asia. This study aimed to dissect the population structure, temporal genetic divergence, and ancestral composition of Min pigs, trace their evolutionary origin, and identify trait-linked functional genes, providing information regarding their evolutionary history and conservation. We analyzed 61 Min pigs sampled across nearly 20 years and 701 reference pigs comprising other Chinese indigenous breeds, Western commercial lines, and Chinese wild boars, using PCA, NJ phylogenetic analysis, Admixture, TreeMix, D-statistic, f4-ratio, and combined selection signature scans (sliding-window F_ST, XP-EHH, and π-ratio). Clear genetic stratification was observed among Min pig subpopulations, reflecting long-term divergence under natural and artificial selection. PCA and Admixture (K = 2-4) separated East Asian indigenous and Western ancestral components, verifying an admixed Northeast Asian origin with a dominant ancient East Asian component and a Western component. Compared with early-2000s Min pigs, contemporary individuals are genetically closer to Western breeds and exhibit a more scattered structure due to shifted ancestral component proportions, further confirmed by D-statistic and f4-ratio values. We identified 321 differentiated SNP loci based on the Animal QTL database, corresponding to core candidate genes (AKT3, ACACA, MAP3K5, FGFR4, C3, and SERPINC1) enriched for meat quality, growth, reproduction, immunity, energy metabolism, and MAPK/PI3K-Akt/AMPK pathways. This study reveals Min pigs' admixed origin and temporal divergence, clarifying their Northeast Asian evolution and providing molecular markers for genetic monitoring and conservation.

Wattles are finger-like appendages on the ventral neck of goats, serving as a distinctive morphological marker for breed identification that serves potential implications for production performance. However, their genetic basis remains incompletely characterized. Here, we integrated a genome-wide association study (GWAS) and selection signature analysis to identify candidate genes and genomic regions associated with the wattle trait in goats. Using a linear mixed model, GWAS on 463 goats (23 wattled and 440 non-wattled) identified 385 quantitative trait loci (QTLs) at a 5% false discovery rate, yielding 346 candidate genes. The most significant association signal was observed on chromosome 10 (72.61-73.48 Mb), where the lead SNP (rs636481767) is located within a region containing GJD2, GREM1, and FMN1, showing strong linkage disequilibrium (r² > 0.6) with surrounding loci. Subsequent selection signature analysis (23 wattled and 23 non-wattled) identified 83 genomic regions harboring 119 candidate genes. The strongest signals were detected at MFSD14B on chromosome 8 (F_ST = 0.154, log₂π-ratio = 2.611) and PDLIM7 on chromosome 7 (F_ST = 0.144, log₂π-ratio = 0.806). KEGG pathway enrichment analysis revealed that GWAS-associated genes were involved in glycosylation and immune responses, whereas selection-signature genes were enriched in DNA repair and the Hippo, Notch, and Wnt pathways. Furthermore, cross-species PheWAS revealed that human FMN1 is associated with dermatological, skeletal, and metabolic phenotypes, while porcine FMN1 is associated with backfat thickness and loin muscle depth. Overall, this study provides molecular markers of potential value for goat breeding and pinpoints key candidate genes for future functional validation of wattle development.

<h4>Background</h4>Owl monkeys are nonhuman primate species in the genus Aotus. The Michale E. Keeling Center for Comparative Medicine and Research maintains an Owl Monkey Breeding and Research Resource composed of four species of owl monkeys: Aotus nancymai, A. vociferans, A. azarae, and A. griseimembra. Disease of the hepatobiliary system has not been well characterized in Aotus.<h4>Methods</h4>Necropsy records from a 17-year period were reviewed to characterize the gross and microscopic lesions of liver disease.<h4>Results</h4>316 adult owl monkeys were submitted for necropsy in the review period; 22 of 316 (6.96%) were diagnosed with hepatobiliary disease. Cardiac-associated hepatopathy was the most common diagnosis (6/22). Previously unreported conditions included cystic mucinous hyperplasia (1/22), acute cholangiohepatitis (1/22), idiopathic end-stage hepatopathy (1/22), cholangiocarcinoma (2/22), and acute cholecystitis (4/22). Vitamin E deficiency anemia (3/22) and hemosiderosis (4/22) have been reported in the genus, and these diagnoses were noted.<h4>Conclusions</h4>The spectrum of hepatobiliary pathology in owl monkeys is broader than previous reports have suggested.

<h4>Background</h4>Extracorporeal membrane oxygenation (ECMO) provides life support to patients undergoing cardiopulmonary surgeries when other interventions are ineffective. Unfractionated heparin (UFH) is routinely used during ECMO due to the increased risk of thrombosis. However, in some cases, like antithrombin deficiency, UFH is not efficacious, and heparin resistance (HR) can develop. There is considerable confusion about HR and what additional management strategies should be taken.<h4>Objectives</h4>This study compared clinical and economic outcomes associated with antithrombin concentrate (ATc) + UFH versus bivalirudin in adult patients undergoing ECMO.<h4>Methods</h4>This retrospective cohort study used data from the 2014-2023 TriNetX database and the 2018-2021 Premier Healthcare Database. Adult patients treated with ATc + UFH or bivalirudin within 3 days of ECMO initiation who did not undergo cardiopulmonary bypass (CPB) or coronary artery bypass grafting (CABG) during their ECMO hospital stay were identified. Propensity score matching was performed to balance the groups. Thromboembolic events, bleeding events, heparin-induced thrombocytopenia, hospital mortality, length of stay, and total hospital cost were compared.<h4>Results</h4>A total of 1601 patients (276 ATc + UFH, and 1352 bivalirudin) were identified from TriNetX, and 1592 (264 ATc + UFH, and 1528 bivalirudin) from Premier. ATc + UFH was associated with lower odds of thrombotic and bleeding events compared to bivalirudin only. Length of stay and total hospital costs trended lower for patients prescribed antithrombin and UFH.<h4>Conclusions</h4>Antithrombin + UFH may be a preferable option for managing ECMO patients, particularly those with heparin resistance.

Beta-coronavirus infection disrupts endoplasmic reticulum (ER) homeostasis; however, the mechanisms by which viral proteins manipulate ER-resident factors remain unclear. Here, we report that the SARS-CoV-2 accessory protein ORF3a binds to the ER chloride channel CLCC1, thereby impairing ER ion homeostasis, activating the unfolded protein response, and inducing endomembrane remodeling. Using newly developed ratiometric reporters, we showed that this interaction exacerbated the basal levels of ER-phagy and nucleophagy. Importantly, CLCC1 counteracted ORF3a by sequestering it within the ER, attenuating its toxicity and suppressing viral replication, which established CLCC1 as a host restriction factor. We further elucidated a spatiotemporal mechanism: during early infection, ORF3a accumulates in the ER, co-assembles with CLCC1 into puncta, and disrupts ER homeostasis; at the later stage, ORF3a overload enables its escape from CLCC1-mediated ER retention and subsequent translocation to lysosomes, facilitating viral egress. This ER-centric function is conserved across diverse beta-coronaviruses, including SARS-CoV-1 and bat- or pangolin-derived strains, revealing a unified pathogenic strategy. Furthermore, in mouse brains, the ORF3a-CLCC1 axis recapitulates key neuropathological features, including ER stress, autophagic dysfunction, neuronal death, and neuroinflammation, providing a mechanistic link to COVID-19-associated neurological symptoms. Our findings identify CLCC1 as an essential host defense protein and establish a conceptual framework to guide future research on antiviral responses at the organelle level and related disease mechanisms.

Along with being a significant disease in dairy cattle which incurs economic loss, mastitis also affects a wide range of parameters that involve milk yield and quality. However, the role of long non-coding RNAs (lncRNAs) in the regulation of mastitis in distinct stages of the disease in Sahiwal cows is yet to be understood. This study characterised lncRNA expression profiles and competing endogenous RNA (ceRNA) networks across healthy, subclinical, and clinical mastitis stages using RNA-Seq data from milk somatic cells. Five biologically relevant ceRNA modules were identified: Five ceRNA modules were identified that are biologically relevant: MSTRG.21615.2, is predicted to sponge bta-miR-760-3p and bta-miR-212, and involved in co-regulation of OAT and SIRT2, leading to dysregulation of urea-cycle and NAD ⁺ -metabolism; MSTRG.19791.1-bta-miR-2382-5p, which activates an extracellular antioxidant network (SOD3, CAT, GPX family, PARK7); MSTRG.1150.15-bta-miR-92b, which regulates the PTEN/PI3K-AKT signalling axis; MSTRG.19363.1-bta-miR-1296, involved in the nuclear import machinery (KPNA7, RCC1, RANBP1, NUP50), with reduced expression in clinical versus subclinical mastitis; and MSTRG.25768.1-bta-miR-2425-5p, linked to extracellular matrix remodelling via the ADAMTS. The binding prediction of all six lncRNA-miRNA pairs were favourable, with the highest predicted affinity observed for MSTRG.1150.15-bta-miR-92b (-15.4 ± 6.9 kcal/mol) and moderate but biologically relevant binding for MSTRG.19363.1-bta-miR-1296 (-7.8 ± 4.2 kcal/mol). Molecular dynamics over 50 ns revealed that the MSTRG.19363.1-bta-miR-1296 complex was the most stable, exhibiting lowest RMSD and radius of gyration values, suggesting that it may be a viable regulator of nuclear transport during the progression of mastitis. These results shed light on the underlying mechanisms of lncRNA-regulated networks associated with dairy cattle mastitis.

Mitochondrial DNA depletion syndrome type 5 (MTDPS5), caused by <i>SUCLA2</i> mutations, is a rare autosomal recessive disorder manifesting as early-onset encephalomyopathy. Clinical diagnosis is challenging due to phenotypic overlap and potentially unremarkable early investigations. We report a male infant of consanguineous parents who presented with progressive hypotonia, global developmental delay, and involuntary nocturnal movements at 3 months. Despite an unremarkable newborn metabolic screen and normal initial brain MRI, the patient demonstrated a significant failure to achieve age-appropriate gross motor milestones, including independent sitting and crawling, by 12 months. Whole-exome sequencing (WES) definitively identified a homozygous likely pathogenic <i>SUCLA2</i> variant (p.Met329Val). Following the initiation of a mitochondrial cocktail, the patient showed substantial clinical improvement. This case emphasizes that MTDPS5 should be considered in infants with unexplained encephalomyopathy, even when initial findings are normal, underscoring the necessity of early WES to guide clinical management.

T-cell acute lymphoblastic leukemia (T-ALL) comprises molecularly diverse subtypes, but robust cross-cohort validations and operational gene-expression definitions are lacking. To establish a gene-expression-anchored framework for T-ALL subtyping, we aggregated 2314 transcriptomes (15 cohorts, age: 0.8-90.8 years). An extended unsupervised approach defined 17 main clusters and 3 subclusters in samples with high blast fractions. Supervised analyses added an overarching immature T-ALL (early T cell precursor [ETP]-like) definition and resolved the LMO2 γδ-like subtype. All clusters contained samples from at least two cohorts. Characteristic genomic driver enrichments were consistent across cohorts, while gene-expression clusters did not correspond exclusively to single driver events but also reflected developmental origins. A machine-learning classifier based on ALLCatchR, our B-cell acute lymphoblastic leukemia (B-ALL) classifier, identified these 20 transcriptomic subtypes and the immature T-ALL (ETP-like) signature with 0.995-1.0 accuracy in a validation set (<i>n</i> = 203). Testing the classifier on a second hold-out data set (<i>n</i> = 265 samples) showed that 92.7% of predictions matched with corresponding driver alterations. Across all samples, 83.2% of cases received high-confidence predictions, 7.3% candidate predictions, and 9.5% remained unclassified, largely because of low blast fractions. We identified a novel gene-expression cluster markedly enriched (P < 0.001) for clonal hematopoiesis mutations (<i>IDH2 R140Q</i>, <i>DNMT3A</i>) and a stem-/progenitor cell-like gene expression. This novel clonal hematopoiesis-related T-ALL subtype was observed in six cohorts and accounted for 8.9% of adults and 39.5% of patients aged >50 years. We extended ALLCatchR into ALLCatchR2, a free R package that now enables B-/T-lineage separation, gene-expression subtyping, blast estimation, and developmental annotation to harmonize T-ALL classification across studies and clinical contexts.

A defining feature of primates is the expansion and increased complexity of the cerebral cortex, processes that are largely associated with evolutionary adaptations of the neural progenitor-enriched outer subventricular zone (OSVZ). Yet, the genetic mechanisms underlying the emergence of the primate OSVZ remain unknown. The tree shrew is a closer outgroup to primates than rodents, its brain exhibits an intermediate phenotype; while being lissencephalic, it nonetheless possesses an OSVZ. Here, we applied laser microdissection technology to conduct comprehensive gene expression analyses of different cortical layers in the tree shrew brain across three key developmental stages. We show that cell cycle-related genes are already involved in the tree shrew OSVZ. In contrast, extracellular matrix-associated pathways appear to be unique to the primate OSVZ. Furthermore, overexpression of ZNF664 significantly increased intermediate progenitor proliferation in the mouse subventricular zone. Together, these results reveal shared molecular mechanisms underlying OSVZ development in primates and tree shrews, providing insights into the origin of primate cortical complexity.

Given the association between early-onset colorectal cancer (eoCRC) and insulin resistance (IR), we investigated the effects of westernized diet (WD)-induced IR on intestinal stem cells (ISCs) by in-depth phenotyping three adolescent mouse models of escalating WD-driven IR. WD-related hyperlipidaemia and hyperglycaemia enhanced ω6-polyunsaturated fatty acid (PUFA) metabolism, while inhibiting glucose metabolism, resulting in colonic epithelial cell (EC) energy deficiency, hypoproliferation, and quiescent ISCs (qISC) expansion. WD-induced systemic IR provoked colonic EC IR, reduced ω6-PUFA metabolism, which was compensated by aberrated triglyceride metabolism and growth factor receptor signalling, causing hyperproliferation of expanded qISCs. Combined with single-cell transcriptomic data and colonic 3D-organoid analysis we revealed that the ω6-PUFA arachidonic acid inhibited glucose uptake, favouring colonic EC IR driven expansion of the lipid-dependent inherently DNA-mismatch prone qISC compartment in expense of the insulin-dependent LGR5⁺ ISCs. Overall, our findings may open new alleys for targeted nutritional interventions in eoCRC prevention.

Neutrophils are double-edged effectors of the innate immune system: while essential for host defense, they can drive significant tissue damage in the context of chronic inflammatory diseases. Identifying strategies to selectively dampen their pathogenic functions without compromising antimicrobial immunity remains a major clinical challenge. Here, we define a conserved IFNAR1-driven circuit mediated by iron homeostasis that sustains neutrophil pathogenicity across distinct mucosal tissues. Using single-cell transcriptomics and a CRISPR-based functional screen, we identified the mitochondrial iron transporter Mitoferrin-1 (SLC25A37) as a key regulator of neutrophil inflammatory programming. Mechanistically, our data indicate that Mitoferrin-1 mediates a feedforward loop in which NET-derived histones activate toll-like receptor 9 (TLR9), which in turn sustains IFN-α/IFNAR1 signaling, rewires mitochondrial metabolism, and ultimately drives tissue damage through markedly increased of NETosis, reactive oxygen species production, and degranulation. Notably, this IFNAR1-driven inflammatory circuit operates entirely independently of antimicrobial function, as pharmacological IFNAR1 blockade fully preserves phagocytic activity against bacterial pathogens in vivo. Targeting this pathway selectively attenuates neutrophil-driven inflammation in both murine models of colitis and acute lung injury, as well as in primary human intestinal organoid systems. Together, these findings identify a targetable metabolic node for selectively disarming pathogenic neutrophil responses in the setting of chronic inflammatory disease.

The cGAS-STING pathway detects cytosolic dsDNA to initiate innate immune responses, serving as a critical surveillance system against infection and cellular stress. Autophagy is an evolutionarily conserved catabolic process that maintains homeostasis by degrading cytoplasmic components. Although these two systems operate through distinct mechanisms, recent studies have uncovered a complex bidirectional regulatory network that intimately links them. On the one hand, STING has an evolutionarily conserved capacity to induce TBK1-independent noncanonical autophagy, and recent studies further link this activity to TFEB-dependent lysosome biogenesis, expanding its functional repertoire beyond classical interferon induction. On the other hand, autophagy restricts cGAS-STING signaling by clearing cytosolic DNA and selectively degrading pathway components, thereby preventing excessive inflammation. Furthermore, mitophagy curtails the release of mitochondrial DNA, a potent cGAS agonist, and recent studies further implicate lysosomes as active signaling platforms associated with mtDNA release, LRRK2 activation, and STING-dependent NF-κB responses. In this review, we discuss recent advances in understanding how the cGAS-STING pathway and autophagy mutually regulate each other at the cellular level, focusing on the molecular mechanisms of this interplay, both canonical and noncanonical, and highlighting how their crosstalk shapes cellular homeostasis and stress adaptation.

Some individuals appear more susceptible to concussion or mild traumatic brain injury (mTBI) and the severity, range, and the persistence of post-concussion symptoms vary considerably between affected individuals. Genetic factors are likely to contribute to this variability. Symptomatic overlap of post-concussion syndrome with neurological conditions such as familial hemiplegic migraine (FHM) caused by rare pathogenic variants in ion channel and synapse protein genes, with high sensitivity to head trauma for some patients, suggests that variation in similar pathways may influence concussion susceptibility and recovery. To investigate this hypothesis, we performed whole exome sequencing in 93 unrelated individuals who had sustained a single or multiple concussions and examined rare protein-altering variants in FHM genes, other neuronal ion channel and transporter genes, and genes involved in neurotransmission. We identified 62 different rare missense variants across 24 genes in 59 participants (63%), with 26 individuals carrying 2 or more variants. The prevalence of specific likely damaging rare variants in the 16 ion channel-related genes that were identified was approximately fivefold higher than that observed from gnomAD population controls (Odds Ratio = 5.44, 95% CI [4.13,7.18], P < 0.0001). Notably, voltage-gated calcium and sodium channel genes, including SCN9A, together with neurotransmission-related genes such as SNCAIP, harboured multiple potentially deleterious variants. These findings suggest that rare deleterious variants in genes involved in ion homeostasis and neurotransmission may contribute to an individual's susceptibility to concussion or more severe post-concussion symptoms. This study provides a foundation for future genetic and functional investigations aimed at improving our understanding of concussion susceptibility and outcomes. Further validation in larger cohorts and mechanistic studies is warranted to determine their utility as biomarkers of concussion risk and prognosis.

<h4>Background</h4>In Huntington's disease (HD), clinical heterogeneity in age at onset and disease progression poses demands on precision medicine approaches. Biofluid biomarkers are needed to monitor disease progression, but may also provide insights into pathogenic mechanisms and support the development of disease-modifying therapies (DMTs).<h4>Objective</h4>To identify dysregulated cerebrospinal fluid (CSF) proteins in HTT gene expansion carriers (HDGECs), and to evaluate their relationship with disease progression.<h4>Method</h4>CSF was collected from a discovery and a validation cohort, both comprising HDGECs and neurologically unaffected controls (HCs). A total of 734 proteins were quantified using a high-sensitive Proximity Extension Assay (PEA) (Olink Neuro Explore panel 1 and 2). Protein association with disease progression was evaluated using the composite Unified HD Rating Scale (cUHDRS). Disease stage was classified according to the HD Integrated Staging System (HD-ISS).<h4>Results</h4>In total, samples included HDGECs (n=61), HCs (n=54), and longitudinal HDGEC samples (n=21). Sixteen proteins were nominally dysregulated in HDGECs vs. HCs. Most pronounced was a twofold decrease of TNFRSF8 (sCD30) in HDGECs, and longitudinally declining levels were found. TNFRSF8 correlated significantly with cUHDRS, after adjustment for age, sex, and CAG. Levels of TNFRSF8 were similarly reduced in the validation cohort, and correlated with Total Functional Capacity (TFC).<h4>Conclusion</h4>TNFRSF8 recently emerged as a biomarker of neuroimmune dysfunction in HD. The results indicate and confirm that it is highly dysregulated in HDGECs and also tracks clinical disease progression. This pathway may be a promising potential target for DMTs.

The role of epigenetic modifiers in astrogenesis is less understood compared to neurogenesis. For example, the Disruptor of telomeric silencing 1 like, DOT1L, conferring H3K79 methylation safeguards neural progenitors from premature differentiation in various brain regions, however, its role in astrogenesis has not yet been reported. Here, we decipher its role during astrogenesis using an Emx1-cre driven DOT1L conditional knock-out during the development of mouse cerebral cortex from E12.5 till P0. We describe two astrocyte types at P0, distinguishable by expression of phagocytosis markers genes. DOT1L loss-of-function (LOF) increased the number of those astrocytes that express fewer phagocytosis marker genes, while it affects a larger number of differentially expressed genes in those astrocytes that strongly express homeostasis marker genes. Our findings establish DOT1L as a barrier between these astrocyte states, limiting differentiation of the astrocytes expressing phagocytosis marker genes. We also identify multiple developmental origins of astrocytes. In addition to apical progenitors (AP), the prevailing stem cell population considered as origin of cortical astrocytes, we identify neural stem cells (NSC) in the dorsal telencephalon differentiating towards astrocytes, and Nkx2.1 -lineage derived astrocytes, originating from the ventral telencephalon. This finding refines the prevailing view that dorsal telencephalon APs are the only source of embryonic cortical astrocytes, and enter astrogliogenesis after neurogenesis. We show that the expression of Emx1 discriminates different astrocyte origins during cortical development, and we describe a new set of transcription factors (TFs) that mark the state conversion between the homeostasis or phagocytosis expression programs in astrocytes. DOT1L orchestrates different TF networks, of which Lhx2, Nfia, Sox6 , and Creb5 drive astrogliogenesis, and Nfia, Nfib , and Tcf4 affect astrocyte state conversion. Importantly, our analysis also shows that NSCs, APs and astrocytes de-differentiate within this lineage trajectory, and lowering the DOT1L-mediated epigenetic barrier strongly favours appearance of astrocytes with limited phagocytosis properties and reactivation of progenitor programs.

In adult mammals, poor functional recovery after spinal cord injury (SCI) is largely due to the very limited capacity to reconstruct damaged neural connections together with neuronal loss. Here, we identify the neuroprotective role of Zdhhc17 as a palmitoyl acyltransferase (PAT) following SCI. Neuron‑specific Zdhhc17 overexpression in vitro and in vivo markedly enhances axon regeneration and functional recovery after SCI in a PAT-activity-dependent manner. Interactome and palmitoylation analyses in cortical neurons identify the karyopherins Kpna2 and Ipo9 as previously unrecognized Zdhhc17 substrates. SCI markedly reduces Kpna2 and Ipo9 protein levels, whereas Zdhhc17‑mediated palmitoylation stabilizes them by suppressing their ubiquitin‑dependent degradation. Functionally, neuronal overexpression of Kpna2 or Ipo9 mimics the therapeutic effects of Zdhhc17, and co‑expression of Zdhhc17 with Ipo9, but not Kpna2, further augments SCI repair. Using H₂O₂‑induced oxidative stress and glutamate‑induced excitotoxicity models, we further show that Zdhhc17 promotes neuronal survival and activates transcription of intrinsic pro‑regenerative genes after injury. These findings suggest that the Zdhhc17-Kpna2/Ipo9 axis is a novel pharmacological target for SCI treatment.

10 million people die of different forms of cancer every year. One of the largest problems is the great variability between patients in drug efficacy, that is partly due to the low specificity of cancer drugs and their consequent severe side effects, which make it difficult to avoid damage to healthy tissues. To address these challenges, several targeted therapies and drug delivery strategies have been developed that can selectively act on cancer cells to improve efficacy and reduce toxicity. One promising approach is the use of graphene or graphene oxide-based materials for this purpose. Here we investigated a graphene oxide (GOX) composite material functionalized with 2 different dendrimers: poly(propylene)imine (DAB-Am-16) and polyamidoamine (PAMAM). These dendrimers were covalently bound to GOX via amide formation between any peripheral amine group of the dendrimers and the carboxyl groups present in GOX. We explored the cellular responses of the materials on BEAS-2B (non-tumoral), LC-HK2 and HepG2 cancer cells. We investigated cell viability, morphological changes in the cytoskeleton organization, the spatial interaction between the materials and the cell nuclei. Our results showed that GOX materials reduced cell viability at higher concentrations, and cellular exposure to these materials was associated with cytoskeletal reorganization, including reduced cytoplasmic and nuclear areas. These findings highlight their potential as promising candidates for biomedical applications.

<h4>Background</h4>Circulating microRNAs have been proposed as minimally invasive markers of cognitive impairment and neurodegenerative disease, but evidence for individual miRNAs in heterogeneous older populations remains limited.<h4>Objective</h4>We examined cross-sectional associations between eight circulating serum miRNAs and continuous cognitive performance in a mixed inpatient-outpatient geriatric cohort.<h4>Methods</h4>This cross-sectional analysis included 65 enrolled participants from a single-center, non-consecutive, mixed inpatient-outpatient geriatric cohort; 63 formed the analytic cohort (32 inpatients, 31 outpatients). Eligibility required written informed consent, availability of a routine-care Mini-Mental State Examination (MMSE) score, administration of Addenbrooke's Cognitive Examination III (ACE-III), fasting serum sampling, and structural brain MRI during the same visit or within 48 hours. Eight predefined serum miRNAs were quantified by RT-qPCR with exogenous spike-in controls, with cel-miR-39-3p used as the normalization reference. The primary outcome was continuous MMSE score. Secondary outcomes were ACE-III total score, the ACE-III attention/orientation subscale, and threshold-defined MMSE and ACE-III impairment categories. Multivariable linear or logistic regression models were adjusted for age, sex, years of education, and recruitment setting. The main analysis used adjusted linear regression for continuous MMSE with Benjamini-Hochberg false discovery rate correction across the eight individual-miRNA MMSE models. Bootstrap was used to assess the stability of the MMSE estimates. Sensitivity analyses for the three strongest MMSE associations included Huber robust regression and miRNA by setting interaction testing.<h4>Results</h4>In multivariable models adjusted for age, sex, years of education, and recruitment setting, higher levels of miR-132, miR-545, and miR-34a were linked to lower MMSE scores. A 1-SD increase in each marker corresponded to an MMSE decrease of about 1.1-1.3 points. These were the strongest single-miRNA findings in the dataset and met the prespecified FDR threshold in the primary models, although bootstrap analyses showed limited precision. Robust regression gave the same negative pattern for all three markers, suggesting that the results were not driven by a small number of extreme observations. Evidence of between-setting differences was seen only for miR-132 and only at the nominal level. No individual miRNA showed a clear nominal association with ACE-III total, and the ACE-III attention/orientation and MRI-adjusted analyses were exploratory. In secondary threshold-based models, no single miRNA remained significant after FDR correction.<h4>Conclusions</h4>In this cross-sectional pilot cohort, higher miR-132, miR-545, and miR-34a were associated with lower MMSE scores in adjusted analyses. However, effect sizes were small, bootstrap analysis showed considerable uncertainty, and the findings are exploratory and hypothesis-generating. These results require evaluation in larger consecutive cohorts and do not establish diagnostic, prognostic, or standalone clinical biomarker utility.

Neurodevelopmental disorders (NDDs), including autism spectrum disorder (ASD) and intellectual disability (ID), are genetically heterogeneous. DEAF1 has emerged as a key NDD risk gene, with pathogenic variants linked to DEAF1-associated neurodevelopmental disorder (DAND), but its role in human neurodevelopment remains unclear. Human cortical organoids (hCOs) provide a physiologically relevant model that recapitulates fetal brain development with an authentic human genetic background. Here, we show that a DEAF1 mutation in human embryonic stem cells disrupts chromatin accessibility at neuronal gene loci, leading to significant transcriptional alterations. In hCOs, this mutation results in aberrant progenitor proliferation, disrupted cortical lamination, and impaired neuronal differentiation. Furthermore, we identify WNT signaling, TGFβ superfamily signaling, and cell cycle regulation as commonly dysregulated pathways across multiple ASD-associated genetic perturbations. Pharmacological inhibition of WNT signaling with a Porcupine inhibitor partially rescues phenotypic defects in DEAF1-mutant hCOs. Our findings identify DEAF1 as a critical regulator of neurodevelopment and support pathway-targeted, mutation-independent therapeutic strategies for ASD and related disorders.

Classic oncological models have largely restricted cancer pathogenesis to autonomous genetic mutations within localized cells. Current research, however, reframes solid tumors as structurally integrated "pseudo-organs" that rely on host neural networks for survival. This review examines the reciprocal signaling networks connecting nervous, immune, and malignant cells within the tumor microenvironment. At the tissue level, we outline how tumors construct de novo neural architecture via axonogenesis, cellular plasticity, and the assembly of functional neuro-neoplastic synapses. Expanding beyond the local niche, we describe the systemic brain-body circuitry. We specifically address how central nervous system pathways respond to tumor-derived cues and how chronic stress signaling disrupts physiological homeostasis. To explain the associated immune evasion, we analyze how sensory neuropeptides, particularly CGRP and, more broadly, Substance P-NK1R signaling, contribute to immune dysfunction, tumor-promoting inflammation, and therapeutic resistance. Finally, we discuss the clinical implications of these pathways. Rather than relying on nonspecific structural denervation, future therapeutic strategies should focus on targeted "neural reprogramming" to uncouple tumors from their neural dependencies and restore antitumor immunity while minimizing neurotoxicity. Ultimately, this review conceptualizes solid tumors as systemically integrated entities, establishing a framework for precision neural reprogramming to overcome immune evasion and therapeutic resistance.

Epigenetic clocks have been widely used to estimate biological age across various tissues, but their accuracy in breast tissue remains suboptimal. Classical models such as Horvath's and Hannum's clocks perform poorly in predicting chronological age in breast tissue, underscoring the need for a tissue-specific approach. In this study, we introduce a Breast Tissue-specific Epigenetic Clock (BTEC), developed using DNA methylation data from 553 healthy breast tissue samples across seven different studies. BTEC significantly outperformed existing clocks, demonstrating superior correlation with chronological age (<i>r</i> = 0.91) and lower prediction errors (MAE = 3.0 y) without requiring dataset-specific regressions adjustments. Notably, BTEC showed consistent performance across ancestry groups, in contrast to existing clocks which exhibited variable ancestry-related biases. We also found that residual-based epigenetic age acceleration, commonly used to correct for prediction bias, is highly dependent on the choice of reference dataset and does not provide a generalizable correction. BTEC's chronological age predictions for tumor-adjacent samples showed distortions, with an average deviation of -2.3 y, which was even more pronounced in tumor samples, where the average difference between predicted and chronological age was -14 to -15 y. When analyzed by molecular subtype, the distortion was greater in the more aggressive HER2+ and TNBC tumors compared to HR+ tumors. Overall, our findings indicate that breast tumors do not generally exhibit accelerated epigenetic aging. The probes used by BTEC were associated with known oncogenes such as JAK1, TP73, PDGFRA, CCND1 and STAT5B, as well as genes involved in cancer-related pathways, including Wnt, Hedgehog and ErbB signaling. Importantly, extreme deviations in epigenetic age, as measured by BTEC-derived epigenetic age acceleration (EAA), were associated with patient survival in HR+ breast cancer patients in an exploratory analysis, highlighting the prognostic value of tissue-specific epigenetic aging measures. These findings demonstrate that BTECs not only improve age prediction in breast tissue but also capture biologically meaningful alterations in tumor epigenetic aging with potential clinical implications.

<i>N</i>-derivatives of pimaricin (<b>PIM</b>) were synthesised as zwitterions or in a non-ionic form, influencing water solubility and toxicity in normal cells (HDF). The introduction of benzyl-<i>O</i>-aliphatic (<b>15</b>) and methylene-nitrophenol (<b>7</b>) moieties into the polyene resulted in high and broad-spectrum antifungal activities, even better than known antifungal agents. The promising antifungal potency of <b>7</b> is attributed to formation of a 6:1 complexes with ergosterol (<b>ERG</b>) as indicated by ITC, FT-IR and NMR studies. The self-assembly of 6:1 complexes into greater structures of polar channels explains antifungal activity of <b>7</b>, as suggested by MOG-PM6 calculations. In contrast, fluorobenzyl derivative <b>3</b> despite weak antifungal potency, revealed unexpected high anticancer activity in PC-3 and SKOV-3 cells, at improved selectivity (IC₅₀ = 0.6 µM; SI ∼ 3-4) and lower toxicity, compared to <b>PIM</b>. The activity of <i>N</i>-derivatives of <b>PIM</b> towards fungi strains or cancer cells, depending on polar head structure, is discussed.

Early diagnosis of hepatocellular carcinoma (HCC), particularly in alpha-fetoprotein (AFP)-negative patients, remains a significant challenge. Characterizing N-glycopeptides with site-specific glycan structural information enables a better understanding of the molecular pathogenesis of liver injury and cancer. Here, we performed an unbiased quantitative analysis of N-glycopeptides from serum in patients with hepatitis B virus (HBV)-related liver diseases using a stable isotope labeling-based glycoproteomic approach. Serum samples from patients with liver cirrhosis (LC) and AFP-negative HCC were compared. A total of 264 unique N-glycopeptides were initially identified, with 30 high-confidence intact glycopeptides retained after stringent quality control. Following immunoprecipitation of ATIII, comparative analysis confirmed a significant downregulation of two ATIII glycopeptides carrying a biantennary disialylated (H5N4S2) glycan at asparagine residues N128 and N224 in AFP-negative HCC compared with LC (p < 0.01). A small AFP-positive HCC group was examined as an exploratory reference. Occupancy of four N-glycosylation sites (N128, N167, N187, N224) was verified, and site-specific glycan heterogeneity was delineated. These findings suggest that site-specific glycosylation changes on ATIII may serve as candidate biomarkers to complement current diagnostic strategies for AFP-negative HCC.

Resistance to <i>Leishmania (L.) major</i> depends on the development of a <i>L. major</i>-specific Th1 response, while Th2 differentiation results in susceptibility. We previously showed that the early microenvironment of infected skin delivers important signals for T cell differentiation. We found increased expression levels of coagulation factor X (F10 and FX for protein, respectively) 16 h after infection in the skin of resistant mice compared to susceptible mice. Activated FX is a ligand of proteinase-activated receptor-2 (PAR2), a modulator of inflammatory responses. To assess the role of PAR2, we analysed the course of <i>L. major</i> infection in PAR2-deficient (PAR2^{^-/^-}) mice on a resistant C57BL/6 background. PAR2^{^-/^-} mice developed significantly larger lesions and harboured more parasites in footpads and draining lymph nodes compared to wild-type mice. In addition, their antigen-specific T cell response was shifted towards Th2. Conversely, early treatment of susceptible BALB/c mice with a PAR2-agonist reduced parasite loads in footpads and spleens and shifted the T cell response towards Th1. This was accompanied by significantly higher expression of the Th1-promoting cytokines IL-6, IL-12, and TNFα in the infected skin. We conclude that PAR2 activation favours Th1-differentiation and resistance in experimental leishmaniasis due to an altered initial microenvironment with increased expression of Th1-promoting cytokines in the infected skin.

<h4>Background</h4>Huntington's disease (HD) is a monogenic neurodegenerative disorder caused by expansion of CAG trinucleotide repeats in the HTT gene, resulting in an aggregation-prone mutant huntingtin protein (mHTT) and progressive neuronal dysfunction. HD pathogenesis involves disturbances in protein homeostasis, mitochondrial function, oxidative stress, epigenetic regulation, and inflammatory signalling. We investigated the effects of curcumin, a naturally occurring polyphenol, on these disease-relevant processes in vitro.<h4>Methods and results</h4>Using established HD cellular models, we examined the effects of curcumin on mHTT aggregation, mitochondrial function, oxidative stress, and endoplasmic reticulum stress. Curcumin reduced mHTT aggregation and PERK-eIF2α signalling in HD cells by elevating autophagic clearance. Curcumin treatment increased ATP levels, partially restored Ppargc1α and Nrf1 expression, reduced mitochondrial as well as cytosolic reactive oxygen species and H3K27 trimethylation. In HD patient-derived lymphoblastoid cell lines, curcumin treatment abrogated inflammatory responses.<h4>Conclusions</h4>Curcumin alters multiple disease-relevant cellular processes in HD models, providing mechanistic insight into pathways associated with mHTT toxicity.

Microbial phenotypes vary at the single-cell level, shaping key community traits like resilience and adaptability. Yet, current methods either lack resolution (e.g., culturing, sequencing), are too costly, or technically complex, limiting widespread use. To address this gap, we introduce and validate a workflow called DE-SWIRL (Double Emulsion-Sorting Workflow with sImple, Rapid emuLsification), an accessible, low-cost workflow enabling ultrahigh-throughput (~107 microcultures/experiment) screening of individual microbial cells. DE-SWIRL integrates a published droplet-templated emulsification protocol producing uniform double emulsions from monodisperse single emulsions with Fluorescence-Activated Cell Sorting (FACS). We validated that double emulsions of 6 and 24 pL can be reliably formed, with ~45% droplet survival. To address persistent large-particle contaminants, we validated a gating strategy and show it enables accurate screening and sorting. When starting from a monodisperse single emulsion population, oil layer variability is higher for droplet-templated emulsification than for on-chip microfluidics, but maintains a comparably uniform core emulsion while offering substantial time savings. We demonstrate DE-SWIRL's utility by isolating viable strains from a synthetic community with up to 99% sorting purity and isolating droplet cocultures from a mixed community. This workflow provides a fast, accessible, and affordable workflow for screening entire microbiomes at a single-cell level using a fluorescent assay of interest.

Thallium (Tl) is an extremely toxic and strongly bioaccumulative metal increasingly detected in agricultural soils, yet its behavior at the plant-rhizosphere interface remains poorly constrained. Here, we integrated in situ diffusive gradients in thin films (DGT) with multi-omics analyses (transcriptomics, metabolomics, and 16S rRNA sequencing) to elucidate how plant-rhizosphere interactions regulate Tl mobility and detoxification in Brassica rapa. In situ DGT profiling coupled with the European Community Bureau of Reference (BCR) sequential extraction identified the root-soil interface (0-3 cm) as a hotspot of labile Tl dynamics, revealing a dose-dependent shift from rhizosphere-mediated Tl mobilization under moderate exposure to immobilization under high stress. This transition was mirrored by a hormesis-driven plant response, with low Tl levels stimulating growth and uptake (bioconcentration factor, BCF = 4.2), followed by growth inhibition and restricted translocation at higher doses. Multi-omics analyses showed coordinated metabolic and transcriptional reprogramming associated with this shift, including altered central carbon metabolism, glutathione homeostasis, phenylpropanoid biosynthesis, and selective regulation of metal transporters (ZIP downregulation; ABC and MATE upregulation). Key metabolites (L-proline, sinapoyl aldehyde) and genes (e.g., TAT, PRDX6) emerged as integrative regulators linking detoxification, redox balance, and osmoprotection. Concurrently, Tl exposure induced a functional succession of the rhizosphere microbiome toward metal-resistant taxa (e.g., Nitrospira, Microvirga), closely associated with changes in root exudation patterns. Collectively, these findings advance a process-based mechanistic understanding of how rhizosphere biogeochemistry, plant molecular responses and microbial dynamics jointly control Tl mobility and detoxification, informing Tl risk assessment and plant-microbe-assisted management.

The non-canonical inflammasome is a protein complex involved in bacterial infections, and its activation leads to excessive inflammatory responses during sepsis. The precise regulation of the non-canonical inflammasome in the body remains unclear. Here, we found that some chemicals that chelate zinc ions positively regulated the activation of the non-canonical inflammasome. These chemicals acted by inhibiting the activity of dipeptidyl peptidase 3 (DPP3). DPP3 cleaved phosphatidylethanolamine binding protein 1 (PEBP1) to generate an N-terminal fragment, which could bind to caspase-4/11 and inhibit the response intensity of the non-canonical inflammasome. PEBP1 N terminus appeared in the serum of mice and patients with sepsis. DPP3-deficient mice exhibited stronger inflammatory cytokine responses and had poor survival in the LPS-induced sepsis model. Promoting the activity of DPP3 effectively constrained the response intensity of sepsis in mice and increased their survival. Our findings provide a perspective for understanding the molecular regulatory process of the non-canonical inflammasome in sepsis.

MicroRNAs (miRNAs) have emerged as critical regulators in the pathogenesis of polyglutamine (PolyQ) diseases-a group of fatal neurodegenerative disorders caused by CAG repeat expansions, such as Huntington's disease, spinocerebellar ataxias, dentatorubral-pallidoluysian atrophy, and spinal and bulbar muscular atrophy. This review synthesizes recent advances in miRNA dysregulation across all nine PolyQ diseases, focusing on studies published since 2019. We examine how specific miRNAs modulate core pathogenic cascades-including mutant protein aggregation, transcriptional dysregulation, mitochondrial dysfunction, and apoptosis-and then link these molecular events to disease-relevant motor, cognitive, and psychiatric phenotypes. The review highlights therapeutic progress, including the preclinical efficacy of adeno-associated virus (AAV)-delivered artificial miRNAs and emerging exosome-based platforms that target mutant transcripts such as HTT, ATXN1, ATXN3, and ATXN7. AAV5-miHTT has advanced to a first-in-human trial for Huntington's disease (NCT04120493)-a key milestone in clinical translation. Circulating miRNAs in plasma and cerebrospinal fluid show diagnostic potential as minimally invasive, stage-specific biomarkers, but challenges persist in normalization, cross-biofluid concordance, and clinical validation. Despite substantial progress, translational barriers remain-including off-target effects, delivery optimization, immunogenicity, and patient heterogeneity. Overcoming these barriers will require integrative approaches that combine single-cell transcriptomics, engineered delivery systems, machine learning, and longitudinally phenotyped clinical cohorts. This review integrates mechanistic insights, biomarker discovery, and therapeutic development to move miRNA-based strategies toward disease-modifying interventions for PolyQ disorders.

Huntington's disease (HD) is a rare, autosomal dominant neurodegenerative disorder marked by progressive motor, cognitive, and behavioral symptoms. Although most cases present in adulthood, approximately 1-15% occur before age 20, termed juvenile Huntington's disease (JHD). JHD often follows a more rapid and variable clinical course, with diverse presentations and unique radiological findings. It's rarity and heterogeneous clinical course make diagnostic and management challenges. This case series highlights the clinical and neuroimaging features of three genetically confirmed pediatric patients with JHD from the same family. Ages of symptom onset ranged from 6 to 12 years, with all patients showing expanded CAG repeats (71-83) in the HTT gene. While all presented with cognitive impairment and motor symptoms, their clinical presentations varied. One patient had prominent dystonia and agitation, while another developed significant bradykinesia and altered mental status. Brain MRI findings were consistent across cases, demonstrating severe bilateral caudate and lentiform nuclei atrophy with corresponding T2/FLAIR hyperintensities and T1 hypo intensities. Susceptibility-weighted imaging revealed focal mineral deposition in the globus pallidus in two patients. Notably, mild thoracic spinal cord thinning was observed in one patient, suggesting potential spinal involvement. This case series provides new insight into the phenotypic variability and also demonstrates the less commonly described imaging observations of JHD, including spinal cord changes and SWI abnormalities. It underscores the importance of early imaging, genetic testing, and multidisciplinary care. Support for affected families is critical, as JHD imposes profound emotional and caregiving burdens.

Metal dyshomeostasis and oxidative stress are implicated in the progression of cancer, and neurodegenerative and peripheral aggregation-related disorders. In this study, we investigated primaquine (PQ), a clinically used antimalarial drug, and PQ-lactobionic acid conjugate (LAPQ), a newly synthesized and characterized derivative obtained through conjugation with lactobionic acid, designed to improve physicochemical and biological properties. Both compounds were evaluated for their copper-coordination ability, antioxidant properties, interaction with amyloid-β (Aβ), and capacity to modulate reactive oxygen species (ROS)-induced cellular damage. As a complementary biological line of investigation, their in vitro antiproliferative activity in the presence and absence of copper was also evaluated. The compounds directly interact with Aβ as demonstrated by surface plasmon resonance (SPR) studies. Biological studies revealed marked differences between the two molecules. PQ displayed intrinsic cytotoxicity, whereas LAPQ exhibited enhanced aqueous solubility and substantially reduced antiproliferative activity under the tested conditions, highlighting the impact of sugar conjugation. In cellular oxidative stress models, LAPQ showed a protective effect, preserving cell viability under ROS-generating conditions. In summary, by interacting with Aβ and exhibiting antioxidant activity, two properties relevant to several neurodegenerative and peripheral aggregation-related disorders, our novel compound LAPQ may provide a potential starting point for the development of therapies targeting Aβ-associated disorders.

A pregnant woman with hereditary antithrombin deficiency was managed with adjusted-dose LMWH throughout pregnancy, monitoring D-dimer and anti-Xa activity, successfully preventing thrombosis and hemorrhage.

<h4>Background</h4>Diabetes of the exocrine pancreas (DEP) is a form of diabetes that arises in the presence of an exocrine pancreas disease and is mainly associated with acute and chronic pancreatitis. Therapeutic management of DEP has not changed over the past decades, and treatment is mostly insulin based. However, type 2 diabetes (T2D) treatment guidelines have changed with the aim of normalizing glucose levels and preventing diabetes-related comorbidities.<h4>Objectives</h4>The aim of this study was to evaluate real-word practice in the treatment of diabetes associated with pancreatitis, with a focus on individuals who had already developed cardiovascular and/or renal diseases.<h4>Designs</h4>We conducted a cross-sectional observational study on adult patients admitted to a tertiary-level hospital in Italy between 2018 and 2022 with a prevalent diagnosis of pancreatitis and diabetes.<h4>Methods</h4>Disease-related real-world data were collected from Hospital Information Systems using automated data extraction strategies and through the implementation of a patient-centered data repository. A multiple source real-world evidence-based methodology was used to identify patients with diabetes and to investigate changes in glucose-lowering treatments through hospitalizations.<h4>Results</h4>We identified 2146 hospitalized patients with pancreatitis from 2018 to 2022, 675 (31.4%) of whom also had diabetes. Data on glucose-lowering therapies were available for 444 of these patients. Individuals with diabetes and pancreatitis were predominantly male (65.2%) and older than those without diabetes (<i>p</i> < 0.001). At hospital admission, 41.9% were on insulin therapy and 44.4% on other drugs, with metformin being the most prescribed. After hospitalization, insulin use increased significantly (61.2%, <i>p</i> < 0.001) while metformin prescriptions decreased (from 44.7% to 26.2%, <i>p</i> < 0.001). Patients with cardiorenal comorbidities showed similar therapeutic patterns, with a pronounced post-discharge reduction in metformin use.<h4>Conclusion</h4>This real-world study provides updated evidence on the treatment of diabetes associated with pancreatitis in a tertiary care setting, showing a predominant use of insulin and metformin regardless of comorbidities. Despite emerging evidence on the benefits of non-insulin therapies in T2D, such approaches are not yet reflected in the management of DEP, underlining the need for specific clinical guidelines and dedicated trials.

Hepatocellular carcinoma (HCC) is characterized by high malignancy and widespread drug resistance at advanced stages, posing formidable challenges in clinical diagnosis and treatment. Mitochondrial dysfunction is a core driver of HCC initiation and drug resistance. However, how its plasticity dynamically regulates tumor growth under various conditions remains to be systematically elucidated. This review systematically delineates the critical roles of mitochondrial dysfunction in HCC pathogenesis, drug resistance, and targeted therapy. Regarding pathogenesis, we integrate four major pathways, namely aberrant oxidative phosphorylation (OXPHOS) coupled with mitochondrial DNA (mtDNA) mutations, glutamine metabolic reprogramming, oxidative stress imbalance, and dysregulated ferroptosis, to elaborate the molecular basis driving HCC malignant progression. In terms of drug resistance mechanisms, mitochondrial plasticity is highlighted as a central adaptive hub that synergistically promotes resistance through multifaceted mechanisms, including remodeling of the tumor immune microenvironment, dynamic regulation of OXPHOS levels, and modulation of ferroptosis sensitivity. With respect to therapeutic strategies, we summarize the current major intervention directions, encompassing ROS/OXPHOS modulators, inhibitors targeting mitochondrial metabolic enzymes and transporters, ferroptosis inducers, as well as nanodelivery systems and combination therapies. Of note, the efficacy of these interventions often depends on the mitochondrial phenotypic state of the tumor; single-target approaches tend to yield limited long-term benefits, whereas combination regimens or dynamic adjustments based on mitochondrial plasticity status hold greater promise for clinical application. Collectively, this review consolidates the evidence positioning mitochondrial dysfunction as a convergence point linking HCC progression, drug resistance, and targeted therapy, and proposes that "targeting mitochondrial plasticity" may represent a novel direction for overcoming drug resistance and achieving precision therapy, thereby providing a theoretical reference for future basic research and clinical translation.

Preeclampsia is a pregnancy-specific multisystem disorder in which placental stress is translated into maternal endothelial dysfunction, hypertension, and end-organ injury. This narrative review summarizes how six regulated cell death modalities-apoptosis, necroptosis, pyroptosis, ferroptosis, autophagic cell death, and cuproptosis-participate in this process at the maternal-fetal interface. Hypoxia, oxidative stress, endoplasmic reticulum stress, metabolic disturbance, and altered metal homeostasis converge on trophoblasts, decidual immune cells, and placental endothelial cells. Apoptosis reduces trophoblast mass and increases apoptotic debris; necroptosis releases DAMPs via the RIPK1/RIPK3/MLKL axis; pyroptosis activates inflammasome-caspase-gasdermin signaling and releases IL-1β and IL-18; ferroptosis generates iron-dependent lipid peroxidation products; autophagy and mitophagy determine whether stressed cells recover or progress to death; and cuproptosis links copper overload to mitochondrial proteotoxic stress. These death-related products, including syncytiotrophoblast debris, extracellular vesicles, cytokines, oxidized lipids, and mitochondrial danger signals, may disseminate from the placenta into maternal organs and amplify systemic vascular inflammation. By comparing the triggers, execution mechanisms, products, and clinical implications of each cell death modality, this review proposes an integrative model in which a self-amplifying placental cell death network drives the clinical syndrome of preeclampsia. We also discuss candidate biomarkers and therapeutic strategies, while emphasizing that future studies should define temporal dynamics, disease subtype specificity, and maternal-fetal safety before clinical translation.

Across various cancer entities, a wide array of subtypes of B cells that infiltrate tumors can be identified. These subtypes confer these cells with high adaptability in their functions and significant involvement in tumor progression. Tumor-infiltrating B cells act as distinctive microenvironmental mediators that build tertiary lymphoid structures, secrete diverse factors, and generate a wide antibody repertoire, thereby offering a valuable indicator of immunotherapy response. Recent clinical trials have demonstrated that intervening in tumor-infiltrating B cells can be an effective treatment strategy for tumors, including common types such as breast cancer, colorectal carcinoma, and non-small cell lung cancer. These findings suggest that tumor-infiltrating B cells should not be overlooked as therapeutic targets, as they offer significant potential to reduce the incidence of nonresponse, drug resistance, and immune-related adverse effects commonly seen in current treatment techniques. In this review, we explore the historical development of B cells in the context of tumors. We then discuss the various subtypes of tumor-infiltrating B cells, analyzing their diverse roles in tumor development. Additionally, we delve into the signaling pathways of tumor-infiltrating B cells, intercellular crosstalk, and multilevel regulatory mechanisms. Finally, based on immune checkpoints and therapeutic targets on the surface of tumor-infiltrating B cells, we summarize the current understanding which can lead to the development of new intervention methods targeting tumor-infiltrating B cells in the future. This review aims to provide a comprehensive summary of B-cell classification and therapeutic potential, emphasizing their critical role in guiding clinical treatment strategies.

Src-family kinases (SFKs) regulate proliferation in colonic epithelial cells (CECs), but the mechanisms that restrain their activity remain poorly defined. We identify Src-like adaptor protein (SLAP), a negative regulator of receptor tyrosine kinase signaling, as a key suppressor of SFK activity in the colon. Constitutive and inducible epithelial-specific Slap deletion using a villin-CreERT2 model increases CEC proliferation and accelerates tumorigenesis in the azoxymethane/dextran sodium sulfate model. Slap deficiency also enhances SFK-dependent expansion of normal and tumor-derived colonic organoids. Mechanistically, we identify the receptor tyrosine kinase EPHB2 as a critical upstream activator of SFKs and a direct target of SLAP-mediated regulation. Loss of Slap increased EphB2 protein abundance and tyrosine phosphorylation, and enhanced its association with active SRC. Pharmacological inhibition of EPHB2 suppressed SRC activation and reversed the hyperproliferative phenotype induced by Slap deficiency. Together, these findings uncover a non-genetic mechanism driving SFK activation during colonic transformation and establish SLAP as a tumor suppressor that constrains oncogenic EPHB2-SFK signaling in the colonic epithelium.

RNA-binding protein TAR DNA-binding protein 43 (TDP-43) can form liquid-like nuclear assemblies whose phase behavior is thought to influence its aggregation propensity and neurotoxic activity. The mechanisms that govern the liquid-to-solid phase transition of TDP-43 remain poorly defined. Here, we combined chemical and genome-wide genetic screens to identify cellular factors that modulate the phase behavior of an RNA-binding-defective TDP-43 mutant. Our screens uncovered multiple cellular processes, including RNA splicing, protein translation, proteostasis imbalance, and nuclear export as TDP-43 phase regulators. We also developed a semi-permeabilized cell system that partially recapitulates the TDP-43 phase transition in vitro, and showed that nuclear export inhibition reshapes the nuclear environment to favor RNA-dependent liquid-liquid phase separation (LLPS) of TDP-43, which mitigates its aggregation. Nuclear export inhibition in a brain organoid model bearing an ALS-associated mutation reduces pathogenic phospho-TDP-43 accumulation. These findings identify multiple modulators of TDP-43 phase transitions in a sensitized model system and establish a framework for further dissecting the link between nuclear transport and TDP-43 phase dynamics.

Eukaryotic tailless complex polypeptide 1 ring complex/Chaperonin containing tailless complex polypeptide 1 (TRiC/CCT) chaperonin is typically considered a cytosolic machine mediating polypeptide folding and assembly of protein complexes. Here, we investigated the nuclear role of TRiC/CCT. Use of a TRiC/CCT temperature-sensitive allele revealed increased production of nascent RNA leading to the accumulation of noncoding transcripts. TRiC/CCT was associated with RNA polymerase II (RNAPII) in vitro and in vivo, including when bound to DNA. Heat treatment of the TRiC/CCT ts chaperonin stabilized the RNAPII complex association and binding to the actin and tubulin substrates. Expression of the Huntingtin protein Htt correlated with lowered RNA production and a decreased association between the RNAPII and TRiC/CCT complexes. Together, our presented data support a model where TRiC/CCT regulates the global activity of RNAPII in reaction to the status of proteostasis. Overall, our work reveals an avenue by which TRiC/CCT contributes to homeostasis by regulating the activity of nuclear RNAPII.

Cell death is essential for maintaining internal stability and metabolic balance in the human body and plays a key role in regulating physiological and pathological processes. Advances in understanding the link between pathological mechanisms and cell death have provided new insights into diagnosis and treatment. A novel, pivotal form of regulated cell death, termed disulfidptosis, has recently been identified. This is characterized by glucose deprivation, elevated expression of solute carrier family 7 member 11 (SLC7A11), accumulation of disulfides such as cystine, disulfide stress, and cell death. These distinctive characteristics indicate that disulfidptosis is mechanistically distinct from other known forms of cell death. Its unique induction mechanism implies its therapeutic potential and warrants further investigation. This cell death process is primarily triggered by intracellular NADPH exhaustion and subsequent actin cytoskeleton collapse, and it exhibits complex crosstalk with ferroptosis, cuproptosis, pyroptosis, necroptosis, apoptosis and autophagy. The core execution relies on the Rac-WRC-Arp2/3 pathway, with emerging involvement of ER stress and p38 MAPK signaling. In this review, we discuss the molecular mechanisms and current status and prospects of targeted therapy for disulfidptosis, as well as its role in cancer and other diseases, including but not limited to neurodegenerative, cardiovascular, autoimmune, and infectious conditions. We hope to provide a comprehensive and systematic framework, including mechanistic analysis, application strategies, clinical translation, and future research directions, to guide in-depth investigation.

Mitochondrial dysfunction is one of the earliest pathological features of Alzheimer's disease (AD), preceding overt neurodegeneration and cognitive decline. Amyloid-β (Aβ) accumulation has long been considered a central pathogenic event in AD, yet how Aβ toxicity is mechanistically linked to mitochondrial impairment during early disease stages remains incompletely understood. To address this, we combined multi-omics with in vivo and in vitro genetic interventions. Here, we show that malic enzyme 3 (Me3) links Aβ aggregation to mitochondrial dysfunction in APP/PS1 mice and neuronal cells. At ultra-early and early AD stages (3 and 6 months), Me3 was upregulated and accumulated within mitochondria, where it colocalized with Aβ42 and physically interacted with it, an association linked to oxidative stress and impaired mitophagy. Knockdown of Me3 reduced mitochondrial reactive oxygen species, improved mitochondrial morphology, and alleviated mitophagy defects in both cellular and mouse models, with statistical significance across these functional measurements (<i>p</i> < 0.05). These results suggest that Me3 functions not only as a metabolic responder to Aβ-associated stress but also as a contributor to early mitochondrial pathology. By identifying the Aβ-Me3 axis, this study provides mechanistic insight into early mitochondrial dysfunction in AD, while further validation in human AD samples remains necessary.

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screening has become a central technology in functional genomics, enabling genome-scale interrogation via pooled perturbations. Early CRISPR screens employed survival or simple phenotypic readouts to identify essential genes and drug resistance mechanisms. However, as biological questions have shifted toward understanding regulatory networks, cellular heterogeneity, and context-dependent gene functions, there has been increasing demand for screening strategies capable of capturing complex cellular phenotypes beyond cell fitness. Recent advances in single-cell sequencing, high-content imaging, and spatial transcriptomics have expanded the resolution of CRISPR screening by enabling multidimensional phenotypic characterization following genetic perturbation. By integrating pooled perturbations with diverse readouts, these approaches systematically map targeted gene edits to transcriptional states, cellular phenotypes, and microenvironmental contexts. Meanwhile, innovations in library design, delivery, and computational pipelines have further improved the robustness and interpretability of high-content screening platforms. This review synthesizes the methodological evolution of CRISPR screening, emphasizing advances in perturbation strategies, delivery systems, and multimodal readouts. Representative applications spanning oncology, immunotherapy, developmental biology, neurobiology, and infectious diseases are delineated to demonstrate refined gene network annotations. Additionally, existing technical bottlenecks, such as scalability, cost constraints, and in vivo limitations, are critically assessed. Finally, future directions are proposed to facilitate the development of precise medicine.

<h4>Background</h4>Thrombosis is a multifactorial disorder with a substantial genetic component, yet gene-gene interactions contributing to thrombotic risk remain insufficiently characterized in Southeast Asian populations. Identifying both individual genetic determinants and interaction effects may improve risk stratification, particularly in families with early-onset thrombotic disease.<h4>Objective</h4>This study aimed to explore individual and interactive genetic contributions to thrombotic events in Vietnamese pedigrees with early-onset venous thromboembolism (VTE) using an integrated analytical framework combining Multifactor Dimensionality Reduction (MDR) and SHapley Additive exPlanations (SHAP).<h4>Methods</h4>A pedigree-based study was conducted among 88 individuals from families with a history of early-onset VTE (<50 years), including 19 affected cases. Genetic variants were identified using next-generation sequencing. Random Forest modeling with SHAP was used to quantify global and local feature contributions, while MDR was applied to detect potential non-linear gene-gene interactions beyond single-locus effects. The Random Forest model was not explicitly adjusted for familial clustering or relatedness; results were therefore interpreted as exploratory.<h4>Results</h4>SHAP analysis identified rs1045642 (ABCB1) as the strongest contributor to model-predicted baseline thrombotic risk. MDR analysis suggested that this background susceptibility may be further modified by a two-locus interaction between rs12041331 (PEAR1) and rs1800790 (FGB). Individuals classified within the high-risk MDR genotype combination showed increased odds of thrombosis (OR = 8.00, 95% CI: 1.67-74.96; p = 0.0028), compared with the corresponding single-locus model (OR = 3.73, 95% CI: 0.93-21.48; p = 0.0405). Confidence intervals were wide, reflecting limited precision.<h4>Conclusion</h4>In this exploratory pedigree-based study, rs1045642 (ABCB1) was associated with baseline thrombotic susceptibility, while a potential interaction between PEAR1 and FGB may further modulate risk in specific genotype combinations. The integration of MDR and SHAP provides an interpretable framework for investigating complex genetic architectures. However, given the small sample size and limited number of events, these findings should be considered hypothesis-generating and require validation in larger independent cohorts.

Nuclear delivery of human papillomavirus (HPV) requires infected cells to undergo mitosis. Incoming HPV DNA is protected by a transport vesicle structure, which becomes unstable post-mitosis. We have previously shown that HPV genome egress takes on average 4 hours post-mitosis completion; thus, we propose that an enzymatic process is involved in degrading this transport vesicle to allow genome egress. Several phospholipase C (PLC) isoforms, including PLCδ3, PLCζ1, and PLCL1, have been previously implicated in HPV infection in large-scale siRNA screens, and the parvovirus VP1 capsid protein has phospholipase enzymatic activity. Therefore, we hypothesized that cellular phospholipases residing in the nucleus mediate egress of the HPV genome from transport vesicles. We utilized siRNA-mediated knockdown and CRISPR/Cas9 knockout techniques to target specific PLC isoforms in both HeLa and HaCaT cell lines. We found that targeting PLC isoforms PLCβ1, β4, δ3, δ4, and γ1 significantly decreased HPV infection in both HeLa and HaCaT cells, as measured using a luciferase-based reporter assay. We also discovered a novel interaction between HPV16 minor protein L2 and PLCs β1, β4, δ1, δ3, and δ4. Furthermore, we observed that knockout of PLCβ4 and δ4 delayed egress of the HPV genome from nuclear membrane-bound vesicles after infection of HeLa cells. We propose that HPV utilizes phospholipase Cs to achieve genome egress from its protective vesicular structure after nuclear delivery. <h4>Importance</h4> The cellular factors involved in the later stages of HPV entry remain to be elucidated. Previous data gleaned from large-scale siRNA screens implicated the involvement of phospholipases in HPV infection. We expanded upon these findings and discovered that the individual phospholipase C (PLC) isoforms β4 and δ4 are important for HPV infectivity. We further linked the loss of infectivity in PLCβ4- and PLCδ4-deficient cells to a defect in HPV genome release from its transport vesicle, which we demonstrated using our differential staining technique. Furthermore, we discovered a novel interaction between the minor protein L2 of HPV16 and PLC isoforms β1, β4, δ1, δ3, and δ4.

Huntington disease is a fatal neurodegenerative disorder caused by CAG repeat expansion encoding polyglutamine in the HTT gene. Recent studies have shown that loss of CAA/CCA interruptions within polyglutamine-coding CAG tracts and adjacent polyproline-coding region are linked to earlier disease onset. It has been hypothesized that somatic repeat instability, influenced by these interrupted CAG tracts, may mediate this effect. Here we demonstrate that HTT CAA/CCA-loss variant linked to early disease onset exacerbates mutant HTT toxicity in cellular models through mechanisms independent of somatic repeat instability. The CAA/CCA-loss variant exhibits significantly higher toxicity than canonical HTT sequences in both transient expression and stable cell line models. Notably, this enhanced toxicity persists in knockout cells of the mismatch repair gene MSH3 where somatic instability is blunted, with a consistent toxicity hierarchy (CAA/CCA-loss > CCA-loss > CAA-loss > canonical HTT) in both wildtype and MSH3 knockout cells. Furthermore, the CAA/CCA-loss variant generates elevated levels of repeat-associated non-AUG (RAN) translation products. In HEK293-based cellular models, these results suggest that the disease-accelerating effects of HTT CAA/CCA-loss variants involve intrinsic properties of the altered sequence context, highlighting the importance of understanding sequence-specific mechanisms in HD pathogenesis beyond polyglutamine length and somatic instability.

Utilizing key developmental cues and refining their orchestrating role in degeneration represents a promising strategy for understanding and treating intervertebral disc (IVD) degeneration, a major cause of chronic lower back pain. Here, we focus on notochordal cells (NCs), which originate from the embryonic notochord and reside in the developing nucleus pulposus. These distinctly vacuolated cells exhibit robust regenerative effects and hold promise for new therapeutic approaches. Dogs, like humans, suffer from the consequences of IVD degeneration. As the IVD matures and degenerates, NCs are replaced by smaller non-vacuolated NP cells (NPCs). The dog was employed as a model to capture, at the single-cell level, the heterogeneity of resident cells by studying the nucleus pulposus tissue at three stages (i.e., juvenile, young adult and degenerate adult). Here, we integrated transcriptomic data with repressive histone H3 lysine 27 trimethylation (H3K27me3) profiles at the single-cell level to assess changes in chromatin states and gene expression across this IVD degeneration-associated cell phenotypic transition. H3K27me3 enrichment on key genes involved in IVD development and homeostasis, such as Brachyury (TBXT), aligns with the observed attenuation during ageing and degeneration seen in both dog and human IVDs. This study further demonstrates that eliminating repressive histone marks, together with CRISPR-mediated gene transactivation, enhances TBXT gene expression in human NPCs derived from degenerated aged discs. Our findings underscore how extensive insights gained through single-cell omics can lead to the identification of crucial cellular cues that may enable degenerate NPCs to regain a healthier phenotype.

Squaramides are well-known charge-neutral bioisosteres of many nitrogenous functional groups, including those found in the side chains of amino acids (<i>e.g.</i>, Arg, Asn, Glu, and Lys). However, there are very few reports of squaramide-functionalisation of these side chains. Herein, we report a convenient three-step synthesis for Boc, Alloc, or Cbz protected squaramide-functionalised Fmoc amino acids (SqAAs) derived from Dap, Dab, Orn and Lys in good yields and high enantiopurity (>95% ee), in milligram to multigram scale (38-93% over three steps for Boc-protected amino acids). These amino acids were found to be compatible with a variety of commonly used amide coupling reagents and green solvents, as well as being extremely resistant to epimerisation in solution-phase coupling. Orn and Dab derivatives (7 and 8) were able to be incorporated into solid-phase Fmoc SPPS workflows in the construction of four tetrapeptides, Arg/Asp-SqAA-Gly-Leu, in good yield (23-57%) and with no significant byproduct formation. We propose that these squaramide-functionalised amino acids could serve as convenient building blocks for constructing bioisosteres of bioactive peptides.

Triple-negative breast cancer (TNBC) is a highly aggressive and treatment-resistant subtype, lacking HER2, progesterone, and estrogen receptors. Systemic chemotherapy remains the primary treatment due to the absence of molecular targets, often leading to poor prognosis and high recurrence. This study used an integrated <i>in silico</i> transcriptomic and network-based bioinformatics approach to identify TNBC key genes (TKGs) and explore potential therapeutic candidates. Four TNBC microarray datasets (GSE36295, GSE38959, GSE45827, and GSE65194) were analyzed with the LIMMA algorithm in GEO2R, revealing 315 TNBC-shared differentially expressed genes (TSDEGs). Eight strongly connected TKGs were identified via topological analysis and protein-protein interaction (PPI) network construction across the STRING and IMEx databases from TSDEGs: CDK1, TOP2A, CCNB1, CDK2, FN1, UBC, PRKDC, and PARP1. Enrichment analysis of biological processes, molecular functions, cellular components, and KEGG pathways, combined with regulatory network analysis involving transcription factors and microRNAs, identified pathogenic roles of these genes. CDK1 and CDK2 were identified as the top pharmacological targets for molecular docking studies. Finally, TKG-guided top-ranked two drug molecules (Alsterpaullone and BLU-222) emerged as promising repurposed therapeutic candidates for TNBC. The absorption, distribution, metabolism, excretion, and toxicity (ADMET) and drug-likeness analyses showed these molecules have favorable pharmacokinetic properties. Molecular dynamics simulations over 100 nanoseconds demonstrated stable binding interactions and favorable behavior for the complexes CDK1-Alsterpaullone and CDK2-BLU-222, as indicated by root mean square deviation, fluctuation, and molecular interactions generalized Born surface area. Overall, these findings may aid in diagnosing and treating TNBC, providing valuable resources for future therapeutic strategies.

Autophagy is induced by nutrient starvation to recycle intracellular constituents; however, its activity must subsequently be attenuated during prolonged nutrient deprivation. The mechanisms underlying this attenuation in mammalian cells remain incompletely understood. Here, using complementary HaloTag-based assays, we show that autophagic activity declines during prolonged starvation in HeLa cells. A genome-wide CRISPR/Cas9 knockout screen designed to identify cells that sustain autophagic activity under these conditions identified protocadherin 17 (PCDH17) as a regulator of autophagy attenuation. PCDH17 depletion maintained autophagic activity during prolonged starvation without detectably altering mTORC1 signaling, ULK1 abundance, or the proximal machinery of autophagosome formation. Instead, PCDH17 depletion increased lysosomal abundance, acidification, and proteolytic activity, whereas PCDH17 overexpression produced reciprocal effects. We further identified a lysosome-associated PCDH17 subpopulation that is supplied predominantly through the biosynthetic ER–Golgi pathway. This pool undergoes proteolytic processing and lysosomal turnover, with starvation preferentially accelerating degradation of the C-terminal fragment while preserving a comparatively stable N-terminal fragment. Together, these findings identify PCDH17 as an unexpected negative regulator of lysosomal function and demonstrate that modulation of lysosomal degradative capacity contributes to autophagy attenuation during prolonged starvation.

Arteriovenous fistula (AVF) is the preferred vascular access for patients with end-stage renal disease; however, its failure is primarily due to neointimal hyperplasia. Five patients who underwent initial AVF surgery served as the control group, and another five patients with failed AVF surgery served as the experimental group. Herein, we employed mass spectrometry (MS)-based quantitative proteomics coupled with tandem mass tag labeling to screen differentially expressed proteins (DEPs) in the anastomotic cephalic vein, followed by bioinformatics analyses and verification experiments. A total of 121 DEPs were identified in the failed AVF group. GO analysis was primarily enriched in protein binding, nucleic acid binding, enzyme binding, mRNA binding, cadherin binding, catalytic activity, and cell adhesion molecule binding. KEGG pathways were mainly enriched in cell aggregation and adhesion, actin cytoskeleton, extracellular matrix-receptor interaction, PI3K-Akt signaling pathway, complement and coagulation cascades, and cholesterol metabolism. Protein-protein interaction network consisted of 86 (71.07%) DEPs, including complement VII (C7), factor IX (F9), SERPINC1, microfibril-associated glycoprotein 4 (MFAP4), complement C1s subcomponent, complement C1q subcomponent subunit A, complement C1q subcomponent subunit B, tissue factor, and von Willebrand factor, which interacting with numerous other proteins. In the expanded validation for different patients, C7, F9, SERPINC1, and MFAP4, were verified by immunohistochemical staining and Western blotting, which were consistent with the proteomics results. Collectively, this study identifies a series of potential diagnostic biomarkers, and explores the underlying mechanisms associated with AVF dysfunction.

Major depressive disorder (MDD) is a highly prevalent psychiatric disorder that remains challenging to treat effectively. Difficulty predicting who will develop the disorder and which individuals will respond to treatment is most likely due to our limited mechanistic understanding and the lack of reliable biomarkers. The current literature suggests that lipid profiles differ in MDD; however, most studies do not integrate other omics layers to elucidate their potential causal relationship at the molecular level. Using phenotypic, genetic, epigenetic and mass-spectrometry lipid data from a Scottish-based cohort of approximately 1,000 participants, we examined associations with individual lipid species. We identified 77 common genetic variant associations (β range = -0.46 to 0.69, p < 3.1x10 -10 ) and 82 genome-wide DNA methylation associations (β range = -1.88 to 1.39, p < 2.2x10 -10 ). We identified 23 lipid species associated with lifetime MDD (β range = - 0.23 to 0.2, p < 0.05), including lipid classes such as lysophosphatidylcholine (LPC), diacylglycerol (DG) and phosphatidylcholine (PC). Our integration analysis showed that genetic variants and DNA methylation at sites annotated to the FADS gene cluster were linked with lipid species that were also associated with lifetime MDD status, including LPC(14:1) (β = 0.15, p = 0.03) and PC(19:0)/(20:4) (β = -0.14, p < 0.05). Our findings provide further support for the role of the FADS gene cluster in lipid metabolism and its association with MDD, while introducing DNA methylation as an additional molecular layer supporting this link.

Survival of extremely preterm infants has improved owing to advances in perinatal and neonatal care. However, bronchopulmonary dysplasia (BPD) has remained unabated, and research into preventive measures for the broader spectrum of prematurity-associated lung disease (PLD) has met with limited success. We summarize emerging therapeutic strategies with the potential to favorably influence the lifelong trajectory of lung health of preterm infants, especially the most immature. This narrative review examines recent preclinical and clinical studies on therapies for BPD prevention, focusing on mechanism-based interventions, biologic strategies, and innovative translational tools. Preclinical evidence converges on actionable targets in alveolarization, inflammatory pathways, redox and mitochondrial homeostasis, vascular and matrix remodeling. Novel molecules and drug-repurposing strategies are being tested in animal models, while patient-derived platforms may enable personalized therapeutic approaches. Mechanism-based approaches are progressing through Phase 1-2 clinical trials; pending efficacy analyses, insulin-like growth factor-1 replacement may become the first pharmacological therapy specifically approved to reduce or attenuate BPD. In parallel, cell-based therapies and extracellular vesicle strategies offer a complementary paradigm, acting through pleiotropic biological reprogramming of the injured lung. Ethical-regulatory complexity and the challenges of standardizing and scaling cell-based and cell-free therapies may limit clinical translation.<h4>Conclusion</h4>A growing repertoire of therapies for BPD holds promise for modifying early respiratory illness and the long-term burden of PLD. Realizing this potential will benefit from both mechanism-informed combinatorial approaches and pleiotropic strategies, supported by patient-derived translational tools and adequately powered efficacy evaluations. Equally essential are tailored neonatal-first regulatory pathways linking the continuum of BPD to the later respiratory morbidity of PLD, a life-course condition with significant public health relevance beyond the neonatal period.<h4>What is known</h4>• Bronchopulmonary dysplasia (BPD) is the current clinical and regulatory endpoint for neonatal pulmonary therapies. • BPD definitions capture the most severe manifestations of prematurity-associated lung disease (PLD), a broader continuum of respiratory morbidity reaching into adulthood; attenuating BPD may improve the whole spectrum of PLD.<h4>What is new</h4>• Therapies targeting alveolar, inflammatory, oxidative, vascular, and remodeling pathways are in clinical development; pending late-stage trials, IGF-1 replacement may become the first disease-modifying therapy to reduce BPD. • Novel targets, experimental platforms, and repurposed drugs hold promise for precision therapies in preterm infants.

<h4>Purpose of review</h4>Obesity reshapes systemic iron handling, yet the resulting iron phenotype is frequently misclassified at the bedside and in population studies. This review examines how excess adiposity disturbs iron metabolism and complicates iron-status assessment, with balanced attention to biology and measurement problems across adults and children.<h4>Recent findings</h4>Adipose tissue is an endocrine, inflammatory, and iron-handling organ. In obesity, interleukin-6 drives hepatic hepcidin through JAK/STAT3 signaling, while leptin, adipose hypoxia, and adipose hepcidin expression may contribute additional signals. Hepcidin degrades ferroportin, reducing duodenal iron export and limiting macrophage iron release, producing hypoferremia and iron-restricted erythropoiesis despite normal or elevated ferritin. We and others have shown that women with obesity may have higher hepcidin, ferritin, and inflammatory markers but lower serum iron. Stable-isotope and intervention studies suggest weight loss reduces inflammation and hepcidin and improves iron absorption, whereas the World Health Organization and Biomarkers of Nutrition for Development frameworks recommend interpreting ferritin relative to inflammation. Iron status in obesity spans a continuum from absolute or functional iron deficiency to sufficiency and, in some individuals, hyperferritinemia with possible dysmetabolic iron overload. Reliable assessment requires a multi-marker strategy: ferritin interpreted alongside an inflammation marker such as C-reactive protein and supported by transferrin saturation, soluble transferrin receptor, reticulocyte hemoglobin, or other context-appropriate indices. Management should address reversible inflammation through weight loss and metabolic risk reduction, use oral or intravenous iron according to the likelihood of true deficiency and hepcidin-mediated oral refractoriness, and avoid reflexive phlebotomy for dysmetabolic hyperferritinemia without confirmed overload.

Synapses, prototypic sites for neuronal communication, are key to brain function. Their organization and properties are instructed by synaptic cell adhesion molecules (sCAMs) that may operate independently or in coordination through yet unknown linker proteins. Here, we used multi-epitope affinity-purifications combined with quantitative mass spectrometry and immuno-EM to comprehensively map synaptic protein networks in the mouse brain. We identify a pre-synaptic core-module assembled from the major sCAMs, Neurexins1-3 and LAR-type receptor protein-tyrosine-phosphatases (PTPRs), and the previously uncharacterized tetraspanins T178A/B. These ternary Neurexin-T178-PTPR complexes form through their trans-membrane domains and assemble during biogenesis in the ER. Loss of T178B leads to module destabilization, accompanied by strong reduction of LAR-PTPRs and re-distribution of synaptic Neurexins. At synapses, the Neurexin-T178-PTPR module recruits stable trans-synaptic protein networks thereby interlinking machineries of the pre-synaptic active zone and establishing stable associations with post-synaptic neurotransmitter receptors. This work uncovers a widely distributed core-module for synaptic adhesion and trans-synaptic signaling in the mammalian brain.

In the gastrointestinal tract, Wnt and BMP signals control Lgr5⁺ stem cell activity during homeostasis, whereas injury elicits an Lgr5-independent, fetal-like regenerative program driven by YAP. Helicobacter pylori (H. pylori) infection activates YAP, but whether fetal-like reprogramming contributes to gastric pathology, and what drives it, has remained unclear. Here we show that H. pylori-induced gland hyperplasia is accompanied by YAP-dependent fetal-like transcriptional response and loss of epithelial BMP signaling. Epithelial BMP inhibition alone is sufficient to induce this program in vivo, through an epithelial-immune-stromal cascade: BMP-deficient epithelial cells secrete chemokines that recruit IL-1β-producing immune cells, and IL-1β drives enrichment of pro-regenerative fibroblasts producing prostaglandin E2. In gastric epithelial-stromal assembloids, IL-1β elicits stromal prostaglandin E2 production and subsequent epithelial YAP activation. Stromal deletion of the IL-1 receptor abrogates H. pylori-driven reprogramming and pathology. These data define a cascade that converts BMP loss into a fetal-like regenerative state and shapes H. pylori-associated gastric disease.

Rare diseases (RDs) are individually uncommon but collectively affect a large global population, and the vast majority still lack effective disease-modifying therapies. With advances in genomics and data-sharing platforms, research has increasingly shifted from a single-disease perspective to the search for convergent molecular pathways that might be shared across clinically distinct entities. In this context, the purinergic P2X7 receptor (P2X7R) has emerged as a putative "shared molecular platform" due to its central role in inflammation amplification, cell death and immune regulation. P2X7R is an ATP-gated ion channel with unique structural and functional features: under high extracellular ATP, it not only forms a non-selective cation channel but can also dilate into a "large pore" permeable to macromolecules, thereby triggering Ca²⁺overload, NLRP3 inflammasome assembly, reactive oxygen species (ROS) production and apoptotic/necrotic-like cell death. This review briefly outlines the epidemiology of RDs and the structural-functional characteristics of P2X7R, then systematically summarizes current evidence linking P2X7R to multiple rare diseases, including Charcot-Marie-Tooth disease, Guillain-Barré syndrome, amyotrophic lateral sclerosis, Huntington's disease, multiple sclerosis, and selected inflammatory and metabolic RDs (CAPS, familial Mediterranean fever, Systemic sclerosis, Dravet syndrome and Gaucher disease). By comparing P2X7R expression and functional alterations, downstream signaling pathways and pharmacological data from animal models across these conditions, we propose that a P2X7R-dependent network centered on a "Ca²⁺-NLRP3-inflammation/cell death axis" may constitute a common pathogenic backbone for diverse RDs. At the same time, disease-specific spatiotemporal expression patterns of P2X7R in central vs peripheral nervous systems and in immune vs target organ cells confer marked context dependence and "double-edged sword" properties. Finally, we discuss opportunities and challenges for P2X7R-targeted strategies, including the impact of disease stage and sex differences on therapeutic efficacy, and key bottlenecks in translating preclinical findings into clinical benefit. A deeper understanding of both shared and disease-specific roles of P2X7R may provide a conceptual framework and therapeutic entry point for precision stratification and multi-target interventions in rare diseases.

CCCTC-binding factor (CTCF) is a key architectural protein in the three-dimensional (3D) genome, yet how its loss reshapes chromatin structure and transcription at single-cell resolution remains unclear. Using HiRES, which jointly profiles chromatin contacts and RNA from the same nucleus, we examined genome-wide effects of CTCF depletion. Topologically associating domain (TAD)-like domains (TLDs) across single cells remained largely unchanged in number and size after CTCF loss, but their boundaries became more variably positioned, and pseudobulk analyses revealed reduced interactions within A compartments. We also developed SALTAFinder to identify Spatially Aggregated Long-distance TLD Assemblies (SALTAs), clusters of TLDs occupying shared 3D space within single cells. A subset of SALTAs is enriched for highly expressed genes and super-enhancers and declines upon CTCF depletion. This structural reorganization coincided with a global reduction in per-cell RNA output, as indicated by HiRES and orthogonal measurements. Together, these findings suggest that CTCF contributes to the coordinated regulation of chromatin organization and transcriptional capacity and is associated with stabilization of long-range active chromatin clusters.

Breast density and estrogen exposure are two major risk factors for breast cancer; however, the underlying biological mechanisms remain incompletely understood. Here, we investigated the extracellular proteomic landscape of normal human breast tissue in situ to define the microenvironment associated with these risk factors. Forty-two postmenopausal women with nondense or dense breasts and 19 premenopausal women underwent microdialysis. We quantified 461 inflammatory proteins in breast tissue and matched subcutaneous fat, enabling discrimination between local and systemic alterations. Breast density was assessed using magnetic resonance imaging. Dense breast tissue exhibited a distinct protein signature characterized by immune-related signaling, altered lipid metabolism, and the extracellular presence of intracellular proteins, consistent with cellular stress and immune modulation, with limited changes in angiogenic and extracellular matrix remodeling proteins. In contrast, estrogen-exposed breasts displayed a proteomic profile dominated by pro-inflammatory cytokines, angiogenic factors, extracellular matrix remodeling proteins, and complement activation, indicative of a dynamic and pro-tumorigenic microenvironment. These findings demonstrate that breast density and estrogen exposure are associated with fundamentally distinct breast microenvironments, both permissive for tumor progression. These breast-specific protein signatures provide mechanistic insight into how these risk factors contribute to cancer development and suggest that effective prevention strategies may require differential targeting.

<b>Background:</b> Radiotherapy eliminates most tumor cells but spares persister tumor cells that evade cell death and drive relapse. Increasing evidence suggests that stromal components of the tumor microenvironment influence treatment responses, yet whether macrophages actively reprogram tumor-intrinsic stress responses to promote radioresistance remains unclear. Here, we investigated the mechanisms by which macrophage-tumor cell interactions regulate ferroptosis and tumor survival after irradiation. <b>Methods:</b> We used macrophage-tumor cell coculture systems, Transwell separation assays, and 3D microfluidic models to examine contact-dependent effects on tumor survival following irradiation. Kinome-wide small interfering RNA screening, RNA sequencing, lipidomic profiling, and quantitative proteomic analysis of secretomes were performed to identify signaling pathways and metabolic changes. Genetic and pharmacological perturbation of Ephrin receptor b4 (Ephb4) signaling were evaluated in vitro and in syngeneic mouse tumor models. Clinical relevance was assessed using transcriptomic analyses and immunohistochemical staining of patient tumor specimens. <b>Results:</b> Macrophage contact reduced lipid peroxidation and cell death in irradiated tumor cells in a contact-dependent manner. Kinome screening identified Ephb4 as a key mediator induced by irradiation in tumor cells. Ephb4 engagement with ephrinb2 on macrophages initiated bidirectional signaling that increased expression of ferroptosis-protective genes (solute carrier family 7 member <i>11</i> [<i>Slc7a11</i>], solute carrier family 3 member 2 [<i>Slc3a2</i>], and glutathione peroxidase 4 [<i>Gpx4</i>]) in tumor cells while activating the toll-like receptor 2- nuclear factor-kappa B pathway and interleukin-6 (IL-6) production in macrophages. Macrophage-derived IL-6 further sustained ferroptosis resistance in tumor cells, and Ephb4-driven secretion of cathepsin S amplified macrophage IL-6 production through a feedforward loop. Genetic or pharmacological inhibition of Ephb4 restored lipid peroxidation and markedly enhanced radiosensitivity in vitro and in vivo. Analysis of patient datasets demonstrated increased EPHB4 expression following radiotherapy and an association between high EPHB4 expression, reduced ferroptosis signatures, and poor treatment response. <b>Conclusions:</b> These findings identify a macrophage-driven ferroptosis evasion program that enables tumor cell survival after irradiation and demonstrate that Ephb4 coordinates bidirectional tumor-macrophage signaling to sustain this resistance. Targeting the Ephb4-ephrinb2 axis represents a potential strategy to enhance ferroptosis and improve radiotherapy efficacy in resistant tumors.

<h4>Background</h4>Lung cancer (LC) and venous thromboembolism (VTE) are closely associated, with VTE contributing to morbidity and mortality among patients with LC. We aimed to identify and characterize a mitochondrial-related transcriptomic signature shared between LC and VTE and to explore its association with immune microenvironment features.<h4>Methods</h4>We applied a multiomics approach focused on mitochondrial-related signaling pathways. Publicly available transcriptomic datasets were analyzed using differential expression profiling and weighted gene co-expression network analysis to identify key regulatory genes. These genes were intersected with a mitochondrial gene set and subjected to functional enrichment analysis. Least absolute shrinkage and selection operator (LASSO) regression was used to identify candidate diagnostic genes validation. Immune cell infiltration was quantified, and associated regulatory mechanisms were explored.<h4>Results</h4>Thirty-nine shared crosstalk genes were identified and were primarily enriched in mitochondrial metabolic processes. LASSO regression identified a five-gene candidate signature (<i>ACAA1</i>, <i>HSD17B10</i>, <i>MTIF2</i>, <i>THOP1</i>, and <i>PDE2A</i>). The model exhibited promising discriminatory performance (area under the curve > 0.9 in LC dataset and 0.7-0.9 in VTE dataset). These genes were significantly dysregulated and were associated with altered immune cell infiltration, particularly in dendritic cell and T cell subsets.<h4>Conclusion</h4>We identified a mitochondrial-related gene signature reflecting shared transcriptomic correlates between LC and VTE. The signature showed variable performance across disease contexts and correlative associations with immune features, supporting its role as a candidate biomarker for further investigation. Prospective validation in independent clinical cohorts is required before any translational application.

Newly emerging and re-emerging viruses are of significant global concern. Several viruses cause significant respiratory distress, including human coronaviruses (hCoVs), influenza A viruses (IAVs), and respiratory syncytial virus (RSV). Viruses usurp and modify host cellular processes to successfully replicate. We used quantitative label-free mass spectrometry-based proteomic analyses of transformed human lung A549 cells infected with representative seasonal hCoV, IAV, and RSV at 24 h post-infection to delineate comparative virus-induced host cell alterations. Numerous proteins involved in lipid metabolism, molecular transport, cell death and survival, and inflammatory responses were dysregulated. RSV induced a more profound response in cellular protein quantities than either CoV or IAV. The most significantly affected proteins were enzymes, ion channels, kinases, transcription regulators, and transmembrane receptors. Many canonical pathways also were differentially affected by each virus. The altered expression of proteins and pathways may provide insights into future therapeutic interventions for these viral infections.

<h4>Background</h4>Alzheimer's disease (AD) is the most common neurodegenerative disease worldwide, imposing a substantial economic burden. As the population ages and cases rise, interest in early identification, prevention, and treatment has intensified. After establishing that pathology involves neurofibrillary tangles, neuroinflammation, and neuronal loss, interventions have been tested in laboratory animals to slow disease progression. One commonly tested intervention is physical exercise, an economical approach. Laboratory animal models of AD aim to demonstrate how different types and intensities of exercise affect disease pathology. This review aims to categorize and elucidate the mechanisms and their outcomes.<h4>Materials and methods</h4>Original manuscripts concerning the impact of exercise, with or without medicinal intervention, on Alzheimer's dementia progression modeled in laboratory animals from 2020 onwards were included. The routes through which exercise exerts its impact at molecular, behavioral, and histomorphological levels were categorized.<h4>Results</h4>The most frequent modeling approach was Aβ injection into the hippocampus of Wistar rats, followed by Streptozocin and other chemicals, and transgenic models. Exercise mainly consisted of treadmill or swimming. Interventions were categorized into short-, medium-, and long-term protocols, ranging from 3 to 24 weeks, with most at 4 weeks, 30-60 min per session, 5 days/week. Molecular tests primarily measured Reactive Oxygen Species (ROS), while the most common behavioral test was the Morris Water Maze (MWM), assessing learning and memory. Histopathological assessments focused on β-Amyloid plaque formation, dark cells, and glial activation. Most studies agreed on AD modeling methods, but exercise protocols varied in intensity, duration, and type. The combined use of molecular, behavioral, and histological tests to assess intervention effects was consistent.<h4>Conclusion</h4>Physical exercise has been linked to lower oxidative stress and neuroinflammation, enhanced cognitive function, and fewer pathological changes in animal models of AD. However, variability in disease models and exercise protocols makes it difficult to identify the best exercise approach.

Somatic mutations arise throughout life, with functional consequences tied to the cell populations in which they occur. Genome-wide studies measure somatic variations in bulk tissue, whereas single-cell approaches resolve cell identity but provide limited sensitivity for complex alleles. Here we developed SniffCell, which uses DNA methylation carried on native long reads to assign somatic variant-supporting molecules to methylation-resolvable cell types. SniffCell builds cell-type-discriminatory methylation signatures across eight tissues, assigns long reads to cell types, and provides cell-type-specific variant calling. Across peripheral blood mononuclear cells and brain benchmarks, SniffCell recovered sorted cell identities and validated cell-type-specific variant assignments using purified immune-cell, neuronal, and oligodendrocyte fractions. In blood, SniffCell recovered lineage-restricted antigen receptor rearrangements and localized a somatic tandem-repeat expansion to T cells. In the frontal cortex, SniffCell identified recurrent neuron-specific tandem-repeat expansions in genes including FGF14, LRRC7 and SH3RF3 . Across three brain cohorts comprising 172 donors, recurrent neuron-associated expansions were enriched for GAA-rich motifs. In donors with matched blood, and diverged more strongly from the inherited repeat length, whereas oligodendrocyte-associated alleles more often tracked it. SniffCell transforms native bulk long-read genomes into a cell-type-aware resource for somatic variant discovery and reveals recurrent somatic instability in human tissues at cell-type resolution.

Mechanisms aimed at recovering from heat-induced damage are closely associated with the organism's ability to survive extreme temperature exposure. In such a scenario, we show that autophagy, as a cytoprotective mechanism, ensures recovery and viability after induced heat stress in <i>Saccharomyces cerevisiae</i>. Our findings indicate that heat shock triggers the targeted degradation of ubiquitinated protein aggregates, mediated by the macroaggrephagy receptor Cue5. Moreover, heat stress induces the turnover of the aggrephagy receptor Cct2 and the polyglutamine repeats of the HTT (huntingtin) protein (polyQ-HTT). Notably, even though Cct2 and polyQ-HTT degradation is vacuole-dependent, it is mediated autonomously of canonical autophagy pathways. Collectively, this study demonstrates a novel role of autophagy in maintaining protein homeostasis after heat stress in yeast and provides insights into the potential medical applications of heat treatment.<b>Abbreviations</b>: Atg1: AuTophaGy related 1; Atg7: AuTophaGy related 7; Atg8: AuTophaGy related 8; Atg13: AuTophaGy related 13; Atg15: AuTophaGy related 15; Cct2: Chaperonin Containing TCP-1 2; CMA: chaperone-mediated autophagy; Cue5: Coupling of Ubiquitin conjugation to ER degradation 5; GFP: green fluorescent protein; HS: heat shock/stress; HTT: huntingtin; Pep4: carboxyPEPtidase Y-deficient 4; polyQ: polyglutamine; RFP: red fluorescent protein.

Not available.

DNA replication fidelity depends on the integrity of the replication fork to prevent DNA damage and preserve genome stability. Disruptions to this process can trigger replication stress, leading to the accumulation of single-strand DNA (ssDNA) and double-strand breaks (DSBs), which drive mutagenesis and ultimately contribute to disease. While the roles of core histones and their post-translational modifications in this context have been more well-studied, far less is known about how linker histones regulate the replication stress response. Here, we demonstrate that the S-phase-phosphorylated form of the linker histone H1 (pH1) plays a key role in DNA damage repair at collapsed replication forks, both in vitro and in cells. Using phosphomimic and phosphonull H1 mutants, we show that phosphorylation enhances H1 assembly with ssDNA. Utilizing intein chemistry for the site-specific incorporation of a photocrosslinker to the C-terminus of H1, we map the direct interactors of H1. We identify phosphorylated H1 at replication forks, where it engages replication machinery and DNA damage response factors, including Histone PARylation Factor 1 (HPF1). We further demonstrate that ssDNA induces pH1-HPF1 interactions that promote liquid-like assemblies, correlating with reduced DNA damage and histone PARylation. Consistent with this role, reduction of total H1 increases cellular sensitivity to DNA damage, a phenotype that is partially rescued by reintroduction of H1.4. Together, these findings establish pH1 as a new regulator of DNA damage repair at collapsed replication forks through the controlled sequestration of repair factors.

African zebu cattle (Bos indicus) exhibit remarkable adaptations to extreme thermal conditions, yet the genomic basis of this resilience remains incompletely characterized. Ethiopia provides a unique natural setting in which closely related zebu populations have adapted divergently to dry-hot (DHETZ) and humid-hot (HHETZ) climates. In this study, we reanalyzed publicly available whole-genome sequencing datasets from 46 Ethiopian zebu cattle from five populations and compared them with Asian zebu, Sudanese zebu, African taurine, and European taurine breeds. By integrating genome-wide SNP analysis, population genetic structure assessment, and multiple selection scans (iHS, Hp, XP-EHH, and XP-CLR), we identified distinct and shared selection signatures between DHETZ and HHETZ. We detected 33.7 million and 34.2 million biallelic autosomal SNPs in DHETZ and HHETZ, respectively. Ethiopian zebu clustered closely with Sudanese zebu but showed clear divergence from Asian zebu and taurine breeds. DHETZ and HHETZ exhibited very low genetic differentiation (FST = 0.0063), consistent with their shared ancestry; however, each group displayed unique selection signals. In DHETZ, iHS and Hp detected 298 and 113 candidate regions, respectively, whereas in HHETZ, they detected 244 and 138 regions, respectively. Cross-population XP-EHH and XP-CLR analyses identified 163 and 227 divergent regions between DHETZ and HHETZ, respectively. Integration of the four selection scans identified 19 high-confidence candidate regions in DHETZ and 13 in HHETZ. DHETZ showed strong selection in genes involved in oxidative stress regulation, protein folding, mitochondrial function, and vascular remodeling, including SESN2, DNAJC8, GRPEL2, ABLIM3, and AFAP1L1. In contrast, HHETZ displayed signatures in genes associated with immune responses, energy metabolism, and angiogenesis inhibition, including MYD88, PRKACA, PRKACB, and WIF1. Several genes, including VEGFC, TNIP3, and DMXL2, were under selection in both groups, suggesting conserved mechanisms of thermotolerance and reproductive adaptation. The shared VEGFC signal and the HHETZ-specific WIF1 signal may indicate a distinct vascular regulatory mechanism in the dry-hot and humid-hot environments. Our results reveal a dual pattern of genomic adaptation in Ethiopian zebu cattle and provide candidate loci for future validation and climate-resilient livestock breeding.

Deletion of chromosome 5q [del(5q)] is the most common cytogenetic abnormality in myelodysplastic neoplasms (MDS) and results in haploinsufficiency of multiple genes, including CSNK1A1. Recurrent CSNK1A1 mutations, predominantly affecting the E98 hotspot, occur almost exclusively in del(5q) MDS and are associated with adverse outcomes, yet their impact on CK1ɑ function remains unclear. Using integrated transcriptomic, (phospho)proteomic, and kinome activity profiling in hematopoietic stem and progenitor cells (HSPCs), combined with in vivo serial transplantation assays, we show that Csnk1a1 E98V represents a change-of-function rather than a loss-of-function mutation. Unlike Csnk1a1 haploinsufficiency, Csnk1a1 E98V preserves long-term hematopoietic reconstitution and does not enhance clonal expansion in vivo. Instead, the mutation induces suppression of kinase signaling networks, leading to coordinated repression of ribosomal gene expression, protein translation, and cell cycle programs. This signaling rewiring is accompanied by metabolic reprogramming characterized by reduced mitochondrial respiration, increased glycolytic flux, and an inability to adapt to metabolic challenges, creating a stress-tolerant but inflexible cellular state. Notably, Csnk1a1 E98V cells exhibit impaired megakaryopoiesis and increased vulnerability to iron overload, as well as RSL-3-mediated ferroptosis. Analysis of del(5q) MDS patients confirmed that CSNK1A1 mutations are associated with distinct clinical features, including thrombocytopenia, elevated myeloblasts, and reduced bone marrow iron levels. Together, our findings support a two-step model in which del(5q)-associated CSNK1A1 haploinsufficiency drives clonal expansion, followed by acquisition of CSNK1A1 mutations that promote stress tolerance rather than increased proliferation. This adaptive rewiring exposes metabolic and iron-dependent vulnerabilities that may be therapeutically exploited.

Cells are the basic unit of life. In multicellular organisms, cells are organized into tissues. This enables a division of labor where tissues perform complex tasks via coordinated actions of specialized cell types. Accordingly, the cellular collective determines tissue function. A comprehensive overview of which cell types exist in solid tissues is lacking. Using skeletal muscle as a model, we discuss basic principles for cell type classification, summarize 62 unified definitions of cell types in terms of lineage, molecular signatures, and function from the literature, and discuss how these cell types contribute to muscle function. For cell types for which quantitative data was available, we compare abundances in immunohistology and single cell and single-nucleus RNA sequencing data, revealing cell types that are commonly over- or underrepresented in each method. The result is a cell type resource that will serve as a benchmark for single-cell studies of skeletal muscle.

Aging is a complex biological process that leads to a growing burden of age-related diseases (ARD). Cellular senescence, which involves irreversible cell cycle arrest and the release of senescence-associated secretory phenotype (SASP), plays a central role in aging and the development of ARD. Natural extracellular vesicles (NEVs) refer to vesicles that derived from cells, organs and body fluids without artificial interventions. In recent years, NEVs have gained attention as mediators of intercellular communication, and showed multifunctional potential in regulating cellular senescence and ARD. On the one hand, these vesicles are capable of spreading senescence signals by transferring molecules such as miRNAs, proteins, and DNA fragments. On the other hand, NEVs derived from animal cells, body fluids, plant cells, and bacteria have demonstrated potential anti‑aging effects. This review synthesizes research findings from the past two decades, with a particular focus on studies published in the last five years that elucidate the molecular mechanisms by which NEVs regulate cellular senescence. This review aims to summarize various applications (biomarkers, drug delivery vehicles, direct therapeutic agents) of NEVs in major ARD, including neoplasms, cardiovascular diseases, neurodegenerative disorders, bone diseases, and diabetes. Several challenges including standardizing production, targeted delivery, mechanism exploration, and clinical translation are also discussed. Combining bioengineering, multi-omics technologies, and improved model systems could be important to fully utilize the potential of NEVs, and offer precise, safe, and effective strategies for ARD therapy.

<h4>Background</h4>Most mutations causing hypertrophic cardiomyopathy (HCM) affect sarcomeric proteins. Mutations in junctophilin-2 (JPH2) are also implicated, but the underlying mechanisms remain unclear. An A405S variant in JPH2 was identified in a male adolescent patient with interventricular septal (IVS) hypertrophy. The corresponding mouse variant (A399S) produces comparable IVS hypertrophy, establishing causality. Prior data indicated that altered intracellular Ca²⁺ handling is unlikely to be the primary driver.<h4>Methods</h4>We generated a CRISPR knock-in mouse model carrying the JPH2-A399S variant. Co-immunoprecipitation mass spectrometry and STED nanoscopy were used to identify JPH2 binding partners. Reactive oxygen species (ROS) were assessed with dihydroethidium in isolated myocytes. Adeno-associated virus serotype 9 (AAV9) was employed to overexpress peroxiredoxin 6 (PRDX6) in mutant hearts.<h4>Results</h4>PRDX6 was identified as a novel and abundant JPH2-interacting protein. PRDX6 expression was selectively downregulated in the IVS of JPH2-A399S mice and was also reduced in human failing hearts. JPH2-A399S mice exhibited increased ROS levels specifically in IVS myocytes. AAV9-mediated PRDX6 overexpression reversed the IVS hypertrophy phenotype.<h4>Conclusions</h4>These findings identify PRDX6 downregulation and consequent oxidative stress as a key mechanism driving JPH2-A399S-associated HCM. The results reveal a previously unrecognized role for JPH2 in cardiometabolic regulation and suggest that restoring PRDX6 levels may represent a therapeutic strategy for this form of HCM.

Recent large-scale studies have enabled new knowledge about genetic underpinnings of morphological and electrophysiological alterations of the retina. Variation in retinal traits, often of neurodevelopmental origin, have been linked to major psychiatric disorders (MPDs). Here, we investigate the genetic overlap between MPDs and key retinal traits to identify underlying molecular mechanisms. We obtained genome-wide associations studies data for bipolar disorder (BD), major depression (MD), schizophrenia (SCZ), and the retinal traits retinal nerve fibre layer thickness (RNFL), ganglion cell inner plexiform layer thickness (GCIPL), and vertical cup-disc ratio (VCDR). We estimated the number of trait-influencing variants shared between traits with MiXeR and identified shared genetic loci with condFDR. Subsequently, we examined the biological pathways of the genes mapped to shared loci. This revealed that GCIPL shared the most genetic variants with MPDs (~60%), followed by RNFL (~40%), and VCDR (~20%). The genetic variants shared between retinal traits and MPDs showed disorder-specific patterns with more pronounced overlaps of SCZ and BD with RNFL, and MD negatively correlated with GCIPL. Gene-pathway analysis highlighted the importance of GABAergic neurotransmission and a two-stage neurodevelopmental process in SCZ, whereas the role of mitochondria and a weaker developmental component were observed in BD. The results also implicated synaptic functioning and gene-expression processes in MD. Furthermore, polygenic analysis suggested that the genetic architecture of retinal traits can distinguish between MPDs. Our findings indicate shared genetic underpinnings between retinal traits and SCZ, BD, and MD, implicating altered neurodevelopment and neurotransmission underlying the retinal link to major psychiatric disorders.

Idiopathic pediatric uveitis (IPU) is a leading cause of irreversible vision loss in children; however, the genetic and molecular mechanisms underlying this condition remain unclear. Herein, trio-based whole-exome sequencing was performed in 28 affected families and targeted sequencing was performed in 1953 sporadic cases from a Han Chinese cohort. A rare missense mutation, A773E in intraflagellar transport 122 (IFT122), was identified in one trio and absent from sporadic cases. Functional assays showed that deleterious IFT122-A773E increased inflammatory factor secretion and exacerbated barrier function damage both in vivo and in vitro. Further studies using proteomics demonstrated that IFT122-A773E increased AP-1 transcription factor subunit (FRA1) expression. The IFT122-A773E substitution enhanced the interaction with IFT43 and up-regulated calcium channels, and in turn led to activation of the MEK/ERK signaling axis. Collectively, our findings suggest that IFT122-A773E may increase susceptibility to IPU through activation of the MEK/ERK/FRA1 axis.

<h4>Introduction</h4>The literature provides limited evidence on the effectiveness of psychological interventions for individuals living with Huntington's Disease (HD). This study aims to examine the role of psychological support in improving the well-being of individuals and families affected by HD.<h4>Methods</h4>The study methodology builds on previous work conducted by the European Huntington Association (EHA). The study includes individuals across three categories: individuals at risk of HD, premanifest HD, or with an HD-negative genetic test. Eligible participants were offered eight online individual cognitive behavioural therapy (CBT) sessions followed by two group sessions. Assessments were performed at pre-intervention, after the individual sessions and after the group sessions. The primary outcomes were anxiety, depression, positive and negative affect, and quality of life.<h4>Results</h4>Twenty-one individuals participated in the study, of whom 33% (n = 7) had an HD-negative genetic test, 29% (n = 6) were at risk, and 38% (n = 8) had premanifest HD; the cohort was 57% women (n = 12). Analyses showed significant reductions in depression and anxiety, with medium to large effect sizes. Negative affect also decreased significantly with a large effect, while positive affect showed a non-significant trend toward improvement. Quality-of-life outcomes (physical and mental health components) did not change significantly. Overall, the strongest effects were observed for mood-related outcomes (depression, anxiety, and negative affect).<h4>Conclusion</h4>Our data shows that a short-term CBT intervention led to improvements in anxiety, depression and negative affect. These findings enhance the evidence base available supporting the efficacy of psychological interventions that may be beneficial for individuals affected by HD. The article also outlines key lessons regarding intervention tailoring and study design to inform future research exploring the efficacy of psychological interventions for this population.

The MYC family oncoproteins, including MYC, MYCN, and MYCL, are potent drivers of tumorigenesis across a broad range of human cancers, frequently linked to aggressive tumor behavior, poor prognosis, and therapy resistance. They function as master transcriptional regulators that orchestrate gene expression programs governing nearly all aspects of tumor development. Despite their pivotal oncogenic role, they have long been considered "undruggable" due to their intrinsically disordered structure, lack of enzymatic activity, and the difficulty of targeting their protein-DNA and protein-protein interactions with conventional small-molecule approaches. Recent advances are beginning to overcome these challenges through innovative molecular strategies that either directly inhibit MYC activity or exploit MYC regulatory networks. Importantly, the first direct MYC inhibitor evaluated in humans, OMO-103, recently demonstrated promising antitumor activity in phase I clinical trials. Indirect approaches have focused on suppressing MYC transcription, translation, or stability by targeting upstream signaling pathways, as well as by exploiting MYC-associated cofactor interactions and synthetic lethal vulnerabilities to improve therapeutic specificity. In this review, we highlight the multifaceted roles of MYC in different cancer types and provide a comprehensive overview of current therapeutic strategies targeting MYC with a particular focus on epigenetic modifiers and metabolic vulnerabilities.

Oncogenic KRAS mutations exhibit a striking tissue-restricted tropism, occurring with high frequency in pancreatic, colorectal, and lung adenocarcinomas while remaining rare in other lineages. The molecular basis for why these specific tissues are uniquely permissive to KRAS transformation, and how this context shapes therapeutic vulnerabilities, remains poorly defined. Here, we utilized CRISPR-mediated genome engineering to generate endogenous, conditional KRAS-mutant isogenic cell line models across three primary permissive lineages (lung, colon, and pancreas) and the nonpermissive breast lineage. Integrated genome-wide CRISPR fitness screens and comparative transcriptome analyses revealed that KRAS-driven synthetic lethal (SL) dependencies are profoundly shaped by their tissue of origin. Strikingly, we observed minimal overlap in SL hits across lineages, with only three genes shared among the permissive lines, suggesting that the <i>KRAS</i> oncogene operates through divergent, context-specific genetic networks. Mechanistically, we show that KRAS activation induces a universal MYC-driven metabolic signature, but the specific machinery required to sustain this state is lineage-restricted. We identified a dependency on the diphthamide synthesis pathway to maintain translational fidelity amid a KRAS-induced hypertranslational state. These findings demonstrate that even when driven by the same oncogene, tumors exhibit distinct regulatory landscapes and unique genetic vulnerabilities. Our results provide a framework for developing lineage-aware therapeutic strategies, moving beyond universal KRAS inhibition toward targeted interventions tailored to a tumor's specific tissue context.

Neuropathic pain remains a major unmet clinical challenge, as current therapies provide limited efficacy and poor tolerability. A major obstacle to developing effective analgesics is the pronounced cellular and molecular heterogeneity of the dorsal root ganglion (DRG), which integrates neuronal, glial, immune, and stromal responses to injury. Recent advances in single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) have enabled cell-type-resolved analyses of these responses and revealed disease-associated cellular states that are obscured by bulk tissue profiling, thereby offering new opportunities to redefine disease mechanisms and therapeutic priorities. This review synthesizes evidence from single-cell studies of peripheral nerve injury, diabetic painful neuropathy, and chemotherapy-induced neuropathy. Although these conditions share common biological processes, including neuronal state remodeling, disrupted glial homeostatic support, context-dependent immune responses, and extracellular matrix reorganization, they do not converge on a single conserved molecular program. Peripheral nerve injury is characterized by neuronal injury and repair states, reactive and metabolic glial remodeling, and neuroimmune recruitment and crosstalk. Diabetic neuropathy is associated with altered sensory coding, impaired satellite glial lipid support, and neurodegenerative remodeling. Chemotherapy-induced neuropathy exhibits substantial agent-specific heterogeneity, including subtype-selective neuronal vulnerability and metalloproteinase-dysregulated satellite glial states following paclitaxel treatment, as well as sex-associated fibrotic remodeling following bortezomib treatment. These cellular states may serve adaptive, maladaptive, or degenerative functions. However, because most available studies rely on single-time-point or repeated cross-sectional sampling, they cannot directly establish temporal progression or causal relationships. We further examine how human DRG atlases bridge experimental models and human disease by determining whether candidate molecular targets and cell states are conserved and by identifying their cellular localization. These datasets also highlight species-specific differences in neuronal organization and non-neuronal transcriptional programs that may limit the direct translation of findings from rodent models. Consequently, the primary translational value of DRG single-cell studies lies in elucidating disease mechanisms and prioritizing candidate therapeutic targets for further investigation. Advancing these discoveries toward clinical application will require complementary evidence from human genetics, human DRG transcriptomic datasets, functional validation studies, and clinical pharmacology. Collectively, these complementary approaches may facilitate the development of mechanism-based and cell-type-informed analgesic strategies.

<h4>Purpose</h4>A growing body of research indicates that Vacuolar Protein Sorting 45 (VPS45) is essential for the development of tumours and the advancement of cancer. Its biological role and expression profile in HCC are still mostly unknown, nevertheless.<h4>Materials and methods</h4>Using a variety of datasets, including the Tumour Immune Estimation Resource (TIMER) and Gene Expression Profiling Interactive Analysis (GEPIA), among others, we examined the existence and prognostic importance of VPS45. Functional enrichment analysis was employed to elucidate the potential mechanisms through which VPS45 may influence HCC. Additionally, we assessed the relationship between VPS45 expression and immune cell infiltration using single-sample gene set enrichment analysis (ssGSEA) and Estimation of Stromal and Immune cells in Malignant Tumour tissues using Expression data (ESTIMATE). The Tumor Immune Dysfunction and Rejection (TIDE) algorithm was leveraged to assess the sensitivity of predicted gene expression to immunotherapy. Utilizing assays including CCK-8, wound healing, and Transwell migration and invasion methods, we performed in vitro experiments on the MHCC-97H cell line to further investigate the biological significance of VPS45 in the progression of hepatocellular carcinoma (HCC).<h4>Results</h4>According to our research, VPS45 expression is markedly elevated in HCC, and elevated VPS45 levels are associated with a worse prognosis for patients. Functional enrichment analysis showed that elevated VPS45 expression is closely linked to the activation of various oncogenic pathways. VPS45 silencing decreases HCC cell invasion, metastasis, and proliferation, according to in vitro investigations. Interestingly, it was shown that VPS45 levels showed a positive link with immunosuppressive cell populations and related genes, but a negative correlation with a number of anti-tumor immune cell types. Immunotherapy resistance is more common in patients with increased VPS45 expression.<h4>Conclusion</h4>This study clarifies the biological role of VPS45 in hepatocellular carcinoma (HCC) and, through comprehensive database analyses combined with in vitro experiments, supports its potential as a diagnostic and predictive biomarker for HCC. In addition to its value for disease detection and prognosis, VPS45 may help predict patient response to immunotherapy.

<h4>Background</h4>Schizophrenia is considered a neuro-developmental disorder leading to disastrous lifelong disability of the patients and their families. There is a lack of data regarding pharmacogenomics of schizophrenia in Sudan. This study aimed to identify different genes affecting the treatment outcomes in Sudanese patients with schizophrenia.<h4>Methods</h4>A case-control study was conducted on seven families having more than one member diagnosed with schizophrenia. This was a small exploratory family-based sequencing study involving 18 affected individuals and 8 controls from seven families. Ethical clearance and informed consent were obtained. Demographic data were collected using a standardized data collection sheet. DNA was extracted from blood samples collected from patients and control groups. Then, whole-exome and genome sequencing were performed. Sixty-six genes associated with schizophrenia, treatment, and treatment resistance were selected from the variant calling file. Variants showing single-nucleotide polymorphisms (SNPs) were identified. These variants were then classified based on their impact on the protein-coding sequence into high- and moderate-impact. Moreover, indel mutations were also identified.<h4>Results</h4>Twelve variants of seven genes (COMT, FMO1, LPL, CYP2E1, ABCC1, GRM3, CYP2C9) were identified as genes with impact and potential association with schizophrenia (p-value=0.006632). Forty-three genes had a moderate impact, and they showed a potential association with schizophrenia (p-value=0.0004436). Two variants were indel mutations (CYP2D6, DTNBP1) and showed association with schizophrenia (p-value=0.004741). The p-values were generated from different databases.<h4>Conclusion</h4>This exploratory family-based sequencing study identified several potentially relevant pharmacogenomic and schizophrenia-associated variants in Sudanese families, warranting validation in larger and ethnically diverse cohorts.

Many endangered species rely on ex situ management for survival when external threats exist on the landscape. Yet, ex situ settings pose challenges through space limitation, altered environment, and diet. This can lead to environmentally determined inbreeding depression, where ex situ animals exhibit reduced reproductive fitness compared with their in situ counterparts, despite originating from the same genetic stock. We investigated epigenetic differences as a potential mechanism underlying environmentally determined inbreeding depression in black-footed ferrets (Mustela nigripes), a North American endemic species reliant on ex situ conservation. More specifically, we explored how environmental context may influence sperm DNA methylation in samples collected from 12 ex situ and 5 in situ males. Average sperm DNA methylation was significantly higher in ex situ individuals. We additionally identified more than 500 differentially methylated regions between ex situ and in situ sperm samples that were enriched for gene ontology terms pertaining to reproduction and development. Putative genes of interest included NPR2, WEE2, SLC15A1, PDE10A, PIP5K1B, CACNA1E, and CACNA1A, all of which have previously been linked to spermatogenesis, sperm motility, or fertilization in mammals. Results suggest that environmental conditions may alter sperm DNA methylation in black-footed ferrets, with possible links to decreased reproductive success in ex situ settings. These findings provide valuable insights into the molecular mechanisms underlying environmentally determined inbreeding depression in black-footed ferrets and other conservation-reliant species, and can serve as a foundation for future research on improving reproductive health in endangered wildlife.

<h4>Background</h4>In patients with colorectal cancer (CRC), intestinal dysbiosis has been observed, with abnormal colonization of the colonic mucosa by pathogenic Escherichia coli strains producing a cyclomodulin named cytotoxic necrotizing factor (CNF). Cyclomodulins are bacterial toxins capable of altering the cell cycle of the infected cell. One of the mechanisms involved in host defense against pathogens and in carcinogenesis is autophagy. Here, we aimed at investigating the role of autophagy in colorectal carcinogenesis in the context of CNF-producing E. coli, designated as CyPEC (cytotoxic necrotizing factor-producing E. coli), infection.<h4>Methods</h4>ApcMin/+ mice predisposed to CRC development with autophagy deficiency specifically in intestinal epithelial cells (ApcMin/+/Atg16l1ΔIEC) were infected with the clinical 21F8 strain or the mutant 21F8Δcnf1, which does not produce CNF1.<h4>Results</h4>In ApcMin/+ mice, infection with 21F8 or 21F8Δcnf1 did not have an impact on the number and the size of colonic tumors. However, in ApcMin/+/Atg16l1ΔIEC mice, infection with 21F8 increased number and size of colonic tumors, compared with uninfected or 21F8Δcnf1-infected condition. This increase was CNF1-dependent as it was not observed upon infection with the 21F8Δcnf1 mutant. Mechanistically, the increase in tumorigenesis in ApcMin/+/Atg16l1ΔIEC mice upon 21F8 infection was associated with enhanced proliferation and decreased apoptosis of colonic epithelial cells.<h4>Conclusions</h4>Our results show that autophagy deficiency could be a genetic susceptibility for the development of CRC in patients with abnormal colonization by CyPEC strains.

The mutation that causes Huntington's disease is a CAG repeat expansion in exon 1 of the huntingtin gene (HTT) that leads to an abnormally long polyglutamine tract in the huntingtin protein (HTT). Mutant CAG repeats are unstable and increase in size in specific neurons and brain regions with age, a phenomenon that constitutes the first step in the pathogenesis of the disease. In the presence of an expanded CAG repeat, cryptic polyadenylation (polyA) sites in intron 1 of the HTT pre-mRNA can become activated leading to the polyadenylation of a prematurely terminated transcript, HTT1a. This encodes the HTT1a protein, which is known to be very aggregation-prone and highly pathogenic. Given that the longer the CAG repeat the more HTT1a is generated, could the production of HTT1a be the mechanism through which somatic CAG repeat expansion exerts its pathogenic consequences? Resolving this issue is very important for the design of therapeutic approaches to lower huntingtin levels. We have used a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 approach to prevent the production of HTT1a in a knock-in mouse model of Huntington's disease. All potential cryptic polyA sites were deleted from Htt intron 1 in HdhQ150 mice and colonies were established that were heterozygous for the intron 1 deletion on a mutant allele (HdhQ150ΔI) and heterozygous for the deletion on a wild-type allele (WTΔI). The CAG repeat sizes in the HdhQ150 and HdhQ150ΔI colonies were well-matched at approximately 195 CAGs. As predicted, the deletion of the cryptic polyA sites from Htt intron 1 prevented the generation of the Htt1a transcript in the HdhQ150ΔI mice. However, very low levels of the HTT1a protein were detected, which resulted from a Htt readthrough product of exon 1 and exon 2, that had retained the deleted intron and terminated at a cryptic polyA site in intron 2. HdhQ150, HdhQ150ΔI, wild-type and WTΔI mice were studied until 17 months of age. Immunohistochemical and homogeneous time-resolved fluorescence analysis showed that HTT aggregation in both HdhQ150 and HdhQ150ΔI brains contained HTT1a, but the dramatic decrease in soluble HTT1a levels in HdhQ150ΔI brains delayed the appearance of aggregated HTT1a by several months. Although this delay in aggregate pathology only partially reversed transcriptional dysregulation, the biomarkers neurofilament light polypeptide (NEFL) and breast regression protein 39 (BRP39) (YKL40) remained at wild-type levels in HdhQ150ΔI mice at 17 months of age. These data demonstrate that the production of HTT1a initiates HTT aggregation and that it is important to target HTT1a in huntingtin-lowering therapeutic strategies.

Breast and prostate cancers are both hormone-driven adenocarcinomas that undergo analogous invasion programs. Using lightsheet microscopy on intact tumors, we identified transitional junctions between precancerous and invasive regions. We then developed a multimodal serial-section workflow integrating volumetric reconstruction with spatial transcriptomics. Analysis of 319 spatial assays from 51 cases revealed gene expression features and novel structural insights defining the shift from precancer to invasive disease. In breast cancer, loss of MGP and PLAT was associated with invasive transition and promoted tumorigenesis in functional assays. In prostate cancer, GDF15, ALDH1A3, ANPEP, and FASN were upregulated along invasive progression, and their knockdown in PC-3 cells suppressed proliferation and migration. Enrichment of tumor-associated macrophages (SPP1+ and MS4A6A+) along non-triple-negative breast cancer breast cancer transitions highlights immune involvement as a potential driver of invasiveness.<h4>Significance</h4>Our method of defining precise spatial locations of invasive transition allows for the direct interrogation of transition drivers, presenting new therapeutic targets for the two most prevalent cancers and providing a framework for studying spatially defined mechanisms of tumor progression. See related commentary by Jing and Li, p. 1720.

Severe burns provoke a systemic "genomic storm," yet cell states associated with divergent outcomes remain unclear. We profiled blood cells by single-cell RNA-Sequencing (73 014 cells) from adult patients with burn injuries within postburn day 17 (n = 4) and healthy donors (n = 5), integrated data with bulk signatures of burn size, inhalation injury, and mortality, and evaluated clinical associations in the American Burn Association National Burn Repository. Burn was associated with emergency hematopoiesis marked by expansion of hematopoietic stem/progenitor-like cells, immature neutrophils, and plasmablast/plasma cell states, alongside depletion of naïve CD4+/CD8+ T cells and dendritic cells. Larger burns (>20% TBSA) showed enrichment of humoral transcriptional programs, including plasmablast/plasma cell activation and suppression of cytotoxic CD8+ T-cell states. In multivariable models, inhalation injury was a stronger predictor of death (adjusted odds ratio [OR] 1.9) than burn size (adjusted OR 1.1) and shared greater overlap with the most perturbed single cells in non-survivors; 55% of co-perturbed cells were neutrophils, implicating granulocyte dysregulation as a common lethal axis. We identified a neutrophil-specific 5-gene panel (OLFM4, RETN, LCN2, ARG1, and BTNL3) that discriminated survivors vs non-survivors after burns (area under the curve [AUC] > 0.9) and generalized to trauma (n = 158; AUC 0.81) and intensive care unit COVID-19 (n = 103; AUC 0.75), providing information orthogonal to conventional biomarkers and severity scores. Cytomorphology corroborated transcriptomic immaturity, with ~2-fold higher band neutrophils and larger neutrophil size in a fatal case. Computational drug-reversal analysis highlighted galectin-1 inhibition as a candidate modulator of mortality-associated neutrophil programs. Together, our findings suggest that immature neutrophils represent a shared immune feature across severe burns and other forms of critical illness.

<h4>Background</h4>Iron overload (IO) disorders, including thalassaemias, hereditary haemochromatosis, and transfusion-dependent anaemias, represent a growing clinical challenge with widespread systemic implications. Reproductive dysfunction remains severely underappreciated despite its high prevalence. Hormonal changes due to iron toxicity are frequently reported, yet are seldom the focus of reproductive medicine, causing fragmented knowledge, inconsistent clinical approaches, and a lack of consensus guidelines.<h4>Objective and rationale</h4>This review synthesizes evidence on the impact of IO on male and female reproductive function, including gonadal dysfunction, impaired fertility, sexual dysfunction, and endocrine-metabolic complications. By addressing gaps in study design, diagnostic criteria, and management, we aim to provide the first comprehensive, expert-driven synthesis on the topic, integrating clinical, translational, and mechanistic insights to establish a structured framework for future research and patient care.<h4>Search methods</h4>A systematic literature search was conducted across PubMed, Scopus, and Web of Science, including studies up to May 2025. Search terms included 'iron overload', 'thalassemia', 'hemochromatosis', 'hypogonadism', 'fertility', 'spermatogenesis', 'ovarian insufficiency', and 'pregnancy'. Quantitative synthesis involved pooling data on prevalence rates of hypogonadism, semen abnormalities, primary and secondary amenorrhoea, age at menarche, and pregnancy outcomes.<h4>Outcomes</h4>Gonadal dysfunction primarily arises from iron deposition within the hypothalamic-pituitary-gonadal axis, coupled with oxidative damage to Leydig and Sertoli cells in males, disrupting testosterone synthesis and spermatogenesis, and to ovarian follicles and granulosa cells in females, causing reduced ovarian reserve and altered hormonal signalling. Iron-induced hypogonadism is the most frequent endocrine complication, significantly impacting reproductive health and quality of life. Our analysis of 1201 men and 2134 women indicated hypogonadism, reflecting impaired testicular endocrine function, in 47.0% of men; among those specifically assessed for spermatogenesis, over half presented azoospermia (17.6%) or other sperm abnormalities (37.5%). In women, primary amenorrhoea was reported in 45.7%, secondary amenorrhoea in 20.0%, and the weighted mean age at menarche was delayed (14.4 ± 2.1 years). Sexual dysfunction, notably erectile dysfunction, commonly accompanies hypogonadism, further impairing quality of life. Female sexual health has not been investigated at all. Pregnancy is increasingly achievable, but remains clinically challenging. Across 3536 reviewed pregnancies, ART was required in ∼20%, miscarriage occurred in 11.2%, and caesarean section was used in ∼80%. Mean gestational age at delivery was 37.1 ± 3.1 weeks, and mean birth weight was 2.64 ± 0.68 kg. Besides gonadal damage (direct or pituitary-related), systemic iron-related endocrine and metabolic disturbances, including hypothyroidism, growth hormone deficiency, diabetes mellitus, and cardiovascular disease, further aggravate reproductive impairments. Although effective iron chelation therapy reduces the systemic iron burden and is effective in preventing endocrine complications when initiated early, evidence supporting the reversal of established reproductive dysfunction remains limited, highlighting the need to optimize iron control from a young age to preserve reproductive health.<h4>Wider implications</h4>This review underscores the critical need for standardized gonadal screening to facilitate personalized reproductive care and early intervention in subjects with IO disorders. We propose an integrated clinical framework, combining early endocrine monitoring, fertility preservation protocols, and reproductive counselling. Future multidisciplinary research should prioritize prospective studies with clearly defined reproductive endpoints and explore optimized chelation strategies to safeguard reproductive potential. Addressing these gaps will fundamentally reshape clinical management, bridging haematology, endocrinology, and reproductive medicine.

<h4>Abstract</h4>Iron overload is a significant cause of transfusion-related morbidity and mortality. Deferiprone (DFP) is an established oral iron chelator approved for the treatment of transfusional iron overload in patients with thalassemia, sickle cell disease (SCD), or other anemias. Although DFP was initially approved as a thrice daily formulation, a twice daily formulation was developed to improve patients' treatment adherence. Here, we assess the safety of DFP twice daily in patients with thalassemia, SCD, or other anemias in real-world clinical practice in the United States. Data for patients referred for DFP twice daily between 1 July 2020 and 31 August 2023 were obtained from the Ferriprox Total Care Registry. The frequencies of adverse events (AEs), serious AEs (SAEs), AEs of special interest, and fatal outcomes were assessed. Of the 425 patients referred to the registry, 348 received ≥1 DFP shipment, and was designated the safety analysis group. The primary diagnosis was a thalassemia syndrome in 129, SCD in 189, and other anemia in 30 patients. Mean (standard deviation) DFP exposure was 343.0 (281.9) days. In total, 1021 AEs and 369 SAEs were reported. As seen with the thrice daily formulation, the most common AEs associated with DFP twice daily were predominantly gastrointestinal or related to underlying disease. Cases of neutropenia and agranulocytosis were rare, reported in 1.7% and 0.6% of patients, respectively, and all cases recovered. None of the 26 fatal outcomes reported were assessed as related to DFP treatment. DFP twice daily was well tolerated in real-world clinical practice, and its safety profile was consistent with DFP thrice daily. No new safety concerns were observed.

Macrophages (Mϕ) constitute a dominant and functionally diverse immune population within the microenvironment of pancreatic ductal adenocarcinoma (PDAC), yet how Mϕ heterogeneity contributes to the tumor remains poorly defined. In an institutional cohort of 145 PDAC specimens, we identified a population of multinucleated giant cells (MGC) of Mϕ origin, an entity previously described in chronic inflammation but rarely in cancer. CD68+ MGCs were present in 28% of tumors, enriched in squamous, nonglandular regions, and more frequent after neoadjuvant chemotherapy. By integrating spatial transcriptomics and quantitative imaging, we defined the features of these cells, which, compared with MGCs in nonneoplastic inflammatory lesions, lacked canonical polarization markers (HLA-DR and CD163) and displayed a distinctive transcriptional program characterized by upregulation of the POLR2K, TUBA8, COX5B, and VDAC1 genes, which encode proteins involved in DNA repair, oxidative stress, and MYC signaling. Spatial analyses revealed activation of hypoxia and extracellular matrix-remodeling pathways in MGC-associated niches, and experimental hypoxia promoted MGC formation in vitro. Consistent with these data, we found that in the The Cancer Genome Atlas (TCGA) Pancreatic Adenocarcinoma (PAAD) dataset a Mϕ MGC gene signature was enriched in the squamous PDAC subtype and correlated with poorer overall survival (P = 0.018). Morphometric and immunofluorescence analyses further showed increased 53BP1+Ki67+ nuclei and nuclear atypia in MGCs, indicating ongoing proliferation despite DNA damage. Together, these data identify MGCs of Mϕ origin as an immune cell state shaped by hypoxia and stress signaling, associated with aggressive tumor phenotypes, and potentially exploitable as an immune classifier in PDAC.

The <i>Drosophila</i> telomere is one of the best-studied examples of active transposable elements (TEs) benefiting, rather than harming, the host genome. All <i>Drosophila</i> species lack telomerase, and most species instead have telomeres composed of head-to-tail arrays of specialized retrotransposons. These TEs ostensibly act as mutualists by elongating chromosome ends, but evidence from species closely related to <i>Drosophila melanogaster</i> suggests that telomeric transposons may also antagonize their host genome. Importantly, the limited number of <i>Drosophila</i> species characterized thus far has precluded our ability to delineate idiosyncrasies from universal evolutionary forces and genetic mechanisms that shape the history of these TEs. Here, we have surveyed long-read genome assemblies of more than 100 species of <i>Drosophila</i>, identifying a total of 396 telomeric TE families. Our findings show that these telomere-specialized elements evolve dynamically and also undergo striking convergent evolution: The complete loss of telomeric TEs has occurred repeatedly across the genus, whereas individual telomeric TE lineages have repeatedly lost one of their two protein-coding genes. These elements have also repeatedly undergone horizontal transfer between distantly related <i>Drosophila</i> lineages and have repeatedly captured host gene fragments that promote their selfish suppression of host TE-silencing systems. Furthermore, telomere specialization itself appears to have evolved convergently, as some nontelomeric families have gained the ability to target their insertions to telomeres. These results provide unprecedented resolution into the evolution of these unusual TEs and highlight several novel mechanisms by which they evolve in conflict both with each other and their host genome despite the essential telomere function they provide.

Tumor masses often exhibit heterogeneity, including escape variant clones that lack antigen-presenting machinery and/or tumor antigens, which poses a major challenge to immunotherapy. Ferroptosis, a form of regulated cell death driven by iron-dependent lipid peroxidation, has been shown to effectively induce cell death in various tumor cells. Recent studies have reported that IFNγ suppresses the expression of system Xc-, thereby enhancing the induction of ferroptosis. Based on this, we hypothesized that combining immunotherapy with ferroptosis inducers could enhance antitumor effects against both antigen-positive and antigen-negative tumor cells. We found that combining RSL3, a ferroptosis inducer, with MART-1-specific T-cell receptor-engineered T cells eradicates a heterogeneous tumor model consisting of human melanoma cells and their β2-microglobulin knockout counterparts. In NOD.Cg-PrkdcscidIl2rgtm1Sug/ShiJic mice, this combination therapy demonstrates a significant antitumor effect against tumors with heterogeneity. These findings suggest that integrating ferroptosis inducers with immunotherapy could overcome the limitations imposed by escape-variant tumor clones, offering a promising strategy for cancer treatment.

<h4>Objective</h4>Posttransplant type 1 diabetes (T1D) is typically attributed to transferred donor autoimmunity. We investigated a distinct etiology in a patient who developed diabetes following haploidentical hematopoietic cell transplantation, assessing whether autoimmunity originated from donor or residual host cells.<h4>Research design and methods</h4>Leveraging HLA disparity between the haploidentical donor and recipient, we used HLA class II tetramers to enumerate islet-specific CD4+ T cells in peripheral blood restricted by shared versus recipient-only HLA alleles.<h4>Results</h4>Tetramer analysis revealed an expanded population of islet-specific T cells in the recipient. The donor showed no such expansion. Despite 99% donor T-cell chimerism, >80% of the islet-specific T cells were restricted by recipient-unique HLA alleles, suggesting they originated from the residual host fraction.<h4>Conclusions</h4>T1D in this patient was most likely driven by residual recipient-derived T cells. Their survival despite myeloablative conditioning and repeated immunotherapy underscores the remarkable durability of established islet autoimmunity.

Only ∼1%-2% of the human genome directly codes for proteins. The remainder consists of non-coding DNA, often referred to as the 'dark genome'. This includes regulatory elements, transposable and repetitive sequences, structural genomic features, pseudogenes, intronic and intergenic regions, and non-coding RNA (ncRNA) genes. These components are increasingly recognized as major regulators of gene expression, cell identity, and disease susceptibility. Currently, dark genome elements, particularly ncRNAs are increasingly recognized as important regulators of cardiovascular health and disease. Advances in genome analysis technologies have greatly improved our understanding of these non-coding regions and revealed clearer connections between the dark genome and cardiovascular traits. This review highlights major parts of the dark genome involved in cardiovascular disease, with emphasis on those for which mechanistic understanding and translational relevance are beginning to emerge. As mechanistic insight into individual and collective components of the dark genome advances, it increasingly enables the development of new opportunities for targeted therapeutics for cardiovascular prevention and disease management.

<h4>Background and aims</h4>Metabolic dysfunction-associated steatotic liver disease (MASLD) is a progressive liver disease that ranges from simple steatosis to inflammation, fibrosis and cirrhosis. To address the unmet need for new MASLD biomarkers, we aimed to identify candidate biomarkers using publicly available RNA sequencing (RNA-seq) and proteomics data.<h4>Methods</h4>An approach involving unsupervised gene clustering was performed using homogeneously processed and integrated RNA-seq data of 625 liver specimens to screen for MASLD biomarkers, in combination with public proteomics data from healthy controls and MASLD patients. Additionally, we validated the results in the MASLD and healthy cohorts using enzyme-linked immunosorbent assay (ELISA) of plasma and immunohistochemical staining (IHC) of liver samples.<h4>Results</h4>We generated a database (https://dreamapp.biomed.au.dk/NAFLD/) for exploring gene expression changes along MASLD progression to facilitate the identification of genes and pathways involved in the disease's progression. Through cross-analysis of the gene and protein clusters, we identified 38 genes as potential biomarkers for MASLD severity. Up-regulation of Quiescin sulfhydryl oxidase 1 (QSOX1) and down-regulation of Interleukin-1 receptor accessory protein (IL1RAP) were associated with increasing MASLD severity in RNA-seq and proteomics data. Particularly, the QSOX1/IL1RAP ratio in plasma demonstrated effectiveness in diagnosing MASLD, with an area under the receiver operating characteristic (AUROC) of up to 0.95 as quantified by proteomics profiling and an AUROC of 0.82 with ELISA.<h4>Conclusions</h4>We discovered a significant association between the levels of QSOX1 and IL1RAP and MASLD severity. Furthermore, the QSOX1/IL1RAP ratio shows promise as a non-invasive biomarker for diagnosing MASLD and assessing its severity.

Nucleos(t)ide analog (NA) therapy suppresses hepatitis B virus (HBV) DNA, but residual hepatocellular carcinoma (HCC) risk persists. O-glycosylated hepatitis B surface antigen glycan isomer (HBsAgGi) uniquely reflects virion burden. We evaluated the impact of tenofovir alafenamide (TAF) versus entecavir (ETV) on 48-week HBsAgGi kinetics and its utility in stratifying HCC risk in 90 NA-naïve patients (ETV: 73; TAF: 17). A favourable response was defined as a reduction or maintenance of low HBsAgGi levels. TAF independently predicted a favourable response compared with ETV (penalised odds ratio 3.60, p = 0.030). A significant interaction occurred between HBeAg status and HBsAgGi response in relation to HCC development (p = 0.038). In HBeAg-positive patients, a poor response was associated with increased HCC risk (hazard ratio 7.34, 95% confidence interval [CI] 1.96-20.35, p = 0.001); however, the exact magnitude warrants cautious interpretation due to wide CIs. This association was absent in HBeAg-negative cases. Adding HBsAgGi response to the aMAP score improved long-term HCC prediction exclusively in the HBeAg-positive cohort. As the assay targets genotype C-specific O-glycosylation, the findings cannot be generalised to genotypes A, B, or D. In conclusion, on-treatment HBsAgGi response is a specific surrogate marker for stratifying residual HCC risk in HBeAg-positive patients, and TAF is more effective than ETV in inducing these favourable kinetics.

<h4>Background</h4>Heart failure with preserved ejection fraction (HFpEF) accounts for over half of all heart failure hospitalizations, with obesity increasingly recognized as a central driver through expansion of epicardial adipose tissue, chronic low-grade inflammation and obesity-related haemodynamic overload culminating in ventricular hypertrophy.<h4>Methods</h4>We synthesized high-impact evidence from pivotal randomized trials and mechanistic studies published between 2015 and 2026, focusing on cardiometabolic outcomes, adipose tissue remodelling and haemodynamic effects of both drug classes.<h4>Results</h4>Tirzepatide produced substantial weight loss (> 20%), significant reductions in epicardial and paracardiac adipose tissue and a 19.5-point improvement in Kansas City Cardiomyopathy Questionnaire scores in the SUMMIT trial. SGLT2i, exemplified by empagliflozin, demonstrated foundational benefit through natriuresis, improved myocardial energetics and reduced heart failure hospitalization independent of diabetes status, with additional renal-protective effects. Because SUMMIT and EMPEROR-Preserved differed in design and populations, direct comparison between agents is not statistically valid; each should be assessed on its own evidence base.<h4>Conclusion</h4>Tirzepatide and SGLT2i act through distinct, physiologically complementary pathways converging on the cardio-renal-metabolic axis. While their combined use is mechanistically promising, it has not yet been tested in a dedicated randomized controlled trial, and prospective studies are needed to establish safety and long-term efficacy in obesity-related HFpEF.

Although exposure to violence has been linked to executive functioning impairments, the biological mechanisms underlying this association remain unclear. This study examined CpG sites associated with executive functions in 78 young adults and their links to family (psychological and physical) and community violence during childhood. The scales used were Conflict Tactics (family violence); Things I Have Seen and Heard (community violence); Barkley Deficits in Executive Functioning. Mean differences (Mann-Whitney-U/Kruskal-Wallis) and Spearman correlations were assessed. An epigenome-wide association study identified differentially methylated CpGs, and linear regressions examined associations with violence exposures. Girls reported more family and psychological violence. Black participants experienced higher family violence. Executive dysfunction (ED) correlated with psychological family and community violence, and family violence forms were interrelated. Differentially methylated positions highlighted genes tied to key biological processes: MRGPRD (sensory neurons development); DPPA3 (germ-cell development); UNKL (cell differentiation); GPR6 (learning and memory); DUSP5 (cell proliferation and differentiation); and CAPN13 (signal transduction, cytoskeletal remodeling and cell differentiation). Candidate gene analysis found associations between executive dysfunction and methylation levels in these genes. Exploratory pathway analysis suggested enrichment of neurodevelopment and synaptic plasticity, intracellular trafficking and cytoskeletal remodeling, genome stability and cellular stress responses, and neuroimmune and inflammatory signaling. These findings suggest associations between ED, childhood violence exposure, and differential DNA methylation in cognition- and health-related genes, suggesting that epigenetic markers may be associated with pathways linking early adversity to later outcomes, although no causal inference can be drawn.

Postmenopausal osteoporosis (PMOP) is linked to iron accumulation. Melatonin has iron-chelating and antioxidant properties, but its mechanism against osteoclastogenesis remains unclear. This study investigated whether melatonin suppresses osteoclast formation by targeting the iron/ROS-CREB-PGC-1β-mediated mitochondrial biogenesis pathway. In vitro, bone marrow-derived macrophages (BMMs) were treated with RANKL and melatonin (10-1000 nM). Melatonin concentration-dependently inhibited osteoclast differentiation, reduced intracellular ferrous and total iron levels, decreased ROS and oxidative stress markers, and suppressed mitochondrial biogenesis. Mechanistically, melatonin indirectly suppressed PGC‑1β expression via inhibition of CREB phosphorylation, without affecting PGC‑1α expression. The CREB activator forskolin reversed melatonin's effects, whereas the CREB inhibitor 666-15 mimicked them. In vivo, ovariectomized (OVX) mice received weekly injections of iron dextran to model moderate iron overload, with or without oral melatonin. Melatonin ameliorated iron‑induced bone loss, improved bone microarchitecture and biomechanical properties, reduced tissue iron stores and bone ROS levels, and suppressed osteoclast mitochondrial biogenesis and the CREB/PGC-1β pathway; these effects were counteracted by forskolin. In conclusion, melatonin prevents osteoclastogenesis and counters bone loss due to iron accumulation in estrogen‑deficient conditions by chelating iron, scavenging ROS, and blocking the iron/ROS‑activated CREB/PGC‑1β axis, thereby suppressing mitochondrial biogenesis. This study offers a mechanistic explanation for using melatonin as a possible treatment for PMOP, particularly when accompanied by iron overload.

<h4>Purpose</h4>Liver fibrosis and cirrhosis represent critical stages in the progression of chronic liver disease, yet their key molecular features remain incompletely understood.<h4>Experimental design</h4>We performed large-scale Olink-based proteomic profiling in over 40,000 participants from the UK Biobank with a median follow-up of 15.6 years to elucidate disease pathophysiology and identify pre-diagnostic biomarkers. Cross-sectional analysis included 66 prevalent cirrhosis cases, and prospective analysis identified 224 incident cirrhosis cases. Machine learning and Mendelian randomization (MR) were applied. An independent cohort was used for validation.<h4>Results</h4>Distinct dysregulated proteins were observed in compensated cirrhosis (CC) and decompensated cirrhosis (DC). In the prospective analysis, 696 proteins were associated with disease onset. A proteomic panel based on these markers achieved an AUC of 0.832 for predicting incident cirrhosis, outperforming established fibrosis scores including FIB-4, APRI, and NFS, and demonstrated robust performance across CC and DC populations. The protein panel showed predictive value (AUC = 0.743) for disease progression in an independent cohort. MR identified 66 proteins with putative causal roles, including 11 potential therapeutic targets.<h4>Conclusions and clinical relevance</h4>These findings provide novel molecular insights into cirrhosis development and support integrated proteomic biomarkers as a discovery and prioritization framework for early risk stratification.

Phosphomannomutase deficiency (PMM2-CDG), the most common congenital disorder of glycosylation (CDG), is characterized by multisystem involvement and a lack of disease-modifying therapies. While previous transcriptomic studies have uncovered disrupted cellular pathways, the functional consequences of these alterations remain poorly understood. To further investigate PMM2-CDG pathophysiology, we integrated proteomic and metabolomic profiling of patient-derived fibroblasts with previously published transcriptomic data. Proteomic analysis was performed using Tandem Mass Tag-based mass spectrometry, while metabolomics was conducted via Nuclear Magnetic Resonance spectroscopy. Multi-omics integration was performed using principal component analysis-based dimensionality reduction, incorporating clinical metadata as supplementary variables. Proteomic analysis identified 43 significantly altered proteins, with enrichment in the retinoic acid synthesis pathway, wound healing, and cytoskeletal organization. Metabolomic profiling revealed altered amino acid levels and elevated concentrations of UDP-GlcNAc, consistent with perturbation of the hexosamine biosynthesis pathway, and increased levels of myo-inositol. Notably, myo-inositol levels showed a strong association with disease severity in the integrated analysis. RT-qPCR confirmed the upregulation of GFPT2. This integrative multi-omics study identifies consistent alterations in the retinoic acid synthesis pathway and hexosamine biosynthesis in PMM2-CDG patient-derived fibroblasts and reveals an association between intracellular myo-inositol levels and disease severity. These findings provide new insights into PMM2-CDG-associated molecular alterations and illustrate the value of multi-omics integration for hypothesis generation in rare diseases.

Alzheimer's disease (AD) is a multifactorial neurodegenerative disorder characterized by progressive cognitive decline, amyloid-β (Aβ) accumulation, oxidative stress, and excitotoxicity. Ferroptosis and N-methyl-D-aspartate (NMDA) receptor activity,may be interconnected in the pathogenesis of Aβ accumulation and associated neurodegeneration in AD. However, the interplay between these pathways remains poorly understood and underexplored for therapeutic intervention against the AD. The review aims to explore the shared molecular triggers of ferroptosis and NMDA receptor overactivation, including the roles of iron, glutamate overload, calcium dysregulation, and reactive oxygen species (ROS) accumulation. We further highlighted the convergent consequences of these processes on mitochondrial dysfunction, lipid peroxidation, and their impact on Aβ pathology. Particular attention is given to P-glycoprotein (P-gp), an efflux transporter involved in the Aβ clearance at the blood-brain barrier, whose expression and function may be modulated by oxidative stress, iron homeostasis, and NMDA receptor signaling. Emerging evidence indicated that ferroptosis and NMDA receptor activity may disrupt P-gp function, thereby impairing Aβ clearance and promoting its accumulation in the AD. Overall, the review elucidates the molecular mechanisms linking ferroptosis and NMDA receptor overactivation and their impact on P-gp-mediated Aβ transport in the AD, providing integrated mechanisms and harnessing their potential for AD therapeutics.

Neurological disorders (NDs) are characterized by substantial loss of specific neurons, with Alzheimer's and Parkinson's diseases being the most frequent NDs and nearly 99% of all "foreign substances" are prohibited from entering the brain by the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (CFB). These barriers, while crucial for brain protection, pose significant challenges for drug delivery, as they restrict the entry of many therapeutic agents into the brain, and this represents the primary manifestation of the absence of pathogenesis-targeting therapeutics. With significant success across multiple cell transplantation research efforts, stem cell therapy has been utilized for decades to treat neurological disorders. They work by replacing injured or lost cells directly, releasing proliferation and neurotrophic factors through autocrine and paracrine actions, suppressing neurological inflammation, and activation of endogenous brain progenitor cells. Nanocarriers derived from stem cells represent an innovative and promising therapeutic strategy for combating neurological diseases, combining faculties of regeneration of stem cells with the precision of nanotechnology. These nanocarriers possess natural biocompatibility and can efficiently cross the BBB to transport the therapeutic agents directly to affected neural tissues, thereby promoting enhanced treatment efficacy while reducing off-target effects. However, key challenges remain in large-scale production, standardization, and long-term safety. Thus, this review has examined the potential applications of stem cell extracellular vesicle (EV)-nanocarriers, mainly exosomes and recent development in the treatment of neurological diseases.

Redox imbalances and mitochondrial dysfunction are key contributors to age-related declines in skeletal muscle and may contribute to impaired exercise responsiveness. Here, we investigated the influence of aging on skeletal muscle redox at rest and in response to acute exercise, examining how mitochondrial quality and quantity relate to skeletal muscle redox status. Skeletal muscle biopsies were obtained from 12 young (22 ± 4 years) and 10 older adults (66 ± 7 years) before and immediately after 60-min of high-intensity knee-extension exercise. We assessed mitochondrial respiration, mitochondrial DNA (mtDNA) copy number and deletion mutation frequency at baseline, while skeletal muscle redox proteomics was performed on pre- and post-exercise biopsies in a subset of participants. Mitochondrial respiration was preserved with age (max respiration, p = 0.123). However, the older adults had a lower mtDNA copy number (p = 0.046) and higher mtDNA deletion frequency (p = 0.001), with widespread remodeling of the skeletal muscle redox proteome, including altered thiol occupancy of proteins involved in metabolism, immune function, and extracellular matrix organization. In response to exercise, young skeletal muscle exhibited predominantly reversible peptide reductions, whereas preferential oxidation of mitochondrial antioxidant proteins, including PRDX3, occurred in older muscle. Both mtDNA deletion frequency and mitochondrial respiration were strongly associated with exercise-induced redox modifications in mitochondrial proteins. These findings suggest that aging alters both the regulation and resolution of exercise-induced redox signaling, with mitochondrial genomic instability and respiration shaping redox responsiveness.

Tau self-assembly and intracellular deposition are associated with a group of neurodegenerative diseases called tauopathies, which include Alzheimer's disease (AD) and Pick's disease. Here, we measured the proteome response in human neuronal cells (differentiated SH-SY5Y) following the addition of a spontaneously amyloidogenic region of tau known as dGAE (tau297-391), which forms AD-like paired helical filaments in vitro, and proteomic analysis showed increased endogenous tau expression. Further interactome analysis uncovered increased association between tau and proteins associated with nuclear chromatin, the nucleolus, and the spliceosome, as well as the thiol-peroxidase, PRDX6, alongside an increase in reactive oxygen species. The present work highlights a method to identify proteome pathways that may play an important role in the development of tau pathology and reveals an oxidative stress response to dGAE.

<h4>Background & aims</h4>Given the likely underestimation of hepatic sarcoidosis, the aims were to evaluate long-term cirrhosis risk in patients with sarcoidosis without documented hepatic involvement at baseline and to assess the impact of multi-organ disease and immunomodulatory therapies on cirrhosis risk.<h4>Methods</h4>A retrospective cohort study using the Clalit Health Services database in Israel included adults with sarcoidosis without documented hepatic involvement or prevalent cirrhosis and age- and sex-matched controls. The primary outcome was incident cirrhosis, analysed with inverse-probability-of-treatment-weighted (IPTW) Fine-Gray modelling with death as a competing risk. Secondary analyses evaluated multi-organ involvement using landmark methods and immunomodulatory therapies as time-varying exposures.<h4>Results</h4>Among 5489 sarcoidosis patients and 6016 controls (142 002 person-years), cirrhosis occurred in 112 (2.0%) and 38 (0.6%), respectively (incidence rate 1.61 vs. 0.53 per 1000 person-years; rate ratio 3.05). In the primary IPTW-weighted Fine-Gray analysis, sarcoidosis was associated with incident cirrhosis (subdistribution hazard ratio [sHR] 2.84, 95% confidence interval [CI] 1.97-4.11, p < 0.001); the weighted 20-year cumulative incidence was 2.8% versus 1.1%. The association was consistent across IPTW-weighted cause-specific, covariate-adjusted and follow-up-truncated analyses (hazard ratio [HR]/sHR range 2.71-2.84). Among patients with confirmed organ involvement, 1057 had single-organ and 87 had multi-organ disease. In the 5-year landmark analysis (n = 940; 18 events), multi-organ involvement was associated with cirrhosis in the parsimonious model (HR 5.33, 95% CI 1.90-15.00, p = 0.002). No immunomodulatory therapy was significantly associated with cirrhosis.<h4>Conclusions</h4>Sarcoidosis was independently associated with increased cirrhosis risk despite no hepatic involvement, particularly in multiorgan disease. This supports a risk-stratified spectrum from heightened clinical awareness to closer hepatic surveillance in selected patients.

Mitochondrial function depends on tight coordination between mitochondrial and nuclear genomes, which requires long-term coevolution to maintain mitonuclear coadaptation. While mitonuclear incompatibility is typically studied in the context of hybridization, other evolutionary scenarios that may disrupt coadaptation between the two genomes remain less explored. Here, we propose that extreme ecological niche shifts may disrupt mitonuclear coadaptation, which we test in carnivorous Miletinae butterflies with an extreme dietary transition. By generating high-quality genome assemblies, we found that Miletinae exhibit extensive chromosomal rearrangements. Comparative phylogenomic analyses revealed a striking asymmetric mitonuclear evolutionary response: Miletinae exhibit elevated mitochondrial nucleotide substitution rates compared to phytophagous relatives, whereas nuclear rates remain stable. This shift reverses the typical lepidopteran pattern where nuclear rates exceed mitochondrial rates. Interestingly, this mitochondrial acceleration is driven primarily by relaxed purifying selection rather than positive selection. To sustain mitochondrial function, the nuclear genome of Miletinae underwent pervasive, multilayered compensatory evolution. We detected strong signatures of positive selection and accelerated evolution in nuclear genes directly interacting with mitochondrial components across oxidative phosphorylation (OXPHOS) complexes, the mitochondrial translation, and replication and transcription machinery. Furthermore, this nuclear compensatory response extends to systems governing mitochondrial homeostasis, including protein quality control and RNA degradation and stabilization. Our results support a model in which extreme ecological transitions can disrupt ancestral mitonuclear coadaptation and promote the emergence of a new coadapted state through systemic nuclear compensation. This study broadens the conceptual framework of mitonuclear coevolution and highlights its role in facilitating evolutionary persistence after major ecological shifts.

Colorectal cancer (CRC) prognosis remains challenging due to tumor heterogeneity and immune evasion driven by aberrant inflammatory signaling, and robust multicohort validated prognostic tools with clear therapeutic implications are still lacking. To address this gap, we integrated five independent CRC cohorts (TCGA, GSE17536, GSE17537, GSE29621, and GSE38832; n = 931) and applied single-sample gene set enrichment analysis (ssGSEA) across 15 inflammation-related signaling pathways to construct a prognostic risk score model via multicohort Cox regression and LASSO-Cox regression. The immune landscape was systematically characterized using immune infiltration algorithms, Cancer Immunity Cycle scoring, CellChat analysis, and single-cell RNA sequencing (scRNA-seq) data (GSE166555). The functional role of the candidate gene PCOLCE2 was further validated through in vitro migration and adhesion assays, and confirmed in a syngeneic C57BL/6 mouse subcutaneous tumor model in which MC38-shPCOLCE2 cells were inoculated and treated with anti-PD-1 antibody (200 μg per mouse, every 3 days) alone or in combination to evaluate the synergistic antitumor effect. The resulting 12-gene inflammation-related risk model achieved robust survival stratification across all five independent cohorts, with AUC values of 0.70-0.85 and C-indices consistently outperforming existing prognostic models, demonstrating strong generalizability as a clinical prognostic tool. High-risk patients exhibited significantly worse overall survival (p < 0.001), an immunosuppressive tumor microenvironment, and elevated pro-stromal remodeling signaling, patterns that were corroborated by single-cell resolution analysis. Mechanistically, PCOLCE2 knockdown suppressed CRC cell migration and enhanced matrix adhesion in vitro. Critically, in vivo combination of PCOLCE2 knockdown with PD-1 blockade produced a synergistic antitumor effect superior to either monotherapy (p < 0.01), highlighting PCOLCE2 as a promising therapeutic target to enhance immunotherapy response. Together, these findings present a clinically actionable, multicohort validated inflammation-related prognostic model for CRC risk stratification, and position PCOLCE2 as a driver of invasion and immune evasion whose combination with PD-1 blockade offers a rational therapeutic strategy to improve immunotherapy efficacy in high-risk CRC patients.

Understanding how small molecules modulate protein aggregation is pivotal for developing strategies against amyloid-related disorders. Polyphenols are promising modulators, yet the connection between their molecular photophysics and aggregation inhibition remains largely unresolved. In this article, femtosecond transient absorption spectroscopy has been employed to unravel the excited-state dynamics of some commonly used polyphenols, namely, Baicalein, EGCG, and Myricetin across varied environments-neat solvents, micelles and reverse micelles, and in the presence of globular and intrinsically disordered protein (in its native and aggregated forms). Excited-state intramolecular proton transfer (ESIPT) emerges to be highly sensitive to hydrogen-bonding, polarity, and local confinement, serving as a subpicosecond reporter of polyphenol-environment interactions. In protein systems, modulation of ESIPT dynamics signals environment-specific embedding of polyphenols within evolving protein landscapes. A unique tri-parameter correlation-linking binding affinities of the polyphenols to the native proteins, ESIPT lifetimes, and aggregation kinetics-reveals that inhibitory potency arises not from tight native binding but from selective engagement with aggregation-prone species. This work posits ESIPT as a sensitive molecular signature that links ultrafast photophysics to the functional inhibition of protein aggregation, providing new insights into how small molecules navigate complex biomolecular environments to regulate self-assembly pathways.

Studies have demonstrated that the nervous system plays an important role in cancer progression. Nevertheless, the effect of mesenchymal stem cells (MSCs) on cancer has yielded conflicting results. In this study, we discovered that cancer-derived MSCs (CA-MSCs) not only exhibit the properties of neural stem cells (NSCs) but also express a significant amount of neural-related products. These products include neurotrophic factors, neuropeptides, synapse-related products, and axon guidance factors. These identified products exhibit similar or different expression status among human NSCs, cancer and fat-derived MSCs. CA-MSCs expressed more BDNF, GDNF, and NGF than NSCs. Based on these neural-related products expressed by CA-MSCs, expression-prognostic analysis was conducted. It revealed that the expression of MDK and MANF was higher in cancer, and the expression of BDNF, NPTX1, and NGF increased as the tumor stage advanced. For ten neuropeptides, their expression levels were lower in cancer. Thirty synapse-related products displayed their respective expression status between cancer and normal tissues. SYPL1, SYBP2/3, and STX4 had a significant impact on tumor prognosis. Higher levels of syntaxin-4, neugrin, and other axogenesis products were associated with a worse prognosis in cancer. Moreover, immunohistochemistry revealed that NSC-like CA-MSCs were abundant in cancer and exhibited a clear growth-promoting effect on cancer. Briefly, CA-MSCs, which are densely distributed in cancer and have the characteristics of NSCs, express a large number of nerve-related products. These products may have significant effects on tumor progression and prognosis.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway, as a key DNA sensor, plays a significant role in the regulation of innate immune responses. This pathway can be activated by sensing abnormal DNA, and is of great significance for resisting the invasion of pathogenic microorganisms and maintaining tissue homeostasis. In addition, the cGAS-STING pathway plays a dual role in cancer, and oncogenic viruses can cause cell carcinogenesis in the body through multiple mechanisms, thereby affecting human health. This manuscript reviews the vital role of cGAS-STING in the immune process, as well as the fact that viruses causing human tumor lesions can activate cGAS-STING, leading to virus inhibition and further preventing the occurrence and development of related cancers, and the paradoxically promoted progression of related cancers by viruses through cGAS-STING. And summarize the agonists and inhibitors that act in different ways based on the activation mechanism of cGAS-STING.

Single-cell RNA sequencing (scRNA-seq) reveals the transcriptional heterogeneity of cells, revolutionizing our understanding of cellular processes. However, the static snapshots obtained from scRNA-seq fail to reveal the time-resolved dynamics of transcription, which impedes critical insights into various biological processes, such as cellular differentiation, embryonic development, disease progression, and responses to external stimuli. Here, we describe Well-TEMP-seq, a protocol for massively parallel profiling of the temporal dynamics of single-cell gene expression. Well-TEMP-seq combines metabolic RNA labeling with a microwell-based scRNA-seq method, Well-paired-seq, to distinguish newly transcribed RNAs marked by T-to-C substitutions from pre-existing RNAs in each of thousands of single cells. Well-TEMP-seq is high-throughput, cost-effective, accurate, and provides a low cell loss rate and high single cell/bead pairing efficiency. More importantly, Well-TEMP-seq can be easily set up in other labs, and the loading of cells and beads can be easily accomplished by an optical microscope and a pipette. We believe that Well-TEMP-seq will be widely adopted and help researchers perform transformative research to unveil the dynamics of single-cell gene expression in diverse biological processes. © 2026 Wiley Periodicals LLC. Basic Protocol 1: Well-paired-seq chip fabrication Basic Protocol 2: Well-TEMP-seq sample processing Basic Protocol 3: Bioinformatics analysis.

Regulation of mitochondrial health is critical for maintaining cellular homeostasis in the nervous system. Damaged mitochondria can have detrimental effects on neuronal health and are thought to be key contributors to the progression of neurodegenerative disorders including Parkinson's disease and amyotrophic lateral sclerosis. To mitigate this damage, multiple quality control mechanisms have evolved to eliminate aged or damaged mitochondria. One such quality control process is autophagy, a process that involves turnover of mitochondria at presynaptic sites and the axon terminal under basal conditions. This highly conserved mechanism sequesters mitochondria from the cytosol within autophagosomes followed by degradation upon fusion with a lysosome. Acute mitochondrial damage activates a selective form of autophagy called mitophagy that involves receptor-mediated engulfment and degradation of the damaged organelle. Multiple mechanisms have been shown to drive efficient mitophagy in neurons and glia, including PTEN induced kinase 1 (PINK1)/Parkin-dependent mitophagy and receptor-mediated mitophagy. Genetic, pathological, and experimental evidence all implicate defects in the removal of damaged mitochondria in the onset or progression of neurodegenerative disease. Both the initiation of PINK1/Parkin-dependent mitophagy and deficits in the removal of damaged mitochondria are linked to activation of neuroinflammatory pathways, including NF-κB and cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) signaling. In this Review, we discuss the molecular pathways governing mitophagy in neurons and glial cells and how deficits in these pathways may lead to neurodegeneration. We also highlight emerging therapeutic strategies aimed at restoring mitophagy to preserve neuronal homeostasis and function.

γδ T cells are a subset of lymphoid cells that, unlike their αβ lineage counterparts, express a heterodimeric TCR that mostly operates in an MHC-independent manner. γδ T cells are abundant in barrier tissues, where they continuously monitor epithelial cells for signs of stress or damage. Thus, γδ T cells are among the first responders to pathophysiological conditions, including viral infection and oncogenesis. Human γδ T cells can be classified based on TCR γ and δ chain usage into three main subsets: (a) Vγ9+Vδ2+ cells, accounting for most circulating γδ T cells; (b) Vδ1+ cells, which are common in epithelial linings, and (c) Vδ3+ T cells, which are fairly rare but exhibit unique specificities. Moreover, both human and murine γδ T cells can assume a spectrum of states with divergent phenotypic and functional properties. Accumulating evidence demonstrates that γδ T cells can mediate robust anticancer effects or support tumor progression and resistance to therapy, depending on numerous variables, including functional state and tumor type. Here, we critically discuss the context-dependent interaction between γδ T cells and cancer, focusing on recent developments and the challenges facing current efforts to manipulate this versatile lymphocyte subset for therapeutic purposes.

Crohn's disease is an inflammatory bowel disease (IBD) commonly treated through anti-TNF blockade. However, most patients still relapse and inevitably progress. Comprehensive single-cell RNA-sequencing (scRNA-seq) atlases have largely sampled patients with established treatment-refractory IBD, limiting our understanding of which cell types, subsets, and states at diagnosis anticipate disease severity and response to treatment. Here, through combining clinical, flow cytometry, histology, and scRNA-seq methods, we profile diagnostic human biopsies from the terminal ileum of treatment-naive pediatric patients with Crohn's disease (pediCD; <i>n</i> = 14), matched repeat biopsies (pediCD-treated; <i>n</i> = 8) and from non-inflamed pediatric controls with functional gastrointestinal disorders (FGIDs; <i>n</i> = 13). To resolve and annotate epithelial, stromal, and immune cell states among the 201,883 baseline single-cell transcriptomes, we develop a principled and unbiased tiered clustering approach, ARBOL. Through flow cytometry and scRNA-seq, we observe that treatment-naive pediCD and FGID have similar broad cell type composition. However, through high-resolution scRNA-seq analysis and microscopy, we identify significant differences in cell subsets and states that arise during pediCD relative to FGID. By closely linking our scRNA-seq analysis with clinical meta-data, we resolve a vector of T cell, innate lymphocyte, myeloid, and epithelial cell states in treatment-naive pediCD (pediCD-TIME) samples, which can distinguish patients along the trajectory of disease severity and anti-TNF response. By using ARBOL with integration, we position repeat on-treatment biopsies from our patients between treatment-naive pediCD and on-treatment adult CD. We identify that anti-TNF treatment pushes the pediatric cellular ecosystem toward an adult, more treatment-refractory state. Our study jointly leverages a treatment-naive cohort, high-resolution principled scRNA-seq data analysis, and clinical outcomes to understand which baseline cell states may predict Crohn's disease trajectory.

Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a bat-originated alphacoronavirus that causes devastating enteric disease in neonatal piglets and possesses significant potential for cross-species transmission. While the early stages of the coronavirus life cycle have been extensively characterized, the host factors indispensable for virion assembly and subsequent export remain largely enigmatic. Here, by performing a genome-wide CRISPR-Cas9 knockout screen using a recombinant icSADS-CoV-GFP reporter virus, we identified the small GTPase Rab10 as a critical host dependency factor for SADS-CoV infection. Viral life cycle analysis revealed that Rab10 is not required for viral attachment, entry, or initial genome replication, but is essential for the virion transport and non-lytic egress. Rab10 deficiency markedly reduced the extracellular release of viral RNA, viral proteins, and infectious progeny, as well as the secretion of SADS-CoV virus-like particles. Confocal imaging showed that Rab10 and viral protein-positive intracellular structures were associated with LMAN1, TGN46, and LAMP1 positive compartments. These findings support a model in which Rab10 coordinates a virus-containing vesicles trafficking pathway associated with ERGIC-TGN-lysosome compartments. Mechanistically, Rab10 facilitates the loading of the viral envelope (E) protein into transport vesicles derived from the ERGIC. Rab10 associates with the SADS-CoV E protein, and mapping analyses implicated the C-terminal PDZ-binding motif, particularly residue V75, in efficient Rab10 association and viral release. Collectively, our findings identify Rab10 as a host regulator of SADS-CoV non-lytic egress and highlight the E-Rab10 interaction and the vesicular trafficking machinery as a potential target for developing antiviral strategies.

<h4>Background</h4>Huntington's disease (HD) is a progressive neurodegenerative disease affecting motor and cognitive abilities, as well as exhibiting psychiatric manifestations. Studies in people with HD (PwHD) consistently report atrophy of the caudate and putamen as an early pathological event and is therefore considered an early biomarker. Investigating whether similar phenotypic features are apparent in rodent HD models is important since it could have translational potential in evaluating the efficacy of novel therapeutic strategies.<h4>Methods</h4>We used high-resolution anatomical images to longitudinally investigate brain morphology in the zQ175DN heterozygous mouse model (HET) and wildtype littermates (WT) at 3, 6, and 10 months of age (M), which reflect different stages of phenotypic progression. We investigated volumetric alterations using semi-automatic segmentations of HD on relevant regions-of-interest (striatum, cerebellum, corpus callosum, cerebral cortex, ventricles, and total brain volume) and whole brain voxel-wise Tensor Based Morphometry (TBM) analysis.<h4>Results</h4>The striatum showed the earliest progressive lower absolute volume in HET mice compared to WT, starting from 3 M, followed by lower absolute volume of cortex and corpus callosum concomitantly at 6 and 10 M. TBM highlighted lower relative local volume in the rostral-medial striatum at all ages, and in cerebral cortex in HET mice at 6 and 10 M. A bigger relative local volume in the cerebellum was observed at all ages in HET mice, and in the globus pallidus, substantia nigra, at 6 and 10 M.<h4>Conclusions</h4>Overall, this study revealed key structural abnormalities that resemble the natural history of PwHD. Hence, analysis of brain structure through MRI in the zQ175DN heterozygous mouse model holds potential for testing disease-modifying treatments expected to slow down or prevent structural degeneration.

Biomolecular phase separation has emerged as a key organizing principle in macroautophagy (hereafter autophagy). In mammalian cells, phase-separated condensates not only serve as substrates for selective degradation, but also act as dynamic platforms for cargo recognition, signaling integration, and autophagosome assembly. The material state of these condensates is an important determinant of autophagic fate. Condensates exist along a continuum ranging from liquid-like droplets to gel-like and solid assemblies, and their progressive maturation can alter accessibility to autophagic machinery. Scaffold proteins and selective autophagy receptors further organize these assemblies into degradation-competent mesoscale reaction fields that couple cargo recognition with phagophore formation. Dysregulation of this phase separation-autophagy axis is increasingly implicated in neurodegeneration, cancer, aging, and stress-associated degenerative disease. Here, we propose a multiscale framework in which molecular accessibility, mesoscale organization, and condensate state transitions collectively shape autophagic outcome, providing a conceptual basis for predictive models and therapeutic strategies aimed at restoring condensate degradability.

RAD51 paralogs (RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3) are evolutionarily conserved essential proteins for cell survival and genome maintenance. RAD51 paralogs were originally identified to play a role in homologous recombination-mediated repair of DNA double-strand breaks (DSBs). However, investigations over the last decade have uncovered new roles of RAD51 paralogs beyond DSB repair in replication stress responses, including replication fork progression, fork stability, and its restart. Recent structural studies have not only uncovered the molecular architecture of previously known RAD51 paralog complexes but also identified novel paralog complex assemblies, providing mechanistic insights into their various genome-maintenance functions. Additionally, a role for RAD51 paralogs in resolving R-loops has been identified, and studies with cancer-associated variants suggest that RAD51 paralogs are potential determinants of cancer susceptibility and therapeutic responses. In the present review, we highlight the recently deciphered structures and novel functions of RAD51 paralog complexes and discuss the clinical and therapeutic implications.

Long-term persistence of managed species will depend, in part, on whether the species harbors the physiological or genetic potential to adjust to warming temperatures, and whether relevant genetic variation is modified by management practices. The critically endangered Delta Smelt (<i>Hypomesus transpacificus</i>) is intensively managed, but little is known about the presence of genetic variation for resistance to elevated temperature. Using a pedigree and whole genome sequencing data, we characterized the genetic basis of CTMax (as a metric of upper thermal tolerance) across control and elevated rearing temperatures, alongside covarying traits (body size and degree of hatchery ancestry). Warmer rearing temperatures increased CTMax through acclimation but also resulted in reduced additive genetic variation for the trait. We observed modest heritability for CTMax at rearing temperatures of 15°C and 18°C (0.26 and 0.16, respectively), but only a limited number of loci were identified that had consistent effects on CTMax across rearing temperatures. Instead, the genomic basis of thermal tolerance was highly dependent on rearing temperature (i.e., many loci detected with a GxE effect). This temperature-dependent genomic architecture is consistent with our finding that additive genetic variation for CTMax was reduced under warmer rearing conditions, indicating a potential constraint on adaptive evolutionary change. The influence of domestication selection was indicated by changes in allele frequency, and divergence in upper thermal tolerance and plasticity, between low and high hatchery ancestry groups. Minimal overlap between loci associated with domestication and CTMax suggests that these traits possess separate genetic underpinnings. Knowledge of genetic variation supporting ecologically relevant physiological variation may be useful for captive management and may inform supplementation of fish to the wild in an ever-warming environment.

Community carriage of extended-spectrum <i>β</i>-lactamase (ESBL)-producing <i>Escherichia coli</i> represents an important reservoir of antimicrobial resistance. However, the genomic diversity and population structure of ESBL-producing <i>E. coli</i> circulating in community settings remain poorly characterized. This study aimed to characterize ESBL-producing <i>E. coli</i> isolated from fecal samples of residents in Ecuador, with an emphasis on the diversity and genomic context of ESBL genes. ESBL-producing <i>E. coli</i> was isolated from fecal samples obtained from 55 residents using MacConkey agar supplemented with cefotaxime. Whole-genome sequencing of the isolates was performed using a hybrid approach combining long- and short-read platforms. Plasmids and <i>β</i>-lactamase genes were identified using DFAST and PlasmidFinder. Bacterial identification and antimicrobial susceptibility testing were conducted by MALDI-TOF MS and the broth microdilution method, respectively. ESBL-producing <i>E. coli</i> were isolated from 35 of 55 fecal samples (63.6%). Complete circular genomes were obtained from 31 isolates. All isolates harbored <i>bla</i> _CTX-M genes, predominantly belonging to the <i>bla</i> _CTX-M-1 group, whereas 65.7% carried <i>bla</i> _TEM, mainly <i>bla</i> _TEM-1, and related variants. Although <i>β</i>-lactamase genes were predominantly plasmid-borne, chromosomal integration was detected in 40% of the isolates. Notably, 87.5% of the isolates harbored IncF plasmids with multiple replicons. Conserved IS<i>26</i>-flanked transposons carrying <i>bla</i> _CTX-M and <i>bla</i> _TEM were frequently identified in the plasmids. Phylogenetic analysis revealed substantial genomic diversity across seven phylogroups, together with closely related isolates detected within and between households. These findings provide high-resolution genomic insights into the ESBL determinants circulating in community residents and reveal region-specific patterns of ESBL genomic diversity.

Lineage plasticity is a major mechanism by which prostate cancer adapts to therapeutic pressure, particularly following sustained inhibition of androgen receptor signaling. A central mediator of this process is BRN2, a POU3F2 neural lineage transcription factor that is normally suppressed in AR-dependent luminal prostate epithelium but becomes aberrantly activated during progression to castration-resistant and neuroendocrine (NE) prostate cancer. This review examines how BRN2 connects developmental neurogenesis to therapy-induced tumor reprogramming. We integrate insights from neurodevelopment and direct lineage reprogramming to describe how BRN2 maintains stem-like and NE transcriptional states. Mechanistically, we discuss how AR suppression, cooperating oncogenic drivers, epigenetic remodeling, and post-translational modification activate BRN2 and reshape transcriptional networks involving SOX2, Wnt signaling, N-Myc, and EZH2. We also evaluate strategies to target BRN2 and its associated dependencies, including direct inhibition of BRN2 DNA binding and pharmacologic disruption of cooperating epigenetic complexes such as BRD4, EZH2, and LSD1, though EZH2 inhibition may paradoxically promote further NE differentiation. We additionally assess BRN2 as a potential circulating biomarker, detectable in extracellular vesicles, though no clinical-grade assay has been validated. This review highlights BRN2 as a key transcriptional regulator of lineage plasticity in prostate cancer, with important implications for overcoming therapy resistance in advanced prostate cancer.

Mitochondrial CLPP has emerged as an unusual therapeutic target because both increasing and decreasing its proteolytic activity can be beneficial, depending on the cellular and disease context. Pharmacological CLPP hyperactivation drives broad degradation of mitochondrial proteins and can selectively collapse mitochondrial fitness in susceptible tumor cells, an approach now clinically validated by the approval of dordaviprone for mutant diffuse midline glioma. Conversely, reduced CLPP activity can preserve respiratory-chain components and promote adaptive metabolic and redox remodelling in selected models of mitochondrial disease, neurodegeneration and metabolic dysfunction, with emerging potential in ischaemia-reperfusion injury. These opposing outcomes reflect the broader role of CLPXP in controlling mitochondrial translation, respiratory-chain integrity and metabolism rather than acting simply as a general protein quality-control system. In this review, we discuss the physiological functions and substrate selectivity of CLPXP, the mechanistic basis and clinical development of CLPP inhibitors and activators, and the growing evidence that therapeutic responses depend strongly on tissue identity, metabolic state and the nature of the underlying mitochondrial defect. Together, these findings position CLPP as a context-dependent therapeutic switch whose activity may need to be tuned in opposite directions to either preserve mitochondrial resilience or selectively dismantle mitochondrial fitness.

Glaucoma is the leading cause of irreversible blindness worldwide, primarily driven by the progressive loss of retinal ganglion cells (RGCs) under pathological high intraocular pressure (ph-IOP). Despite the established role of ferroptosis in RGC degeneration, specific molecular targets that can be used for clinical intervention still need to be optimized, and the slow onset of conventional gene therapy vectors is incompatible with the acute clinical course of glaucoma. Here, we integrate single-cell RNA sequencing and spatial transcriptomics to profile the dynamic transcriptomic landscape of the rat retina across acute, subacute, and chronic stages of ph-IOP injury. Through ferroptosis-focused screening of an early-activated RGC gene cluster, we identify the lipid metabolism regulator phosphatidylethanolamine-binding protein 1 (PEBP1) as a candidate mediator of RGC ferroptosis. We demonstrate that Pebp1 is specifically upregulated in injured RGCs with a trajectory mirroring ferroptosis pathway activation, and that AAV-mediated Pebp1 knockdown suppresses ferroptosis through the GPX4/ACSL4 signaling axis, thereby preserving RGC survival, retinal structure, and visual function. To overcome the critical time-window bottleneck-the several weeks delay required for AAV-mediated silencing versus the rapid, irreversible RGC loss in acute glaucoma-we engineer Exosomes-siPebp1, a mesenchymal stem cell-derived exosome system loaded with siPebp1, which enables immediate single-dose intervention post-injury. This system exhibits efficient RGC uptake, prolonged intraocular retention, and robust target gene silencing, and, in a head-to-head comparison, significantly outperforms unloaded exosomes, liposomal formulations, and AAV vectors in RGC protection, without detectable acute systemic or local toxicity. Collectively, this study implicates Pebp1 in ph-IOP-associated RGC ferroptosis and supports exosome-mediated siRNA delivery as a rapid, cell-free intervention strategy for acute glaucomatous injury.

Fungal pathogens are an escalating global public health concern, particularly in the context of invasive and opportunistic infections. Cryptococcosis, primarily caused by Cryptococcus neoformans var. grubii, can affect multiple organs and often leads to life-threatening meningitis in immunocompromised individuals. Given limited antifungal therapies and emergence of resistance and toxicity-related constraints, the development of novel anti-cryptococcal agents remains an urgent priority. A library of innovative 3-hydroxypyridin-4(1H)-one-based hybrids (5a-f) was synthesized and evaluated for antimicrobial activity against clinically relevant Gram-positive and Gram-negative bacteria, as well as fungal species Candida albicans and C. neoformans var. grubbi. Safety was assessed through cytotoxicity studies in HEK293 and HepG2 cells and hemolytic evaluation, while iron-chelating capacity and lipophilicity were also investigated. All compounds formed stable iron(III) complexes and displayed no significant toxicity up to 25 μM. Series 1 compounds (5a-c) were less lipophilic than Series 2 (5d-f), mainly due to regioisomeric position of hydroxyl group on 2-methyl-4-pyridone scaffold, whereas fluorination increased lipophilicity in both series. Notably, compounds 5c-f emerged as potent, selective, and nontoxic antifungal agents against C. neoformans var. grubii (MIC < 16 µg/mL; CC₅₀ > 32 µg/mL; HC₁₀ > 32 µg/mL), highlighting the potential of 3-hydroxypyridin-4(1H)-one-based hybrids as a promising approach for cryptococcal meningitis therapy.

Longgu (fossilized bone), a mineral medicine in traditional Chinese medicine, has been used for centuries to treat symptoms resembling modern anxiety and insomnia. This review evaluates the hypothesis that Longgu's therapeutic effects arise from a synergistic network of its constituent metal ions. We synthesized evidence from chemical analyses, pharmacological studies of Ca, Mg, Zn, Fe, Cu, and Mn, and clinical trials of Longgu-containing formulations. The metal ions form an integrated network exhibiting three emergent properties. First, ionostatic balance is achieved through Ca/Mg-mediated regulation of neuronal excitability. Second, cross-pathway synergy occurs where zinc concurrently modulates neurotransmission and promotes neuroplasticity via BDNF. Third, systemic integration involves zinc and manganese co-regulating the gut-brain axis. Clinical studies demonstrate that Longgu-containing formulations improve outcomes in depression, anxiety, and insomnia, particularly for comorbid conditions, and show synergistic effects when combined with conventional antidepressants. This network pharmacology framework repositions Longgu as a model for developing multi-target therapeutics, offering a strategy to overcome limitations of single-target psychiatric drugs.

Melittin, the cytolytic peptide of honeybee venom, exhibits potent anticancer activity in triple-negative breast cancer (TNBC), yet the molecular programs underlying its cytotoxic effects remain incompletely defined. To address this gap, MDA-MB-231 TNBC cells were exposed to melittin at half-maximal inhibitory concentration(half IC50) and IC50 across early(0.5, 1, and 2 h), mid(3, 4 h), and late (12, 24 h) time windows. Proteomic profiling was performed using label-free data-independent acquisition(DIA) parallel accumulation-serial fragmentation(PASEF). Approximately 5800 proteins were quantified, revealing distinct dose-dependent stress responses. An integrative exploratory framework combining time-resolved log2 fold-change trajectories, area-under-the-curve(AUC) based temporal prioritization, and independent heatmap visualization identified proteins associated with melittin-induced stress remodeling. Half IC50 exposure showed a transient stress-adaptive signature characterized by chromatin remodeling(HMGN2, H2AZ1), structural and RNA-associated buffering(LRRC7), and indirect mitochondrial quality-control signaling(CPAMD8, SPATA4), which progressively weakened over time. In contrast, IC50 treatment induced rapid chromatin remodeling dominated by histone H1 variants(H1.4, H1.2), early RNA instability(LRRC7), and late-stage cytoskeletal disassembly marked by MICAL3 induction, consistent with progression toward apoptosis. These trajectories paralleled dose-dependent apoptotic phenotypes. Overall, data suggest that melittin elicits dose- and time-dependent proteomic stress responses in TNBC cells and identify candidate trajectory-associated proteins and pathways linked to adaptive stress remodeling or progression toward cytotoxic collapse.

Androgenetic alopecia (AGA) is the most common form of non-scarring hair loss, driven by genetic factors and increased sensitivity of scalp hair follicles to dihydrotestosterone (DHT), which causes progressive miniaturization of dermal papilla cells and shortens the hair growth phase. The precise molecular pathogenesis of AGA remains incompletely understood. The study aimed to elucidate the regulatory mechanism between secreted frizzled-related protein 2 (SFRP2) and follistatin-like 1 (FSTL1), explore their impact on DHT-induced mitochondrial dysfunction and senescence in dermal papilla cells (DPCs), elucidate the effect of the SFRP2-FSTL1 axis on oxidative stress-related DPCs changes, and identify new therapeutic targets for AGA treatment. In the study, SFRP2 was highly expressed in the DPCs of AGA patients. Knocking down SFRP2 improved hair regeneration and follicle morphology within AGA model mice, promoting proliferation, migration, and invasion of DPCs in vitro. SFRP2 knockdown also alleviated inflammation, senescence, mitochondrial dysfunction, and oxidative stress. SFRP2 was found to bind to FSTL1, thereby promoting FSTL1 protein stability. Knocking down FSTL1 counteracted the negative effects of SFRP2 overexpression in vitro. In conclusion, SFRP2 emerges as a critical regulator in AGA by promoting stability and activity of FSTL1, establishing a novel SFRP2-FSTL1 axis that exacerbates DHT-driven pathogenic changes in DPCs. These findings identify SFRP2 and FSTL1 as key mediators of androgen-induced cellular dysfunction and suggest that disrupting this axis could offer a promising therapeutic strategy for slowing or reversing the progression of AGA.

Intestinal polyps may recur after endoscopic removal, and postoperative surveillance is important for reducing the risk of colorectal neoplasia. Although several studies have evaluated recurrence after conventional polypectomy or electrosurgical resection alone, evidence regarding recurrence after argon plasma coagulation (APC) combined with electrosurgical resection remains limited. This study aimed to identify factors associated with intestinal polyp recurrence after APC combined with electrosurgical resection. This single-center retrospective cohort study was conducted at the Affiliated Hospital of Southwest Medical University from March 2019 to April 2024. We included 330 adult inpatients who underwent argon plasma coagulation combined with electrosurgical resection for intestinal polyps and completed postoperative colonoscopic follow-up. The primary outcome was postoperative intestinal polyp recurrence detected by follow-up colonoscopy. Demographic, clinical, laboratory, and endoscopic characteristics were collected, and factors associated with recurrence were evaluated using univariable and multivariable logistic regression analyses. Among the 330 included patients, 230 were male (69.7%), the mean age was 56.89 ± 11.25 years, and the mean follow-up duration was 15.58 ± 10.63 months. According to postoperative colonoscopic follow-up, 225 patients were classified into the recurrence group and 105 into the non-recurrence group. In multivariable logistic regression analysis, male sex (odds ratio [OR]: 2.195, 95% confidence interval [CI]: 1.236-3.899, P = .007), polyp number ≥ 4 (OR: 2.117, 95% CI: 1.160-3.862, P = .015), and history of cholecystectomy (OR: 3.699, 95% CI: 1.184-11.554, P = .024) were independently associated with postoperative recurrence of intestinal polyps. In this single-center retrospective cohort study, male sex, polyp number ≥ 4, and history of cholecystectomy were independently associated with postoperative intestinal polyp recurrence after argon plasma coagulation combined with electrosurgical resection. These findings may help inform postoperative surveillance strategies, but further prospective studies are needed for validation.

Reliable determination of ionization energies (IE) and electron affinities (EA) is essential for understanding energy-level alignment in solar cells. Here, we benchmark cyclic voltammetry (CV) and electrochemical voltage spectroscopy (EVS) for IE and EA determination against ultraviolet photoelectron spectroscopy (UPS) for IE and the combination of UPS and photothermal deflection spectroscopy (PDS) for EA. The investigated materials are FA0.8Cs0.2Pb(I1-yBry)3 perovskite thin films with varying bromide fraction y = 0-1. Onset potentials and energies were reproducibly extracted using tangent-based scripts, avoiding data smoothing and minimizing user bias. The optical band gap scales linearly with bromide fraction. The IE remains approximately constant up to y = 0.4 (EVS) and y = 0.6 (CV), increasing upon further substitution. All investigated methods consistently reveal a decrease in EA with increasing bromide content. Finally, we highlight practical considerations relevant to the application of EC methods and UPS.

In populations with low genetic diversity, the capacity to adapt to environmental change is limited. While epigenetic mechanisms can generate phenotypic plasticity, their role in mediating adaptive life-history transitions in the wild is largely unknown. Here, we show that in adult Namibian cheetahs (Acinonyx jubatus jubatus), a species with remarkably low genetic diversity, the transition from a floater to a male holding a territory is associated with widespread DNA methylation changes, with a predominance of hypermethylation in territorial males. Using paired blood samples collected from the same individuals before and after becoming territorial, we identified differentially methylated genes which function in pathways related to anatomical and head development, brain development and neurodevelopment, cAMP biosynthesis and ion channel transport, consistent with the observed phenotypic shifts in morphology, spatial movement and aggressive behaviour. These findings suggest that DNA methylation mediates this life-history transition at the molecular level. More broadly, they provide rare evidence that epigenetic regulation is relevant in phenotypic diversity within species with low genetic diversity, potentially supporting adaptive potential at the population level.

<h4>Introduction</h4>A higher incidence of dementia, including Alzheimer's-like pathology, is observed in aged people living with human immunodeficiency virus-1 (HIV-1). However, mechanisms linking HIV-1 to Alzheimer's disease (AD) pathology remain unclear, due to the lack of animal models that allow for concurrent studies of HIV-1 and AD.<h4>Methods</h4>We created a novel amyloid precursor protein (APP) (Swedish mutation) knock-in (KI) AD mouse on an immunocompromised NOG background, NOG/APP^KM670,671NL/IL-34 (NAIL). Following CD34+ hematopoietic stem cell (HSC) reconstitution, humanized hNAIL mice develop human microglia-like cells in the brain and human immune system in the periphery. This allows, for the first time, studies of progressive brain HIV-1 replication in an AD brain. Four-month-old HSC reconstituted mice were infected with the HIV-1_ADA strain, and evaluated at 8 weeks post infection to study the role of brain HIV-1 replication on AD-like pathologies.<h4>Results</h4>HIV-1 replication increased amyloid-beta (Aβ) load in the brain and reduced synaptic and neuronal integrity. Cell type-specific spatial transcriptomic analysis demonstrated that Aβ and HIV-1 drive distinct transcriptional patterns, whereas dual pathology amplified AD-like pathology. Neurons showed the highest transcriptional change, with genes linked to neuroinflammation, protein trafficking, and synaptic dysfunction.<h4>Discussion</h4>The hNAIL mice enable interrogation of HIV-AD comorbidities, with a future potential for the development of novel therapeutic interventions.

<i>Mycobacterium abscessus,</i> a non-tuberculous mycobacterium, is a cause of severe respiratory infections, notably in individuals with underlying lung conditions. Its high levels of intrinsic and acquired antimicrobial resistance make it particularly difficult to treat and horizontally acquired genetic elements may facilitate the spread of resistance. A small number of plasmids have been identified in this species, but their distribution, transmission dynamics across subspecies and clonal lineages remain poorly characterized. We analysed short-read genomic data from 3,060 <i>M</i>. <i>abscessus</i> isolates, including longitudinal samples, to characterize plasmid diversity and dynamics. Using a graph-based pan-genome approach, we identified 28 plasmids, including 15 previously unreported plasmids, mapped their distribution onto the species phylogeny and assessed their functional potential. Overall, 23.1% of isolates carried at least one plasmid, with higher prevalence in dominant circulating clones (DCCs) compared with non-DCCs. Plasmid carriage varied across subspecies and clonal backgrounds, and plasmids encoded numerous genes which may be linked to bacterial adaptation. Several plasmids persisted across multiple time points within individual patients, suggesting they can be highly stable over the course of a chronic infection.

Protein-drug conjugates (PDCs) constitute a rapidly growing group of precise anticancer agents. Receptor-mediated endocytosis is a critical step in PDC action, which ensures the delivery of PDCs into the interior of cancer cells. Here, we report TriFHS-MMAE, the first multivalent PDC specifically targeting heparan sulfate proteoglycans (HSPGs) overexpressed by pancreatic cancer cells, which induces ultrafast and highly efficient aggregation-dependent endocytosis (ADE) of HSPGs. Using high-content screening with a near-kinome-wide library of inhibitors, we identified signaling pathways that govern ADE of HSPGs and discovered cascades that selectively operate in pancreatic cancer cells versus healthy cells. We show that priming cells with identified endocytic chemical modulators improves the targeting of the TriFHS-MMAE conjugate in vitro and in vivo. Overall, these findings provide insights into the interplay among signaling, endocytosis, and PDCs and support the development of specific therapies for pancreatic cancer.

Sarcopenia, the age-related loss of muscle strength and mass, contributes to adverse health outcomes in older adults. Exercise engages calcium (Ca²⁺)- and redox-dependent signaling pathways that enhance muscle performance and adaptation, whereas aging disrupts Ca²⁺ and redox homeostasis. CaMKII is a key transducer of both signals, raising the possibility that sustained CaMKII signaling becomes maladaptive with aging. Here, we show that CaMKII protein abundance is increased in aged mouse skeletal muscle and that sustained CaMKII activation in young muscle is sufficient to impair contractile function before substantial atrophy develops and, with prolonged activation, to promote progressive muscle loss. Sustained CaMKII activation also disrupted mitochondrial organization and shifted the young-muscle transcriptome toward an aged profile characterized by inflammatory and stress-response pathways. Inhibiting canonical NF-κB signaling partially preserved contractile force during prolonged CaMKII activation without preserving muscle mass, and mediation analysis implicated heme/iron-related transcriptional remodeling in the muscle-mass-independent decline in force. Conversely, expression of CN19o, a peptide inhibitor of CaMKII, in aged muscle improved contractile function and shifted the transcriptome away from an aging-associated profile without inducing hypertrophy. Together, these findings identify sustained CaMKII signaling as a contributor to age-associated muscle dysfunction and support a context-dependent shift from adaptive CaMKII signaling in youth to maladaptive signaling in aging, consistent with antagonistic pleiotropy.

<h4>Background</h4>Intrahepatic cholangiocarcinoma (ICC) is a highly heterogeneous and aggressive malignancy originating from the intrahepatic bile duct epithelial cells. Despite advances in surgery, chemotherapy, targeted therapy, and immune checkpoint inhibitor (ICI)-based regimens, durable disease control remains limited by primary or acquired resistance and marked biological heterogeneity.<h4>Main body</h4>Immunotherapy, primarily based on ICIs, has shown clinical potential for advanced ICC; however, its activity is constrained by a highly immunosuppressive tumour microenvironment (TME), including desmoplastic stromal barriers, myeloid-cell-mediated suppression, impaired antigen presentation, cytokine- and chemokine-driven immune exclusion, and metabolic remodeling. This review summarizes the tumour immune microenvironment (TIME) of ICC, with emphasis on cellular crosstalk among stromal, myeloid, and lymphoid compartments, extracellular matrix remodeling, metabolic constraints, and immune-escape processes. We critically appraised first-line chemoimmunotherapy evidence from advanced biliary tract cancer (BTC) trials and interpreted its relevance to ICC according to clinical evidence, tumour location, molecular subtype, and biomarker context. Biomarkers are discussed according to their current level of clinical validation, and molecular alterations, such as FGFR2 fusions, IDH1 mutations, and BAP1/ARID1A alterations, are considered in relation to immune phenotypes and mechanism-guided combination hypotheses. Finally, we evaluated emerging approaches, including next-generation checkpoint inhibitors, antibody-drug conjugates, bispecific antibodies, CAR-T/CAR-macrophage (CAR-M)/CAR-natural killer (CAR-NK) cell therapies, cancer vaccines, and dendritic cell-based interventions, with particular attention paid to antigen heterogeneity, stromal trafficking barriers, manufacturing complexity, and the limited maturity of ICC-specific clinical evidence.<h4>Conclusions</h4>By integrating the mechanism, clinical evidence, molecular subtype and translational limitations, this review provides a balanced framework for biomarker-guided precision immunotherapy in ICC.<h4>Key points</h4>The clinical evidence and limitations of immunotherapy for intrahepatic cholangiocarcinoma are critically evaluated, with emphasis on ICC-specific interpretation of broader biliary tract cancer trials. The ICC tumour immune microenvironment is integrated into functional immune axes involving stromal, myeloid, lymphoid and metabolic regulation. Molecular subtypes and immune-related biomarkers provide a framework for biomarker-guided precision immunotherapy in ICC. Current ICC immunotherapy trials remain limited by heterogeneous designs, small ICC-specific cohorts and immature clinical evidence. Emerging immunotherapies, including next-generation checkpoint inhibitors, antibody-drug conjugates, cellular therapies and vaccines, require ICC-specific evaluation of biological barriers and clinical feasibility.

<h4>Rationale</h4>Circular RNAs (circRNAs) are stable, tissue- and developmental-stage-specific regulators of gene expression and candidate disease biomarkers. Their expression profile in abnormal lung development in congenital diaphragmatic hernia (CDH) is unknown.<h4>Objective</h4>To evaluate circRNA expression profile in CDH-associated abnormal lung development.<h4>Methods</h4>We profiled circRNAs in rat CDH and control lungs at embryonic day (E)15 and E21 by microarray. We validated identified circRNAs using back‒splice junction amplicon sequencing, RT-qPCR and in situ hybridisation. We modified a circRNA function prediction tool to predict circRNA::micro(mi)RNA::messenger(m)RNA interactions and compared these with Oxford Nanopore RNA sequencing and existing human CDH datasets.<h4>Measurements and main results</h4>Microarrays revealed a unique circRNA biosignature during CDH lung development. CircAnp32e was expressed in a sex-specific and spatiotemporal expression pattern in the epithelium at E15. The predicted mature sequence of circAnp32e overlapped 90% with its human orthologue. circRNA::miRNA::mRNA interaction networks at E15 and E21 revealed enrichment in inflammation/infection, smooth muscle cell function, cell proliferation/cell cycle regulation and response to hypoxia pathways. Parental genes of differentially expressed circRNAs at E15 enriched pathways linked to cell proliferation/cell cycle/cancer, while at end-gestation, inflammation and cardiovascular processes were also overrepresented. Rat and human CDH lungs showed overlapping pathways with additional enrichment for RNA processing and protein binding/modification in humans. In a human bronchial epithelial (BEAS-2B) nitrofen-injury model, ANP32E and circANP32E were downregulated, and the predicted let-7 target was significantly dysregulated.<h4>Conclusion</h4>A unique circRNA signature during abnormal lung development in CDH may mediate inflammatory responses, smooth muscle cell function and cell proliferation regulation via miRNA sponging. Overlap of downstream pathways in rat and human CDH suggests conserved functions across species. This circRNA biosignature defines strong candidate biomarkers for CDH and a basis for future prospective prenatal investigation.

<h4>Background</h4>The piriform cortex (PIR) is a key region of the olfactory system and is increasingly implicated in Alzheimer's disease (AD). However, the cellular heterogeneity of principal neurons (PN) in this region and their selective vulnerability in AD remain poorly understood.<h4>Methods</h4>Here, we integrated single-cell multi-omics datasets to systematically characterize PIR PN heterogeneity. We identified four transcriptionally and epigenetically distinct PN subtypes with different spatial distributions across the anterior and posterior PIR.<h4>Results</h4>In AD mouse models, the abundance of the Reln-positive superficial-layer (Reln+ SL) subtype was spatially associated with amyloid-β pathology, and its reduced proportion was confirmed by multiplex immunofluorescence. Cross-region comparison revealed that Reln+ SL neurons shared a Gpc5-high/Sort1-low molecular signature with vulnerable layer II excitatory neurons in the entorhinal cortex (EC). In human AD, this vulnerability-associated signature showed progressive enrichment in EC layer II neurons across Braak stages I-III. Furthermore, A Sort1-containing co-expression module captured regional differences between lateral and medial EC layer II populations and was enriched for proteostasis-related pathways.<h4>Conclusions</h4>These findings reveal shared molecular features associated with selective vulnerability across PIR and EC layer II neurons and identify a proteostasis-related program linked to regional differences in neuronal vulnerability during AD.<h4>Key points</h4>Single-cell multi-omics reveals molecular heterogeneity among piriform cortex principal neurons. Reln+ superficial-layer neurons show selective vulnerability in AD. Cross-region comparison identifies a shared Gpc5-high/Sort1-low signature in vulnerable layer II neurons of EC and PIR.

Metabolic reprogramming and immune evasion are two key mechanisms that facilitate tumor progression. Tumor cells achieve their energy requirements for rapid proliferation through metabolic reprogramming, such as glucose metabolism, lipid metabolism, and amino acid metabolism. Tumor cell metabolic reprogramming can influence immune cell function in the tumor microenvironment through various mechanisms, ultimately facilitating tumor immune escape. Targeting tumor cell metabolism and combining metabolic regulation with immunotherapy can enhance anti-tumor immune cells, inhibit the function of immunosuppressive cells, improve anti-tumor therapy, and overcome immune escape. This review elucidates the regulatory mechanisms by which glucose, lipids, amino acid metabolism, nucleotide metabolism, oxidative phosphorylation, lactate, and hypoxia modulate the proliferation, differentiation, and function of anti-tumor immune cells and immunosuppressive cells. Regulatory strategies to restore the anti-tumor function of immune cells in response to metabolic changes were discussed. These strategies are expected to improve the effect of immunotherapy and are used in combination with other treatments to provide new ideas for cancer management.

<h4>Background</h4>Variants in STX1B/syntaxin-1B are linked to a spectrum of fever-associated epilepsy syndromes. While studies in murine models have provided mechanistic insights, their relevance to human disease in a heterozygous context may be limited.<h4>Methods</h4>We investigated two pathogenic STX1B variants using isolated single neurons and neuronal network cultures derived from patient-specific induced pluripotent stem cells. These carried either a de novo p.G226R variant, associated with severe developmental epilepsy, or an InDel variant (p.K45delinsRCMIE/p.L46M) linked to a transient familial seizure syndrome. Synaptic function and network excitability were assessed using patch-clamp and multi-electrode array recordings, alongside morphological and transcriptomic profiling.<h4>Findings</h4>G226R exhibited both gain- and loss-of-function characteristics, with increased miniature excitatory postsynaptic current frequency in networks but not in autapses, and synaptic failure during sustained high-frequency stimulation. For the InDel variant, the predicted loss-of-function phenotype based on reduced syntaxin-1B levels was not detectable at the single-cell level, likely masked by compensatory synaptic upregulation. At the network level, however, both variants were associated with neuronal hyperexcitability, characterised by more frequent and prolonged bursting activity, with a much stronger phenotype in G226R-containing networks. Transcriptomic profiling revealed a differential dysregulation of synaptic and other neuronal genes.<h4>Interpretation</h4>The divergence between morphological, electrophysiological and transcriptomic findings suggests that compensatory mechanisms may contribute to network hyperexcitability. Initially engaged to maintain homoeostasis, they may ultimately contribute to a pathological network state. The graded severity of network alterations across STX1B variants correlates with the clinical phenotypes.<h4>Funding</h4>BMBF (Treat ION-01GM2210A, SNAREopathies-01EW1809A), 2023 FEBS Summer Fellowship, Fortüne programme (2610-0-0), EKFS college precise.net, Open Access Publishing Fund of University of Tübingen.

<h4>Background</h4>Single-cell and single-nucleus RNA sequencing have transformed our understanding of human skeletal muscle biology, yet reproducibility and cross-study comparison remain limited by the lack of a unified reference framework and consistent cell-type annotation.<h4>Methods</h4>We systematically searched for scRNA-seq and snRNA-seq datasets from adult human skeletal muscle. Seven eligible studies were retrieved and harmonised. We benchmarked multiple integration strategies to construct a joint reference atlas and derived modality-aware marker panels. Selected findings were validated by immunofluorescence in muscle biopsies.<h4>Findings</h4>We generated a harmonised atlas comprising 122,000 cells and 630,000 nuclei from 88 healthy individuals and resolved 17 major skeletal muscle cell populations, spanning mononuclear compartments and multinucleated myofibers. Cross-modality analysis identified tissue- and modality-aware marker panels and nominated both established and previously unrecognised markers. NOVA1 emerged as a selective marker of fibro-adipogenic progenitors and was validated at the transcript and protein levels. Focusing on myonuclei, pseudotime modelling reconstructed differentiation trajectories from quiescent muscle stem cells to mature type I and type II myofibers and revealed lineage-specific programs, including transient activation of protocadherin-γ genes during type I myofiber differentiation. We further provide an interactive web application for marker-based cell-type prediction using the reference atlas.<h4>Interpretation</h4>This integrated reference atlas and accompanying annotation tool establish a standardised framework for human muscle transcriptomics, promoting consistent cell-type assignment and providing a baseline for future studies of muscle development, ageing, and disease.<h4>Funding</h4>Else Kröner-Fresenius-Stiftung and the German Research Foundation.

Follistatin-like 1 (FSTL1) has been considered a vital adipokine. Several lines of evidence have shown that FSTL1 is essential for white adipose tissue (WAT) adipogenesis, brown adipose tissue involution, and thermogenesis. However, the roles of FSTL1 in WAT browning and its underlying mechanisms are not fully understood. Our data show that FSTL1 overexpression induces WAT browning, enhances energy metabolism, and promotes glucose uptake in male mice at ambient room temperature. Overexpression of FSTL1 specifically in WAT prevents metabolic dysfunction in mice with diet-induced obesity. Mice with FSTL1 deficiency exhibit cold intolerance and are prone to obesity. FSTL1 cannot directly induce adipocyte browning in vitro. FSTL1 promotes sympathetic innervation both in vivo and in vitro. Furthermore, sympathetic denervation in adipose tissue abrogates FSTL1-induced browning, enhancement of energy metabolism, and regional glucose uptake. Notably, FSTL1 is endocytosed by sympathetic neurons primarily via tropomyosin-related kinase B (TrkB) in vitro. Blocking TrkB suppresses WAT browning and sympathetic innervation induced by FSTL1 overexpression in adipose tissue. Our findings shed new light on the role of FSTL1 in WAT browning and identify FSTL1 as an adipose-derived 'neurotrophic factor' adipokine.

Neutrophils exhibit substantial functional heterogeneity shaped by both developmental stage and cellular maturation, but their mechanisms remain incompletely defined. CD11c (ITGAX) is traditionally considered as a cell surface marker of dendritic cells. We recently reported that it is highly expressed intracellularly in neutrophils and contributes to their maturation; however, its role at the cell surface is unclear. Here, we showed that surface CD11c expression level was heterogeneous across neutrophils, enabling classification into CD11c^hi and CD11c^-/lo populations. CD11c^hi neutrophils display enhanced phagocytic capacity and features of neutrophil aging, including increased CXCR4 and reduced CD62L expression. Bulk RNA sequencing across the three pediatric age groups revealed that differentially expressed genes (DEGs) between CD11c^hi and CD11c^-/lo neutrophils were observed most in infants followed by preschool age children. Infant neutrophils exhibited reduced phagocytic capacity and distinct gene expression patterns compared to older children. Functional interrogation of candidate genes (ANXA2, COL6A3, DCN) demonstrated their role in regulating phagocytosis without affecting reactive oxygen species production. Machine learning analysis further identified age-dependent ontology of the DEGs associated with cell surface CD11c expression, including pathways related to adhesion, extracellular matrix interactions, stress responses, and metabolic regulation. Integrative network analysis positioned CD11c linked to cytoskeletal remodeling and phagocytosis. Collectively, these findings support a model in which developmental age establishes the baseline transcriptional landscape of neutrophils, while CD11c serves as a marker of neutrophil aging rather than acting as a primary driver of transcriptional reprogramming.

<i>Citrus nippokoreana</i>, a traditional Korean citrus species, is rich in flavonoids and polyphenols with antioxidant properties; however, its neuroprotective potential remains unclear. Neurodegeneration is closely associated with oxidative stress, neuronal apoptosis, and impaired synaptic signaling. In this study, we investigated the effects of <i>C. nippokoreana</i> peel extract (CNE, 30% ethanol) using <i>in vitro</i> and <i>in vivo</i> models. CNE significantly protected HT22 hippocampal neurons against glutamate-induced oxidative stress by reducing intracellular reactive oxygen species (ROS) and apoptosis, while regulating AKT/Nrf2-associated antioxidant pathways and enhancing BDNF/CREB signaling. In a scopolamine-induced mouse model, oral administration of CNE (50 or 100 mg/kg/day) improved cognitive performance in behavioral tests. CNE also restored the expression of BDNF, CREB, and HO-1, and reduced acetylcholinesterase activity in the hippocampus. Histological analysis confirmed reduced neuronal damage. Furthermore, serum metabolomics revealed modulation of tryptophan metabolism, phosphatidylcholine species, and redox-related pathways. To explore constituents potentially contributing to the neuroprotective effects of CNE, major compounds identified by phytochemical profiling were screened in glutamate-induced HT22 cells. Several phytochemicals exhibited protective effects, suggesting that the biological activity of CNE may result from the combined actions of multiple constituents rather than a single compound. These findings suggest that CNE attenuates oxidative stress-induced neuronal damage and improves cognitive function through coordinated regulation of antioxidant defense, neurotrophic signaling, and cholinergic function.

<b>Background:</b> Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality, with resistance to immunotherapy posing a major clinical challenge. Natural killer (NK) cells exhibit impaired infiltration and cytotoxicity in HCC; however, the mechanisms underlying NK cell-mediated immune evasion are still poorly understood. This study investigated how NOP2/Sun RNA methyltransferase 2 (NSUN2), a 5-methylcytosine (m⁵C) RNA methyltransferase, induces metabolic reprogramming and immunosuppression to drive HCC progression. <b>Methods:</b> We conducted a genome-wide CRISPR screen in HCC cells cocultured with NK cells. To delineate the downstream mechanisms, we integrated profiling of the m5C epitranscriptome, transcriptome, and chromatin landscape with metabolic characterization. The impact of <i>NSUN2</i> on histone lactylation and programmed cell death 1 ligand 1 (PD-L1) transcription was further investigated. Functional assays in vitro and in vivo using syngeneic murine models and pharmacological inhibition validated these findings. Clinical relevance was assessed using patient tissues, The Cancer Genome Atlas dataset, and immunotherapy cohorts. <b>Results:</b> Genome-wide CRISPR screening in HCC cell-NK cell coculture models identified <i>NSUN2</i> as a key suppressor of NK cell-mediated cytotoxicity. Mechanistically, NSUN2-mediated RNA m⁵C modification enhanced the messenger RNA stability and expression of glycolytic enzymes, including enolase 1 (<i>ENO1</i>), pyruvate kinase M1/2 (<i>PKM</i>), and lactate dehydrogenase A (<i>LDHA</i>), thereby increasing lactate production. Accumulated lactate promoted histone H3 lysine 18 lactylation (H3K18la), which enhanced chromatin accessibility at the <i>CD274</i> (encoding PD-L1) promoter and recruited signal transducer and activator of transcription 3 (STAT3) to drive PD-L1 expression, ultimately inhibiting NK cell-mediated cytotoxicity. Clinically, high NSUN2 expression was associated with elevated PD-L1 levels, poor prognosis, and immunotherapy resistance in patients with HCC. In vivo, <i>NSUN2</i> knockout increased NK cell infiltration and suppressed tumor growth, while the STAT3 inhibitor TTI-101 combined with anti-PD-L1 therapy enhanced NK cell cytotoxicity and inhibited HCC progression. <b>Conclusions:</b> Our data demonstrated that NSUN2 drove immune evasion in HCC by coupling m⁵C-dependent glycolytic reprogramming with H3K18la-mediated epigenetic activation of PD-L1. These findings suggest that NSUN2 could represent a critical nexus between m⁵C RNA methylation and immunosuppression, providing a therapeutic rationale for combination immunotherapy in HCC.

<h4>Background</h4>To determine whether urine 5-HIAA (u5HIAA) concentrations correlate with health-related quality of life (HRQL) in patients with neuroendocrine neoplasms (NEN).<h4>Methods</h4>A retrospective single-centre cross-sectional study was conducted including 386 patients with histological or radiological diagnosis of NEN. Patients completed standardized questionnaires at clinic appointments. Spot u5HIAA concentrations were correlated with Patient Health Questionnaire-9 (PHQ-9) depression scores, Generalized Anxiety Disorder-7 (GAD-7) anxiety scores, European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) scores, and the Quality of Life Questionnaire Gastrointestinal Neuroendocrine Tumour Module 21 (QLQ-GINET21) scores. Correlation was evaluated with the Spearman rho test.<h4>Results</h4>258 patients had completed HRQL data and corresponding u5HIAA available. Unknown primary sites and sites which do not typically secrete 5-HIAA (26 cases) were excluded in the analysis making a total of 232. One hundred and nineteen (51.3%) were male, 145 (62.5%) had a small intestine primary. Median age was 66 years and median u5HIAA was 7.8 µmol/mmol. Males had significantly higher u5HIAA (median:10.0, IQR: 5.6 - 15.4) compared to females (5.6, IQR: 3.3 - 11.3) (p=0.001). Males reported significantly higher Global Health Status (p=0.018) and lower GINET21 (p=0.001), GAD-7 (p=0.001) and PHQ-9 (p=0.009) scores. Within the overall cohort, there was a significant association between high u5HIAA and low GAD-7 (p=0.032); and between high u5HIAA and low GINET21 GI symptoms (p=0.024). No significant association was found between u5HIAA and PHQ-9 (p=0.383), QLQ GINET21 endocrine symptoms (p=0.197), EORTC QLQ-C30 Global Health Status (p=0.746) or diarrhoea score (p=0.305).<h4>Conclusion</h4>High circulating concentrations of serotonin may be associated with less anxiety in patients with NEN, further prospective studies are required to corroborate these results. Males had higher u5HIAA and fewer symptoms than females. The sex differences in 5-HIAA values and HRQL in this cohort warrant further investigation.

Congenital dyserythropoietic anemia is a group of hereditary disorders characterized by erythroid hyperplasia and ineffective erythropoiesis, resulting in anemia of varying severity. Congenital dyserythropoietic anemia Type 1 (CDA-1) is classically associated with biallelic mutations in the CDAN1 gene. Here, we report the first case of compound heterozygous CDAN1 mutations p.(D1043V) and p.(S1036F) in clinical practice, presenting in a 24-year-old woman with mild, asymptomatic macrocytic anemia and hyperferritinemia. These variants are currently classified as variants of uncertain significance; however, this report represents the first clinical case of this compound heterozygous CDAN1 variant combination in a patient with a phenotype consistent with CDA-1. As the phenotypic boundaries of CDA-1 continue to expand, clinicians should consider CDA-1 in the differential diagnosis of unexplained macrocytic anemia, even in the absence of severe anemia.

Corn silk (CS), an abundant agricultural waste, is typically converted into carbon dots (CDs) through energy-intensive methods such as hydrothermal treatment and microwave-assisted synthesis. In contrast, this study introduces a novel, simple, non-thermal (non-synthetic), and chemical-free extraction strategy to produce CDs directly from corn silk at ambient conditions. The properties of the resulting CDs were comprehensively characterized using transmission electron microscopy (TEM), Fourier transform infrared (FTIR) spectroscopy, and photoluminescence (PL) spectroscopy. While existing literature extensively reports biomass-derived CDs as probes for Fe³⁺ detection, their application for Cu²⁺ sensing remains limited and often requires complex surface functionalization. Remarkably, our non-synthetic extracted CDs exhibit high sensitivity toward Cu²⁺ ions with a competitive limit of detection (LOD) of 33 nM. Furthermore, to elucidate the impact of processing conditions, we systematically investigated the hydrothermal carbonization of corn silk across a temperature gradient. Our findings reveal that the preparation method acts as a functional switch: the non-synthetic route enables Cu²⁺ sensing, while hydrothermal treatment optimizes Fe³⁺ detection at 200 °C and enhances antimicrobial activity at 240 °C. This work establishes the first energy-free, non-synthetic approach to transform corn silk into high-performance Cu²⁺ nanoprobes, providing a sustainable and tunable platform for converting biomass into multifunctional nanomaterials.

<h4>Purpose</h4>Low back pain (LBP) is a heterogeneous pain condition with a measurable genetic contribution, but the genes, brain cell types, and spatial tissue contexts through which inherited risk is expressed remain unclear. We aimed to define cell-type-specific and spatially contextualized genetic mechanisms underlying LBP.<h4>Methods</h4>FinnGen R12 LBP GWAS summary statistics (42,521 cases and 353,224 controls) were integrated with brain single-nuclei eQTL data across eight major brain cell classes. We evaluated genome-wide polygenic signal using LDSC, prioritized genes using MAGMA and PoPS, and performed brain cell-type-specific eQTL-anchored Mendelian randomization, primarily based on single-instrument Wald ratio estimates, followed by Bayesian colocalization. Spatial genetic mapping was conducted using gsMap in an E16.5 murine embryonic atlas and two adult human lumbar spinal cord Visium sections. Selected candidates were assessed by RT-qPCR in neuronal-like and astroglial-like inflammatory cell models.<h4>Results</h4>LDSC supported interpretable polygenic signal for LBP. MAGMA and PoPS showed partial gene-level convergence, with TCF4 and TMEFF2 supported by both approaches. Across 1641 tested gene-cell type exposures, significant eQTL-anchored MR associations were concentrated in excitatory neurons, oligodendrocytes, inhibitory neurons, and astrocytes. Integrated eQTL-anchored MR, colocalization, and gene-prioritization evidence highlighted CLEC18A, QPRT, and GMPPB as higher-priority non-MHC candidates with moderate, but not strong, colocalization support. gsMap localized LBP-associated enrichment to neuroaxis-related embryonic regions, including brain, spinal cord, sympathetic nerve, and dorsal root ganglion, and to neuronal-like niches in adult lumbar spinal cord. RT-qPCR showed model-dependent expression changes, with QPRT and LGI4 preferentially responsive in neuronal-like SH-SY5Y cells and GMPPB and DPYSL5 responsive in astroglial-like U251 cells.<h4>Conclusion</h4>These findings support neuronal and glial regulatory programs as plausible contributors to LBP genetic susceptibility and highlight CLEC18A, QPRT, and GMPPB as higher-priority non-MHC candidates with moderate colocalization support. The results provide a spatially contextualized framework for candidate prioritization in LBP, while emphasizing the need for larger cell-type-specific eQTL resources and functional validation before therapeutic or mechanistic conclusions can be drawn.

<h4>Introduction</h4>Metabolic dysfunction-associated steatotic liver disease (MASLD) has been established to classify steatotic liver disease more accurately. We investigated the association between MASLD and long-term kidney transplant outcomes.<h4>Methods</h4>Kidney transplant recipients who underwent national health screening programs from 2009 to 2017 and with follow-up until 2021 were identified using the Korean National Health Insurance Service data. MASLD was defined based on cardiometabolic criteria and the fatty liver index.<h4>Results</h4>Among 8326 kidney transplant recipients, 2027 (24.3%) had MASLD, 25 (0.3%) had MASLD with increased alcohol intake, and 419 (5.0%) had MASLD with other liver diseases. The incidence rates of graft failure (17.5 vs. 11.3, <i>P</i> < .001) and mortality (12.5 vs. 8.9 per 1000 person-years, <i>P</i> < .001) were significantly higher in the MASLD group than in those without steatotic liver disease. After adjusting for demographics, lifestyle, comorbidities, estimated glomerular filtration rate, proteinuria, and acute rejection, the adjusted hazard ratios for graft failure and mortality were 1.23 (95% confidence interval, 1.03-1.46) and 1.03 (95% confidence interval, 0.85-1.25), respectively.<h4>Conclusions</h4>This study identified MASLD as a significant risk factor for graft failure in kidney transplant recipients, highlighting the need for MASLD screening and management in kidney transplant recipients.

High-grade serous ovarian carcinoma (HGSC) responds poorly to immune checkpoint blockade, partly due to a macrophage-dominated immunosuppressive microenvironment. We integrated single-cell spatial proteomics and spatial transcriptomics across 50 HGSC tumors and applied SPACEstat to resolve higher-order immune communities and their transcriptional programs. We identified six immune community types, with macrophage-dominated Myelonets representing the predominant spatial pattern of immune organisation. In chemotherapy-exposed tumors, Myelonets showed coordinated lipid metabolism–immunosuppression and inflammation–MHC-II macrophage transcriptional programs, with SPP1, C1Q, VEGF, MMPs, and CCL18 linked to immunosuppressive states and fibroblasts emerging as key mediators of macrophage communication. Chemotherapy contracted large Myelonets while increasing CD8+ T-cell organization into Lymphonets. Persistent macrophage dominance within Myelonets was associated with adverse outcomes among patients who achieved a complete response to treatment. Together, we identify Myelonets as clinically relevant, multicellular immunoregulatory niches sustained by spatiotemporally coordinated macrophage programs and stromal crosstalk.

Disseminated cancer cells (DCC) in non-metastatic carcinoma patient bone marrow (BM) are predictive of metastasis. Those detected by epithelial cytokeratin or EpCAM expression have poorly-characterized transcription profiles due to their extreme rarity: 1∼2 cells per 2x 10 6 BM cells in every third non-metastatic patient. We here characterize the transcriptomes of DCCs. Single-cell RNA-sequencing (scRNA-seq) of 864 EpCAM-positive cells (from 1,151 cancer patients) in BM or lymph nodes (LN) revealed plasma, immune, myeloid, erythroid progenitor cells and two candidate DCC populations, termed M0-DCC and M1-DCC. M0-DCC, mostly from non-metastatic M0-stage patients, displayed the highest known adult stemness scores, and were transcriptomically reminiscent of human cleavage-stage, preimplantation embryos. M1-DCC represented cancer cells undergoing the epithelial-mesenchymal transition (EMT), corresponding to later, implanting and gastrulating embryos. Detection of early-embryo-like DCC categorised patients at highest risk for metastatic progression. Furthermore, high M0-DCC scores predicted the metastatic potential of human cell lines from the Cancer Cell Line Encyclopedia. M0-DCC gene expression profiles can be reversibly induced from M1-DCC-like cells in vitro . The close correspondence between gene expression profiles in immediate early embryonic development and metastatic founder cell candidates provides strong evidence that the onset of cancer and metastasis recruits mechanisms employed in fertilization.

Neighbourhood deprivation is one of the few potential policy-modifiable risk factors for psychiatric and neurological disorders, but the neurobiological pathways underlying these associations remain unclear. We investigated these relationships across three cohorts spanning the life span: the Healthy Brain and Child Development (HBCD) Study (n = 84, aged 0–4 weeks postnatal), the Adolescent Brain Cognitive Development (ABCD) Study (n = 4,792, aged 9–10 years), and the UK Biobank (UKB; ∼ 500,000 adults, aged 44–87 years). Neighbourhood deprivation was associated with elevated disease risk, and individual lifestyle factors accounted for only a small fraction of this burden, indicating that the much larger residual effect reflects broader contextual characteristics of deprived environments rather than individual behaviours alone. Across all cohorts, greater deprivation consistently predicted lower cortical and subcortical brain volume, with effects detectable in early development and substantially stronger in adulthood. Across disorders, regional brain volume emerged as a consistent neuroanatomical mediator linking neighbourhood deprivation to neuropsychiatric disease. We further showed that deprivation preferentially affects brain regions intrinsically vulnerable to neuropsychiatric disorders. Spatial decoding analyses implicated dopaminergic and serotonergic neurotransmitter systems together with specific excitatory and inhibitory neuronal classes. Importantly, both the deprivation–disease associations and their neuroanatomical mediation patterns replicated across independent populations. Our study delivers a translational framework linking neighbourhood deprivation to brain health which could inform public health policies and preventive interventions.

Hormone-producing cells in the pancreas and intestine regulate whole-body metabolism and are dysregulated in obesity and diabetes. A deeper understanding of endocrine lineage formation (endocrinogenesis) in these organs is essential to elucidate the healthy state and disease trajectories. Here, we performed cross-species (mouse and human) and cross-system (in vivo and in vitro) comparisons of pancreatic endocrine progenitors (EPs), complemented by cross-organ (pancreas and intestine) analyses, using single-cell transcriptomics and epigenomics, together with mouse bulk proteomics. We uncovered conserved and distinct gene regulatory networks (GRNs) and cell-cell communication patterns during lineage allocation across species, systems, and organs. Additionally, we identified diabetes-associated genes restricted to embryonic pancreatic EPs but largely absent in the adult pancreas, suggesting that dysregulation during development may predispose to diabetes. Furthermore, our multi-source EP profiling resolved the transcriptional programs underlying the aberrant, in vitro-enriched enterochromaffin cell formation during differentiation toward human islet cells, suggesting that these cells may not correspond to a naturally occurring cell type present in vivo development. Finally, we revealed conserved, dynamic shifts in the cell cycle and cytoskeletal organization, alongside divergent mRNA translation, during endocrinogenesis. Collectively, this multi-source EP profiling establishes a resource mapping the molecular landscape of endocrinogenesis across two major metabolic-endocrine organs.

Fission increases the number of crypts in the intestine during neonatal growth and also restores crypt density after injury by bifurcation of a pre-existing parent crypt into daughter crypts. While fission is typically symmetric in healthy crypts, it is more asymmetric in diseases, and the relationship between parent crypt shape and daughter crypt (a)symmetry is difficult to study as crypt budding and fission are stochastic in organoid models and difficult to control in vivo. Here, a photoresponsive hydrogel is introduced to spatiotemporally control daughter crypt emergence from mature parent crypts in intestinal organoids, enabling longitudinal tracking of crypt bifurcation in vitro. Variation of the photopatterned dimensions tunes parent crypt shape and reveals that both fission efficiency and crypt symmetry depend on parent crypt geometry. Epithelial boundary analysis identified parent crypt curvature as a key factor influencing daughter crypt symmetry. High-curvature or narrow crypts yielded symmetric daughter crypts, whereas wider parent crypts with lower epithelial curvature generated progressively more asymmetric crypts. Mechanistically, non-muscle myosin IIA acts as one key regulator of crypt symmetry. Overall, this work introduces a reproducible and spatiotemporally controllable in vitro model of crypt fission, allowing identification of mechanical determinants of fission that influence intestinal regeneration and development.

Huntington's disease (HD) is a neurological condition with limited treatment. Its hallmarks are progressive loss of neurons, chorea, cognitive, motor, and metabolic impairments. The current study uses 3-nitropropionic acid (3-NP) to cause HD-like indications in rats. L-theanine (L-TH), an active component of Camellia sinensis, has already been shown to possess anti-inflammatory and anti-oxidant effects. The impact of the isolated phytoconstituent, L-TH, on 3-NP-induced changes in three distinct brain regions was assessed in the current study. The animals received a 7-day pretreatment consisting of normal saline, extract of C. sinensis (100 and 200 mg/kg b.w., i.p.), L-TH (10 mg/kg b.w., i.p.), and standard Tetrabenazine (TBZ, 25 mg/kg, b.w., i.p.). The administration of 3-NP (10 mg/kg b.w., i.p.) to the treatment groups began on the 8th day of the protocol and continued until the 21st day. The brain homogenates underwent biochemical and neurochemical assessments. In conclusion, L-TH improved behavioral outcomes and attenuated 3-NP-induced neurochemical alterations in brain homogenates. Histopathological analysis revealed lower levels of tissue damage in the L-TH-treated group compared to the 3-NP group. Molecular docking studies further indicated favorable interactions of L-TH with the HTT protein. These findings suggest that L-TH may attenuate 3-NP-induced Huntington's disease-like alterations in rats. However, additional studies using genetic HD models and molecular mechanisms are required to confirm its therapeutic efficacy and underlying mode of action.

Spinocerebellar ataxia type 2 (SCA2) is a neurodegenerative disorder that shows cerebellar glucose hypometabolism, systemic hypermetabolism and weight loss. This study aimed to explore novel molecular mechanisms of disease for SCA2, based on the assessment of blood glucose homeostasis. A case-control and correlational study was conducted in 79 normoglycemic Cuban patients with SCA2 and 83 sex- and age-matched control subjects during a fasting state. A subset of 20 patients with SCA2 and 19 control individuals underwent an oral glucose tolerance test (OGTT). Several indices for assessing blood glucose homeostasis, derived from the fasting state or the OGTT, were included in the study. Fasting glucose levels showed a small increase, whereas the QUICKI index for insulin sensitivity was slightly decreased among patients. Markers of glucose homeostasis derived from the OGTT were no different between patients and controls. Fasting insulin levels, QUICKI, McAuley´s, HOMA2-%S, HOMA2-IR, and HOMA2-%β indices showed weak to moderate correlations with markers of body composition. McAuley´s, TyG, HOMA2-IR, and HOMA2-%β indices correlated with the age at onset, progression rate, or INAS count. Patients with SCA2 with mild-to-moderate ataxia do not have any major alteration in blood glucose homeostasis. Markers of glucose homeostasis associate with body composition and have nominal modifying effects on disease severity and progression rate in patients with SCA2, with McAuley's index showing effects more consistently. Further studies are needed to describe the changes in blood glucose homeostasis in the different stages of disease, and to delve into its pathophysiological relevance.

Excipients are widely used to suppress the self-association of therapeutic proteins, yet their mechanisms of action are not well understood and often assumed to be nonspecific. Here we show that excipient-mediated solubilization of therapeutic antibodies is markedly molecularly specific. Using a high-throughput combinatorial droplet microfluidic platform, we systematically quantify the effects of common pharmaceutical excipients across a diverse panel of monoclonal antibodies (mAbs). Although all studied excipients enhance solubility, their effects can vary significantly between antibodies, spanning dynamic ranges from approximately 7-fold to over 200-fold. Integrating experimental solubilization measurements with sequence- and structure-derived molecular descriptors, we identify interpretable physicochemical determinants underlying excipient responses; for example, the histidine effect is strongly dependent on mAb dipole moment. Our findings reveal trends that highlight the molecular specificity and complexity of antibody-excipient interactions, as well as the limitations of purely generic formulation rules. Overall, this study provides a quantitative framework for analyzing excipient effects across diverse antibodies and supports the development of predictive approaches for rational formulation design. The integration of high-throughput experimentation with molecular feature analysis offers a foundation for improving our understanding and prediction of antibody-specific formulation behavior.

<h4>Background</h4>HLA-B27 is strongly associated with a spectrum of immune-mediated diseases, including axial spondyloarthritis (axSpA), psoriatic arthritis (PsA), inflammatory bowel disease (IBD), and acute anterior uveitis (AAU). However, the shared molecular mechanisms and cross-disease diagnostic biomarkers remain poorly defined. This study aimed to identify common gene signatures and potential diagnostic biomarkers across HLA-B27-related diseases using bioinformatics analysis and experimental validation.<h4>Methods</h4>Gene expression datasets for axSpA, PsA, ulcerative colitis (UC), Crohn's disease (CD), and AAU were retrieved from the Gene Expression Omnibus (GEO) database. Disease-specific diagnostic genes were identified via differential expression analysis, least absolute shrinkage and selection operator (LASSO) regression, and receiver operating characteristic (ROC) curve analysis (AUC > 0.9). Shared biomarkers were determined by intersecting differentially expressed genes (DEGs) across all five diseases. Protein-protein interaction (PPI) networks, functional enrichment analyses, and single-sample gene set enrichment analysis (ssGSEA) were performed to explore biological pathways and immune infiltration patterns. Competing endogenous RNA (ceRNA)-transcription factor (TF) regulatory networks were constructed, and potential therapeutic agents were predicted using the Drug-Gene Interaction Database (DGIdb). Finally, the expression of candidate genes was validated by RT-qPCR and Western blot in peripheral blood mononuclear cells (PBMCs) from patients with ankylosing spondylitis (AS) and UC.<h4>Results</h4>A total of 2485, 380, 2682, 2232, and 481 DEGs were identified for axSpA, PsA, UC, CD, and AAU, respectively, with ABCD2 being the only shared biomarker across all five diseases. Disease-specific diagnostic gene panels were established (3 genes for axSpA, 6 for PsA, 15 for UC, 9 for CD, and 4 for AAU). Two major gene clusters were uncovered: an arthritis-related cluster (LAG3, IL15, PRF1, TBX21, IL2RB) and an extra-articular disease-related cluster (IL6, FN1, F2R, HIF1A, ANGPT2). Immune infiltration profiles varied significantly across diseases. Regulatory network analysis revealed complex ceRNA-TF interactions and identified several candidate drugs. In PBMCs, ABCD2 was upregulated in AS and UC, PRF1 in AS, and FN1, IL-6, and F2R in UC.<h4>Conclusion</h4>This study identifies ABCD2 as a potential cross-disease biomarker and two context-dependent key gene clusters for HLA-B27-associated immune-mediated diseases, providing exploratory candidates for future research.

Autophagy plays a critical role in maintaining cellular homeostasis and regulating stem cell fate. Although microRNAs (miRNAs) are being increasingly recognized for their ability to modulate autophagy and pluripotency in human pluripotent stem cells (hPSCs), the underlying molecular mechanisms remain incompletely understood. This study defines a role for miR-150 deficiency in biasing ectoderm-mesoderm lineage specification in hPSCs and identifies SOX2 as a key downstream node regulated primarily through altered mRNA stability, with additional involvement of an autophagy-associated pathway. miR-150-deficient hPSCs were generated by CRISPR/Cas9-mediated deletion of the miR-150 locus. A range of molecular and cellular techniques were used to investigate how miR-150 regulates SOX2 and KLHL20 expression, modulates the KLHL20-ULK1 interaction, and influences autophagy-mediated SOX2 degradation. Directed differentiation protocols were used to determine the role of miR-150 in germ layer lineage specification. The loss of miR-150 led to increased SOX2 expression by stabilizing its mRNA, resulting in elevated SOX2 protein levels in hPSCs. Concurrently, miR-150 deficiency increased KLHL20 mRNA stability, disrupted ULK1-mediated autophagy, and inhibited SOX2 degradation. Functional assays demonstrated that miR-150 fine-tunes SOX2 expression through dual regulation of mRNA stability and autophagy via the KLHL20-ULK1 axis, maintaining a balanced commitment to the mesodermal and ectodermal lineages. These findings establish miR-150 as a regulator of SOX2 activity and autophagy in hPSCs. By targeting both SOX2 and the KLHL20-ULK1 axis, miR-150 coordinates SOX2 turnover through complementary mechanisms, ensuring precise mesodermal and ectodermal lineage commitment. This multiregulatory strategy provides new insights into how miRNAs integrate intracellular signaling pathways to direct stem cell fate decisions.

Phycocyanobilin, a bioactive compound derived from Arthrospira platensis C1, was investigated for its potential role in systemic lupus erythematosus (SLE) based on its structural similarity to bilirubin, with a Tanimoto score of 93%. Molecular docking revealed favorable binding affinities between phycocyanobilin and several protein targets, including EGFR, FYN, HLA-B, LCK, LYN, and TP53. Target prediction further identified LYN kinase as a key candidate. Molecular dynamics simulations demonstrated stable binding of the phycocyanobilin-LYN complex, with interaction profiles comparable to those of the native ligand, staurosporine. Binding free energy and residue-level analyses supported strong and stable interactions, highlighting key contributions to complex stability. Overall, these findings provide mechanistic insight into the interaction between phycocyanobilin and LYN, suggesting that this compound may modulate LYN-associated signaling pathways and warrants further investigation in the context of SLE.

Neurons actively shape immune responses that maintain central nervous system integrity. We identify SPP1 (secreted phosphoprotein 1) as a neuron-derived signal that reprograms microglia into a neuroprotective, homeostatic state after injury and during neurodegeneration. In mouse models of glaucoma and optic nerve damage, neuronal SPP1 enhances microglial autophagy, debris clearance, and anti-inflammatory activity, preserving neuronal survival and visual function. SPP1 is elevated in neurons of human and primate glaucomatous retinas, where SPP1⁺ cells show increased resilience. In Alzheimer's disease brain, neuronal SPP1 correlates with neuronal survival, while microglia around Aβ plaques display defective autophagy. In human iPSC co-cultures, SPP1 enhances microglial Aβ clearance and prevents neurodegeneration. Thus, SPP1 defines a protective neuron-microglia axis in glaucoma and possibly other neurodegenerative diseases.

Endothelial dysfunction is recognized to contribute to chronic tissue remodeling in the lung, yet endothelial-derived mechanisms driving these processes remain largely undefined. Here, we show that endothelial inactivation of the LATS1 and LATS2 kinases, key suppressors of the transcriptional regulators YAP and TAZ, elicits distinct responses depending on endothelial identity within different pulmonary vascular beds. Our data indicate that LATS1/2 inactivation in general capillary endothelial cells induces a reactive capillary injury-like state, whereas a separate endothelial population located within veins adopts a distinct profibrotic state. We show that expansion of this reactive venous endothelial cell population, which is marked by EBF1 and ACKR1 expression, is associated with fibroblast activation, macrophage accumulation, and epithelial remodeling, collectively generating a microenvironment characteristic of fibrotic lung disease. We further demonstrate that pharmacologic inhibition of YAP/TAZ-TEAD signaling prevents stromal and immune remodeling and fibrotic lesion formation following endothelial LATS1/2 inactivation. These findings identify LATS1/2-mediated restraint of YAP/TAZ as essential for lung endothelial homeostasis and highlight a distinct venous cell response as a direct contributor to fibrotic lung remodeling.

Metastatic relapse can arise years after apparently successful treatment because disseminated cancer cells (DCCs) persist in reversible states of cellular, angiogenic, or immune-mediated dormancy. The strongest direct evidence that acute viral inflammation can disturb this equilibrium currently comes from experimental breast-cancer lung-dormancy models in which influenza A virus or SARS-CoV-2 promoted DCC cell-cycle re-entry and metastatic expansion, with interleukin-6 (IL-6) required for the initial awakening phenotype. Human findings reported in the same study are observational and do not establish that respiratory viral infection causes metastatic relapse. This hypothesis-generating Perspective therefore separates direct viral-dormancy evidence from direct inflammation-dormancy evidence, established metastatic-niche biology, mechanistic extrapolation, and unvalidated clinical hypotheses. We critically evaluate IL-6/JAK/STAT3 signaling, the time-dependent balance between antiviral and antitumor immunity, neutrophil extracellular trap-mediated matrix remodeling, epithelial RNA sensing, endothelial and perivascular responses, and selected extracellular-vesicle mechanisms. The evidence does not establish a universal infection-to-niche-to-relapse pathway across cancers or organs. Instead, it supports a context-dependent model in which respiratory infection may transiently perturb the DCC-niche equilibrium in susceptible settings. Priority studies should independently replicate the pulmonary phenotype, define causal timing, distinguish local lung injury from systemic effects, test cross-cancer and cross-organ generalizability, and prospectively evaluate human associations. No post-viral biomarker panel, intensified imaging strategy, or pathway-directed intervention is currently validated for routine clinical use.

Members of the Wiskott-Aldrich syndrome protein (WASP) family orchestrate cytoskeletal reorganization that modulates B-cell receptor (BCR) signaling and B-cell fate decisions. WHAMM, a WASP-family nucleation-promoting factor member that associates with actin, membranes and microtubules, has not been functionally characterized in B cells. To investigate the role of WHAMM in B-cell development and function, we analyzed a conditional mouse model in which Whamm was deleted in the B-cell lineage. WHAMM-deficient mice exhibited altered splenic B-cell composition, characterized by an accumulation of splenic transitional B cells and a reduction of follicular B cells. Meanwhile, WHAMM deficiency altered the spatial organization and kinetics of proximal BCR signaling, and modified the dynamics of BCR-induced actin remodeling. However, WHAMM-deficient B cells showed largely preserved BCR internalization, antigen presentation, PI3K-AKT-mTOR signaling, ROS production, and mitochondrial membrane potential. Together, this work indicates that WHAMM helps shape splenic B-cell populations and regulates proximal BCR signaling and actin dynamics, while several downstream functional responses remain largely preserved in this study.

Respiratory syncytial virus (RSV) causes acute lower respiratory tract infection in young children, the elderly, and immunocompromised hosts, and no RSV vaccine or therapy exists for infants, beyond prophylaxis. PIWI-interacting RNAs (piRNAs), small non-coding RNAs acting through P element-induced wimpy testis (Piwi) proteins, remain largely unexplored. We characterized PIWIL4 expressions and function during RSV infection or poly(I:C) stimulation, a proxy of viral infection. PIWIL4 was expressed in primary and immortalized small airway epithelial cells following RSV infection. While PIWIL4 knockdown did not affect RSV replication, it decreased cytokine and growth factor secretion, altered gene and piRNA expression, and modulated pathways involved in cytokine production, metabolism, and airway remodeling. Limited overlap between piRNA targets and transcriptomic changes suggests PIWIL4 regulates airway epithelial cell responses partly independently of piRNAs. Our study positions PIWIL4 as a key regulator of airway epithelial cell responses to viral infections.

<h4>Background</h4>METTL5, an N6-methyladenosine (m6A) RNA methyltransferase, has been implicated in tumor progression, but its prognostic value and non-invasive prediction in lung adenocarcinoma (LUAD) remain unclear. This study aimed to develop a pathomics-based machine learning model to predict METTL5 expression from histopathological images and evaluate its prognostic significance in LUAD.<h4>Methods</h4>A total of 327 LUAD patients from The Cancer Genome Atlas (TCGA) with matched hematoxylin and eosin (H&E) slides, transcriptomic, and clinical data were included and randomly divided into training and validation sets (7:3). Quantitative histopathological features were extracted using PyRadiomics. Feature selection was performed via maximum relevance minimum redundancy (mRMR) and recursive feature elimination (RFE), followed by construction of a Gradient Boosting Machine (GBM) model. A pathomics score (PS) was generated to assess prognostic relevance. Survival analyses, gene set variation analysis (GSVA), tumor mutational burden (TMB), immune infiltration analysis, and in vitro functional assays were conducted.<h4>Results</h4>METTL5 overexpression was independently associated with poor overall survival [hazard ratio (HR) =1.637, P=0.007]. The model achieved good predictive performance [area under the curve (AUC) =0.847 in the training set and 0.752 in the validation set]. High PS was significantly associated with worse survival and remained an independent prognostic factor (HR =1.563, P=0.03). Elevated PS correlated with altered metabolic pathways, increased TMB, and immune microenvironment changes. METTL5 knockdown reduced proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) in A549 cells.<h4>Conclusions</h4>The pathomics-based model accurately predicts METTL5 expression and provides prognostic stratification in LUAD, supporting its potential as a practical imaging-derived biomarker.

γδ T cells have raised interest as effector cells in cancer immunotherapy, due to their broad and HLA-independent reactivity towards multiple tumor entities. Despite promising ongoing clinical studies, there is a need to improve the effector function and thereby the efficacy of γδ T cells. In addition to its role as anti-oxidant, Vitamin C is an epigenetic modifier and exerts multiple effects on T-cell differentiation. While Vitamin C enhances <i>in vitro</i> proliferation, cytokine production and cytotoxicity of γδ T cells, the associated signaling pathways have so far not been elucidated. Here we demonstrate that Vitamin C enhances the cytotoxic effector function of unmodified as well as anti-CD19 chimeric antigen receptor (CAR) and TCR fusion construct (εTRuC)-transduced zoledronate-expanded γδ T cells. This was associated with increased upregulation of CD25 and IFN-γ secretion. Vitamin C increased calcium influx and nuclear translocation of NFAT and NF-κB in short-term expanded γδ T-cell lines. Although Vitamin C did not alter CD3ζ phosphorylation following stimulation with anti-CD3 antibodies, it reduced phosphorylation of ERK1/2 without affecting p38 MAP kinases. Further analysis showed that Vitamin C did not modulate the expression of selected immune checkpoint molecules on γδ T cells. Our results identify important signaling events where Vitamin C enhances the effector activity of human γδ T cells.

Mammalian tissue function depends on the balance between proliferative, mitotic cell populations and irreplaceable post-mitotic cells. Throughout life, aging progressively disrupts cell type balance and thus tissue function through opposing forces on different cell types. Aging promotes the aberrant proliferation or entry into pro-inflammatory senescent states of mitotic cells, while contributing to the functional erosion and, in vulnerable populations, loss of post-mitotic irreplaceable cells. The divergent age-associated vulnerabilities of mitotic and post-mitotic cells caused by aging generate a case of <i>homeostatic conflict</i>, in which tissue maintenance simultaneously demands the elimination of damaged mitotic cells and the preservation of irreplaceable ones. We examine the concept of <i>homeostatic conflict</i> using the aging brain as an example, and illustrate how the senescence-prone glia expand and acquire SASP-rich transcriptional states, while post-mitotic neurons undergo widespread loss of molecular identity and cell death, resulting in cell type imbalance and consequently tissue dysfunction. Based on the observed <i>homeostatic conflict,</i> we then devise potential context-sensitive therapeutic strategies by combining interventions that protect from ferroptosis to selectively protect irreplaceable cells with interventions that eliminate maladaptive ones to resolve the age-associated cell type imbalance and thus the underlying <i>homeostatic conflict</i>. We present <i>homeostatic conflict</i> as a concept that goes beyond the reductionist view in which numerous isolated processes promote aging in isolation, and provides a logical framework that integrates multiple processes and explains how their interaction drives age-associated dysfunction in a manner instructive to the development of pharmacological combination strategies.

Depressive disorders (DD) are highly heterogeneous psychiatric illnesses. Their complex pathogenesis, coupled with suboptimal response rates to current clinical treatments, poses significant challenges to global mental health. High-throughput RNA sequencing (RNA-seq) has emerged as a powerful tool for dissecting the molecular mechanisms of DD at the transcriptomic level. This paper reviews recent advances utilizing bulk RNA-seq, single-cell RNA-seq (scRNA-seq), and single-nucleus RNA-seq (snRNA-seq) to explore the pathogenesis of major depressive disorder (MDD), perinatal depression (PND), perimenopausal depression (PMD), and post-stroke depression (PSD). Current evidence indicates that immune-inflammatory activation, impaired synaptic plasticity, and mitochondrial energy metabolism dysfunction constitute shared core pathological mechanisms across these disorders, validating classical pathogenic hypotheses at the molecular level. Furthermore, distinct clinical subtypes exhibit significant transcriptomic heterogeneity; for instance, MDD shows marked sexual dimorphism and immunophenotypic characteristics. Specific molecular pathways also distinguish different DD categories: PND and PMD mainly involve imbalances in neuroendocrine-immune interaction networks driven by hormonal fluctuations, whereas PSD is characterized by the interaction between innate immune responses and autophagy flux impairment. Although RNA-seq has significantly advanced our understanding of DD, challenges such as restricted sample sources, high data heterogeneity, and insufficient clinical translation remain. Future research should prioritize single-cell and spatial transcriptomics to resolve tissue heterogeneity, promote multi-omics integration to construct causal regulatory networks, and strengthen cross-species comparative studies. These efforts will accelerate the development of precision diagnosis and personalized treatment strategies.

<h4>Background</h4>The Ordos fine-wool sheep is a high-quality fine-wool breed in China, renowned for its excellent wool quality, meat production, and adaptability to the arid and semi-arid regions of Inner Mongolia. Body weight and wool traits are important economic characteristics in sheep breeding. This study aimed to identify genetic loci associated with body weight (BW), wool length (WL), and wool fineness (WF) in Ordos fine-wool sheep.<h4>Methods</h4>A genome-wide association study (GWAS) was conducted in 388 Ordos fine-wool sheep genotyped using the GenoBaits^® Ovine 40K SNP panel. Single nucleotide polymorphisms (SNPs) associated with BW, WL, and WF were identified, and candidate genes located near the SNPs reaching the suggestive threshold were subjected to functional annotation and enrichment analysis.<h4>Results</h4>A total of 22 SNPs were identified as potentially associated with BW, WL, and WF traits, corresponding to 27 annotated genes. Functional annotation highlighted six potential candidate genes, including LAMA2, ARHGAP18, IGFBP2, IGFBP5, CA10, and AXIN1, which may play important roles in regulating body weight and wool growth in sheep.<h4>Conclusions</h4>The identified genes provide valuable candidate loci for BW, WL, and WF traits in Ordos fine-wool sheep. The results of this study provide preliminary references for further exploration of the genetic mechanisms of wool traits in Ordos fine-wool sheep and the development of molecular breeding markers.

The H295R assay (OECD TG456) is used to screen chemicals for interference with steroidogenesis, measuring testosterone and estradiol levels as endpoints. Focusing on these two hormones, however, may overlook informative changes in other steroid classes, particularly progestogens and glucocorticoids. Here, we extended the effect-based H295R-CALUX® read-out to include progestogens and glucocorticoids using PR- and GR CALUX® assays. First, we evaluated whether PR- and GR-CALUX® could be used as H295R read-outs by determining whether control samples produced adequate hormone levels and whether responses to control compounds forskolin and prochloraz met predefined criteria. Progestogenic and glucocorticoid activity was consistently quantifiable, forskolin induced both modalities above the predefined threshold, and prochloraz inhibited glucocorticoid activity while increasing progestogenic activity, consistent with upstream pathway inhibition. The extended H295R-CALUX® read-out was then applied to analyse a set of conazoles. Each conazole was first pre-screened for ERα/AR/PR/GR agonism or antagonism and for cytotoxicity to avoid interference from direct receptor activation and cytotoxicity in the H295R read-out. In the H295R assay, all conazoles except 1,2,4-triazole, affected at least one hormone modality, with a common pattern of reduced androgenic, estrogenic and glucocorticoid activity and compound-specific increases or decreases in progestogenic activity. Further, targeted LC-MS/MS steroid profiling mirrored the effect-based results obtained with the H295R-CALUX® read-out and supported the hypothesis of CYP17A1-related perturbations for some conazoles. Overall, the addition of progestogen and glucocorticoid read-outs, alongside androgen and estrogens, enhances the resolution of the H295R assay, hence improving its relevance for endocrine safety assessment.

<title>Abstract</title> <p>Schizophrenia (SCZ) is a highly polygenic disorder and is associated with several comorbidities, including depression, cardiovascular disease, and diabetes; yet the biological pathways underlying this clustering remain poorly understood. This study examined associations between a polygenic risk score for SCZ (PRS-SCZ) and circulating protein levels in 44,661 participants of the UK Biobank. We assessed 1,459 proteins (with ≤ 10% missingness) spanning neurological, inflammatory, cardiometabolic, and oncology panels. After adjustment for technical (population structure, genotyping chip), individual (age, sex, lifestyle), and clinical (comorbidities and medication) variables and multiple testing correction, the standardised PRS-SCZ was significantly negatively associated with C2, RNASET2, BTN2A1, CDSN, HLAE, LTA, MICA-MICB, CNTN3, FCER2, INHBC and CPVL concentrations, and positively with ICAM3, BTN3A2, IL5RA, SLAM7 concentrations. These protein biomarkers may provide insights into the biological pathways underlying schizophrenia and/or its comorbidities.</p>

<title>Abstract</title> <p> <bold>Introduction:</bold> Gestational adaptations require central autonomic recalibration via the hypothalamic paraventricular nucleus (PVN), yet how chronic hypertension alters its transcriptomic response during pregnancy remains unknown. We investigated PVN transcriptomic and cardiovascular autonomic profiles in pregnant normotensive (Wistar) and spontaneously hypertensive rats (SHRs) to uncover mechanisms underlying gestational hypothalamic adaptations to hypertension. <bold>Methods</bold> : Radiotelemetry-equipped Wistar rats and SHRs were evaluated for blood pressure, short-term variability, and baroreflex sensitivity alongside PVN bulk RNA-sequencing across virgin and late-pregnant states. <bold>Results</bold> : Both strains exhibited physiological gestational blood pressure drops. However, pregnant SHRs displayed blunted chronotropic reactivity, elevated cardiac sympathetic drive (increased LF HR and LF/HF ratio), and a doubling of spontaneous baroreflex sensitivity. Transcriptomically, pregnant Wistars showed a minimal homeostatic response (33 DEGs: e.g. <italic>Cish, Igfbp3, Irf7</italic> ). In stark contrast, pregnant SHRs underwent massive transcriptomic reprogramming (951 DEGs), marked by loss of astrocytic potassium buffering ( <italic>Kcnj10</italic> , <italic>Kcnb1</italic> ), chaperone depletion ( <italic>Hspa1a</italic> , <italic>Hspa1b</italic> ), excitatory neuropeptide elevation ( <italic>Hcrt</italic> , <italic>Trh</italic> ), and neuroimmune activation ( <italic>Zap70</italic> , <italic>Fcrl2, Itgam, Cmklr1</italic> ). Crucially, intersecting baseline hypertensive DEGs with gestational shifts in SHRs unmasked a significant inverse correlation, where 95.9% of hypertension-altered genes (140/146) underwent active directional reversal during pregnancy ( <italic>Hspa1a, Hspa1b, Ptgds, Per1</italic> ), enriching for translational, ribosomal, and bioenergetic pathways. <bold>Conclusion:</bold> Pre-existing chronic hypertension converts gestational PVN adaptation from a subtle homeostatic adjustment into an extensive genomic requirement (951 vs 33 DEGs). This massive transcriptomic remodeling represents an active counter-regulatory program in SHRs to sustain pregnancy to term, mitigating autonomic risk via heightened baroreflex sensitivity </p>

Long considered transcriptional noise, noncoding RNAs (ncRNAs), including microRNAs and long noncoding RNAs (lncRNAs), are now recognized as central regulators of cellular function, acting as scaffolds, structural elements, and regulators of gene expression. This expanding functional landscape is reshaping our understanding of cancer biology, immune regulation, and the limits of translation itself. Beyond gene regulation, lncRNAs may be implicated in the earliest stages of oncogenic transformation, orchestrating molecular reprogramming and remodeling of the immune microenvironment in premalignant lesions. Their functional analysis has further exposed an unexpected translational dimension with some lncRNAs harboring open reading frames encoding micropeptides. These cryptic micropeptides constitute an underexplored dimension of the immunopeptidome that could shape T-cell development and antitumor immunity, with implications for cancer immunosurveillance and therapeutic targeting. Collectively, these findings call for a revised molecular dogma in which the noncoding genome is recognized as a major regulator of cellular function, oncogenic transformation, and immune surveillance.

Osteoclastogenesis is driven by tightly coordinated transcriptional programs downstream of receptor activator of nuclear factor-κB ligand (RANKL) signaling, in which reactive oxygen species (ROS) function as essential secondary messengers. Although extracellular peroxiredoxin 1 (PRDX1) has been implicated in the suppression of osteoclast differentiation, the role of intracellular PRDX1 in regulating osteoclastogenic signaling remains poorly understood. Here, we identify intracellular PRDX1 as a redox-sensitive negative regulator of osteoclastogenesis and bone resorption. PRDX1 deficiency markedly enhanced osteoclast differentiation, bone resorption activity, and osteoporotic phenotypes in vivo. Transcriptomic and mechanistic analyses revealed that PRDX1 attenuates osteoclastogenic gene transcription by blocking the nuclear factor kappa B (NF-κB)/p65 signaling axis. Loss of PRDX1 increased intracellular ROS accumulation, promoted p65 nuclear translocation and promoter occupancy, thereby amplifying osteoclastogenic transcriptional programs. Mechanistically, PRDX1 underwent TNF receptor associated factor 6 (TRAF6)-mediated Lys67-dependent ubiquitylation and lysosomal degradation in response to RANKL signaling. In addition, RANKL-induced Src activation promoted phosphorylation of PRDX1 at Tyr194, which enhanced TRAF6-mediated ubiquitylation without substantially altering overall TRAF6 binding. Together, these findings reveal a PRDX1-ROS feedback loop that modulates NF-κB activity during osteoclastogenesis and identify regulated PRDX1 degradation as a mechanism by which RANKL signaling amplifies osteoclastogenic responses. These findings establish intracellular PRDX1 as an important modulator of skeletal homeostasis, suggesting its potential relevance as a pharmacological target in diseases characterized by excessive bone resorption.

Intervertebral disc (IVD) degeneration, a leading cause of chronic lower back pain, is associated with loss of vacuolated notochordal cells (NCs) and fibrotic remodeling of the nucleus pulposus. Emerging therapies increasingly focus on NCs, which are rare but therapeutically relevant cells for regenerating degenerated IVDs. In this study, we used CRISPR-based transactivation (CRISPRa) to direct the differentiation of human induced pluripotent stem cells (iPSCs) into the NC lineage. We tested CRISPRa-mediated activation of NOTO, TBXT, FOXA2, SOX5, SOX6, and SOX9, coupled with single-cell sequencing of Aggrecan-2A-mScarlet reporter iPSCs. This approach identified the SOX5/6/9 combination (SOX-trio) as critical for promoting NC lineage commitment. The SOX-trio yielded the largest cell population expressing a range of genes previously associated with NC identity, including SHH, FOXA1, FOXA2, FOXJ1, FN1, ALCAM, KRT8, and KRT18. Our study demonstrates the integration of CRISPRa with single-cell technologies as a powerful platform for investigating and enriching iPSC-derived NCs, supporting future regenerative strategies across various fields.

<h4>Objective</h4>This study aimed to establish and verify reference intervals (RIs) for coagulation factor XII (FXII) and antithrombin III (ATIII) in an adult population from the Jiangsu region of China and to evaluate the influence of analytical platform on RI interpretation.<h4>Methods</h4>We retrospectively analyzed data from 301 apparently healthy adults attending the Health Examination Center of Nanjing Drum Tower Hospital. Plasma FXII and ATIII activities were measured in parallel using two automated coagulation analyzers, the Sysmex CS-5100 and CS-6500. Statistical analysis followed CLSI EP28-A3 guidelines. Distribution normality was assessed to determine the appropriate method for RI establishment: the non-parametric percentile method for non-normally distributed data and the parametric method for normally distributed data. Analyzer-related agreement was further assessed using Bland-Altman analysis. The derived RIs were verified using an independent sample set of 20 apparently healthy adults.<h4>Results</h4>FXII showed a non-Gaussian distribution, whereas ATIII showed an approximately Gaussian distribution. For the CS-5100 analyzer, the RI was 32.2%-109.1% for FXII and 84.1%-104.7% for ATIII. For the CS-6500 analyzer, the corresponding RIs were 37.1%-111.1% for FXII and 86.1%-106.7% for ATIII. The newly established RIs differed from the manufacturer-provided ranges currently used in routine practice. No clinically meaningful sex-based partitioning was identified. Although analyzer-related differences were statistically significant, Bland-Altman analysis showed modest mean biases between the CS-6500 and CS-5100 platforms, with mean differences of +5.09% for FXII and + 1.17% for ATIII. All verification samples fell within the proposed RIs.<h4>Conclusion</h4>This study established and verified regional RIs for FXII and ATIII in healthy adults from Jiangsu, China. Although analyzer-related differences between the CS-5100 and CS-6500 platforms were modest, the findings highlight the importance of local RI verification when applying manufacturer-derived intervals or changing analytical platforms. These locally derived RIs may improve the interpretation of FXII and ATIII results and reduce unnecessary follow-up testing caused by inappropriate generic intervals.

We describe a 1-year-2-month-old boy with <i>KMT2A</i>-rearranged leukemia presenting with concurrent acute myeloid leukemia in bone marrow and B-lymphoblastic leukemia/lymphoma in a right calf mass. Both lesions harbored an identical <i>KMT2A::MLLT10</i> fusion, confirming common clonal origin. This represents the first documented case of spontaneous "spatial lineage divergence" at diagnosis, distinct from therapy-induced lineage switch. The finding highlights the potential need for multi-site biopsy and molecular profiling in extramedullary disease.

The emergence of significant viral pathogens and their escalating threats to human and animal health have intensified scientific interest in bats as disease vectors and natural reservoir hosts. Among chiropteran species, the Egyptian rousette (Rousettus aegyptiacus) assumes particular epidemiological significance as the established natural reservoir for orthomarburgviruses, encompassing the two recognized members Marburg virus and Ravn virus. While contemporary research predominantly emphasizes the detection and characterization of specific zoonotic agents within these populations, fundamental knowledge regarding their baseline pathology and the comprehensive impacts of disease on morbidity remains limited. Pathological investigations of free-ranging bat populations face inherent methodological constraints, frequently hampered by insufficient sample sizes or selection bias toward compromised specimens (e.g., moribund or deceased animals). This study addresses these limitations through comprehensive post-mortem examination and histologic evaluation of 69 free-ranging Egyptian rousettes from Uganda. These specimens were procured during systematic efforts to identify filoviral reservoir hosts, thereby providing an unprecedented opportunity to investigate spontaneous disease processes within an ostensibly healthy, representative population. Gross pathological examination identified lesions in 85.5% (59/69) of specimens, including scarring and fibrosis of the plagiopatagium and dactylopatagium (85.5%; 59/69), focal fibrotic nodules (17.4%; 12/69), and lacerations (17.4%; 12/69) within wing membranes. Histopathological analysis demonstrated the universal presence of microscopic alterations (100%; 69/69), with a cumulative total of 794 discrete lesions documented across 28 organ systems. The liver emerged as the most frequently affected organ, along with multiorgan perivascular cellular aggregates (both 81.2%; 56/69). This investigation represents the first comprehensive characterization of naturally occurring pathological processes in free-ranging Egyptian rousettes from Uganda. Elucidating the intrinsic factors that modulate reservoir species health is fundamental to understanding zoonotic disease spillover dynamics and developing evidence-based strategies for mitigating future outbreak risks in human and susceptible animal populations.

<h4>Background</h4>Dysregulation of multivesicular bodies (MVBs) in Alzheimer's disease (AD) contributes to aberrant tau secretion via extracellular vesicles (EVs). This may potentially explain our previous paradoxical observation of elevated free-form p-tau217 alongside reduced p-tau217⁺ EVs in plasma. This study aimed to investigate the mechanisms underlying the reduction of p-tau217⁺ EVs to uncover AD therapeutic targets.<h4>Methods</h4>By integrating hippocampal spatial transcriptomics of human brain with EV proteomics of cerebrospinal fluid, we identified key regulators of p-tau217⁺ EV release. Subsequently, we investigated the mechanisms underlying the synthesis and secretion of p-tau217⁺ EVs. The regulatory roles of these candidate proteins were systematically evaluated through shRNA knockdown and interference with a synthetic peptide in both Aβ₄₂-treated cells and AD model mice.<h4>Results</h4>Heat shock protein family A member 8 (HSPA8) was identified as a crucial regulator of EV biogenesis and release, mediating the Aβ-SNAP29 interaction to disrupt SNARE complex assembly and impair p-tau217⁺ EV secretion. In AD models, HSPA8 inhibition with shRNA rescued p-tau217⁺ EVs and improved cognitive function. Additionally, blocking the Aβ-SNAP29 interaction with a selective peptide inhibitor for HSPA8 reversed the decline in p-tau217⁺ EV and cognitive deficits.<h4>Conclusions</h4>These findings reveal a role of HSPA8 in regulating the MVB-mediated EV release and tau propagation, and highlight HSPA8 as a promising therapeutic target for modifying AD progression.

Heart and kidney diseases frequently coexist, but the genetic basis of this relationship remains unclear. We analyzed genetic data from large-scale studies to investigate how kidney function (estimated glomerular filtration rate, eGFR) and six common cardiovascular diseases share genetic risk factors. Using MiXeR method, and conjunctional false discovery rate (conjFDR) to identify overlapping genetic regions, we found 478 shared genomic loci between eGFR and cardiovascular diseases. These shared genes are involved in tissue development and structure. We also identified 29 genes that could be targeted by existing medications approved by the US Food and Drug Administration, such as PRKAG2, PDE1A, and IGF1R. Among these, genetically predicted higher level of IGF1R expression is associated with a higher eGFR, which reflects good kidney function and is protective against cardiorenal diseases, such as atrial fibrillation, and myocardial infarction. These findings reveal genetic overlap between kidney function and cardiovascular diseases, highlighting potential targets for understanding and treating cardiorenal syndrome.

<h4>Background</h4>Breast cancer remains a leading cause of female mortality worldwide, with therapeutic benefit limited by tumor heterogeneity and drug resistance. Identification of novel therapeutic targets through integration of genetic causality inference and functional validation is urgently needed.<h4>Methods</h4>Plasma protein quantitative trait loci (pQTL) from the Fenland cohort (~10,700 individuals) and the FinnGen R10 SomaScan subset (n = 828) were integrated with breast cancer genome-wide association study data from the Breast Cancer Association Consortium (122,977 cases and 105,974 controls). Causal protein-disease relationships were inferred using summary-data-based Mendelian randomization (SMR), with colocalization and HEIDI tests. Multi-level validation was performed using TCGA-BRCA transcriptomic data. Functional validation included CCK-8, EdU, wound healing, Transwell invasion, and flow cytometry apoptosis analysis evaluated <i>CPNE1</i> knocdown and/or overexpression of <i>the MST1 gene</i> which encodes macrophage-simulating protein (MSP).<h4>Results</h4>SMR identified 23 proteins in Fenland and 10 in FinnGen at FDR < 0.05. MST1/MSP and CPNE1 were supported in both datasets with PP.H4 ≥ 0.80 and non-significant HEIDI tests. Genetically predicted circulating MSP was positively associated with breast cancer risk, whereas circulating CPNE1 showed an inverse association. TCGA-BRCA showed higher <i>CPNE1</i> mRNA in tumors (P = 2.57×10⁻²⁵) and lower <i>MST1</i> mRNA (P = 3.04×10⁻²³). <i>CPNE1</i> expression was highest in triple-negative breast cancer and correlated positively with clinical stage (ρ = 0.211), whereas <i>MST1</i> expression was lowest in triple-negative breast cancer and correlated inversely with stage (ρ = -0.164). In T47D and MDA-MB-231 cells, <i>CPNE1</i> knockdown and <i>MST1</i> overexpression each reduced proliferation, migration, and invasion and increased apoptosis; the combined group showed greater changes than either single intervention.<h4>Conclusion</h4>This study prioritizes <i>the MST1 gene</i>, which encodes MSP, and <i>CPNE1</i> as candidate proteins for further investigation in breast cancer. However, the circulating-protein associations, tumor-mRNA patterns, and cell-autonomous perturbations represent distinct and directionally discordant biological contexts. The findings therefore support context-dependent candidate roles and justify mechanistic, in vivo, and formal interaction studies, but do not yet establish therapeutic efficacy or synergy.

CFTR modulators like elexacaftor-tezacaftor-ivacaftor (ETI) improve morbidity in cystic fibrosis (CF), but their systemic effects on young children are not yet clear. We hypothesize that ETI has anti-inflammatory effects and reverse disease-associated molecular signatures in children with CF. This exploratory pilot study evaluates the cellular mechanisms of ETI on peripheral immune cells in children (<12 years) with CF using a multi-omics approach. Seventeen children with CF (median age 8.7 years, 26% female) and 12 controls (median age 9.6 years, 42% female) were included for blood RNA-sequencing, proteomics and phosphoproteomics. Baseline pathway enrichment analysis revealed systemic inflammation, transmembrane transporter deficiencies, reduced protein kinase, and GTPase activity. Three months post-ETI, anti-inflammatory effects, epigenetic modulation, and increased protein kinase activity were observed, indicating partial reversal of cellular abnormalities. ETI modified systemic inflammatory, epigenetic, and phosphorylation pathways in young children with CF, offering insights into CF pathology and potential biomarkers for treatment monitoring.

Identification of gene expression changes in postmortem brain tissue of Alzheimer's disease donors compared to controls has implicated numerous biological pathways for Alzheimer's disease pathophysiology. Nonetheless, there is still limited understanding of how gene expression dysregulation underpins specific proteinopathies core to Alzheimer's disease. Here, we investigate brain transcriptomic changes in a well-characterized cohort of Alzheimer's disease donors to identify genes and networks that associate with Alzheimer's disease endophenotypes, including neuropathology measures (Braak stage, Thal phase and cerebral amyloid angiopathy score) and Alzheimer's disease-related brain protein levels (Apolipoprotein E, Amyloid-β 40, Amyloid-β 42, tau and phospho-Tau). Bulk transcriptome measures were collected from the temporal cortex tissue of 477 Alzheimer's disease donors. Following quality control, transcriptome-wide association studies were performed for each endophenotype. We used weighted gene co-expression network analysis to build co-expression networks and integrated transcriptome with epigenetic and genetic data from the same donors. We detected a total of 5740 Bonferroni-significant temporal cortex gene associations with Alzheimer's disease endophenotypes, most of which were with brain tau levels. We discovered tau-associated co-expression modules enriched in known and novel Alzheimer's disease pathways. We found that a <i>beneficial (or neutral)</i> brain biochemical state of higher total tau and lower phospho-Tau is associated with increased levels of synaptic, DNA damage/repair, nucleic acid metabolism and myelin processes. In contrast, in a detrimental state with lower total and higher phospho-Tau, there is upregulation of vascular and immune, and downregulation of mitochondrial and myelin pathways. There are brain gene expression perturbations that are associated with Alzheimer's disease endophenotypes. While some of these associations are common across multiple endophenotypes, many are distinct for different Alzheimer's disease-related proteins. Based on these findings, we propose a hypothetical model of <i>dynamic brain gene expression changes</i> that track with progressive Alzheimer's disease proteostasis. These expression changes hold potential to serve as dynamic, precision biomarkers of brain Alzheimer's disease progression. This study demonstrates the potential of integrative multi-omics and deep Alzheimer's disease endophenotypes in well-characterized brain tissues to precisely uncover the complex biology of Alzheimer's disease.

<h4>Objective</h4>Depression is a heterogeneous psychiatric disorder and a growing public health concern, characterized by its high prevalence, recurrence rate, and association with suicide. There is evidence suggesting that both genetic susceptibility and environmental factors can regulate gene expression through DNA methylation, thereby influencing the occurrence and development of depression. The olfactory sensory neuropeptide 1 (<i>OLFM1</i>) protein is a risk factor for mental disorders. However, there are no reports yet regarding the correlation between the <i>OLFM1</i> gene and depression, nor have there been any studies on the association between <i>OLFM1</i> gene DNA methylation and depression.<h4>Methods</h4>Genomic DNA was extracted from peripheral blood samples of patients with depression (<i>n</i> = 100) and healthy controls (<i>n</i> = 100) using the QIAamp DNA Blood Mini Kit. Subsequently, the extracted genomic DNA was subjected to bisulfite treatment using the EZ DNA Methylation-Gold™ kit. DNA methylation levels of 107 CpG sites in six fragments of <i>OLFM1</i> exon 1 and its downstream were detected by the Illumina HiSeq platform using MethylTarget™ technology.<h4>Results</h4>Methylation levels across the overall <i>OLFM1</i> CpG island and its six fragments (<i>OLFM1</i>-1 to <i>OLFM1</i>-6) were significantly reduced in the depression group relative to controls. Analysis of the <i>OLFM1</i> gene fragments revealed that 84 of 107 CpG sites were significantly hypomethylated in depressed individuals. When patients were divided by sex, male patients displayed hypomethylation at 65 CpG sites, substantially more than the 37 sites found in females.<h4>Conclusion</h4><i>OLFM1</i> hypomethylation is associated with depression and may serve as a potential epigenetic biomarker.

<h4>Objective</h4>To identify plasma protein biomarkers associated with incident non-fatal major adverse cardiovascular events (MACE) in diabetic kidney disease (DKD) patients.<h4>Research design and methods</h4>We analyzed 317 DKD patients from the UK Biobank. Plasma proteomics and clinical data (demographics, metabolism, renal function) were integrated. In an exploratory discovery phase, three sequential Cox regression models (crude, socio-demographic-adjusted, socio-demographic-metabolic adjusted) screened non-fatal MACE-associated proteins. To prevent information leakage, the cohort was then randomly split into training (70%) and testing (30%) sets; machine-learning feature selection, hyperparameter optimization, and final model development were performed exclusively within the training set. The associated proteins were input into the four-step machine-learning pipeline (LASSO-Cox, random survival forest, Boruta, XGBoost-Cox). Predictive performance was validated using Kaplan-Meier survival analyses, longitudinal trajectory modeling, and ROC benchmarking. An interactive web application was deployed for clinical implementation.<h4>Results</h4>Of 1,463 plasma proteins, 561 were associated with non-fatal MACE across Cox models, with 14 overlapping proteins. Nine core proteins (ANG, IL1R1, CXCL14, ESAM, PTGDS, HAVCR1, FGFR2, IGSF8, CCL3) were validated: ANG showed the strongest non-fatal MACE association (HR = 3.88, 95%CI 2.33-6.48, p<0.001), and all high-expression groups had elevated non-fatal MACE risk. GO/KEGG enrichment highlighted inflammatory-immune pathways like positive regulation of MAPK cascade, Cytokine-cytokine receptor interaction and PI3K-Akt signaling pathway as key mechanisms. The model integrating proteins, demographic factors, and clinical variables achieved the highest predictive performance across non-fatal MACE (AUC = 0.768), myocardial infarction (MI) (0.808), and stroke (0.816) outcomes, with superior stability in cross-validation. CoxBoost + Elastic Net framework was selected as the optimal framework via benchmarking of 101 algorithms. The model demonstrated favorable calibration in high-risk patients and yielded positive net clinical benefit across decision thresholds of 5% to 45%. The web tool (https://jiangli2941.github.io/MACE-prediction-v2/) enables input of 28 variables, outputs non-fatal MACE risk status, risk probability, and highlights abnormal indicators.<h4>Conclusion</h4>Plasma proteomics combined with machine learning identifies robust non-fatal MACE predictors in DKD.

<h4>Background</h4>Lung cancer remains a leading cause of cancer incidence and mortality globally. Metabolic reprogramming promotes tumor progression and shapes an immunosuppressive tumor microenvironment. Galactose metabolism is involved in multiple malignancies, but its prognostic value in lung adenocarcinoma (LUAD) remains unclear. This study aimed to develop and internally validate a galactose metabolism-related multigene prognostic model for LUAD.<h4>Methods</h4>A retrospective prognostic model development and internal validation study was performed using RNA sequencing (RNA-seq) and clinical data from 585 LUAD patients in The Cancer Genome Atlas (TCGA). Differential expression, functional enrichment, univariate and multivariate Cox regression were applied to construct a prognostic gene signature. Internal validation was performed using bootstrap resampling. Model performance was evaluated by time-dependent receiver operating characteristic (ROC), C-index, calibration, and Kaplan-Meier analysis. Associations between the model and immune infiltration, immunotherapy responsiveness, and tumor stemness were also analyzed.<h4>Results</h4>A six-gene prognostic model (<i>GALT, GANC, PGM1, GALM, B4GALT1, PGM2</i>) was developed. The model showed good discrimination with 1-, 3-, and 5-year area under the curve (AUC) values of 0.719, 0.693, and 0.684, respectively. The low-risk group exhibited significantly longer survival, increased antitumor immune infiltration (CD8+ T cells, M1 macrophages, activated CD4+ memory T cells), higher expression of T cell proliferation-related genes, lower immune checkpoint expression, better predicted immunotherapy response, and lower tumor stemness compared with the high-risk group.<h4>Conclusions</h4>We developed and internally validated a six-gene prognostic model for LUAD based on galactose metabolism. The model shows moderate prognostic performance and is associated with antitumor immunity and tumor stemness. It may be used for prognostic risk stratification and to guide personalized immunotherapy in LUAD.

Glutathione peroxidase 4 (GPX4) is a selenocysteine (Sec)-containing antioxidant enzyme and the only known mammalian enzyme capable of directly reducing membrane-embedded phospholipid and cholesterol hydroperoxides. It is recognized as a central regulator of ferroptosis, modulating cellular redox balance and influencing cell fate under oxidative stress. Despite a decade of research, critical gaps remain. Existing reviews largely focus on isolated diseases or single targeting strategies, and few provide an integrated framework that spans molecular regulation, physiological function, and clinical translation. The regulatory networks that control GPX4, from transcription to post-translational modifications and protein interactions, remain incompletely defined, and its ferroptosis-independent functions are underexplored. Moreover, the context-dependent and bidirectional roles of GPX4 across different diseases have not been systematically analyzed to guide appropriate therapeutic strategies. To address these gaps, this review delineates the structural basis and isoform-specific functions of GPX4, maps its multilayered regulatory network, and defines its roles across key physiological and pathological processes, including cancer, neurodegeneration, ischemia-reperfusion (I/R) injury, and autoimmune diseases. We also evaluate GPX4-targeted chemical strategies and analyze four core translational barriers: target specificity, systemic toxicity, acquired resistance, and tissue delivery, with evidence-based solutions for each. We conclude by identifying unresolved mechanistic questions and outlining priorities to accelerate clinical translation.

Although advances have been made in tumor immunotherapy, the immunosuppressive tumor microenvironment and treatment resistance still limit durable clinical benefit. Rather than functioning exclusively as terminal cytotoxic phenomena, regulated cell death (RCD) pathways generate surface cues, soluble mediators and cellular cargo that reshape immune responses. Here, we review the reciprocal interactions linking tumor-associated macrophages (TAMs) with apoptosis, necroptosis, pyroptosis, autophagy-related regulation, ferroptosis and cuproptosis. We examine how death execution, the kinetics and magnitude of tumor cell death, membrane integrity, and signal accessibility shape RCD-derived signals, and how macrophage lineage, functional state and signal-processing capacity determine their inflammatory and antitumor consequences. These reciprocal interactions regulate corpse clearance, antigen presentation, metabolic adaptation, tumor progression, and therapeutic resistance, while macrophages can in turn alter the susceptibility of viable tumor cells to death. We further discuss crosstalk among RCD pathways, extracellular vesicle-mediated communication, microbiota-related regulation and spatiotemporal reconfiguration during tumor progression and treatment. Finally, we evaluate nanomedicine strategies that coordinate tumor-selective RCD with macrophage regulation while emphasizing limitations in cell-type specificity, safety, patient selection and clinical evidence. This framework may guide more precise intervention along the RCD-macrophage axis.

In this study, a new series of nimesulide-derived ureas (<b>3a-j</b>) was designed and synthesized to investigate structural modifications of nimesulide and to evaluate their biological activity. The compounds were obtained in high yields (68-88%) under organotin-catalyzed conditions, and their structures were unequivocally established using comprehensive spectroscopic techniques, including ¹H NMR, ¹³C NMR, FTIR, and HRMS. Following structural confirmation, the antioxidant properties of the synthesized derivatives were evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, where compounds <b>3e</b>, <b>3f</b>, and <b>3i</b> displayed notable radical-quenching activity. The nimesulide derivative <b>3i</b> exhibited notable cytotoxicity across multiple breast cancer models, including the triple-negative MDA-MB-231 and 4T1 cell lines, as well as the luminal MCF-7 subtype. Consistent with its enhanced potency, <b>3i</b> triggered a robust shift from cell viability to programmed cell death, characterized predominantly by a dramatic accumulation of late-apoptotic populations following 72 h of treatment. This apoptotic signature was especially pronounced in Triple-Negative Breast Cancer (TNBC) cells, in which viable cell fractions were nearly eliminated. These findings suggest that structural modification of the nimesulide derivative can substantially improve its in vitro anticancer efficacy. Taken together, the observed cytotoxic and apoptosis-inducing effects suggest that compound <b>3i</b> may represent a potential NSAID-based anticancer candidate. In line with its pronounced cytotoxic and pro-apoptotic profile, compound <b>3i</b> also demonstrated strong inhibition of the MetAP2 enzyme, further reinforcing its potential as a multifunctional nimesulide-based anticancer agent. To obtain insight into the binding pose and binding energy of all synthesized compounds (3a-j) were docked into the active site of the MetAP2 enzyme. The computational inhibition constant values were correlated with the experimental values. To test the dynamic behavior of MetAP2-inhibitor complexes, a molecular dynamics (MD) simulation was also carried out for a 200 ns duration. MD revealed that the drugs bind to the active site of the MetAP2 enzyme, as indicated by stable RMSD and RMSF plots. In conclusion, <i>in silico</i> results and in vitro studies suggest that the nimesulide derivatives may be novel, potential NSAID-based anticancer drug candidates for treating breast, prostate, gastric, and glioblastoma.

Understanding the molecular mechanisms driving selective neuronal vulnerability to different neurodegenerative disorders remains a crucial, unsolved question. Here, we explored the case of Huntington's disease (HD), where the striatum and, specifically, dopamine receptor 1 (D1R) and dopamine receptor 2 (D2R) medium-sized spiny neurons (MSNs) exhibit differential susceptibility to the HTT CAG-repeat expansion mutation, with D2R-neurons being impacted earlier and more significantly. To unravel differences between D2R and D1R MSNs, we employed a multidimensional approach, integrating genomic, transcriptional, pattern distribution, and somatic instability analyses. Specifically, we used Htt CAG knock-in mouse models harboring 18 (Htt^Q20: "control") or ~190 (Htt^Q175: "HD") consecutive CAG repeats, expressing tdTomato and EGFP under the control of Drd1 and Drd2 promoters, respectively. First, comprehensive genomic and transcriptomic analyses following fluorescence-activated cell sorting (FACS) of dissociated striatal neurons revealed distinct gene expression profiles with a significant upregulation of oxidative phosphorylation and translation pathways in D1R-positive neurons already at the pre-symptomatic stage. These transcriptional changes were not accompanied by major copy number variations, as shown by parallel genomic analysis. Secondly, histological analyses revealed a greater proportion of D1R-positive neurons compared to D2R-positive neurons in HD mice, particularly in the ventral-medial neostriatum, with D2R-positive neurons presenting an increased nuclear accumulation of mutant huntingtin aggregates. In summary, our integrative study suggests that the distinct vulnerability of MSNs in HD might result from a combination of an early transcriptional compensatory response of D1R neurons together with specific susceptibility of D2R neurons.

<h4>Clinical problem</h4>Preeclampsia is a heterogeneous hypertensive disorder of pregnancy characterized by new-onset hypertension and end-organ dysfunction, affecting 5% to 10% of pregnancies globally. It is associated with significant maternal and fetal morbidity and mortality, and delivery remains the only definitive treatment. Understanding the underlying mechanisms is essential for the development of effective therapies.<h4>Recommendations</h4>Given the heterogeneity of preeclampsia and the absence of a single experimental model that fully recapitulates the human disease, the purpose of this review is to evaluate current animal models of preeclampsia according to the pathophysiological mechanisms they represent. We discuss the strengths, limitations, and translational relevance of these models, identify gaps in the existing experimental framework, and highlight opportunities for future model development and therapeutic discovery.<h4>Summary</h4>Current models reproduce distinct features of preeclampsia, including abnormal placentation, placental ischemia, angiogenic imbalance, endothelial dysfunction, immune activation, and metabolic dysregulation. Despite mechanistic insights, translational success has been limited. Future progress will depend on mechanism-driven model selections, development of models that better capture early placentation defects and disease heterogeneity, multi-hit approaches, including the incorporation of maternal comorbidities, and integration of human-relevant experimental systems to improve clinical translation.

Associations between various genetic variants and the risk of hepatocellular carcinoma (HCC) have been extensively explored but produced contradictory results. The aim of the present systematic meta-analysis was to determine and validate genetic variants that are associated with HCC risk. Two-step literature searches of PubMed, Embase, Web of Science, and Google Scholar databases and various meta-analyses were performed, and a comprehensive field synopsis and epidemiological evidence were provided. A total of 20,081 publications were identified, of which 830 were deemed eligible for inclusion. Eventually, 36 variants in 27 genes were identified to be associated with HCC risk. Moreover, cumulative epidemiological evidence of an association was graded as moderate for nine variants in eight genes (<i>ESR1</i> rs2234693, <i>GRP78</i> rs430397, <i>HLA-DP</i> rs3077, <i>HLA-DQ</i> rs2856718, <i>MnSOD</i> rs4880, <i>TNFα</i> rs361525, <i>HFE</i> rs1800562 and rs1799945, and <i>UGT1A7</i> High/Low) and strong for three variants in three genes (<i>IL-1B</i> rs1143627, <i>COL18A1</i> rs7499, and <i>NQO1</i> rs1800566); <i>HFE</i> rs1800562 was deemed to have a false-positive association. Thus, 11 variants in 11 genes were identified to be associated with HCC risk. This synopsis helps elucidate the mechanisms of carcinogenesis of HCC and provides insights into the early diagnosis and novel treatments of HCC by targeting those potential genes.

<h4>Background</h4>Glioblastoma (GBM) is characterized by high invasiveness and metabolic heterogeneity. Protein Tyrosine Phosphatase Receptor Type Z1 (PTPRZ1) has been implicated in glioma stemness and tumor grade progression, while the molecular mechanism remains unknown.<h4>Methods</h4>Integrated bioinformatics analysis of TCGA dataset and TMA IHC profiling were used to explore clinical relevance of PTPRZ1, and potential molecular mechanisms of PTPRZ1 involved glioma progression. Ivy Glioblastoma datasets analysis and multiregional GBM IHC were performed to spatially characterize the expression patterns of PTPRZ1 in GBM tissues. Zebrafish xenograft model was established to elucidate the role of PTPRZ1 in driving glioblastoma cell migration. Western blotting, mitochondrial functional assays, Transwell migration assays, and human GBM tissue multiplexing were employed to investigate the mechanism of PTPRZ1 mediated glioma cell mobility via maintaining mitochondrial functions.<h4>Results</h4>Spatial profiling revealed that PTPRZ1 is significantly enriched in the infiltrating areas and peritumoral margins of GBM, and acts as a poor prognosis factor in lower grade astrocytoma, proneural and mesenchymal GBM. Mechanistically, PTPRZ1 expression was found to be strongly correlated with cell adhesion molecules (CAMs) and mitochondrial metabolism across multiple glioma subtypes. Knockdown of PTPRZ1 triggered a shift from elongated to fragmented mitochondrial morphology by specifically downregulating the mitochondrial fusion protein OPA1. This mitochondrial dysfunction led to a metabolic shift toward glycolysis and a marked accumulation of ROS, which suppressed N-cadherin expression, thereby impairing glioma cell migration. These findings were corroborated in vivo, where PTPRZ1 depletion significantly reduced disseminated tumor foci in a zebrafish model.<h4>Conclusion</h4>Our study demonstrates that PTPRZ1 drives glioblastoma migration by sustaining an OPA1-dependent mitochondrial fusion program, which suppresses mitochondrial ROS levels and subsequently stabilizes a pro-migratory phenotype via N-cadherin expression. Targeting the PTPRZ1/OPA1/ROS axis represents a promising therapeutic strategy to inhibit the invasive expansion of GBM cells.

<h4>Background</h4>Alzheimer's disease (AD) is a progressive neurodegenerative disorder of late life that causes cognitive and functional decline and substantial mortality. Machine learning (ML) is increasingly used to discover patterns in clinical and biomarker data that support earlier and more accurate AD detection.<h4>Objectives</h4>This scoping review addresses three core research questions. First, we investigate recent trends in using machine-learning techniques to detect Alzheimer's disease using blood biomarkers. Second, we identify the blood biomarkers involved in Alzheimer's detection and evaluate how machine learning has been applied to improve the diagnostic capabilities of these biomarkers. Third, we highlight significant challenges associated with using machine learning for blood biomarker data in Alzheimer's detection and examine proposed advancements or solutions to handle these problems.<h4>Methods</h4>In June 2025, we searched six academic databases to identify relevant papers on blood biomarkers and ML methods for Alzheimer's Disease. Search queries were developed based on our predefined research questions. Papers were then screened using defined inclusion and exclusion criteria, where titles, abstracts, and full texts of articles were systematically reviewed.<h4>Results</h4>Following the screening approach, we selected 36 papers that fulfilled our inclusion and exclusion criteria. Through careful examination, we classified blood biomarkers into four types: transcriptomics, proteomics, multi-omic biomarkers, and general elemental blood biomarkers. Across these studies, proteomic blood biomarkers consistently emerged as significant indicators for Alzheimer's disease, including Alpha-2-Macroglobulin (A2M), Apolipoprotein E (ApoE), Eotaxin-3 (EOT3), plasma phosphorylated tau (p-tau 181), and neurofilament light chain (NfL). Furthermore, we explored challenges such as small sample sizes, lack of standardization, heterogeneity, and data imbalance.<h4>Conclusions</h4>This review provides insights into how combining blood biomarkers with ML can enhance AD prediction. The review summarizes key challenges and identifies critical gaps for future research.

<i>HFE</i>-related hemochromatosis (HC) is caused by hepcidin dysregulation and is characterized by excessive iron absorption and accumulation in the liver, heart, and endocrine glands, which leads to complications including arthropathy, cirrhosis, and hepatocellular carcinoma. Standard of care (SoC) typically involves phlebotomy, with iron chelators used less frequently. However, these therapies may cause side effects and negatively impact quality of life, making them troublesome for some patients. Using the Delphi methodology, a survey was developed to identify unmet needs and clinical challenges within the current therapeutic landscape of HC, and to establish consensus statements for its management. Consensus was defined as ≥75% agreement. Thirty-two HC specialists from Europe, Australia and the USA responded to the survey. After three rounds of survey refinement, final consensus statements were compiled. The Delphi process identified key unmet needs in HC, including lack of alternatives to phlebotomy, persistent symptoms, and burden related to the use of current SoC. The consensus process helped establish definitions for high phlebotomy treatment burden, intolerance, and suboptimal response to phlebotomy. The process also identified the subgroup of patients overly burdened by phlebotomy. Patient-reported outcomes were considered key to assessing phlebotomy's impact, although they are rarely measured in clinical practice. The Delphi study highlighted the limitations of phlebotomy, with respondents identifying a high unmet need for patients who cannot be managed with or do not tolerate this approach. The study suggested exploring alternative therapy options for patients who experience high treatment burden or intolerance to phlebotomy.

Stroke is a major cause of long-term disability with variable recovery. While clinical factors such as initial severity play a role, genetic factors are increasingly recognized as important contributors to stroke recovery. Genotype studies are generally focused on a single post-stroke behavioural domain, but some genes might relate to broad mechanisms of plasticity. This study therefore aimed to identify cross-phenotypic genetic variants associated across two or more stroke recovery domains. DNA from Stroke, Stress, Rehabilitation, and Genetics study participants was genotyped, resulting in 9 814 610 variants. In order to examine cross-phenotypic results, we first conducted genome-wide association studies on the six recovery domains: motor (grip force), cognition (Telephone Montreal Cognitive Assessment), depression (Patient Health Questionnaire-8), stress (Primary Care Post-Traumatic Stress Disorder Screen), functional status (Stroke Impact Scale-Activities of Daily Living), and disability (modified Rankin Scale 0-2 versus 3-6), some of which were tested longitudinally, yielding nine phenotypes. Models were adjusted for age, sex, initial severity (NIH Stroke Scale score), and ancestry. Cross-phenotype associations were identified by evaluating single nucleotide polymorphisms (SNPs) associated (<i>P</i> < 5e-5) with multiple phenotypes. To determine how these genetic variants may relate to biological mechanisms of recovery, we conducted gene enrichment analyses. Participants (<i>n</i> = 565, 59% male) had mild-moderate initial stroke severity (median acute NIH Stroke Scale score = 4). After accounting for the correlation structure among the nine phenotypes, we observed 319 cross-phenotypic SNPs, 3.45 times the expected number. Five of the cross-phenotypic SNPs were linked to genes relevant to neural development, function and plasticity, e.g. <i>ERICH1</i> (rs11778883-C), <i>FOX3</i> (rs55726768-G), <i>LIFR-AS1</i> (rs76401391-T), <i>RPS6KA2</i> (rs113518460-C) and <i>TUBGCP2</i> (rs147150392-C), as were enrichments in <i>RAB5-EEA1</i>, <i>CTNNA1-CTNNB1</i>, <i>CIN85-SH3GL2</i> and <i>ELMO1-DOCK2</i> complexes. Multiple gene enrichments were found, e.g. Stroke Impact Scale-Activities of Daily Living and Patient Health Questionnaire 8 at 3 months were enriched for <i>CREB</i> phosphorylation, which is important for long-term potentiation. We identified cross-phenotypic SNPs associated with multiple behavioural domains of stroke recovery. Some of these genes encode, or regulate, druggable proteins. These genetic factors are not well captured by clinical or neuroimaging assessments and so provide a unique window into stroke recovery. These findings, if validated, suggest that some genes may be broadly important to stroke recovery.

<b>Background</b> Coagulation factor VIII (FVIII) is a critical component of the intrinsic coagulation pathway. While elevated FVIII levels are an established risk factor for venous thromboembolism (VTE), genetic variants in the <i>F8</i> gene directly causing such elevations remain scarce. Here, we report a novel complete <i>F8</i> tandem duplication identified in a female patient with splanchnic venous thrombosis (SVT). <b>Methods</b> We performed genetic testing using a thrombophilia panel targeting 35 genes involved in thrombosis and haemostasis to detect both point variants and copy number variations (CNVs). Family co-segregation analysis and phenotypic assays for FVIII and von Willebrand factor (VWF) were conducted. The structural basis of the identified <i>F8</i> copy number gain was elucidated using optical genome mapping (OGM). Full-length <i>F8</i> mRNA amplification, quantitative PCR, plasma FVIII Western blotting, and X-chromosome inactivation analysis were performed to assess the functional consequences of the duplication. Thrombin generation test (TGT) was employed to assess the hypercoagulable state. <b>Results</b> Genetic testing identified three copies of all 26 exons of the <i>F8</i> gene in the proband, which was also detected in her mother (CNVs = 3) and son (CNVs = 2). One-stage clotting and chromogenic assays confirmed persistently elevated FVIII activity in the proband and her mother, accompanied by increased FVIII antigen levels. The OGM analysis confirmed a 229 kb tandem duplication including the <i>F8</i> gene on one of the proband's X chromosomes. The junction regions exhibited high sequence homology and were rich in repetitive sequences, which precluded precise breakpoint mapping. Full-length <i>F8</i> mRNA amplification revealed no aberrant transcripts, whereas quantitative PCR showed increased <i>F8</i> mRNA expression in all carriers. Plasma FVIII Western blotting indicated FVIII heavy and light chains of expected molecular weights with increased band intensity in carriers. X-chromosome inactivation analysis in female carriers showed no significant skewing. TGT in two available carriers showed increased thrombin generation compared with a normal control at both low (1 pM) and high (5 pM) tissue factor concentrations. <b>Conclusion</b> We identified a novel complete <i>F8</i> tandem duplication associated with increased FVIII expression and a hypercoagulable phenotype in a female patient with SVT. These findings support <i>F8</i> gene dosage gain as a rare gain-of-function mechanism contributing to elevated FVIII levels and thrombophilia, while variation in VWF levels and acquired risk factors may modify thrombotic penetrance.

A substantial number of genetic variants have been associated with high-altitude adaptation (HAA), yet most of them are located in non-coding genomic regions, leaving their specific functions and underlying mechanisms largely unknown. In this study, we analyze whole-genome and transcriptome sequencing data from a self-established cohort comprising 61 native highlanders (NHs) and 164 acclimatized newcomers (ANs), identifying 6,586 cis- and 34,203 trans-expression quantitative trait loci (eQTLs), along with 130 cell type-specific eQTLs. By further combining these data with a large East Asia (~30% Tibetan) genome-wide association study (GWAS) cohort, we employ colocalization and causal inference analyses to prioritize 85 cis-eQTLs associated with HAA and identify several novel candidate causal genes, including EXOC8, which is experimentally confirmed to regulate erythroid differentiation. Additionally, network analysis of these causal genes uncovers multiple regulatory pathways, mainly involving energy metabolism, autophagy, ubiquitination and inflammation. Our study offers a comprehensive eQTL map and reveals causal chains of "variant-gene-phenotype" for HAA-related traits, which provides new insights into potential regulatory mechanisms and targets for prevention and treatment of altitude sickness.

<h4>Introduction</h4>Critically low blood donation rates and recurrent shortages in the blood supply have become pressing challenges in transfusion medicine. Xenotransfusion using porcine red blood cells (pRBCs) represents a promising alternative; however, risks of immune rejection and infection necessitate the development of genetically engineered porcine models - a process that is time-consuming, costly, and technically complex. In this study, we established an immortalized porcine erythroid progenitor cell line as a platform for genetic manipulation and the evaluation of xenotransfusion potential.<h4>Methods</h4>Porcine hematopoietic progenitor cells (CD117⁺) were isolated from bone marrow mononuclear cells and immortalized via lentiviral transduction with E6/E7 oncogenes from Human papillomavirus type 16 (HPV16) and Sus scrofa papillomavirus (SPV2).<h4>Results</h4>The resulting cells were optimized for long-term culture and exhibited stable proliferation. The immortalized porcine erythroid progenitor cells retained the capacity to differentiate into red blood cells.<h4>Discussion</h4>Our findings demonstrate that E6/E7-mediated immortalization facilitates the generation of porcine erythroid progenitor cells capable of differentiating into red blood cells, offering a valuable tool for genetic engineering and a promising avenue toward safe, sustainable heterologous blood transfusion strategies.

<h4>Background</h4>Gastric intestinal metaplasia (GIM) is a typical precancerous lesion of gastric cancer (PLGC). Previous studies have demonstrated that Xinkai Kujiang formula can effectively alleviate GIM, but its underlying mechanism remains largely unclear.<h4>Methods</h4>The GIM rat model was established using 2% sodium salicylate and 20 mmol/L sodium deoxycholate, and then the rats were treated with Banxia Xiexin Decoction (BXD) and Xinkai Kujiang Decoction (XKD) for 4 weeks. Multi-omics analyses including 16 S ribosomal RNA gene sequencing, transcriptomics, single-cell RNA sequencing, network pharmacology, and component identification were performed to explore the therapeutic mechanisms of Xinkai Kujiang formula on GIM.<h4>Results</h4>In the model rats, severe gastric mucosal atrophy was observed, characterized by disordered glands and goblet cells. Following intervention with BXD and XKD, gastric mucosal thickness was restored, glandular structures became regularly arranged, and the number of metaplastic goblet cells markedly decreased. Microbiota profiling of gastric mucosa revealed significant enrichment of <i>Lactobacillus</i> and <i>Enterococcus</i> in the model group. These abundances were reduced in the BXD group, and short-chain fatty acid-producing bacteria such as <i>Alistipes</i> and <i>Lachnospira</i> were enriched. In the intestine, opportunistic pathogens like <i>Streptococcus</i> and <i>Enterococcus</i> were enriched in the model group, whereas <i>Corynebacterium</i> and <i>Bifidobacterium</i> were enriched in the XKD group. Transcriptomic analysis presented that BXD upregulated innate immune-related genes in the gastric mucosa, and single-cell RNA sequencing (scRNA-Seq) showed that XKD alleviated GIM by inhibiting the VEGF and HIF-1α pathways, reducing angiogenesis, suppressing inflammatory infiltration, and regulating energy metabolism.<h4>Conclusion</h4>BXD and XKD improve gastrointestinal microbiota disorders and metabolic disorders, enhance gastric mucosal immunity, and inhibit the VEGF and HIF-1α pathway. Collectively, these multi-omics data provide novel insights into the therapeutic mechanisms of Xinkai Kujiang formula for GIM.

Rheumatoid arthritis (RA) is a highly prevalent systemic autoimmune disease characterized by a complex and partially understood pathogenesis. The substantial challenges in early identification and marked therapeutic heterogeneity pose a significant burden on affected patients. Despite notable advancements in diagnostic techniques and therapeutic interventions in recent years, optimal patient care remains hindered by several ongoing clinical challenges. To address these limitations, artificial intelligence (AI) has been increasingly integrated into the field of rheumatology. By leveraging its potent capabilities in large-scale data and medical image processing, AI offers diversified and individualized approaches to RA diagnosis and management. Herein, following a review of the fundamental concepts of RA and AI, we explore the multifaceted applications of AI across RA diagnosis, treatment response prediction, <i>de novo</i> drug discovery, and both clinical and basic science research. Furthermore, we provide a comprehensive analysis of the strengths and pitfalls of implementing these models in real-world clinical settings. By summarizing the AI technologies that either hold significant promise for practical clinical application or have already been translated into routine practice, this review aims to catalyze the progress of AI-guided precision medicine and drive the intelligent evolution of RA management.

<h4>Background</h4>Chronically activated T-effector (Teff) cells can play a pivotal role in T-cell-mediated diseases including systemic lupus erythematosus subsets, lupus nephritis and graft-vs-host disease (GvHD). Since T cells require diverse costimulation signals to fully activate, proliferate, and differentiate into Teffs, we hypothesized that modulating two costimulatory pathways (CD28 and OX40) with a single bifunctional molecule would provide better control of pathogenic Teffs in autoimmune diseases than currently available T-cell costimulation modulators. We assessed the effects of dual-pathway blockade by M5542, a novel CD80, CD86, and OX40L antagonist and bifunctional fusion molecule. We determined the advantages of M5542 blocking both CD28 and OX40 pathways over single-pathway blockade in <i>in vitro</i> potency and <i>in vivo</i> studies, to more effectively curb T-cell-mediated inflammation in autoimmune diseases.<h4>Methods</h4>M5542, single-agent comparators CTLA-4Ig (abatacept), anti-OX40L, and both single agents in combination were tested for their ability to inhibit proinflammatory cytokine production in a mixed lymphocyte reaction of activated monocyte-derived dendritic cells and T cells from healthy donors or peripheral blood mononuclear cells from individuals with SLE. The target occupancy of CD80 and OX40L on dendritic cells was assessed by flow cytometry. The effects of M5542 and comparators on T-cell-mediated inflammation were assessed using a keyhole limpet hemocyanin (KLH) peptide-induced delayed-type hypersensitivity (DTH) humanized hOX40/hOX40L mouse model. All molecules were also tested for their efficacy to reduce disease in a humanized xenogenic GvHD (xGvHD) mouse model.<h4>Results</h4>M5542 potently reduced proinflammatory cytokine production and dampened T-cell proliferation compared with CTLA-4Ig or anti-OX40L <i>in vitro</i>. M5542 also neutralized OX40L-mediated T-regulatory cell (Treg) dysfunction <i>in vitro</i> and suppressed Teff proliferation in conjunction with Tregs <i>in vitro</i>. Moreover, M5542 dose-dependently reduced T-cell-mediated ear swelling and anti-KLH antibody production in the KLH-DTH mouse model. In the xGvHD model, M5542 demonstrated improved efficacy over CTLA-4Ig or anti-OX40L alone in suppressing human IFNγ and preventing disease.<h4>Conclusion</h4>In this preclinical study, the bifunctional M5542 blocked CD28 and OX40-mediated T-cell inflammation with greater immunomodulatory activity than monofunctional agents and in some assays comparable to or better than the combination of the monofunctional agents. These findings support further evaluation of M5542 in T-cell-driven autoimmune disease settings.

This paper explores the fabrication of hydroxyapatite-capsaicin nanocomposite coatings on Mg-Ag alloy by the electrophoretic deposition (EPD) method at different deposition times and voltages. The microstructure, surface chemistry, hardness, corrosion behavior, and cell proliferation of the deposited layers at different deposition times and voltages were examined. Based on microstructural observations, the deposited layers became denser at higher deposition time and voltage. In addition, the hydroxyapatite-capsaicin nanocomposite became thicker at higher EPD voltage. FTIR and XPS analyses confirmed the successful incorporation of capsaicin into the HAP coating with partial retention of CAP-derived surface functionalities after annealing. Also, the hardness of the coated layer increased with voltage and time. However, EPD voltage was more effective in increasing the hardness. The maximum hardness of 141 HV was obtained at a deposition voltage of 160 V. The hardness profile of the deposited composite showed greater fluctuations with increasing deposition voltage. Electrochemical polarization tests demonstrated that corrosion resistance improved with increasing deposition voltage and time. Furthermore, cell growth was improved at higher EPD time and voltage. MTT assay results confirmed the enhanced cytocompatibility of the HAP-CAP coatings. Also, the Mg²⁺ ion release rate and pH variation decreased as the deposited layer became thicker and denser. The minimum Mg²⁺ ion release rate of 5.2 ppm h^-1 was obtained at a deposition voltage of 160 V. Compared with conventional HAP coatings, incorporation of capsaicin enhanced the biological performance while preserving the structural stability of hydroxyapatite, which thus provides a promising strategy for the development of multifunctional biodegradable implant coatings.

Neural progenitor cell differentiation is a complex process requiring the proper integration of instructive and permissive factors. Instructive cues including signaling molecules and transcription factor networks have been well studied in this context, but permissive factors such as cell homeostasis have not. Cell homeostasis is critical to support the health and stability of a cell and enable the cell to act on instructive differentiation cues. Our study investigates a homeostasis protein, FAF2, and its function in neural progenitor cells. FAF2 is an adaptor protein involved in endoplasmic reticulum (ER) associated degradation to remove misfolded proteins and restore ER homeostasis. Here we show that knocking out <i>Faf2</i> in neural progenitor cells results in increased ER stress signature at the protein and transcription level, indicating a conserved functional role in neural progenitor cells. Induced neural differentiation of FAF2 deletion cells shows a failure of neurite development but RNA-seq indicates genes that support neural differentiation are induced. Reducing ER stress in FAF2 knockout cells with a small molecule inhibitor can rescue neural differentiation, providing evidence that excess ER stress contributes to the inhibited differentiation. Taken together, these results reveal that FAF2 is a critical protein in neural progenitor cells for the maintenance of ER homeostasis and execution of neural differentiation.

Keloid, as a unique form of pathological scar, involves complex interactions across multiple dimensions in its pathogenesis, including genetic susceptibility, epigenetic regulation, immune microenvironment disorders, metabolic reprogramming, abnormal mechanical force transduction, and fine-tuning by non-coding RNAs. At the genetic level, alleles such as HLA-DRB1*15, HOX gene family, and high-frequency mutation genes including MUC4 constitute the foundation of congenital susceptibility; at the epigenetic level, reduced DNA methylation and imbalanced m6A modification promote fibrosis by regulating target genes such as COL1A1. In core signaling pathways, TGF-β/Smad, MAPK/ERK, PI3K/AKT/mTOR, and Wnt/β-catenin pathways form a cross-talk network, driving persistent activation of fibroblasts and excessive extracellular matrix deposition. Characteristics of the immune microenvironment include M2-like macrophage polarization, Th17/IL-17 axis activation, and chronic low-grade inflammation maintained by inflammatory factor networks such as CXCL12. Regarding metabolic reprogramming, keloid cells exhibit "Warburg effect" features, prioritizing aerobic glycolysis while suppressing oxidative phosphorylation, alongside ferroptosis resistance and abnormal sphingolipid metabolism. Mechanical forces trigger the opening of mechanosensitive PIEZO1 ion channels, inducing abnormal calcium influx and subsequent YAP/TAZ nuclear translocation, which creates a self-amplifying vicious cycle alongside elevated matrix stiffness. Furthermore, lncRNAs, circRNAs, and miRNAs finely regulate the fibrotic process through competing endogenous RNA (ceRNA) networks and m6A modification-dependent mechanisms. The integration of these multidimensional mechanisms provides a theoretical basis for developing multi-target combination therapeutic strategies. Future research should promote the application of precision medicine in keloid management through multi-omics integration and artificial intelligence algorithms. Microenvironment; Metabolic reprogramming; Mechanical force transduction.

Arrestins were originally defined as terminators of G protein-coupled receptor (GPCR) signaling, yet structural and mechanistic advances now reveal them as programmable, spatiotemporal integrators of cellular signaling. Recent cryo-electron microscopy studies have revealed a diverse spectrum of GPCR-arrestin engagement modes, including core-, tail-, loop-, side-engaged, and membrane-anchored conformations, across GPCR classes and arrestin isoforms. These structures reveal that arrestin recruitment operates as a conditional, allosterically regulated process rather than a binary on-off switch. The selection of the arrestin microstate is governed by layered regulatory inputs, including GPCR kinase-dependent phosphorylation barcodes, membrane and lipid cofactors, and isoform-specific mechanics, which together define the signaling geometry, duration, and subcellular localization. This structural logic provides a mechanistic foundation for biased signaling, noncanonical endosomal signaling, and GPCR-independent arrestin functions. Importantly, emerging therapeutic strategies, including intracellular allosteric modulators and molecular glues, demonstrate that arrestin signaling can be reprogrammed by directly sculpting transducer assemblies rather than ligand efficacy alone. Here, we synthesize recent structural, biochemical, and physiological insights to outline how arrestins decode regulatory inputs into signaling outcomes and how this knowledge enables the development of next-generation, structure-guided GPCR therapeutics.

Epigenetics plays a central role in cancer development and progression by governing the expression of genes involved in these processes. Accumulating evidence strongly indicates that these mechanisms not only alter cancer cell-intrinsic properties but also mediate reciprocal interactions between cancer cells and the tumor microenvironment (TME). In the context of cancer cell-intrinsic changes, epigenetic alterations contribute to cancer cell plasticity by regulating stemness, cell state transitions, and therapy resistance. In addition, epigenetic changes are a key driver of the establishment of a tumor-supportive environment by modifying the states of TME components, including fibroblasts, macrophages, and myeloid-derived suppressor cells. Epigenetic reprogramming in cancer cells and TME components is often induced by factors such as hypoxia and the secretion of exosomal long noncoding RNAs (lncRNAs), cytokines, and metabolites. In this review, we discuss various epigenetic mechanisms controlling cancer cell-intrinsic states, cancer cell-TME crosstalk, current epigenetic therapies, and future research directions that may lead to the discovery of new biomarkers and the development of effective epigenetic anticancer therapeutics.

African swine fever remains a major constraint on global pig production because effective treatment is unavailable, and disease control relies primarily on biosecurity and outbreak management. Field observations from endemic regions increasingly suggest that some domestic pigs can survive exposure to ASFV. Therefore, the aim of this study was to characterize genomic differences between Mozambican Landim pigs that survived or died after natural ASFV infection and to identify variants potentially associated with differential disease survival. Whole-genome sequencing was performed on 32 naturally exposed Landim pigs, including 16 survivors and 16 non-survivors, and the resulting reads were aligned to the Sscrofa11.1 reference genome. Variant frequencies were compared between groups, and functional annotation was performed. Genetic structure was assessed using PCA, genetic distance-based clustering, and admixture. The data revealed marked genomic differentiation between survivors and non-survivors. Comparative analysis identified 4,804 polymorphisms that significantly differentiated, of which 45.0% were intergenic. Missense variants were identified in the TTC12 and WWC1 genes, and functional annotation of genes harboring significantly differentiating variants indicated processes related to immune regulation, T-cell activation, intracellular signaling, cytoskeletal organization, vesicular transport, ubiquitin-dependent proteostasis, and lipid metabolism. Given the marked genetic stratification between survivor and non-survivor groups, the observed differences between them cannot be unambiguously separated from underlying population structure. Therefore, although some of the identified variants may contribute to differential survival following ASFV exposure, the findings should be considered exploratory and require validation in larger, independent, genetically comparable populations.

Ferroptosis is a distinct modality of programmed cell death driven by iron-dependent lipid peroxidation, exerting a pivotal regulatory influence on the infection dynamics and immune responses associated with zoonoses. This article systematically elucidates the mechanistic interplay between significant zoonotic pathogens and the core regulatory networks of host ferroptosis, encompassing iron metabolism, the antioxidant defense systems typified by GPX4, and lipid peroxidation. Furthermore, it synthesizes current advancements and challenges regarding small molecules, natural products, and bioactive components of traditional Chinese medicine that target key ferroptotic checkpoints for anti-infective therapy. This review aims to establish a theoretical foundation for the development of novel, precision prevention and control strategies for zoonoses, grounded in ferroptosis modulation within the "One Health" perspective.

Three-dimensional genome organization shapes transcriptional regulation, yet measuring its spatial coordination in situ within intact tissues remains challenging. We present Spatial Hi-C-RNA, a multimodal platform that simultaneously maps genome-wide chromatin contacts and transcriptomes from the same tissue section at near-single-cell resolution. Across the mouse brain, developing embryos, and human melanoma, Spatial Hi-C-RNA generated multimodal maps that aligned with tissue anatomy while revealing complementary chromatin- and RNA-defined spatial patterns. Multiscale features, including A/B compartments, topologically associating domains, and chromatin loops, were associated with region- and cell-type-specific transcriptional programs. In mouse embryos, Spatial Hi-C-RNA resolved coordinated chromatin and transcriptional remodeling during neuronal maturation across developmental stages. In human melanoma, chromatin architecture delineated intratumoral subregions not detected by RNA alone and linked tumor-state transitions to changes in compartments, domain boundaries, and regulatory programs. Spatial Hi-C-RNA thus provides a broadly applicable framework for investigating genome structure-function relationships in development and disease within native tissue environments.

Splenomegaly is a defining feature of myelofibrosis, yet the contribution of splenic mesenchymal stroma to disease progression remains unclear. We combined spatial and single-nucleus transcriptomics of patient spleens with spatial and single-cell transcriptomics, as well as imaging analyses, of murine spleens to map extramedullary hematopoiesis niches. Activated red pulp reticular cells localize near hematopoietic stem and progenitor cells, and early disease is characterized by marginal zone disruption with lymphoid depletion preceding stromal remodeling. Trajectory analyses reveal a shift in reticular cells from hematopoiesis-supportive to inflammatory and pro-fibrotic states, driven by macrophage- and megakaryocyte-derived signals that activate complement and induce tumor necrosis factor α (TNF-α), transforming growth factor β (TGF-β), extracellular matrix, and Thbs1 programs. Non-hematopoietic complement component C3 deficiency or pharmacological C3 inhibition suppresses these pathways, restores splenic architecture, and reduces splenomegaly and bone marrow fibrosis. These findings identify complement-dependent stromal reprogramming as a mechanism governing hematopoietic niches and as a targetable axis in myelofibrosis.

<h4>Background</h4>Mendelian susceptibility to mycobacterial disease (MSMD) is a subset of inborn errors of immunity (IEIs) predisposing to clinical infections from less virulent mycobacterial species. Variants in 22 genes, particularly in IL-12Rβ1, have been identified that impair interferon-gamma (IFN-γ)-mediated immunity. We report a new rare MSMD patient with a homozygous <i>IL12RB1</i> variant who presented with severe and persistent ascites.<h4>Case report</h4>We investigated a 30-month-old girl who developed an abscess and lymphadenopathy (LAP) in the left axilla after receiving a BCG vaccine. She was diagnosed and treated for BCG-osis. She presented with ascites, which gradually worsened, along with symptoms of liver dysfunction. Her portal hypertension (PHT) was considered the most likely cause of ascites after exclusion of other common causes. The whole exome sequencing (WES) revealed a homozygous rare variant (c.635G>A; p.Arg212Gln) in the <i>IL12RB1</i> gene. Sanger sequencing analysis confirmed that both parents were heterozygous carriers of the variant. She made a good recovery after receiving the appropriate antimycobacterial and ascites treatments, with significant resolution of her symptoms. Because disseminated <i>M. bovis</i> BCG infection occurred in the setting of IL-12Rβ1 deficiency, prolonged susceptibility-guided antimycobacterial therapy with adjunctive IFN-γ1b and close follow-up by infectious disease and immunology, was planned.<h4>Conclusion</h4>This case highlights severe persistent ascites as an unusual manifestation of PHT in IL-12Rβ1 deficiency associated with MSMD. Evaluation for PHT should be considered in MSMD patients with abdominal distension, gastrointestinal symptoms, hepatosplenomegaly, abnormal liver function tests, growth failure, or unexplained free abdominal fluid.

Pyrazoles and chalcones have been extensively studied over time due to their broad range of therapeutic potentials. In this study, a new series of pyrazole-carboxylate derivatives were synthesized, characterized, and evaluated for their antibacterial, anticancer and anti-inflammatory activities. Among the synthesized derivatives, compound 5a exhibited significant percentage inhibition in colony counting assay. Compound 5c demonstrated significant cytotoxic activity with an IC₅₀ value of 9.91 µg mL^-1, while also exhibiting lower cytotoxicity towards non-cancerous HEK-293T cells (IC₅₀ = 36.31 µg mL^-1), indicating favourable selectivity. Mechanistic studies, including DAPI staining and flow cytometric analysis, indicated that compounds 5c and 5f inhibited cancer cell growth predominantly by inducing apoptosis rather than cell cycle arrest. Anti-inflammatory activity was determined using protein denaturation assay where compound 5f demonstrated promising activity with an IC₅₀ value of 59.37 ± 0.149 µg mL^-1. Furthermore, molecular docking analysis further provided insights into the binding interactions of the derivatives and the targeted protein. In addition, drug-likeness evaluation using swissADME indicates that the compounds satisfied Lipinski's rule of five.

Diabetic nephropathy (DN) is a major microvascular complication of diabetes. Hypoxia-inducible factor 1-alpha (HIF1A) and ferroptosis contribute to the progression of DN, but the circulating biomarkers associated with these pathways are still unknown. In this study, the whole-blood transcriptomic datasets in the Gene Expression Omnibus were combined with ferroptosis-related genes from the FerrDb database and published literature. The differentially expressed genes of HIF1A and of DN were intersected, and then the feature selection with Boruta and SVM-RFE was performed, followed by external validation, immune infiltration analysis, prediction of regulatory network, drug-target prediction, and RT-qPCR validation. We identified 773 HIF1A-related genes, 1445 DN-related genes, and 22 overlapping candidate genes. Using machine-learning analysis, nine core genes were identified, including SLC7A5 and SLC2A14, which consistently upregulated in both training and validation datasets and corroborated by RT-qPCR. Both genes were positively associated with activated natural killer cells and negatively associated with monocytes in estimated circulating immune-cell composition analysis. Regulatory-network and DrugBank analyses generated hypotheses regarding upstream miRNAs and potential compound interaction. These findings identify SLC7A5 and SLC2A14 as candidate circulating biomarkers related to HIF1A expression and ferroptosis-related gene signatures in DN. Further large-cohort and functional studies are required to confirm their diagnostic and mechanistic relevance.

<h4>Background</h4>T-lineage acute leukemias are aggressive malignancies for which treatment options are limited.<h4>Objectives</h4>This study evaluated the efficacy and safety of a non-cytotoxic regimen combining venetoclax, azacitidine, and dexamethasone (VAD) as first-line induction therapy for T-cell acute lymphoblastic leukemia (T-ALL) and T/myeloid mixed-phenotype acute leukemia (T-Myeloid MPAL).<h4>Design</h4>We retrospectively analyzed the clinical parameters and survival data of 11 patients with newly-diagnosed T-ALL or T-Myeloid MPAL who received the VAD regimen.<h4>Methods</h4>The complete remission (CR) rate and adverse events were assessed. Kaplan-Meier curves were constructed to evaluate survival outcomes.<h4>Results</h4>Among 11 patients (7 with early T-cell precursor [ETP] ALL, 2 with T-Myeloid MPAL, and 2 with non-ETP T-ALL), the overall CR rate after one cycle of VAD therapy was 90.9%, with 45.5% achieving measurable residual disease (MRD) negativity by flow cytometry. The historically poor-prognosis ETP-ALL subgroup showed an 85.7% CR rate (6/7), while both T-Myeloid MPAL cases attained MRD negativity. No tumor lysis syndrome or treatment-related mortality occurred. Over a median follow-up of 358 days, all 5 patients who received transplantations remained relapse-free.<h4>Conclusion</h4>The VAD regimen demonstrated a high response rate and favorable safety, enabling effective transplantation bridging with avoidance of conventional cytotoxic therapy.

<h4>Purpose</h4>This propensity score-matched study aimed to compare survival outcomes between preoperative TACE followed by hepatectomy and upfront hepatectomy in patients with resectable hepatocellular carcinoma.<h4>Patients and methods</h4>In this single-center retrospective cohort study at Viet Duc University Hospital, 304 consecutive patients with resectable HCC treated between January 2015 and October 2024 were analyzed: 168 underwent upfront hepatectomy and 136 received preoperative TACE followed by hepatectomy. Propensity score matching (PSM; 1:1 nearest-neighbor, caliper 0.01) was performed to balance pre-treatment covariates between groups. Overall survival (OS) and disease-free survival (DFS) were compared using Kaplan-Meier analysis and the Log rank test. Independent prognostic factors were identified using multivariable Cox regression.<h4>Results</h4>Median follow-up was 58.9 months. After PSM (113 matched pairs), median OS was not reached in the upfront hepatectomy group versus 62.5 months in the preoperative TACE group (P = 0.322), and median DFS was 32.0 versus 29.2 months (P = 0.056). In multivariable Cox analysis of the matched cohort, preoperative TACE was not independently associated with OS (HR 1.15, 95% CI 0.74-1.79; P = 0.526) or DFS (HR 1.19, 95% CI 0.92-1.75; P = 0.130), whereas advanced TNM stage (OS: HR 2.45, 95% CI 1.08-5.52, P = 0.032; DFS: HR 2.26, 95% CI 1.14-4.46, P = 0.019) and microvascular invasion (OS: HR 1.48, 95% CI 1.09-2.40, P = 0.007; DFS: HR 1.52, 95% CI 1.01-2.29, P = 0.035) remained independent prognostic factors. Within the TACE group, patients achieving complete pathological necrosis (n = 26) had OS and DFS comparable to upfront hepatectomy, whereas incomplete or partial necrosis was associated with significantly worse survival (P = 0.05 for OS and P = 0.01 for DFS).<h4>Conclusion</h4>In this retrospective, propensity score-matched cohort, preoperative TACE was not associated with improved overall or disease-free survival in patients with resectable HCC and does not appear justified as a routine strategy in patients who are already surgical candidates. Because the analysis was retrospective and residual confounding cannot be excluded, these findings should be regarded as hypothesis-generating. Any role for preoperative TACE in selected settings, such as borderline resectability or when complete tumor necrosis is achieved, requires prospective confirmation.

Upconversion nanoparticles (UCNPs) convert tissue-penetrating near-infrared photons into sharp visible emissions, offering background-free optical readouts for deep-tissue imaging. Here we report multilayer NaLuF₄:Yb,Tm@NaLuF₄ UCNPs that simultaneously act as fluorescent probes and radioisotope precursors. Under 980 nm excitation, the particles generate bright blue and red upconversion signals for real-time tracking. At the same time, their lutetium-rich core can be neutron-activated in a single step to yield β^-- and γ-emitting ¹⁷⁷Lu. The β-particles confine cytotoxic dose within a 2 mm range, whereas the co-emitted γ photons enable quantitative single-photon emission computed tomography (SPECT). To enhance the tumor selectivity of the nanoplatform, we grafted a c-Met DNA aptamer onto the PEGylated shell, guiding the nanoconstructs to target head and neck squamous cell carcinoma (HNSCC) with amplified c-Met receptor levels. The aptamer markedly enhances cellular uptake, improves therapeutic indices, and limits off-target irradiation. Moreover, the optical modality remains non-radioactive, allowing pre-treatment imaging without patient exposure, and supplements the relatively low (∼10%) γ yield of ¹⁷⁷Lu for accurate intra-procedural localization. Collectively, this single nanosystem unites targeted radiotherapy, radionuclide therapy, SPECT, and NIR-mediated optical imaging, delivering a coherent "see-and-treat" strategy for HNSCC. The modular design also provides a versatile platform that can be transferred to other solid tumors bearing actionable biomarkers across diverse clinical oncology imaging and therapy settings.

<h4>Background</h4>Glutamine dependence is a hallmark of tumor cell metabolism, and further molecular classification based on glutamine metabolism in patients with hepatocellular carcinoma (HCC) may provide clinical value. This study thus comprehensively examined the patterns of HCC-specific alterations in glutamine metabolism.<h4>Methods</h4>Consensus clustering analysis was conducted on samples from The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset based on glutamine metabolism-related genes, which was validated in the GSE76427, the Liver Cancer-France (LICA-FR) cohort, and the Liver Cancer-Japan (LIRI-JP) cohort from the ICGC. Somatic mutation features were evaluated with the Maftools package in R. The activity of oncogenic pathways was estimated via gene set enrichment analysis (GSEA) or single-sample GSEA (ssGSEA). The tumor microenvironment was analyzed using both the CIBERSORT algorithm (for immune cell infiltration estimation) and the ESTIMATE algorithm (for stromal and immune score calculation). Drug sensitivity and immune checkpoint blockade (ICB) response were also analyzed, for which a classifier was built via least absolute shrinkage and selection operator (LASSO). Immunohistochemistry (IHC) was performed to validate the protein expression levels of key differentially expressed genes (DEGs). Intracellular glutamine content under different glutamine concentrations was measured. The viability of HCC cell lines under varying glutamine concentrations was assessed via Cell Counting Kit-8 (CCK-8) assays. Cell migration and invasion were evaluated through Transwell assays, and protein expression was analyzed via Western blotting.<h4>Results</h4>HCC samples were classified into two glutamine metabolism-based clusters, with cluster 1 having a more advanced stage of disease and shorter survival than cluster 2. A higher frequency of genetic mutations and stronger activation of oncogenic pathways was found in cluster 1. There were substantial differences in immune cell infiltration and stromal scores between clusters 1 and 2. Cluster 1 exhibited significantly higher infiltration of immunosuppressive cells and lower stromal scores compared to cluster 2. Cluster 1 had a stronger response to ICB due as indicated by a higher tumor mutation burden (TMB) and T cell-inflamed gene expression profile score, immune checkpoints, and Tumor Immune Dysfunction and Exclusion (TIDE)-predicted data. Moreover, the LASSO classifier accurately differentiated the two clusters. The DEGs between the two clusters were validated in clinical samples. IHC confirmed the differential expression of glutamine metabolism-related genes in HCC samples. CCK-8 assays showed no significant effect of glutamine concentration on cell proliferation. However, Transwell assays revealed that glutamine deprivation (0.2 mM) reduced migration and invasion, while high-glutamine conditions (10 mM) promoted them. Western blotting showed increased expression of metabolism-related proteins under high-glutamine conditions and reduced expression under deprivation.<h4>Conclusions</h4>Altogether, these findings indicate the involvement of glutamine metabolism in HCC and may help inform patient stratification and the formulation of precision therapeutics for this population.

<h4>Aim</h4>Episodic Migraine (EM) is a highly prevalent and disabling neurological disorder. Recent therapeutic advances have targeted the calcitonin gene-related peptide (CGRP), a neuropeptide implicated in the pathophysiology of migraine. Promising outcomes have been obtained with monoclonal antibodies and, more recently, with a new class of drugs consisting of small-molecule CGRP receptor antagonists, known as gepants. Atogepant, an oral agent of the gepant class, has demonstrated efficacy in migraine prevention, yet its effects on central brain activity and neurotransmitter circuits remain largely unclear. This exploratory study investigates changes in brain functional connectivity and changes related to key neurotransmitter systems, following 12 weeks of treatment with atogepant in a group of EM patients.<h4>Methods</h4>This is an exploratory single-arm longitudinal study assessing clinical and neuroimaging changes before and after 12 weeks of atogepant treatment. We enrolled patients diagnosed with EM according to ICHD-3 criteria, without prior exposure to anti-CGRP therapies. Participants underwent clinical assessments (monthly migraine days MMD, acute drugs intake MAM) and resting-state functional MRI (rs-fMRI) before and after 12 weeks of treatment with atogepant 60 mg once daily. Longitudinal functional connectivity analyses were performed at the whole-brain level with a region-of-interest analysis. Additionally, longitudinal neurotransmitter-related functional connectivity was investigated within the serotoninergic and the dopaminergic systems.<h4>Results</h4>A total of 15 patients completed the evaluation. A significant group-level reduction in MMD (T = -7.09, <i>p</i> < 0.001) and MAM (T = -6.35, <i>p</i> < 0.001) following 12 weeks of atogepant treatment was observed, accompanied by a reduction of allodynia symptoms, albeit not statistically significant. Patients exhibited significant longitudinally increased functional connectivity, involving the right superior frontal gyrus, bilateral putamen, left pallidum, left anterior and bilateral posterior cingulate cortex. The greater the longitudinal increase in anterior-posterior cingulate cortices connectivity, the larger the improvement in MMD, MAM, and allodynia symptoms after treatment. Additionally, longitudinal connectivity changes were observed within the orbitofrontal cortex in the mesocorticolimbic dopaminergic system.<h4>Discussion</h4>In this exploratory cohort, significant clinical improvement after atogepant treatment was accompanied by longitudinal functional connectivity changes in EM patients. These preliminary findings may reflect either direct or indirect central modulation linked to atogepant treatment. A deeper understanding of the observed central changes may help to clarify the mechanisms underlying anti-CGRP therapies for migraine.

Autoimmune gastritis (AIG) arises from a T-cell-mediated immune attack on the gastric parietal cell and carries a dual neoplastic risk: gastric adenocarcinoma through spasmolytic polypeptide-expressing metaplasia (SPEM) and intestinal metaplasia, and type-1 gastric neuroendocrine tumour (NET) through hypergastrinaemia-driven enterochromaffin-like (ECL) cell hyperplasia. This "one origin, two fates" framework is conceptually established but has never been given a molecular trajectory from an AIG origin, and molecular data for the neuroendocrine endpoint are essentially absent. We integrated public single-cell transcriptomes of the human AIG corpus (GSE271866; three samples, 14,323 cells) and of a histologically staged premalignant cascade (GSE134520; 44,012 cells) with a cross-sectional cohort of approximately 203 AIG patients, treating the two axes asymmetrically: the adenocarcinoma axis was reconstructed at the molecular level, whereas the neuroendocrine axis was clinically and literature anchored. The AIG corpus showed near-absent parietal cells (0.45%) with extensive SPEM (18.6%) and intestinal metaplasia (16.3%). Cascade pseudotime increased monotonically with histological stage (Spearman's <i>ρ</i> = 0.496; unchanged on the unbridged graph) and defined a gastric-to-intestinal progression signature that, transferred to the AIG corpus, tracked the AIG-intrinsic pseudotime (<i>ρ</i> = 0.673). Most AIG metaplastic cells were projected to the strong-metaplasia stage (90.9%; bootstrap 95% CI 84-98%; <i>n</i> = 55), a suggestive rather than definitive placement that is nonetheless consistent with AIG occupying the atrophy-to-metaplasia segment of the adenocarcinoma axis. In the cohort, hypergastrinaemia scaled with corpus atrophy (gastrin-17 versus pepsinogen I/II ratio <i>ρ</i> = -0.413, <i>p</i> = 9.9 × 10^-8), and single-cell analysis confirmed a gastrin-responsive ECL population (<i>HDC</i> ⁺ 93%, <i>CCKBR</i> ⁺ 76%) whose neuroendocrine programme was orthogonal to the intestinal programme of the adenocarcinoma arm. Because AIG is a prototypic autoimmune disease, we frame how the autoimmune infiltrate steers each fate as the central open immunological question. AIG thus enters the metaplasia-to-cancer axis from a corpus, parietal-cell-loss origin and diverges, through a serum-gastrin-ECL bridge, towards the neuroendocrine fate; we provide an interpretable progression signature and a severity-anchored, hypothesis-generating framework-not an outcome predictor, since the cohort is cross-sectional with essentially no neoplastic endpoints-and report the molecular data gap for the neuroendocrine endpoint as a field-level priority.

<h4>Background</h4>Cardiovascular disease represents the primary cause of mortality among individuals with metabolic dysfunction-associated steatotic liver disease (MASLD). We aimed to examine the association between grip strength and the risk of incident major adverse cardiovascular events (MACE) in this population.<h4>Methods</h4>This prospective cohort analysis included participants with MASLD from the UK Biobank. Participants were categorized into high, medium, and low grip strength groups based on sex-specific tertiles. Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for the associations between grip strength and incident MACE and its components (heart failure, myocardial infarction, and stroke).<h4>Results</h4>A total of 119, 004 participants with MASLD were included (mean [SD] age, 57.24 [7.89] years; 69, 421 men [58.3%]). During a median (IQR) follow-up of 13.6 (12.7-14.4) years, 14, 137 (11.9%) incident MACE cases were recorded. Compared with the high grip strength group, the low grip strength group exhibited a higher risk of MACE (HR = 1.35, 95% CI: 1.29-1.41, <i>P</i> < 0.001), heart failure (HR = 1.52, 95% CI: 1.42-1.62, <i>P</i> < 0.001), myocardial infarction (HR = 1.26, 95% CI: 1.19-1.34, <i>P</i> < 0.001), and stroke (HR = 1.33, 95% CI: 1.21-1.45, <i>P</i> < 0.001). A significant interaction between grip strength and sex was observed (<i>P</i> for interaction = 0.001), with the association between low grip strength and MACE being stronger in women (HR = 1.45, 95% CI: 1.34-1.56, <i>P</i> < 0.001) than in men (HR = 1.29, 95% CI: 1.16-1.45, <i>P</i> < 0.001). No significant interaction was observed for age.<h4>Conclusion</h4>In individuals with MASLD, lower grip strength was associated with an increased risk of MACE, with a stronger association observed in women. These findings highlight grip strength as a potential marker of cardiovascular risk among individuals with MASLD.

<h4>Background</h4>Manganese (Mn)-induced neurotoxicity is a major public health concern. The aim of this study is to assess whether retinoic acid, a metabolite of vitamin A, can counteract manganese-induced neurotoxicity in SH-SY5Y cells by counteracting epigenetic dysregulation.<h4>Methods</h4><i>In vivo</i>, we assessed neurobehavioral function, striatal pathology, synaptic ultrastructure, Mn content, dopamine levels, and expression of Trim28, α-synuclein (α-syn), H4K91ac, and DAT in Mn exposed rats. <i>In vitro</i>, we investigated the interactions between α-syn, Trim28 and H4 using immunofluorescence, molecular docking and immunoprecipitation techniques. We used chromatin immunoprecipitation to detect the enrichment of H4K91ac within the DAT promoter region. Additionally, we knocked down α-syn and Trim28 using siRNA to explore their potential molecular mechanisms.<h4>Results</h4>Mn exposure induced behavioral deficits, synaptic damage, and dopaminergic dysfunction in rats. Mechanistically, Trim28 mediated α-syn nuclear translocation. Nuclear α-syn bound histone H4, reducing H4K91ac enrichment at the DAT promoter, suppressing DAT expression, and disrupting dopamine homeostasis. Knockdown of Trim28 or α-syn reversed these effects. Retinoic acid inhibits the nuclear translocation of α-syn by modulating Trim28, and restores the levels of H4K91ac and DAT, thereby alleviating manganese-induced neurotoxicity in cells.<h4>Conclusion</h4>This study elucidates the role of the Trim28/α-syn/H4K91ac axis in manganese-induced dopaminergic dysfunction and identifies Trim28 as a potential target for retinoic acid.

The alpaca (<i>Vicugna pacos</i>) is a domesticated South American camelid of increasing global relevance, valued for its high-quality fiber, cultural significance, and unique biological traits including heavy-chain-only antibodies. Its reproductive physiology differs fundamentally from that of conventional livestock, encompassing an induced rather than spontaneous ovulatory mechanism in females and an unusually viscous ejaculate in males, differences that constrain the direct application of protocols developed for cattle, sheep, and swine and that have historically limited the pace of genetic improvement in the species. This review provides an integrated account of alpaca reproductive anatomy, follicular and neuroendocrine physiology, seminal plasma biochemistry, and semen cryopreservation, together with the current state of artificial insemination, multiple ovulation and embryo transfer, laparoscopic ovum pick-up, <i>in vitro</i> embryo production, and intracytoplasmic sperm injection. By examining female and male reproductive biology within a single mechanistic framework rather than treating each in isolation, this review draws out the regulatory pathways that connect the two systems and identifies the knowledge gaps that a fragmented, single-sex treatment of the subject would otherwise leave unaddressed. Particular attention is given to the biological determinants that most directly limit ART efficiency and to the translational steps required to move current protocols from experimental proof of concept toward standardized, field-applicable practice. Building on prior reviews that have illuminated individual ART modalities and single-sex reproductive biology, this work is intended to support researchers and practitioners advancing reproductive technology in camelids and the broader effort to conserve and genetically improve alpaca populations worldwide.

HFO-153-10mczz-E (decafluoro-3-hexene) is a volatile, nonflammable hydrofluoroolefin (HFO) developed as a dielectric fluid in heat transfer applications, such as two-phase immersion cooling (2-PIC). Its high vapor pressure and low water solubility indicate that inhalation is the primary route of exposure and that if released into the environment, the molecule would primarily partition into the air. The available toxicology dataset primarily comprises studies performed with Good Laboratory Practice (GLP) and under Organisation for Economic Co-operation and Development (OECD) test frameworks. This dataset includes acute, subacute, and subchronic inhalation studies, and reveals low systemic toxicity in rats at concentrations of up to approximately 3,000 ppm, with no treatment-related findings in reproductive or developmental endpoints. An <i>in vitro</i> and <i>in vivo</i> genotoxicity battery yielded no indication of genotoxicity. Additional studies illustrated no cardiac sensitization in dogs up to 5,000 ppm and no evidence of ocular/skin irritation or skin sensitization. Environmental testing across algae, invertebrates, and fish revealed no adverse effects in algae or daphnia, with the exception of acute toxicity in fish. However, interpretation was constrained by substantial volatility-related loss of the test material. Environmental partitioning, modeled bioaccumulation factors, and physicochemical properties collectively indicate a low bioaccumulation potential. Collectively, the empirical data characterize HFO-153-10mczz-E as a highly volatile fluorinated fluid with a well-defined toxicological profile, with available data suggesting a low human-health hazard and limited environmental exposure potential under plausible use conditions, though further studies may be warranted.

Alcohol-associated liver disease (ALD) is a growing global health concern, with alcohol-associated hepatitis (AH) leading to the highest morbidity and mortality. Available therapies are limited and often inadequate. Platelets contribute in a variety of ways to liver disease pathogenesis, but their role in AH remains largely unexplored. In this study, we addressed the hypothesis that platelets contribute to pathological inflammation in AH. Using patient samples and a multiomics approach, we found that platelets undergo proinflammatory transcriptomic and proteomic changes in AH, with 2 alarmins, S100A8 and S100A9, being among the top upregulated genes/proteins. Additionally, the abundance of platelet-derived microparticles containing S100A8 and S100A9 in AH patient plasma was increased and correlated with disease severity (assessed by model for end-stage liver disease sodium [MELD-Na]) and endotheliopathy (assessed by ICAM1, CXCL8, and vWF). We mechanistically linked S100A9 with endotheliopathy via crosstalk between primary human liver sinusoidal endothelial cells and primary human monocytes. We also demonstrated that IL-6 upregulates S100A9 in megakaryocytic cells in a JAK/STAT-dependent manner, modeling changes occurring in the bone marrow in patients with AH. Our studies establish proinflammatory platelets as important contributors to AH pathology. Moreover, antiplatelet agents - or, more specifically, S100A9 targeted drugs - are potential therapeutic strategies in AH.

Despite advances in delineating the molecular diversity and projection patterns of midbrain dopamine (DA) neurons, subtype-specific contributions to motor learning and movement execution remain poorly defined. Here, we applied intersectional ablation and inhibitory chemogenetics to dissect the roles of calbindin-expressing (CALB1⁺) and nonexpressing (CALB1^-) DA neurons in locomotion. Using newly engineered intersectional autocleavable Caspase3 constructs, we ablated CALB1⁺ or CALB1^- DA neurons in the mouse midbrain. CALB1^- DA neuron ablation caused severe weight loss, whereas CALB1⁺ DA neuron ablation produced no overt health impairments. Nonetheless, loss of either subtype led to a bradykinetic-like phenotype on the initiation and vigor of voluntary movements. Only ablation of CALB1^- DA neurons impaired performance on the accelerated rotarod. To test if these phenotypes are the result of DA subtype activity, we silenced either population using the inhibitory DREADD hM4Di. Consistent with ablation, silencing CALB1^- DA neurons impacted the initial performance on the rotarod, whereas inhibition of CALB1⁺ DA neurons did not impact performance on the first day, but prevented across-day improvement. Silencing both populations impaired the initiation and vigor of voluntary movements. We next investigated whether this locomotor phenotype stemmed from reduced DA release in the dorsolateral striatum (DLS). While CALB1^- silencing abrogated DA transients in the DLS, CALB1⁺ silencing unexpectedly resulted in increased transients in DLS. Thus, our results demonstrate that DA transients in the DLS are not invariably coupled with movement execution. Altogether, these findings uncover both distinct and shared roles of molecularly defined DA subtypes in shaping different aspects of locomotion.

Neurological disorders are the leading cause of health loss worldwide. The growing number of patients suffering from such conditions calls for improved strategies for their prevention, diagnosis, and therapy. To better understand human pathologies, relevant models and methodologies must be made available. In this study, we focused on a biomedical model capable of recapitulating the complexity of human pathology, the pig (Sus scrofa). Brain tissue and cerebrospinal fluid samples from a transgenic minipig model of Huntington's disease were subjected to multiple extraction and fractionation steps. A proteomic mass spectrometry (MS) methodology then allowed the generation of a porcine spectral library for 8,321 proteins. Using data-independent acquisition (DIA), we demonstrated that our porcine spectral library substantially enhanced the quantitative potential of this untargeted MS approach, generating reproducible proteome-wide data. The porcine library also provides a comprehensive resource for the development of targeted MS assays, enabling the quantification of selected proteins with a key role not only in neuroscience.

NADPH oxidases (NOXs) have emerged as central hubs that link environmental, metabolic, and immune cues through spatially organized redox signaling. However, their roles across tissues and disease states have not been comprehensively evaluated in an integrated manner. This review integrates recent advances in structural biology, immunometabolism, toxicology, and systems biology to provide an updated, comprehensive, and accessible view of NOX biology. Recent advances in high‑resolution cryo-EM, AlphaFold‑based modeling and molecular dynamics studies have provided new insights into NOX architecture, catalytic sites, post‑translational modifications and regulatory mechanisms, and docking interfaces for RAC1 and p47^phox. Emerging evidence further indicates that cellular NOX-derived ROS can reprogram macrophage and T-cell metabolism, stabilize HIF-1α, and tune the balance between effector and regulatory states, thereby linking NOX activity to checkpoint control and tumor immune escape. A second focus is on how real‑world pollutants converge on NOX isoforms as proximal mediators of redox signaling across lung, vascular, hepatic, renal, and neural tissues. NOX activation during cellular injury may contribute to oxidative stress, mitochondrial dysfunction, inflammasome activation, and fibrotic signaling, through extracellular vesicles, lipid rafts, and noncoding RNAs. Finally, the review evaluates emerging therapeutic strategies, including isoform-selective/pan-NOX/peptide inhibitors, and nanozymes. It also discusses emerging approaches such as exosome-based biomarkers, network pharmacology, and machine learning for patient stratification and pharmacodynamic monitoring. By highlighting key mechanistic gaps and translational opportunities, this review establishes NOXs as actionable nodal regulators at the intersection of immunity, metabolism, environmental exposure, and human disease.

Volumetric MRI of the caudate and putamen is a robust biomarker of striatal degeneration in Huntington's disease (HD). Striatal degeneration may be reflected beyond volume alone, in additional shape and intensity alterations. We evaluated five novel morphometric biomarkers of the caudate and putamen-curvature, flatness, eccentricity, elongation, and focal assessment of divergence estimation (FADE; T1-weighted signal shift)-using cross-sectional and longitudinal MRI data from healthy controls (N = 100), individuals with premanifest HD (preHD; N = 100), and individuals with early manifest HD (earlyHD; N = 100). Geometric measures showed significant cross-sectional group differences with large effect sizes, although volume remained the most sensitive individual biomarker. Longitudinally, most geometric measures detected abnormal rates of change, particularly in the caudate, with medium-to-large effect sizes. The direct comparison between preHD and earlyHD proved challenging for all biomarkers; however, elongation yielded the largest effect size and outperformed volume in this comparison. FADE detected abnormal signal increases in HD groups at baseline but not longitudinally. Power analyses confirmed that volume retains the highest sensitivity, while geometric measures follow closely. Combining volume and geometric measures improved the cross-validated classification accuracy starting at low sample sizes. Correlations between scores and results from atrophy simulations confirmed that HD leaves two signatures on the striatum: loss of volume and loss of shape. While volume remains the strongest standalone biomarker, geometric measures provide complementary information that sharpens disease detection and may enhance biomarker frameworks in future HD clinical trials.

<h4>Background</h4>Type 1 diabetes mellitus (T1DM) is a chronic disease that significantly impacts patients' quality of life. Its prevalence is rising globally each year. This study aims to identify potential biomarkers associated with T1DM through comprehensive bioinformatics analysis, further enhancing T1DM early diagnosis and treatment.<h4>Methods</h4>Transcriptome datasets from T1DM patients and the control group were from the Gene Expression Omnibus (GEO) database. Differentially Expressed Genes (DEGs) were identified and subsequently analyzed using Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and protein-protein interaction (PPI) network analysis. Hub genes were identified using Enzyme-Linked Immunosorbent Assay (ELISA) on clinical samples comprising 17 T1DM patients and 19 controls. Immune cell infiltration was estimated using the Cell-type Identification By Estimating Relative Subsets Of RNA Transcripts (CIBERSORT) algorithm, while the diagnostic performance of the hub genes was evaluated <i>via</i> receiver operating characteristic (ROC) curve analysis.<h4>Results</h4>A total of 20 up-regulated and eight down-regulated DEGs were identified in the GEO database. Functional enrichment analysis showed that immune activation played an important role in T1DM. The expression levels of the hub genes, <i>CTSG</i> and <i>LTF</i>, were further validated in clinical samples. ROC analysis showed moderate diagnostic performance, with AUC values of 0.75 (training set) and 0.67 (validation set).<h4>Conclusions</h4>The results indicate that <i>CTSG</i> and <i>LTF</i> may serve as promising diagnostic biomarkers for T1DM. Our study is positioned as exploratory with moderate diagnostic relevance rather than definitive biomarker discovery. The findings are preliminary and require further validation before any clinical application.

<h4>Introduction and importance</h4>Primary gastric melanoma (PGM) is an exceedingly rare malignancy, especially in Asian populations, that presents major diagnostic and therapeutic dilemmas.<h4>Presentation of the case</h4>We report the case of a 53-year-old Vietnamese man who presented with only vague epigastric discomfort. During a routine esophagogastroduodenoscopy, a small pigmented lesion was incidentally discovered in the stomach. An initial biopsy confirmed the presence of malignant melanoma, supported by immunohistochemical positivity for SOX10. A subsequent comprehensive workup - including dermatological and ophthalmological evaluations and comprehensive systemic imaging - found no other primary melanocytic sites, thereby fulfilling three of Blecker's criteria. A total laparoscopic gastrectomy with D2 lymphadenectomy was performed. Postoperative histopathology showed no residual tumor in the gastric specimen or in any of the lymph nodes examined.<h4>Clinical discussion</h4>The absence of established treatment standards for PGM necessitated a complex multidisciplinary discussion. Key questions centered on balancing oncologic radicality (total gastrectomy, D2 lymphadenectomy) with patient quality of life, especially given the likelihood of complete excision at the time of the initial biopsy. The finding of no residual disease post-resection complicated the indication for adjuvant systemic therapy. The diagnosis was supported by SOX10 staining and the fulfillment of Blecker's criteria (pending completion of the 6-month surveillance period). The upfront radical approach, though potentially excessive, preempted the need for a second operation to confirm pathological clearance.<h4>Conclusion</h4>This report contributes to the limited literature on PGM, highlighting the importance of early detection, multidisciplinary discussion, and long-term follow-up in such rare presentations.

<h4>Background</h4>Huntington's disease is prevalent globally, with approximately 4.88 cases per 100,000 people, based on a systematic review and meta-analysis of 33 studies published between 2010 and 2022. Despite its significant prevalence, no proper treatment is available that directly addresses Huntington's disease. The existing treatments focus on symptom management, such as controlling chorea and psychiatric symptoms. The drugs used for this purpose may also cause side effects, including depression and Parkinson's disease.<h4>Methods</h4>An integrated experimental and computational approach was employed, involving cold maceration extraction of <i>Sesamum indicum</i>, LC-MS/MS based phytochemical profiling, ADMET screening, molecular docking, molecular dynamics simulations, and MMBPSA binding free energy analysis to identify potential inhibitors of Huntington's diseases associated targets.<h4>Results</h4>Among the LC-MS/MS identified compounds, eight compounds satisfied the ADMET criteria. Valdiate (PubChem CID: 129715809) demonstrated favourable multi-target binding with docking score of -7.3 kcal/mol against HDAC4 and -8.9 kcal/mol against HDAC7. Molecular dynamics simulations confirmed stable protein-ligand interactions, while MMPBSA analysis yielded binding free energies of -23.22 ± 2.45 kcal/mol (HDAC4) and -28.27 ± 2.28 kcal/mol (HDAC7), identifying Valdiate as the most promising potential inhibitor.<h4>Conclusion</h4>Valdiate may serve as a potential inhibitor of mutant huntingtin-associated pathological pathways, pending further <i>in-vitro</i> and <i>in-vivo</i> validation.