Gene Literature Dashboard

Congenital antithrombin (AT) deficiency, primarily caused by <i>SERPINC1</i> variants, is a major risk factor for venous thromboembolism (VTE). We previously reported the <i>SERPINC1</i> p.M313T variant in three VTE probands with normal AT activity and antigen levels. This study aims to elucidate its pathogenetic mechanism.Thrombin generation test (TGT), thermal stability, native-urea PAGE, <i>in vitro</i> protein expression, and enzymatic assays were performed. Glycosylation analysis was conducted using glycosidase treatment, and structural analysis was performed through molecular dynamics simulation.AT in probands' plasma samples exhibited reduced thermostability and increased proportions of denatured and latent forms compared with normal pooled plasma. The recombinant AT-M313T protein exhibited increased inhibitory activity, consistent with findings in proband plasma based on AT activity and TGT. Despite this enhanced activity, the mutant protein demonstrated reduced thermostability and a marked tendency to transition into the latent form, potentially predisposing carriers to thrombosis under stress conditions. These characteristics may result from the introduction of abnormal O-linked glycosylation within the breach region, confirmed in both plasma-derived and recombinant AT. Molecular dynamics simulation revealed a less compact structure, with increased spacing in the shutter region and enhanced flexibility of the reactive center loop.The <i>SERPINC1</i> p.M313T variant exhibits dual characteristics of high inhibitory activity and low structural stability, which together contribute to the transient AT deficiency. These findings suggest that AT deficiency may be underdiagnosed and highlight the importance of integrating techniques such as native-urea PAGE into standard diagnostic workflows to identify variants associated with structural abnormalities.

BACKGROUND: Monogenic epilepsies are a group of rare disorders that manifest in early childhood and are usually associated with neurodevelopmental abnormalities and poor drug response. The field of genetic epilepsy is continuously evolving in alignment with the vast advancements in genetic testing in the last decade. METHODS: Recent studies and databases were reviewed to explore the latest updates in monogenic epilepsies. FINDINGS: This literature review reveals the remarkable progress in understanding monogenic epilepsy classification, genetic aetiologies, and treatment over the past decade. The International League Against Epilepsy classification system continues to evolve, providing increasingly precise diagnostic frameworks that incorporate genetic and etiologic information. The genetic landscape of epilepsy has expanded dramatically, with around 2000 genes now associated with seizures and/ or epilepsy pertaining to complex and overlapping neuronal networks. Besides the locus heterogeneity, monogenic epilepsy phenotypes exhibit phenotypic variabilities due to differences in pathogenic causative variants as well as unique genetic backgrounds in affected individuals. Identification of the underlying molecular aetiology permits personalized management approaches which are currently limited approved agents. Genes encoding antiepileptic drug targets, metabolic enzymes and transporters are additionally implicated in the precision medicine. CONCLUSION: Current knowledge in Locus heterogeneity should be addressed in epilepsy phenotypes to avoid extensive diagnostic delays particularly in cases that can be rescued with available management options.

Targeted therapy, which modifies genes and their expression, holds great promise for treating a variety of diseases, including cancer, inborn errors of metabolism, and acute and chronic inflammatory and infectious conditions. However, it also presents challenges related to RNA delivery, immune responses, side effects of delivery vectors, and the need for individualized formulations. To overcome these limitations, the choice of lipids and formulation processes might be re-evaluated, with a focus on eliminating critical components, such as poly(ethylene glycol) (PEG) and ethanol. Thus, a purely water-based formulation for lipid nanoparticles was developed, offering a material-efficient, time-saving process with high reproducibility. Initially, a stealth lipid containing poly(2-methyl-2-oxazoline) (PMeOx) was used, and the formulation was later expanded to include approved lipids. These nanoparticles not only efficiently transfect primary human immune cells but also effectively deliver multiple nucleotides in CRISPR-Cas9 applications. Moreover, an in vivo comparison revealed that the nanoparticles exhibited preferential transfection in extrahepatic tissues. This distinguishes them from conventional cholesterol-rich lipid nanoparticles, which primarily target the liver regardless of the application route.

Spontaneously occurring primary canine pulmonary adenocarcinoma (cPAC) exhibits clinicopathological and molecular similarities to never-smoker human lung cancers. Shared genomic alterations, including point mutations, indel mutations and copy number changes particularly in HER2 signalling, are significant therapeutic targets, especially for HER2 and tyrosine kinase inhibitors. Whilst progress has been made in identifying mutational drivers in canine cancers, the role of somatic gene fusions in cPAC remains poorly understood, despite their importance in other cancers as drivers and therapeutic targets. This study investigates the fusion landscape in cPAC by analysing RNA-seq data from a cohort of 36 primary tumour samples and reports oncogenic fusions with therapeutic potential. Notably, NRG1 fusions were identified in a subset of tumours, including recurrent SDC4::NRG1 events, potentially playing key roles in disease progression. NRG1 fusions, known to activate HER2 signalling, are mutually exclusive with HER2 gene alterations, indicating convergence on the same pathway. Tumours with SDC4::NRG1 fusions also overexpress HER2 pathway-related genes, reinforcing NRG1-driven activation. Similar fusions occur in never-smoker human non-small cell adenocarcinoma lacking other common drivers, underscoring their therapeutic importance. These findings highlight NRG1 fusions as critical contributors to cPAC tumorigenesis and warrant further clinical and comparative investigation. Additionally, novel fusions disrupting the PTEN axis were identified, leading to truncated PTEN and associated protein domains. These disruptions could impair tumour-suppressive pathways, presenting additional therapeutic targets. This research emphasises the broader relevance of fusion-driven mechanisms in cPAC tumorigenesis, advancing the understanding of both canine and human lung cancers for clinical and comparative studies.

<h4>Background</h4>Obstructive hypertrophic cardiomyopathy (HCM) and hereditary hemochromatosis (HH) rarely coexist. The therapeutic role of cardiac myosin inhibitors in this setting has not been described.<h4>Case summary</h4>A 67-year-old woman with known homozygous C282Y HH was referred for an abnormal stress echocardiogram. Cardiac magnetic resonance showed septal hypertrophy, systolic anterior motion of the mitral valve with left ventricular outflow tract obstruction, and iron overload. Owing to beta-blocker intolerance, mavacamten was started, with marked reduction in left ventricular outflow tract gradient.<h4>Discussion</h4>This case highlights the complexity of diagnosing and managing obstructive HCM in the context of HH. To our knowledge, this is the first reported use of mavacamten in this setting, demonstrating a favorable hemodynamic and clinical response.<h4>Take-home messages</h4>Multimodality imaging is essential to differentiate primary obstructive HCM from HH-related cardiomyopathy. In this case, mavacamten was a well-tolerated alternative, though further data are needed to establish broader applicability.

Huntington's disease (HD) is characterized by heterogeneous rates of clinical progression, complicating patient monitoring and clinical trial design. Although speech and language alterations are increasingly recognized as part of the HD cognitive phenotype, their value as short-term prognostic biomarkers of clinically meaningful disease progression and neurodegeneration remains unestablished. In this prospective 12-month longitudinal study, we investigated whether objectively quantified spontaneous speech and language measures predict short-term clinically meaningful progression and relate to biomarkers of neurodegeneration in HD. Eighty-six participants (42 manifest HD, 24 premanifest gene carriers, and 20 healthy controls) underwent baseline spontaneous speech assessment, structural MRI, and plasma neurofilament light chain (NfL) quantification. Clinically meaningful worsening was defined using validated minimal clinically important difference thresholds in the composite Unified Huntington's Disease Rating Scale (cUHDRS). Spontaneous speech and language measures progressively deteriorated across disease stages and were associated with reduced cortico-subcortical gray matter volume in distributed associative and integrative regions. In manifest HD, logistic regression analyses revealed that baseline language integrity independently predicted clinically meaningful worsening at 12 months (OR = 3.840, 95% CI = 1.46-13.33; AUC = 0.783). Combining speech-derived measures with plasma NfL improved discrimination accuracy of individuals with accelerated clinical progression (AUC = 0.807). Spontaneous speech represents an early, accessible and sensitive marker of neurodegeneration in HD. The combination of speech and language derived measures and plasma NfL enables accurate identification of individuals at risk of accelerated, clinically meaningful disease progression, supporting their potential utility as short-term prognostic biomarkers for clinical trials enrichment and stratification.

Circular RNAs (circRNAs) are covalently closed, single-stranded RNAs generated via backsplicing. They are highly stable and evolutionarily conserved, making them promising candidates for cancer therapy and diagnosis. CircRNAs regulate cancer progression by modulating genome instability, angiogenesis, metastasis, stemness, and chemoresistance. They do so through mechanisms including microRNA (miRNA) sponging, protein interaction, translational templating, and transcription/translation regulation. CircRNAs play a critical role in cancer immunotherapy. They modulate immune checkpoint blockade (ICB) responses and cytokine secretion to reshape the tumor immune microenvironment (TME). CircRNAs also serve as stable platforms for neoantigen-based cancer vaccines and improve in vivo chimeric antigen receptor T cell (CAR-T) therapy by replacing unstable linear mRNA. Additionally, circRNAs are potential noninvasive biomarkers due to their abundance in body fluids and differential tumor-normal expression. Despite challenges such as unclear regulatory networks, off-target effects, and inefficient delivery, this review systematically summarizes the biogenesis of circRNAs, their functional mechanisms, their roles in cancer progression, and their applications in cancer immunotherapy. The review also highlights their utility as biomarkers and future translational directions, providing a focused overview of their potential to advance cancer immunotherapy.

Chagas disease is a significant public health problem with benznidazole (BZ) and nifurtimox being the only available treatments. This study evaluated the trypanocidal activity of disulfiram (DS) in Swiss mice infected with the Y and VL-10 <i>Trypanosoma cruzi</i> strains. DS was tested alone or in combination with BZ at different doses for 20 consecutive days during the acute or chronic phase of the infection. Parasitological cure, inflammation in the heart and colon, and serum nitric oxide levels were assessed; in Y strain-infected animals, liver oxidative damage was also evaluated, while in VL-10-infected animals, parasitemia, survival rate, and collagen formation were analyzed. The combination of BZ + DS (100 mg/kg +50 mg/kg) induced 50% parasitological cure, reduced cardiac inflammation in Y strain infection, and decreased cardiac fibrosis in VL-10 strain infection. However, the results suggest a limited adjuvant effect of disulfiram, warranting further investigation.

<h4>Background</h4>Iron deficiency is the most common micronutrient deficiency worldwide, negatively affecting quality of life and health. Young women are at particularly high risk due to increased iron demands during growth, menstrual blood losses, and dietary habits. There are no official guidelines recommending screening for iron deficiency in healthy young women. This study aimed to develop a screening instrument (IRON-5) to predict iron deficiency in this group, with relevance for use in primary care and school health services.<h4>Methods</h4>In 2023, a cross-sectional study was conducted to assess the prevalence of iron deficiency and associated factors in high school girls. A questionnaire assessing dietary habits, menstrual bleeding patterns, iron supplement use, and hormonal contraceptive use was administered. Iron deficiency was defined as serum ferritin <15 µg/L. A post-hoc analysis identified the strongest predictors of iron deficiency and incorporated them into a risk-scoring model.<h4>Results</h4>The final instrument included five predictors: a non-omnivorous diet, heavy menstruation causing discomfort, avoidance of activities due to menstruation, no iron supplementation, and non-use of hormonal contraceptives. A scoring system (0-5) was developed. Using a threshold score of ≥ 3, the model achieved a sensitivity of 74% and specificity of 57% for detecting iron deficiency.<h4>Conclusion</h4>The IRON-5 is a concise and user-friendly screening instrument designed to identify young women at risk of iron deficiency. It demonstrated good sensitivity and may serve as a cost-effective method to prioritize individuals for blood testing. IRON-5 could support identification and prevention strategies in school-based and primary care settings.

Strong evidence has emerged over the last two decades implicating proteasome-dependent and independent protein polyubiquitination in the memory consolidation process. Recently, it was shown that multiple forms of polyubiquitination, including proteasome-dependent K48 and proteasome-independent M1 polyubiquitination, regulate fear memory formation in a sex-dependent manner in the amygdala. However, prior work focused on single time points during the postlearning period, leaving questions about whether these are true sex differences in polyubiquitin modifications that persist throughout the extended consolidation process. Here, using unbiased polyubiquitin-type specific proteomics, we identified the protein targets of K48 and M1 polyubiquitination in the amygdala 2 and 4 h after contextual fear conditioning. Notably, we found that while the sex differences in the targeting of proteins with these polyubiquitin modifications persist for several hours after fear conditioning, the temporal dynamics of these changes vary across males and females. Further, while target protein pathways vary significantly across sexes at every time point examined, there are several notable overlaps between males and females. Together, these data provide the first comprehensive analysis of sex differences in proteasome-dependent and independent protein polyubiquitination in fear memory formation and significantly advance our understanding of the potential sex-specific roles of diverse polyubiquitin modifications in the memory consolidation process.

Disruption of organelle interactions due to metabolic stress is a crucial factor in the pathological processes of many degenerative diseases. Compared with animal cells, the participation of chloroplasts enables plant cells to show defensive adaptation under stress. Therefore, delivering plant-derived photosynthetic systems into animal cells may help to establish a more stable organelle interaction network. Here, we show that plant-derived photosynthetic systems can effectively restore homeostasis of the interaction network of animal organelles. Specifically, plant-derived nanothylakoid units provide energy to animal cells, regulate intracellular Ca²⁺ homeostasis, increase endoplasmic reticulum (ER) lipid unsaturation and global membrane fluidity, reduce abnormal contact between mitochondria and ER, and alleviate mitochondrial dysfunction. By combining implantable light-emitting diodes with wireless charging, we expand photosynthesis therapy, enabling treatments for deeper tissues. This study provides a proof-of-concept for disease treatment based on the regulation of organelle interaction networks by natural photosynthetic systems and establishes a therapeutic approach for treating deep tissues.

The immune features in pediatric tumor are poorly explored. To characterize immune features of pediatric cancer, we performed an immunogenomic analysis of public database (TARGET) for pediatric solid tumor (PST) (n = 423) and pediatric hematological tumor (PHT) (n = 2302). We clustered PST and PHT samples into 5 subtypes (S1-S5) and 4 subtypes (H1-H4), respectively, based on immune features. In the PST cohort, cluster S1 with elevated expression of Wound_CSR (fibroblast core serum response in wound healing) and B cell signatures exhibited the worst overall survival. Conversely, cluster S4 (HR = 0.378, 95% CI: 0.24–0.59, P-value < 0.001) with down-regulated expression of these features was associated with prolonged survival. We also validated the prognostic significance of the S4 immune subtype in an independent neuroblastoma cohort from the ZJUCH (n = 127), which demonstrated favorable patient outcomes. In the PHT cohort, we observed that the relationships between immune clusters and prognosis differed between FLT3-ITD mutation-positive AML (AML-1) and FLT3-ITD mutation-negative AML (AML-2). In AML-1, cluster H2 featured upregulated infiltration of neutrophils, monocytes and antigen processing signatures, possibly leading to the worst overall survival. While in AML-2, cluster H2 exhibited a favorable outcome. The study highlights the potential of immune features as biomarkers for prognosis and treatment planning in pediatric cancers and provides novel insights into their immunological landscape.

PURPOSE: In this eight-year study, children with unclassified inborn errors of immunity (IEI) who have no pathogenic variants in targeted next-generation sequencing (tNGS) were re-evaluated by using singleton whole exome sequencing (sWES) and followed-up at least five years. They were also investigated if they had affected family members or not. In addition, all patients had detailed immunological and clinical work-up. METHODS: In 2017, 108 children with probable diagnosis of IEI had tNGS and pathogenic variants were detected in 38 (35.1%) patients who were not included into the study. The rest 70 patients were continued to be immunologicaly followed-up (67.0 ± 19.7 months) and 15 of them (21.4%) recovered spontaneously. Fifty-five patients were divided into two groups; (i) familial patients (n = 30); (ii) non-familial patients (n = 25). In all of them, sWES analysis was performed. RESULTS: There were 19 affected fathers, 9 mothers and 16 siblings whereas consanguinity rate for parents was 8.5%. In sWES analysis, no genetic alterations were detected in 28 (50.9%) patients and 14 cases showed predefined pathogenic variants which are not associated with IEI. Probable IEI causing variants were found in 13 of 55 cases. However, after genotype/phenotype examinations and segregation analysis by sanger sequencing for their parents, we were able to define only three pathogenic variants in three cases, namely two for IKBKB gene and one for PRKDC in familial cases. Fifty-five cases were screened with sWES and only three pathogenic variants were revealed (3/55, 5.45%). When we add this result to tNGS finding (35.1%), all together 40.55% of unclassifed IEI cases could be defined by both of these genetic investigations. CONCLUSION: Even with NGS-based approaches improving IEI diagnoses, detection rates still remain below 50%. Whole genome sequencing in the near future would offer the advantage of much more comprehensive and accurate variant detections.

As part of the Danish National Genome Centre (DNGC) initiative, the Central Denmark Region has implemented short-read whole-genome sequencing (srWGS) as a first-tier diagnostic tool for suspected monogenetic disorders. Despite increasing adoption of genome sequencing, evidence from large-scale implementation across clinical specialties remains limited. Here, we evaluate the implementation from patient inclusion and srWGS scaling to diagnostic performance. From 2021 to 2024, we sequenced 2317 patients with suspected genetic diseases across a wide range of medical specialities. Following clinical evaluation and informed consent, Illumina srWGS was performed. Patients were categorised into clinical subgroups based on phenotype and age to support targeted variant filtration and germline variant reporting. The primary outcome was diagnostic yield across all disease groups/categories. The project is a public/private partnership co-funded by the Novo Nordisk Foundation. Diagnostic yield ranged from 6% in children with cancerto 60% in patients with skin disorders, with an overall yield of 20%. We observed substantial variation in the clinical use of srWGS as a first-tier diagnostic tool across patient categories. Regional implementation of srWGS within the DNGC framework demonstrates its scalability as a first-tier diagnostic tool for monogenic disorders. Importantly, the combination of expert-guided inclusion criteria for srWGS and mixed public/private funding has ensured equitable access to genetic diagnostics. We identify patient groups with high diagnostic returns well suited for srWGS, as well as groups where alternative strategies could also be applied.

Amyotrophic lateral sclerosis (ALS) is a heritable disorder where rare variants with low-to-moderate penetrance are thought to dominate genetic risk. To identify such rare variants, we harmonized and analyzed exome data from 22 cohorts, totaling 17,919 individuals with ALS and 200,703 controls across discovery and replication phases. Rare variant analyses identified several new risk genes, with replication confirming association of YKT6 and supporting HTR3C, GBGT1 and KNTC1. We also provide strong, independent validation for genes with limited previous evidence: ARPP21, DNAJC7 and CFAP410. Notably, in ARPP21, we identified a new high-effect variant (p.P747L) and confirmed that p.P563L is an ALS-associated variant leading to an aggressive disease course. Beyond new discoveries, our analyses largely recapitulated the known genetic architecture of ALS, identifying risk variants in over 20% of cases and supporting a cumulative oligogenic risk model. These findings highlight new translational targets and show that rare variant analyses capture substantially more genetic risk than common variant genome-wide association studies.

BACKGROUND: The pathogenic role of B cells in pulmonary emphysema remains incompletely understood. This study aimed to delineate the specific B cell subsets and molecular mechanisms driving disease progression. METHODS: Using single-cell RNA sequencing (scRNA-seq), we characterized lung B cell heterogeneity in a murine model of emphysema induced by porcine pancreatic elastase (PPE). Functional roles were assessed via B cell depletion and pharmacological inhibition of a disintegrin and metalloprotease 10 (ADAM10). RESULTS: Emphysema triggered the formation of pulmonary tertiary lymphoid structures (TLS) enriched with proliferating BAFF⁺ B cells. scRNA-seq revealed a selective expansion of B1 cells, primarily the B1b subset, which was accompanied by increased ICOS expression on lung CD4⁺ T cells. B cell depletion attenuated alveolar destruction, abolished TLS, reduced serum autoantibody levels, and improved lung function. Mechanistically, ADAM10 was specifically upregulated in B1 cells. Functional studies demonstrated that ADAM10, through activation of the Notch signaling pathway, was essential for the proliferation of B1 cells and their capacity to activate CD4+ T cells. Pharmacological inhibition of ADAM10 recapitulated the therapeutic effects, reducing B1 cell numbers, suppressing TLS formation, diminishing ICOS⁺ CD4⁺ T cells and autoantibody titers, and ultimately ameliorating lung pathology and function. CONCLUSION: Our findings identify ADAM10 as a critical regulator of B1 cell-driven immunopathology in emphysema, highlighting the ADAM10-Notch axis in B1 cells as a potential therapeutic target.

BACKGROUNDS: Colorectal cancer (CRC) is one of the most common and fatal cancers worldwide, often diagnosed at advanced stages. Iron homeostasis, a crucial metabolic pathway in cancer, significantly impacts tumor progression. RESULTS: This study systematically investigated the function and significance of iron homeostasis-related genes (IHGs) in CRC, leading to the establishment of iron homeostasis-related molecular subtypes (IHMS) and a prognostic model based on their expression profiles of IHGs. Both IHMS and the model showed strong predictive accuracy for prognosis, clinical parameters, and response to immunotherapy. Patients classified as IHMS-B or exhibiting high model risk scores had poorer prognoses, while those categorized as IHMS-C or with low model risk scores were associated with more favorable outcomes and increased sensitivity to immunotherapy across multiple datasets. The model key gene HAMP (encoding hepcidin) was identified as a key oncogenic regulator that promotes CRC progression; elevated levels of HAMP/hepcidin were linked to peritoneal metastasis. Knocking out HAMP resulted in reduced tumor burden and inhibited proliferation and cancer stemness in both in vivo and in vitro models. Additionally, integration of in-house bulk-seq and tissue-array data further substantiated the clinical significance of HAMP/hepcidin and its association with immune landscapes. CONCLUSIONS: In conclusion, both IHMS and the model serve as effective predictors for CRC prognosis and patient stratification. HAMP/hepcidin, a central regulator of iron homeostasis, plays a pivotal role in driving the progression and metastasis of CRC, underscoring the therapeutic potential of targeting the HAMP-mediated iron axis to improve clinical outcomes in CRC.

Antibody labeling represents a critical step for the preparation of application-specific immunoreagents because the process can affect the binder functionality and result in a highly heterogeneous sample. Therefore, site-specific derivatization methods are preferable, and the addition of a cysteine is a common solution because its thiol group enables the reaction with several commercially available reagents. We first showed that the addition of an extra cysteine did not decrease the yield and functionality of nanobodies, indicating that it did not interfere significantly with the natural disulfide bond present in such macromolecules. The major limitation of single amino acid labeling is that the signal might be weak and compromise the sensitivity. Consequently, we evaluated the possibility of modifying a nanobody by introducing multiple cysteines into its sequence. The resulting variants were assessed for their labeling homogeneity and performance as reagents for flow and mass cytometry. The experimental data indicated that multiple cysteines favored the formation of dimers and resulted in multiple degrees of functionalization. However, the worst consequence was that their efficiency as reagents was significantly lower than that of nanobodies possessing a single extra cysteine.

Classic Hodgkin lymphoma (cHL) is composed of rare malignant Hodgkin and Reed-Sternberg (HRS) cells within a T-cell-rich tumor microenvironment (TME). Epstein-Barr virus (EBV) is present in ∼25% of cases, but its contribution to pathogenesis and immunomodulation remains unclear due to technical barriers. Using complementary spatial proteomics and transcriptomics across multi-institutional cohorts, we systematically map key EBV-linked TME reorganization. EBV-positive cHL exhibits distinct immunological features, including memory CD8 T cell enrichment, heightened T cell dysfunction spatially correlated with HRS proximity, and terminally exhausted T cell signatures contrasting with progenitor-exhausted patterns in EBV-negative disease. We identify EBV-encoded LMP1 as a factor in T cell dysfunction through enhanced HRS:CD8 interactions, and its expression level correlates with T cell terminal exhaustion in a distance-dependent manner. This spatial framework dissects viral-mediated immune evasion in the cHL TME, highlighting potential therapeutic opportunities to target virus-associated T cell dysfunction for precision immunotherapy in virus-associated malignancies.

<h4>Objective and background</h4>Postmenopausal osteoporosis (PMOP) significantly increases the risk of fragility fractures and has substantial negative effects on patients. Accumulating evidence suggests that the pathogenesis of PMOP is associated with ferroptosis. In this study, we screened and validated core ferroptosis-related genes (FRGs) in PMOP and investigated their potential links using bioinformatics analysis.<h4>Methods</h4>Differentially expressed genes (DEGs) were identified between PMOP patients and healthy postmenopausal women based on the GSE230665 dataset in the Gene Expression Omnibus (GEO) database. Weighted gene co-expression network analysis (WGCNA) was conducted to sort and extract significant module genes. FRGs were acquired from an online ferroptosis database. By intersecting DEGs, FRGs, and significant module genes, we identified ferroptosis-associated DEGs (FDEGs). We performed Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) analyses on FDEGs. Multiple algorithms within the cytoHubba plugin were employed to detect the core gene from the PPI network. The expression of the core gene was subsequently validated in the PMOP rat model, and the diagnostic efficacy of the core gene was evaluated using receiver operating characteristic (ROC) curves applied to external datasets. GeneMANIA and Gene Set Enrichment Analysis (GSEA) were conducted to explore important functions and pathways of the core gene. We further utilized the CIBERSORTx website to evaluate immune infiltration in PMOP and molecular docking analysis was performed to develop potential drugs for PMOP. Finally, we established a transcription factors (TFs)-mRNA-miRNAs regulatory network for mechanisms investigation.<h4>Results</h4>In this study, we identified phosphatase and tensin homologue deleted on chromosome 10 (PTEN) as the core gene through PPI analysis and various algorithms. qRT-PCR analysis and immunohistochemical staining revealed a significant elevation of PTEN expression in the PMOP rat models, and ROC curves demonstrated the relatively high diagnostic properties of PTEN. Moreover, GSEA analysis indicated that PTEN is involved in functions and pathways associated with MAPK signaling, oxidative phosphorylation, mTOR signaling, <i>etc.</i> We discovered three drugs that bind tightly to PTEN, including CX-5461, UMI-77, and GSK2256098. Ultimately, we constructed a TFs-mRNA-miRNAs network comprising 39 nodes (16 TFs, 22 miRNAs, and 1 mRNA) and 38 edges.<h4>Conclusion</h4>PTEN was preliminarily identified and verified as a key ferroptosis gene in postmenopausal osteoporosis through bioinformatics analysis, and the diagnostic model constructed based on PTEN was of high value. These findings may provide novel perspectives on the pathogenesis of PMOP at the transcriptome level and establish a theoretical foundation for further research.

<i>Hevea brasiliensis</i> (<i>H. brasiliensis</i>) is the principal source of natural rubber, but its productivity is severely threatened by powdery mildew caused by the biotrophic fungus <i>Erysiphe quercicola</i> (<i>E. quercicola</i>). Although circular RNAs (circRNAs) are emerging as key regulators in plant stress responses, their functions in <i>H. brasiliensis</i> immunity remain largely unexplored. Here, we aimed to systematically characterize circRNAs involved in the early immune response of <i>H. brasiliensis</i> to <i>E. quercicola</i>. Transcriptome sequencing and bioinformatic analyses identified 52 and 177 differentially expressed circRNAs (HbcircRNAs) at 1 and 3 days post-inoculation, respectively. Twelve HbcircRNAs with significant expression changes were validated, and nine were confirmed as true circRNAs. Functional assays using spray-induced gene silencing (SIGS) in <i>H. brasiliensis</i> leaves and heterologous overexpression in the <i>Arabidopsis thaliana eds1</i> mutant, which is susceptible to <i>E. quercicola</i>, revealed that HbcircARF3, HbcircSCSA1, and HbcircARFGAP8, together with their homologous linear counterparts (HbLinearRNAs), exert distinct regulatory effects on disease resistance. Silencing of these HbcircRNAs enhanced host immunity, whereas overexpression increased susceptibility. Pathway analyses suggested their involvement in auxin signaling, mitochondrial energy metabolism and vesicle trafficking. Collectively, our findings uncover the differential regulatory roles of circular and linear RNAs in the <i>H. brasiliensis-E. quercicola</i> interaction, providing mechanistic insights and potential molecular targets for breeding disease-resistant <i>H. brasiliensis</i>.

<h4>Introduction</h4>Ulcerative colitis (UC) is driven by mucosal inflammation and epithelial injury. Single cell RNA sequencing (scRNA-seq) enables high resolution profiling of immune, stromal and epithelial compartments in UC, whilst spatial transcriptomic (ST) and proteomic (SP) enable interrogation of cell-cell interactions, niche-specific expression profiles, and spatially restricted pathological processes. Although scRNA-seq, ST, and SP technologies have been rapidly evolved in the past ten years, relatively limited clinical applications have been demonstrated. This systematic review aims at comprehensively analysing current evidence in UC studies as an approach to identifying key limitations and proposing recommendations for strengthening future spatial research towards translations into clinics.<h4>Methods</h4>A comprehensive search of Embase, Medline, and grey literature was conducted to identify studies using scRNA-seq or spatial transcriptomic or proteomic technologies in adult UC cohorts. Outcomes of interest included insights into disease pathogenesis or treatment response.<h4>Results</h4>scRNA-seq studies revealed alterations across the innate and adaptive immune systems, as well as stromal and epithelial compartments in UC colonic tissue. Spatial studies provided insights into: (i) cellular composition of the UC microenvironment; (ii) inflammatory features in treatment responders versus non-responders; and (iii) ligand-receptor interactions as potential spatial biomarkers and therapeutic targets.<h4>Conclusion</h4>Overall, single-cell and spatial studies are deepening our understanding of UC pathogenesis and treatment response. However, they are often limited by small sample sizes and heterogeneous UC phenotypes. Future studies should prioritise robust cohort design and careful sample stratification. This will be critical to generating mechanistically precise, reproducible, and clinically meaningful insights into the heterogeneity of UC pathogenesis and treatment response.<h4>Systematic review registration</h4>https://www.crd.york.ac.uk/PROSPERO/, identifier CRD42024601628.

<h4>Objective</h4>This review article aims to determine the properties, uses, toxicity, and other side effects of crosslinking agents in tissue scaffolds when applied in vitro and in vivo.<h4>Methods</h4>A literature search was performed using the PubMed-NCBI (MEDLINE) database (https://pubmed.ncbi.nlm.nih.gov/) with keywords: <i>crosslinking reagent, collagen, hydroxyapatite</i>, and <i>bone regeneration</i>. GRADE criteria were used to assess the quality of evidence.<h4>Results</h4>A total of six articles were included in the study. Improved mechanical properties of collagen-hydroxyapatite scaffolds with high porosity can be achieved by employing crosslinking methods, including physical dehydrothermal (DHT) treatment, chemical treatment with glutaraldehyde (GA), Microbial Transglutaminase (mTGase), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), or a combination of both DHT and EDAC. Furthermore, the crosslinking of EDAC and DHT can lead to forming ester bonds between activated carboxyl groups and hydroxyl groups.<h4>Conclusion</h4>The combination of DHT and EDAC crosslinking can increase mechanical strength, make the pore size appropriate, make the scaffold more stable, and support cell adhesion so that new cells can grow, and the process of osteogenesis can run more optimally.

<h4>Background</h4>Gastric cancer (GC) is one of the most common malignant tumors in the digestive tract, characterized by a high degree of malignancy and a poor prognosis. Complex disease heterogeneity, late diagnosis, and suboptimal treatment often lead to poor prognoses for patients. In addition to genetic alterations and environmental factors, changes in epigenetic mechanisms also play an important role in the occurrence and progression of GC. Revealing the current research status of epigenetics in GC has important guiding significance for the diagnosis and treatment of GC, as well as for future research directions.<h4>Methodology</h4> In this study, we used specific keywords to search for publications related to gastric cancer epigenetics (GCE) published in the past 20 years in the Web of Science Core Collection database. The VOSviewer software (Version 1.6.20; Center for Science and Technology Studies (CWTS), Leiden University, Leiden, The Netherlands) was used to conduct clustering and co-occurrence analysis on countries, authors, institutions, and keywords in the publications. CiteSpace software (Version 6.3.1R6; Chaomei Chen, College of Computing and Informatics, Drexel University, Philadelphia, PA) was used for burst analysis and timeline clustering analysis.<h4>Result</h4>A total of 4,655 papers on GC in the field of epigenetics were retrieved. China is the country with the largest number of published papers (<i>n</i> = 1,876), while the United States has the highest centrality (0.32). Nanjing University ranks first among all institutions in terms of the number of published papers (<i>n</i> = 175). The author Ushijima Toshikazu from Japan has the highest number of published papers (<i>n</i> = 48). Yuan Yuan is a representative author of the recent research hotspots. In the keyword ranking, keywords related to DNA methylation are relatively numerous. Tumor microenvironment and immune infiltration are key buzzwords with research potential.<h4>Conclusions</h4>Based on bibliometric methods, this study provides new insights into the research progress of GC from the perspective of epigenetics and offers new ideas and inspirations for future GC research.

Dam-calf contact (DCC) rearing is a form of cow-calf contact (CCC) in which dairy cows nurse their calves for some months until weaning while additionally being milked throughout lactation. Growing implementation, consumer demands, and research interest indicate that CCC and DCC are viable systems for the dairy industry. However, machine milking in these systems presents notable challenges with multifactorial drivers. Those are discussed in this review based on current scientific knowledge regarding milking in nursing dams: Machine milk yield (MMY), flow and fat content of the harvested milk are reduced compared to non-nursing controls while protein and lactose levels vary and udder health is unaffected or improved. In systems with full-time DCC, relative MMY reduction can reach 60-70% during the nursing period whereas highly restrictive suckling regimes cause smaller MMY losses but may not meet the calf's nutritional needs. Post-weaning and lactation MMY can be reduced or unaffected. Low udder fills at the start of milking delay milk let-down potentially conflicting with standardized milking procedures and machine settings. A 0.5-1.5% decline in milk fat concentration indicates incomplete milk ejection and incomplete udder emptying which can further reduce MMY. These effects likely reflect a disturbance of the milk ejection reflex, as in nursing dams, suckling is a more potential stimulus for oxytocin (OT) release than milking. Underlying central mechanisms of inhibited OT release at milking in nursing dams may be related to the cow-calf bond affecting the dams' oxytocinergic system but the complexity of required experimental set-ups compromises research in cattle. In bonded dams, multisensory stimulation by the calf may be a stronger trigger for OT release than the milking procedure. Further, separation from the calf during milking and dam-individual variation regarding the ability to release milk in response to machine stimulation may play a role. Zooming out, social scientific studies reveal that DCC farmers see poor milkability as an unsolved challenge. However, it is not limited to nursing dams, as stress affects milk ejection. To quantify incomplete udder emptying, research should refine strip milk fat analysis for on-farm use. Cow-individual milk ejection dynamics must be better understood to assess if breeding for better milk let-down in response to machine milking while maintaining good maternity traits can improve milkability in DCC systems.

The spaceflight environment exposes biological systems to microgravity and cosmic radiation, which are factors known to induce oxidative stress and neurodegenerative processes. As the central nervous system is highly susceptible to disruptions in redox homeostasis, the development of effective strategies to safeguard astronaut health and cognitive function during extended space missions has become imperative. In this study, we investigate cerium oxide nanoparticles (nanoceria, NC) as an antioxidant agent for human neuron-like cells aboard the International Space Station. Nanoceria demonstrated excellent biocompatibility, strong antioxidant properties, and the ability to stimulate neurite extension under both Earth gravity and simulated microgravity. Following the return of the samples to Earth, transcriptomic analyses revealed that nanoceria effectively counteracted the detrimental transcriptional alterations triggered by spaceflight stressors, thereby maintaining neuronal homeostasis. Importantly, the expression of genes involved in antioxidant defense, mitochondrial activity, and dopamine metabolism remained stable in nanoceria-treated neurons, in contrast to the dysregulation observed in untreated controls. These findings position cerium oxide nanoparticles as promising antioxidant neuroprotectants for long-duration space missions and related neurodegenerative conditions.

Sheep production contributes to a secure and diverse food and fibre supply in the United States, with growing ethnic diversity strengthening demand. Katahdin is a composite hair-type sheep breed developed in the United States that has become the most popular breed in many regions of the country and the first one to have genomic selection implemented in its breeding program. Therefore, the main objectives of this study were to estimate variance components of reproductive traits, including number of lambs born (NLB), number of lambs weaned (NLW), age at first lambing (AFL), and interval from first to second lambing (LI), in Katahdin sheep using the AIREML method and the single-step Genomic Best Linear Unbiased Prediction (ssGBLUP) approach, and to identify genomic regions and candidate genes associated with these traits. The datasets used consisted of 127,536 animals in the pedigree, phenotypic records of 56,128 parities from 24,067 ewes, and genomic data from 10,032 animals with 30,308 single-nucleotide polymorphisms (SNP) after quality control. Analyses were performed using the BLUPF90 family of programs. We observed low heritability estimates for all studied traits (0.09 ± 0.00 for NLB, 0.08 ± 0.00 for NLW, 0.09 ± 0.01 for AFL, and 0.08 ± 0.01 for LI). The genetic correlations between the traits ranged from 0.17 ± 0.02 (AFL and LI) to 0.79 ± 0.02 (NLB and NLW). All traits were found to be highly polygenic with all 14 significant SNP on eight (OAR) chromosomes (3, 6, 7, 8, 9, 12, 13, and 15) having small effects on the total variability on the traits. These SNP were located near or within 18 candidate genes: four genes associated with NLB (AAK1, GFPT1, SLC23A2, and GDAP1), four with NLW (ARHGAP18, TTLL2, UNC93A, and GPR31), six with AFL (NAP1L5, FAM13A, HS3ST1, CCDC181, NME7, and BLZF1), and four with LI (TAF4, CDH4, CADM1, and SEL1L). These candidate genes have been previously associated with fertility, embryonic development, growth, disease resistance, and climatic adaptation traits. Our findings indicate that fertility and reproduction traits in Katahdin sheep can be improved through direct genetic selection. Genetic improvement for these traits will benefit from genomic selection as more accurate estimates of breeding values for selection candidates can be obtained at a younger age. Although the studied traits are influenced by a complex interplay of genetic and environmental factors, the candidate genes identified enabled a better understanding of the biological mechanisms underlying reproductive performance in Katahdin sheep.

Polycystic ovary syndrome (PCOS) is an endocrine and metabolic condition affecting reproductive health, marked by disrupted ovulation, excessive androgen levels, and impaired endometrial receptivity. Oxidative stress is an important part of its pathophysiological process. Atractylenolide III (ATIII), a major sesquiterpene lactone derived from Atractylodes macrocephala Koidz., exhibits antioxidant properties; however, its role and molecular mechanisms in PCOS have not yet been explored. A PCOS rat model was established using dehydroepiandrosterone (DHEA) and subsequently treated with ATIII or metformin. Estrous cycle, ovarian morphology, serum hormone levels, glucose metabolism, and endometrial receptivity markers were evaluated. Oxidative stress status was evaluated by detecting the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and antioxidant-related genes and proteins expression. Ovarian transcriptomic sequencing was performed to explore potential molecular pathways associated with ATIII treatment. ATIII administration was associated with restoration of estrous cycle, reduction of cystic follicles, improvement of ovulation, and normalization of serum hormones and metabolic parameters in PCOS rats. Ovarian stromal fibrosis was attenuated, and morphological and molecular markers of endometrial receptivity were improved. ATIII treatment was accompanied by increased SOD activity, decreased MDA accumulation, and restoration of NRF2-related antioxidant genes and proteins. Transcriptomic analysis revealed enrichment of oxidative stress-related pathways, within which Cybb (encoding NOX2) was found as a differentially regulated gene that was upregulated in PCOS ovaries then reduced following ATIII treatment. ATIII improves reproductive and metabolic abnormalities and enhances endometrial receptivity in a PCOS rat model. These results are mediated by attenuation of ovarian oxidative stress and modulation of Cybb-related redox signaling. These data offer mechanistic insight into the pharmacological potential of ATIII in PCOS.

In Parkinson's disease and other synucleinopathies, αSynuclein (αSyn) misfolds and forms Ser¹²⁹-phosphorylated aggregates (pSyn¹²⁹) with the factors controlling this process largely unknown. Here, we used arrayed CRISPR-mediated gene activation and ablation to discover new pSyn¹²⁹ modulators. Using quadruple-guide RNAs (qgRNAs) and Cas9, or an inactive Cas9 fused to a synthetic transactivator, we ablated 2304 and activated 2428 human genes related to mitochondria, trafficking, and motility functions in HEK293 cells. After exposure of cells to αSyn fibrils, pSyn¹²⁹ signals were recorded by high-throughput fluorescence microscopy and aggregates were identified by image analysis. We found that pSyn¹²⁹ was increased by activating the mitochondrial protein OXR1, which decreased ATP levels and altered the mitochondrial membrane potential. Instead, pSyn¹²⁹ was reduced by ablation of the endoplasmic reticulum (ER)-associated protein EMC4, which enhanced ER-driven autophagic flux and lysosomal clearance. OXR1 activation preferentially modulated cellular reactions to fibrils derived from multiple system atrophy (MSA) patients, whereas EMC4 ablation broadly reduced pSyn¹²⁹ across diverse αSyn polymorphs. These findings were confirmed in human iPSC-derived cortical and dopaminergic neurons, where OXR1 preferentially promoted somatic aggregation and EMC4 reduced both somatic and neuritic aggregates. These results uncover previously unrecognized roles for OXR1 and EMC4 in αSyn aggregation, thereby broadening our mechanistic understanding of synucleinopathies.

Lipid nanoparticles (LNPs) have demonstrated their effectiveness as carriers for the delivery of RNA therapeutics. As the core components of the LNP system, ionizable cationic lipids (ICLs) are still the major focus of research in this field. In this study, we reported a class of novel β-triketone-based ICLs synthesized via click chemistry. We demonstrated that the reaction could be completed instantaneously, and a library of ICLs was generated in a short time. In vitro and in vivo screening identified a lipid that was comparable to FDA approved Dlin-MC3-DMA lipid in siRNA LNPs-mediated gene silencing. In addition, we adopted a computational approach to characterizing the click chemistry in lipid synthesis, as well as the binding between siRNA and ICLs. Our work offers a novel approach to synthesizing ICLs of new structural features, which may not only improve our understanding of the structure-activity relationship (SAR) of ICLs but also lead to the development of improved LNPs for more effective delivery of nucleic acid therapeutics.

Transforming growth factor β (TGFβ) signaling pathways are integral for a plethora of biological processes. SMAD2 and SMAD3 are the principal transcriptional effectors of TGFβ superfamily ligands, yet quantitative, genome-wide mapping of their DNA-associated complexes under physiological contexts has remained limited due to the lack of specific, robust models. Here, we generated two versatile epitope-tagged mouse models in which endogenous SMAD2 and SMAD3 proteins are globally tagged with hemagglutinin (HA) and podoplanin (PA) sequences, respectively, enabling high-fidelity profiling of SMAD2 and SMAD3 binding across tissues. To demonstrate the broad application of our models, we exemplified the usage of our lines in ovarian biology, where we defined the transcriptional programs downstream of GDF9, a key oocyte-derived ligand in folliculogenesis from the TGFβ superfamily. By integrating genomic and transcriptomic analyses, we identified direct genes induced by the GDF9-SMAD2/3 axis and identified gene sets suppressed by this signaling cascade, highlighting a previously underappreciated role of GDF9 in attenuating competing pathways to ensure proper ovarian granulosa cell fate transitions. Short-term GDF9 stimulation shifts SMAD2/3 cofactor recruitment toward lineage- and differentiation-associated transcription factors, without significant global changes in H3K27ac landscapes, indicating that GDF9 signals through targeted SMAD recruitment to preacetylated chromatin regions. Network analyses further demonstrated that GDF9-SMAD2/3 direct targets align with luteinizing hormone-driven preovulatory signaling. Together, our study generated epitope-tagged mouse models that provide extensive and applicable in vivo genetic toolkits for tissue-specific dissection of TGFβ family signaling and reveal a comprehensive, direct transcriptional network through which GDF9 coordinates granulosa cell differentiation and follicular maturation.

<h4>Background</h4>Chimeric antigen receptor (CAR) T-cell therapy has demonstrated safety and modest efficacy against diffuse midline gliomas (DMGs), a highly aggressive pediatric brain tumor. However, mechanisms of CAR T-cell resistance in DMG settings remain unknown.<h4>Methods</h4>We compared the efficacy of B7-H3 CAR T-cells between SJ-DIPGX7c (DMG) and U87-MG (adult glioblastoma) patient-derived cell lines and showed impaired efficacy both in vitro and in vivo. We performed live-cell imaging and single-cell RNA sequencing to investigate deficiencies in immune synapse (IS) formation between CAR T-cells and DMGs. Lastly, we genetically deleted <i>RASA2</i>, a negative regulator of T cell activation, and evaluated the resulting impact on IS formation and quality, as well as in vitro and in vivo functionality.<h4>Results</h4>We show that limited efficacy of B7-H3 CAR T-cells is due to DMG-mediated inefficient interaction between CAR T-cells and DMG cells. Specifically, DMG cells impair the IS formation, resulting in poor CAR T-cell activation, cytokine secretion, and limited anti-tumor response in vivo. RASA2 deletion improved CAR T-cell activation through the formation of a more functional IS. RASA2-deleted CAR T-cells exhibited enhanced calcium flux, increased accumulation of activated signaling molecules and lytic granules at the synapse, and increased actin cytoskeleton dynamics, which produced larger synaptic areas and resulted in enhanced migration ex vivo. Further, RASA2-deleted CAR T-cells demonstrated improved in vitro functionality and superior early in vivo anti-tumor responses against DMGs compared with controls.<h4>Conclusions</h4>Our study highlights the importance of understanding tumor-specific factors that limit CAR T-cell response and using this information to design superior next-generation CAR T-cells. Specifically, we identify cytoskeleton remodeling and T cell motility as therapeutically actionable targets for future engineering approaches.

DDX3X, a DEAD-box RNA helicase, has been identified as a risk gene for autism spectrum disorder (ASD). To elucidate the role of DDX3X mutations in ASD pathogenesis, HT22 cell models and mouse models with Ddx3x knockdown specifically in the medial prefrontal cortex were established. Ddx3x knockdown in HT22 cells resulted in slower growth, while in mice, it induced autism-like behaviors. Proteomic analysis in cortex revealed that many down-regulated proteins were involved in synaptic plasticity. The differentially expressed proteins (DEPs) were associated with long-term potentiation, glutamatergic and GABAergic synapses, postsynaptic density, branched-chain amino acid degradation, and the oxytocin pathway. Integrating cellular and cortical omics studies revealed overlaps in the ubiquitin-proteasome system and the 'de novo' protein folding pathway. These pathways were inhibited in the cortex of the model mice. The expression of several important proteins was validated. Confocal microscopy and transmission electron microscopy demonstrated a significant reduction in dendritic spine density and postsynaptic density of mouse models. Electrophysiological experiments showed that the frequency of miniature excitatory postsynaptic currents was significantly reduced. These results suggest that DDX3X deficiency impairs synaptic function, thereby affecting neurodevelopment and social abilities. Abnormal synaptic plasticity may contribute to the pathogenesis of ASD with DDX3X gene mutations.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with diverse genetic and environmental origins, yet whether these factors converge on common molecular pathways remains unclear. This study identifies dysregulation of the Notch signaling pathway as a shared mechanism in both hereditary and nonhereditary ASD models. Aberrant histone deacetylase 3-mediated epigenetic regulation of Notch signaling during embryonic forebrain development disrupts the specification of vasoactive intestinal peptide (VIP + ) GABAergic interneuron subtypes (VIP-INs), which originate in the caudal ganglionic eminence (CGE). CGE-specific ablation of Notch1/2 genes in ASD models restores the loss of VIP-INs, normalizes maladaptive excitatory and inhibitory balance, and selectively improves social behaviors. A single antenatal dose of a γ-secretase inhibitor ameliorates multiple ASD-associated neuronal, behavioral, and transcriptomic changes in adult models. The study indicates a strong convergence of ASD-related factors on Notch signaling dysregulation and establishes this pathway as a promising therapeutic target for developmental and behavioral deficits in ASD.

The past decade has seen remarkable progress in identifying genes that, when impacted by deleterious coding variation, confer high likelihood for autism spectrum disorder (ASD), intellectual disability and other associated developmental disorders. However, most underlying gene discovery efforts have focused on individuals of European ancestry, limiting insights into genetic liability across diverse populations. To help address this, the Genomics of Autism in Latin American Ancestries (GALA) Consortium was formed, presenting here the largest sequencing study of autism in Latin American individuals (n > 15,000, including 4,717 participants with an ASD diagnosis). We identified 35 genome-wide significant (false discovery rate < 0.05) autism-associated genes, with substantial overlap with findings from European cohorts, and highly constrained genes showing consistent signal across populations. The results provide support for emerging (for example, MARK2, YWHAG, PACS1, RERE, SPEN, GSE1, GLS, TNPO3 and ANKRD17) and established autism genes and for the utility of genetic testing approaches for deleterious variants in individuals from diverse backgrounds; the results also demonstrate the ongoing need for more inclusive genetic research and testing. We conclude that the biology of autism is consistent across populations, with no detectable influence of ancestry.

The role of phosphatidylcholine transporters such as Stard7 in intestinal cancer development is unknown. To explore this issue, we generated a mouse model lacking Stard7 in intestinal epithelial cells (IECs). Loss of Stard7 impaired mitochondrial Complex I activity, led to a severe metabolic and lipid reprogramming, enhanced mitochondrial ROS production and potentiated an mTORC1/ATF4 signature. As a result, levels of enzymes involved in serine biosynthesis were enhanced in Stard7-deficient IECs. We next assessed the consequences of Stard7 deficiency in both Wnt-dependent tumor initiation and in inflammation-driven tumor development. Strikingly, despite generating similar molecular signatures, Stard7 deficiency inhibited tumor development in Azoxymethane (AOM)/Dextran Sulfate Sodium (DSS)-treated mice but promoted Wnt-driven cancer initiation in the intestine. Apc^+/Min mice lacking Stard7 in IECs developed more tumors in the distal colon as well as a specific microbiota signature. Collectively, our results suggest that the genetic status critically controls the effects of Stard7 deficiency on intestinal tumor development.

False-positive HIV antigen/antibody screening disrupts medical care and has psychological implications for misdiagnosed patients. From 2018 to 2023, our emergency-department screened 102,369 patients for HIV. Among 698 reactive antigen/antibody screens, 582 (83.4%) were confirmed true positives and 116 (16.6%) were false-positive, defined as a reactive antigen/antibody test with an undetectable viral load. Among a series of 13 patients with false-positive results, we evaluated clinical characteristics and conducted repeat testing. False reactivity resolved in 10 (77%) cases while 3 (23%) cases were persistently discordant. False positivity was seen during immune activation states including acute non-HIV infection, autoimmune disease, hematologic conditions, and liver disease.

OBJECTIVE: Immunometabolic dysregulation is increasingly recognized as a pivotal contributor to systemic lupus erythematosus (SLE). This study aims to identify novel circulating biomarkers within plasma exosomes that link fatty acid metabolic rewiring to SLE pathogenesis. METHODS: Plasma exosomes were isolated from patients with active SLE, inactive SLE, rheumatoid arthritis (RA) and healthy controls by ultracentrifugation. The proteomic and metabolomic profiles were characterized using liquid chromatography-mass spectrometry (LC-MS). Exosomal fibroblast activation protein (FAP) expression was validated by nanoflow cytometry, and fatty acid metabolites were confirmed by targeted metabolomics. RESULTS: Proteomic profiling revealed a marked enrichment of FAP in plasma exosomes from SLE patients, with the highest levels in active SLE. Nanoflow cytometry further validated that plasma exosomal FAP was specifically increased in SLE but not RA. Besides, metabolomics identified a characteristic fatty acid signature in SLE plasma exosomes, featuring increased saturated (palmitic acid, etc) and ω-6 polyunsaturated fatty acids (PUFAs) (arachidonic acid (AA), etc), alongside decreased ω-3 PUFAs (α-linolenic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), etc). Moreover, the AA/DHA and total ω-6/ω-3 ratios were significantly elevated, while the EPA/AA ratio was reduced in active SLE. Integrated omics analysis revealed the close relationship of exosomal FAP with fatty acid metabolism. Notably, exosomal FAP levels correlated positively with SLEDAI-2000 score and the pro-inflammatory fatty acid profiles (AA, total ω-6, AA/DHA ratio), but negatively with the anti-inflammatory profiles (DHA, EPA, total ω-3, EPA/AA ratio). CONCLUSION: We identify plasma exosomal FAP as a novel biomarker linking fatty acid metabolic dysregulation to SLE.

No abstract available.

No abstract available.

Neurodegenerative diseases (NDs), including Alzheimer's disease (AD), Parkinson's disease (PD), dementia with Lewy bodies (DLB), and frontotemporal dementia (FTD), share overlapping clinical and pathological features. We analyzed cerebrospinal fluid (CSF) and plasma proteomes from 2,705 and 3,009 samples, respectively, across these NDs, identifying disease-specific and shared molecular signatures. CSF showed more disease-associated proteins than plasma, with AD and DLB exhibiting the strongest cross-tissue similarity. Pathway analyses revealed shared dysregulation of immune-related processes in CSF and plasma across the NDs, as well as disease-specific impairment of glycosylation and apoptotic pathways in AD; ATF4 and PERK signaling in PD; fibroblast growth factor receptor (FGFR) and interleukin signaling in DLB; and glycoprotein hormones disruption in FTD. We developed disease-specific predictive models showing high accuracy (area under the curve [AUC]: 0.81-0.95 in CSF and 0.80-0.89 in plasma). These findings reveal distinct and convergent mechanisms across NDs, highlighting potential biomarkers and pathways for diagnostic and therapeutic strategies in neurodegeneration.

In this study, we synthesized two novel vinyl-substituted, BINOL-hybridized tricyclic-ladder-type siloxanes (referred to as Vi-TLS-BINOLs) <i>via</i> a B(C₆F₅)₃-catalyzed Piers-Rubinsztajn reaction. Fluorescence studies of the Vi-TLS-BINOLs revealed enhanced fluorescence emission attributed to the restricted intramolecular motion by the ladder-type structure. In comparison to the precursor BINOL, Vi-TLS-BINOLs L1 and L2 exhibit a 5.2-fold and 4.8-fold increase in fluorescence quantum yield in THF, respectively. Furthermore, it was observed that both Vi-TLS-BINOLs could selectively recognize Fe³⁺ through fluorescence quenching. ESI-HRMS analysis confirmed the formation of [L1 + 2Fe] and [L1 + 3Fe] complexes with Fe³⁺. The detection limits for Fe³⁺ were determined to be 10.6 µM and 16.3 µM, respectively.

Glioblastoma (GBM) is a highly aggressive, therapy-resistant brain tumor with inevitable recurrence despite maximal multimodal treatment. Increasing evidence suggests that this intractability arises from coordinated cellular programs rather than a single dominant pathway. Central to these programs are glioma stem-like cells (GSCs), which sustain self-renewal, phenotypic plasticity, and resistance to genotoxic and metabolic stress, and yet the molecular basis of their long-term tumor-propagating capacity remains incompletely understood. Here, we synthesize recent advances to propose an integrated conceptual framework-the Triadic Nexus-in which GSC stemness, telomere maintenance mechanisms, and metabolic reprogramming function as a self-reinforcing regulatory system. We review how telomerase reactivation versus alternative lengthening of telomeres (ALT) differentially shape genomic stability, immune signaling, and metabolic states and how metabolic plasticity feeds back to regulate stemness and telomere-associated stress responses. Drawing on single-cell, spatial, and multi-omics studies, we highlight how these interdependent axes collectively sustain therapy resistance and tumor recurrence. Finally, we discuss the translational implications of the Triadic Nexus, emphasizing rational combinatorial therapeutic strategies and biomarker-guided patient stratification based on telomere and metabolic signatures. By unifying stemness, telomere biology, and metabolism into a mechanistically testable model, this review provides a systems-level framework for understanding GBM persistence and guiding next-generation therapeutic interventions.

The corneal endothelium, a monolayer of nonregenerating cells on the posterior cornea, maintains corneal transparency through active fluid pumping. This review examines how corneal endothelial cells (CEnCs) meet high energy demands and adapt metabolically from quiescence to stress, with particular emphasis on their intrinsic metabolic plasticity. In CEnCs, ATP is generated through both glycolysis and oxidative phosphorylation (OXPHOS), with their relative contributions shifting according to oxygen tension and cellular state; OXPHOS remains relatively stable, whereas glycolysis increases during repair and stress. Environmental factors, including pH and temperature, further modulate metabolic flux and pump activity. CEnCs are also continuously exposed to oxidative stress arising from mitochondrial respiration, NADPH oxidase activity, and UVA exposure. Under physiological conditions, antioxidant networks involving NRF2/DJ-1 and related enzymes limit reactive oxygen species (ROS) accumulation. However, in ageing and endothelial disorders such as FECD and diabetes, these defences become impaired. We discuss how disrupted mitochondrial dynamics and mitophagy contribute to bioenergetic failure and endothelial cell loss across disease contexts. Finally, we critically review emerging therapeutic strategies, including metabolic modulators, gene-based approaches, organelle-targeted interventions, and cell-based endothelial replacement. By integrating these mechanisms, this review highlights how therapeutic modulation of metabolism and organelle quality control, together with strategies to address translational barriers, may protect the corneal endothelium, improve graft survival, and delay the need for corneal transplantation.

Ferroptosis is a redox-driven and iron-dependent type of programmed cell death, with lipid peroxidation as a central and required feature of the process. During ferroptosis, cells exert strong proinflammatory effects, suggesting that ferroptosis may play a role in the regulation of inflammation and immune response. However, very few studies have investigated the process of ferroptosis and lipid peroxidation in microglia, the innate immune cells of the brain. In this review, we summarize the concept of ferroptosis and present a list of 120 ferroptosis-relevant proteins, which includes over twice as many entries as the current Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway for ferroptosis. We compare our manually compiled list with microglial activation signatures reported by us and others, revealing ferroptosis-relevant changes in models for microglial activation. Finally, we highlight a selection of ferroptosis-relevant proteins as potential biomarker candidates for ferroptosis.

<i>Boule</i> is the ancestral member of the <i>Deleted in Azoospermia</i> (<i>DAZ</i>) family and is pivotal for gametogenesis and male fertility in most animals. However, there is a dearth of information on molluscan <i>boule</i>. Here, we identified a counterpart (<i>Pcbol</i>) from the genome of <i>Pomacea canaliculata</i>, which has emerged as a cosmopolitan alien species and notorious pest that causes devastating damage to aquatic biodiversity, freshwater ecosystems and crop production in invaded ranges. This study aimed to investigate the biological roles of <i>Pcbol</i> in male reproduction and to decipher the molecular mechanisms underpinning its modulation via dsRNA-delivered RNA interference (RNAi). The bioinformatic analysis showed that the <i>Pcbol</i> genomic sequence is 12,934 nt in length, harboring an open reading frame of 294 nt that encodes 97 aa residues, with an RRM domain evolutionarily conserved among molluscan orthologues. Spatiotemporal expression profiling indicated the predominant abundance of Pcbol in adult males and testis tissues. dsPcbol, injected at a dose of 4 μg/per snail for 5 days, yielded optimal silencing at both transcript and translation levels of Pcbol, as revealed by qRT-PCR and Western blotting. Immunofluorescence echoed a pronounced reduction in Pcbol signal intensity following RNAi. In addition to the arrested reproductive gland phenotype, the number of sperm cells substantially dwindled upon dsPcbol treatment relative to the dsGFP control. In biochemical and fecundity assays, <i>Pcbol</i> depletion triggered a significant decrease in Te/SP/Arg content and suppressed the number of deposited eggs and hatchability. Furthermore, spermatogenic genes like <i>CDC25/TSSK1/SPATA17/DDX4/Dmrt2/Sox2/Kelch10/SPO11</i> displayed considerable downregulation post <i>Pcbol</i> silencing, with molecular docking predicting a strong affinity between CDC25 and Pcbol. These molecular modules may interact with Pcbol to mediate knockdown effects on spermatogenesis dysfunction. Collectively, our findings not only confirmed that <i>boule</i> was indispensable for spermatogenesis and male fertility in a mollusk, but also highlighted the <i>Pcbol</i>-based male sterile technique (MST), which can be incorporated into precision pest management (PPM) strategies for sustainable control of <i>P. canaliculata</i>.

The management of multiple sclerosis (MS) is shifting from a phenotype-based framework toward a biologically driven precision medicine model, as conventional magnetic resonance imaging (MRI) inadequately captures smoldering inflammation and progression independent of relapse activity (PIRA). This systematic review aimed to synthesize current evidence on the diagnostic and prognostic utility of fluid biomarkers in distinguishing acute inflammatory injury from chronic neurodegeneration. A comprehensive search of Web of Science, PubMed, and Scopus (January 2020-September 2025) identified 28 eligible studies including 7775 participants (6365 MS patients and 1410 controls). Biomarkers derived from serum, plasma, cerebrospinal fluid (CSF), and stool were evaluated in relation to clinical disability measured using the Expanded Disability Status Scale (EDSS) and magnetic resonance imaging (MRI) outcomes. Neurofilament light chain (NfL) consistently predicted acute inflammatory activity, gadolinium-enhancing lesions, and relapse-associated worsening, but levels were reduced by high-efficacy therapies and did not reliably predict PIRA. In contrast, glial fibrillary acidic protein (GFAP) was associated with astrogliosis, disability progression, and retinal thinning, even in patients with low inflammatory activity. Additional CSF, metabolic, and immunologic markers correlated with neurodegeneration and disease severity. Nevertheless, broader clinical use will require greater assay standardization, improved consistency across cohorts, and validation in prospective longitudinal studies. These findings compel a shift toward a multi-biomarker model to guide personalized therapeutic strategies and develop targeted neuroprotective treatments for progressive multiple sclerosis.

<b>Background/Objectives</b>: Multiple myeloma (MM) is an incurable plasma cell neoplasm in which diagnosis and prognostication rely on invasive bone marrow examinations that may not capture biological heterogeneity across different disease sites. There is a clinical need for non-invasive biomarkers that can accurately predict treatment outcomes. <b>Methods</b>: We performed a global proteomic profiling of bone marrow-derived extracellular vesicles (EVs) from nine MM patients and ten controls. A total of 8839 proteins were identified, of which 14 met predefined selection criteria. These candidates were quantified in serum-derived EVs using targeted proteomic analysis. Prognostic relevance of selected proteins was evaluated in newly diagnosed MM (NDMM) patients treated with daratumumab-containing frontline regimens (<i>n</i> = 26) and healthy individuals (<i>n</i> = 60). Progression-free survival (PFS) was analyzed using univariable and multivariable models. <b>Results</b>: IL5RA (<i>p</i> = 0.003) and BCMA (<i>p</i> < 0.001) were significantly elevated in serum EVs from MM patients compared with controls. Higher serum EV-IL5RA and EV-BCMA were associated with a trend toward shorter PFS. Combined assessment of these biomarkers enabled clear stratification of MM patients into three prognostic groups, including a cohort with markedly inferior outcomes, with a 20-month PFS of 0 (<i>p</i> = 0.001). In multivariable analysis, the combined serum EV-IL5RA and EV-BCMA signature suggests an independent prognostic potential (HR = 38.49 [95% CI, 1.51-47.79], <i>p</i> = 0.015). <b>Conclusions</b>: Serum EV-IL5RA and EV-BCMA are novel non-invasive biomarkers, measurable through routine blood testing, with strong potential to improve risk stratification in NDMM patients in the era of daratumumab-based frontline therapy.

Major depressive disorder (MDD) lacks reliable laboratory tests for diagnosis and treatment monitoring, underscoring the need for robust molecular readouts in blood. Beyond symptom-based classification, MDD can also be viewed as a condition involving impaired homeostatic regulation across stress-responsive, immune, metabolic, and neural systems. Consistent with this perspective, altered intron retention (IR) patterns have been observed in peripheral blood in depression-related and treatment-response contexts, supporting the translational relevance of this RNA-processing layer to mood disorders. A key observation underpinning this review is that IR can function as a reversible, intervention-responsive readout of physiological state. In a pre-symptomatic stress-like state in klotho mutant mice (a premature-aging model), widespread IR increases revert toward a healthy pattern upon treatment, suggesting that IR is embedded in a controllable homeostatic layer. Against the backdrop of limited cross-cohort transferability of differential gene expression (DGE) signatures, we propose that IR provides a mechanistically grounded biomarker layer because it reports regulated RNA processing states rather than context-fragile abundance endpoints. We operationalize IR as a post-transcriptional "throttle" on effective gene output, with increased IR/detained intron (DI) states acting as a reversible brake and decreased IR acting as an accelerator that increases translation-competent mRNA supply. Mechanistic exemplars across immune, metabolic, and neuronal systems (e.g., IFNG, OGT, MAT2A, neuronal activity-triggered intron excision, and intron detention-mediated stemness/differentiation switching in adult neural stem cells) show that defined inputs can switch IR/DI states to tune output kinetics. Integrating these findings, we propose an "Intron Retention Homeostat" (IR-Homeostat) model in which cells sense deviations from physiological set points and implement feedback control of gene output through switchable IR/DI regulation. This framework positions IR not only as a robust state readout for stratification, treatment response prediction, and pharmacodynamic profiling, but also as a tractable entry point to identify the molecular sensors and mediators that couple homeostatic signals to RNA processing control.

<h4>Background</h4>The growing prevalence of cardiovascular diseases (CVDs) has created an increasing demand for alternative biologically functional vascular grafts. Among the explored tissue engineering strategies, decellularization provides a means to generate suitable acellular scaffolds from donor tissues with preserved complex extracellular matrix (ECM) architecture and vascular basement membrane (BM), both of which are critical for graft endothelialization and function. To assess decellularization efficiency and quality, proteomic profiling offers high-resolution characterization of tissue composition, enabling systematic analysis of extracellular matrix subgroups and residual proteins with the aid of open-access annotation tools, thus providing critical insights beyond conventional methods.<h4>Methods</h4>Human saphenous vein segments were decellularized using a CHAPS-based protocol, and decellularization efficiency was evaluated by DNA and glycosaminoglycan (GAG) quantification, together with histological evaluation. A subset of samples (n = 9) was further subjected to label-free quantitative proteomic profiling using LC-MS/MS to assess ECM composition, BM integrity, and residual cellular content.<h4>Results</h4>Decellularization led to 99.96% DNA removal and 53.78% GAG retention, with well-preserved tissue morphology. Decellularization efficiency was further assessed by label-free quantitative proteomics, which showed well-preserved ECM composition. Collagens, proteoglycans, and BM proteins were largely well-maintained, whereas ECM glycoproteins and matrisome-affiliated proteins were moderately reduced. The strongest decrease in protein abundance was observed among cellular proteins, and Gene Ontology analysis confirmed the reduction of key cellular protein groups.<h4>Discussion</h4>In this study, we provide a comprehensive proteomic evaluation of decellularized human saphenous veins, demonstrating selective preservation and loss across ECM subgroups and cellular proteins. The findings highlight proteomics as a valuable complement to conventional assays for assessing decellularization efficiency and establishing a reference framework for optimizing protocols in vascular tissue engineering.

Ferroptosis, fundamentally defined as an iron-catalyzed and lipid peroxidation-driven cell death process, constitutes an underlying disease mechanism in autoimmune pathogenesis. Recent studies demonstrate that ferroptosis not only shapes innate and adaptive immune responses-including T and B lymphocytes, macrophages, neutrophils, and dendritic cells-but also remodels inflammatory microenvironments to either preserve tissue integrity or exacerbate damage. Pharmacological modulation of ferroptosis, via iron chelators, GPX4 activation, and lipid peroxidation blockade, hold promise as adjuncts to current immunomodulatory approaches. Collectively, ferroptosis represents both a unifying pathogenic framework and a translational target for precision interventions in autoimmunity.

<h4>Background</h4>Netrin-1, a laminin-related guidance cue protein with emerging immunomodulatory roles, has shown conflicting associations with type 2 diabetes mellitus (T2DM) in human studies. We conducted a systematic review and meta-analysis to evaluate circulating netrin-1 levels in individuals with T2DM compared to healthy controls.<h4>Methods</h4>A comprehensive search of MEDLINE/PubMed, Scopus, and Europe PMC up to March 17, 2025, was performed for studies reporting netrin-1 levels in T2DM patients. Eligible studies were observational and provided extractable quantitative data. Study quality was assessed using the Newcastle-Ottawa Scale and JBI checklist. Standardized mean differences (SMDs) with 95% confidence intervals (CIs) were calculated using a random-effects model. Heterogeneity, subgroup, sensitivity, and publication bias analyses were performed using R software.<h4>Results</h4>Twenty studies involving 1,798 articles were included. Pooled analysis revealed significantly elevated netrin-1 levels in T2DM (SMD = 0.57; 95% CI: 0.03-1.11; p = 0.0393), with substantial heterogeneity (I² = 96.4%). Subgroup analyses indicated geographic and diagnostic criteria as key moderators. Egyptian studies and those using ADA guidelines reported increased netrin-1, while Indian and WHO-based studies reported reductions. The 95% prediction interval (-2.03 to 3.18) reflected wide variability. No significant publication bias was detected (LFK index = -0.49).<h4>Conclusion</h4>This meta-analysis supports a potential link between netrin-1 and T2DM, suggesting its role as a biomarker of metabolic inflammation. However, substantial heterogeneity and study-level differences necessitate further standardized, longitudinal research to clarify its clinical relevance.

Sequestosome 1 (SQSTM1/p62), long established as a selective autophagy receptor and ubiquitin-binding scaffold, is now recognized as an emerging regulatory hub that integrates signaling, metabolism, and stress adaptation in cancer. Beyond its canonical role in proteostatic cargo degradation, recent advances have revealed that p62 orchestrates nutrient sensing, redox control, innate immune signaling, and metabolic reprogramming through highly dynamic, context-dependent mechanisms. A nascent paradigm emerging from recent studies is that p62 function is specified by a hierarchical post-translational modification (PTM) code, with phosphorylation acting as the primary regulatory layer. Site-specific phosphorylation events-together with modulatory PTMs such as S-acylation, arginine methylation, and O-GlcNAcylation-reshape p62 interaction networks, liquid-liquid phase separation (LLPS) behavior, and signaling output. Through these mechanisms, p62 operates as a sophisticated signal-metabolism interface that couples stress signaling pathways, including NFE2L2/NRF2, AMPK, mTORC1, and NF-κB to the systemic rewiring of glucose, lipid, amino acid, and nucleotide metabolism. Notably, a phosphorylation-dependent positive feedback loop between p62 and AMPK has emerged as a key driver of metabolic plasticity, enabling tumor cells to survive and proliferate under the stringent metabolic stress conditions of the tumor microenvironment. This review integrates recent mechanistic insights into the PTMs, phase behavior, and signaling hub functions of p62, highlighting how these principles manifest in distinct oncogenic contexts, such as lung, prostate, and brain tumors. We further discuss emerging therapeutic strategies that seek to modulate p62-centered assemblies rather than indiscriminately inhibit p62 function. Collectively, these findings position p62 as a phosphorylation-governed oncogenic nexus whose precise manipulation may enable new strategies for context-dependent precision oncology.

Drone brood homogenate (DBH), a nutrient-rich bee product, has received limited scientific attention despite its potential immunomodulatory and gut-protective properties. This study evaluated the effects of a dietary DBH supplementation on the intestinal barrier-related gene expression, phagocytic activity, and lymphocyte subpopulations in pigs. Eighteen weaned pigs were assigned to three groups (control, DBH100, DBH200) and fed DBH at 0, 100, or 200 mg/kg feed for 18 days. The gene expression of tight junction markers (occludin, claudin-1) and mucosal integrity-associated proteins (lumican, OLFM4) was assessed in the ileum by qRT-PCR. Phagocyte function and peripheral blood lymphocyte subpopulations were analysed by flow cytometry. DBH200 significantly upregulated the occludin, claudin-1, lumican, and OLFM4 expression, indicating enhanced intestinal barrier support. The phagocytes from both DBH-treated groups exhibited an increased engulfing capacity and an elevated oxidative burst index, though the percentage of active phagocytes was only weakly affected. The DBH supplementation did not alter the total T (CD3⁺) or B (CD21⁺) cells; however, both DBH groups showed a significantly increased CD4⁺ : CD8⁺ lymphocyte ratio, which is consistent with immune stimulation. These findings suggest that DBH may beneficially modulate the gut barrier integrity and selected components of innate and adaptive cellular immunity in pigs.

Water remediation technologies urgently require materials that are sustainable, efficient, and regenerable. Here, we report a dynamic covalent aerogel derived from chitosan and formylated lignin, enabling dual functionality in heavy metal adsorption and heterogeneous catalysis. The aerogel is formed through reversible imine bonds between chitosan and aldehyde-functionalized lignin, synthesized via a mild Reimer-Tiemann reaction. The resulting monoliths exhibit high structural integrity, solvent and pH resistance (pH ≥ 7), and remarkable adsorption capacities for Cu²⁺, Pb²⁺, and Zn²⁺ ions (up to 32.8 mg/g for Cu²⁺). Density functional theory (DFT) calculations reveal tridentate coordination as the dominant binding mechanism. When loaded with Cu²⁺, the aerogels act as effective heterogeneous catalysts for the Chan-Lam crosscoupling reaction in ethanol. They achieve initial yields of up to 92% and retain their catalytic activity over multiple reuse cycles. Importantly, the dynamic covalent framework allows complete recovery and regeneration of all components, underscoring its potential as a circular, bio-based platform for sustainable water detoxification and green catalysis.

Endothelial heterogeneity and plasticity play an important role in lung development, homeostasis, and pathology. In recent years, increasing evidence has demonstrated that endothelial dysfunction contributes to the progression of various lung diseases, such as ADRS, PF, PH, and lung developmental disorders. Therefore, targeting endothelial cells could hold promising therapeutic strategies for preventing disease development. Although significant advances in technology have revolutionized our understanding of endothelial heterogeneity and plasticity, effective and curative treatment options remain limited. Here, we discuss the molecular and functional diversity of lung endothelial cells and their critical role in maintaining lung homeostasis and in lung pathologies. We also briefly describe advanced technologies, such as single-cell RNA sequencing and spatial transcriptomics, to uncover complex cell communication and underlying mechanisms. Furthermore, this review will identify future research questions for developing therapeutic approaches targeting lung endothelial cells.

<h4>ABSTRACT</h4> Advances in DNA sequencing and assembly technologies are spurring a shift from haploid reference genomes to sample-specific diploid assemblies. Here, we generated the first telomere-to-telomere (T2T) diploid reference for the widely used human embryonic stem cell (hESC) line, H9 (WAe009-A). This haplotype-resolved assembly is highly accurate with comprehensive annotation of genes, segmental duplications, methylation, and chromatin conformation. Pangenomic and phased-locus inference point to H9’s mixed ancestry with a predominant European component. H9-specific genomic features include near-perfect telomeres ∼1.65-fold longer than other T2T assemblies, consistent with telomerase activity during pluripotency; chromosome 17 inversions that can predispose offspring to neurological syndromes; and expansions of ncRNA clusters, with overall genomic stability maintained despite extensive culturing. Mapping multi-omic datasets to the genome, we demonstrate the power of this resource for allele-specific, high-precision transcriptomic, genetic, and epigenetic analyses, with far-reaching implications for human development and disease.

G_q/11-coupled GPCRs regulate critical physiological processes through calcium mobilization, making intracellular Ca²⁺ dynamics a key readout for drug discovery and biomarker detection. However, the transient nature of calcium signals necessitates real-time monitoring with specialized equipment, creating barriers for high-throughput screening and limiting accessibility. Here we present CalLuc-2.1, a luminescent calcium biosensor that converts brief Ca²⁺ spikes into persistent luminescence changes readable tens of minutes after stimulation. By eliminating the need for precisely-timed detection, CalLuc-2.1 enables endpoint measurement of GPCR activation using standard plate readers. We demonstrate robust performance (Z' > 0.88) across multiple G_q/11-coupled GPCRs in both agonist and antagonist screening formats. Furthermore, CalLuc-2.1 successfully detects endogenous GPCR ligands directly in human serum, offering a simpler alternative to immunoassays and mass spectrometry for biomarker quantification. This approach makes calcium-based GPCR assays accessible to any laboratory with basic luminescence detection capabilities, potentially accelerating both drug discovery and clinical research applications.

Glycosphingolipids (GSLs) are regulators of membrane organization and trafficking. According to the glycolipid-lectin (GL-Lect) driven endocytosis model, GSLs are part of a mechanism in which they are used by oligomeric lectins from pathogens or cells to build tubular endocytic pits from which clathrin-independent endocytic carriers detach by friction-driven scission. This review revisits the unique architectural logic of GSLs and explores how this underpins mechanisms for endocytic trafficking and signaling. We examine notably how GSLs orchestrate retrograde trafficking from the plasma membrane to the Golgi apparatus and subsequent polarized secretion in specialized contexts such as migrating cells, epithelial tissues, and immune synapses. Despite their importance, GSL functions have remained difficult to explore, because of the scarcity of minimally perturbing, chemically defined tools. We survey recent development in GSL probe design and discuss principles for next-generation tools that retain native function while enabling mechanistic dissection. Finally, we discuss how these advances are reshaping therapeutic strategies, with implications for vaccine delivery, cancer targeting, and lysosomal storage disorders. Collectively, this review reframes GSLs as substrates of membrane organization, with vast untapped potential in both cellular membrane biology and translational medicine.

<h4>Purpose</h4>Lung adenocarcinoma (LUAD), the most common subtype of non-small cell lung cancer (NSCLC), often presents with mild or absent symptoms in its early stage, leading to delayed diagnosis and poor outcomes. This study aimed to elucidate the molecular mechanisms underlying early-stage LUAD and to identify effective biomarkers for early detection and therapeutic intervention.<h4>Methods</h4>A machine-learning framework integrating random forest (RF) and least absolute shrinkage and selection operator (LASSO) was used to identify candidate diagnostic biomarkers. The robustness of the identified marker was evaluated in an independent external dataset. Single-cell RNA sequencing was employed to localize gene expression within the tumor microenvironment. Additional analyses-including cell-cell communication inference, copy number variation (CNV) profiling, and pseudotime trajectory reconstruction-were performed to investigate the functional role of the identified biomarker in LUAD progression.<h4>Results</h4>IGSF9 emerged as a promising diagnostic biomarker and potential therapeutic target for early-stage LUAD, with its diagnostic value validated in an external dataset. Single-cell RNA sequencing located IGSF9 expression primarily in alveolar cells within the tumor microenvironment. Cell-cell communication analyses suggested that IGSF9 contributes to immune evasion and promotes tumor cell migration and invasion. CNV analysis revealed substantial genomic instability in the alveolar compartment. Pseudotime trajectory inference indicated that IGSF9 may drive the differentiation of a stem-like AT2 subpopulation toward a more malignant state.<h4>Conclusion</h4>This study identifies IGSF9 as a robust diagnostic biomarker for early-stage LUAD and elucidates its multifaceted role in malignant transformation. These findings provide valuable insights for early diagnosis and precision oncology in LUAD.

m6A is a pervasive post-transcriptional RNA modification that regulates RNA splicing, stability, localization, and translation in the brain. In this review, we outline the core m6A regulatory machinery and summarize its spatial organization across neurons and glial cells, highlighting established roles in brain development, synapse formation, and axon growth. We then focus on experience-dependent plasticity, synthesizing evidence that neuronal activity and environmental inputs dynamically reshape m6A to regulate immediate-early transcription and local translation at synapses across sensory, cognitive, emotional, and motor domains. With aging, m6A programs are reconfigured in a cell-type-specific manner, a shift associated with reduced plasticity and increased vulnerability. We further survey disease-associated alterations in m6A across Alzheimer's disease, Parkinson's disease, Huntington's disease, stroke-related cognitive impairment, ALS and FTD, as well as metal or toxin exposure, emphasizing convergent effects on dopaminergic and glutamatergic signaling, synaptic integrity, inflammation, and cellular stress responses. Finally, we discuss emerging opportunities and conceptual challenges in targeting m6A enzymes or reader proteins, and outline priorities for future work, including cell-type- and subcellular-resolved mapping, causal perturbation in defined circuits and life stages, and the development of biomarkers and selective modulators. Together, these observations position m6A as a molecular interface linking experience-dependent plasticity, brain aging, and neurodegenerative vulnerability.

Targeted protein degradation can be induced by recruiting a protein of interest to an E3 ligase, resulting in its ubiquitination and subsequent proteasome-mediated degradation. However, only a small number of E3 ligases have been utilized for degradation. Expansion of the repertoire of useful E3 ligases via the identification of ligands to those ligases could broaden the scope and applicability of the degradation paradigm. We have identified KLHL12 as an E3 ligase with higher expression in cancer over normal tissues. We report here the use of NMR-based screening to identify fragments that bind to KLHL12, and X-ray structures of a fragment hit bound to KLHL12. Using this structural information, we optimized the hits, leading to the first reported small molecules that bind to KLHL12 with submicromolar affinity. Derivatives of these compounds may be useful for the construction of PROTACs to selectively degrade protein targets in tumors while sparing normal cells.

<h4>Background and aims</h4>Currently, there are no effective drug treatments for aging. Mendelian randomization (MR) has been widely applied to repurpose existing drugs and identify new therapeutic targets. Our aim is to identify aging-related therapeutic targets and evaluate their potential adverse effects, underlying mechanisms, and actionable drugs.<h4>Methods</h4>We integrated druggable genome data, <i>cis</i>-expression quantitative trait loci (eQTL) data from human blood, and genome-wide association study (GWAS) summary data on aging. Using two-sample MR, we analyzed the potential causal relationships between druggable genes and aging. To ensure the reliability of our results, we conducted sensitivity analyses, Bayesian colocalization analyses, and repeated validations using different blood <i>cis</i>-eQTL data sets. We assessed the side effects of identified drug targets using phenome-wide MR (Phe-MR). Additionally, we performed two-step mediation MR analyses to evaluate the role of plasma proteins in mediating the causal relationships between these genes and aging, exploring potential mechanisms. Finally, we investigated actionable drugs for the target genes using relevant databases.<h4>Results</h4>MR analysis identified five potential druggable genes related to aging: ATP1B3, VKORC1, SLC5A11, HNRNPA1, and SMN2. Phe-MR revealed no significant adverse effects when targeting these genes. Mediation MR identified ten plasma proteins linking these genes to aging, offering mechanistic insights. Drug repurposing analysis supports that cardiac glycosides, Bisacodyl, Olsalazine, and Tegoprazan might be potential therapeutics for aging by inhibiting ATP1B3.<h4>Conclusions</h4>Our research indicates that ATP1B3, VKORC1, SLC5A11, HNRNPA1, and SMN2 are potential targets for antiaging therapies. These findings could help prioritize the development of aging-related drugs.

<b>Background/Objectives</b>: Early-onset high myopia (eoHM), defined as high myopia manifesting before 10 years of age, is largely attributed to genetic defects. This study aimed to investigate the genetic underpinnings of eoHM in a cohort of Chinese patients. <b>Methods</b>: We recruited 64 Chinese patients with eoHM. Comprehensive clinical evaluations were performed, and whole exome sequencing (WES) was conducted to identify potential pathogenic variants. The genetic findings were analyzed and correlated with the clinical phenotypes. <b>Results</b>: A total of 64 unrelated Chinese patients with suspected early-onset high myopia were initially recruited. Following whole exome sequencing (WES) and variant annotation, final 37 patients with variants in known myopia-associated genes were included in the analytical cohort. The mean age of onset for the cohort was 5 years (IQR, 4-7), with a mean spherical equivalent refraction of -7 D (IQR, (-8)-(-6)). Genetic analysis revealed variants in 28 known myopia-associated genes. We identified pathogenic or likely pathogenic variants in 11 of the 37 patients (29.7%, 95%CI: 0.1737-0.4590), while the overall diagnostic yield was 17.2% (11/64, 95%CI: 0.0970-0.2839) in initial 64 recruited patients. These genes included seven well-established eoHM-related genes, such as ARR3, CACNA1F, P4HA2, TRPM1, COL11A1, COL2A1, and PAX6. Additionally, variants of uncertain significance (VUS) in seven other candidate genes were detected in patients with eoHM. <b>Conclusions</b>: Our findings expand the genetic spectrum of eoHM and reinforce the critical role of genetic testing in its etiological diagnosis and clinical management. Observed patterns of genotype-phenotype associations are descriptive and should be considered hypothesis-generating, requiring validation in larger cohorts. Additionally, we identify several candidate genes that may serve as prospective biomarkers, though these findings require validation in larger cohorts and functional studies.

<b>Background</b>: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the gastrointestinal tract and are primarily driven by activating mutations in KIT or PDGFRA. Although tyrosine kinase inhibitors (TKIs), particularly imatinib, have substantially improved outcomes, most patients with advanced disease eventually develop resistance, resulting in disease progression. <b>Methods</b>: We performed a narrative review with scoping approach of interventional clinical trials registered on ClinicalTrials.gov between January 2020 and July 2025 to characterize the contemporary investigational therapeutic landscape in GIST. Eligible studies included clinical trials evaluating novel agents, combinations, or alternative strategies beyond current regulatory approvals. Trial characteristics, therapeutic classes, endpoints, enrollment, and funding sources were analyzed. <b>Results</b>: A total of 27 ongoing trials were identified. Most studies were phase I/II and focused on metastatic or unresectable disease, predominantly in the second-line or later settings. TKIs remained the dominant therapeutic class, included in over 70% of trials, either as monotherapy or in combination. Emerging strategies comprised antibody-drug conjugates, immune checkpoint inhibitors, HIF inhibitors, FGFR inhibitors, and epigenetic modulators. Only four phase III trials were identified, reflecting the difficulty of conducting large, randomized studies in GIST. No trial used overall survival or quality of life as a primary endpoint. <b>Conclusions</b>: The current investigational landscape in GIST is largely focused on overcoming TKI resistance in advanced disease. Molecular stratification and personalized approaches dominate ongoing research, but evidence generation remains limited by small sample sizes and slow recruitment. Future trials integrating innovative therapeutic platforms and patient-centered outcomes are essential to improve long-term disease control and quality of life.

<h4>Background</h4>Evidence on risk factors for a complicated course of lower respiratory tract infections (LRTIs) in primary care remains limited and often consensus based. While socioeconomic status (SES) and migration background have been linked to complicated LRTIs in population-based studies, their predictive value in primary care remains unclear. Consequently, these factors are not incorporated within current guidelines, which may contribute to health inequalities. Therefore, we aimed to evaluate the added value of SES and migration background as predictive factors of a complicated course of LRTIs in primary care.<h4>Methods</h4>Routine care data from Dutch general practices participating in the Extramural LUMC Academic Network database (Leiden-The Hague-Zoetermeer region) from 2014 to 2023, excluding COVID-19 years, was linked to sociodemographic and hospital claims data from Statistic Netherlands. Adults presenting with LRTI complaints were included (n = 145,445). Multivariable logistic regression models were constructed to predict 30-day hospitalisation or death following LRTI. Models included conventional risk factors with SES and migration background subsequently added.<h4>Results</h4>In this study we show that after adjusting for conventional clinical factors, SES is a strong predictor of a complicated course of LRTI, whereas migration background is not. Patients in the lowest SES category have an adjusted odds ratio of 1.46 (95%CI: 1.31 - 1.62) for a complicated course compared to the highest.<h4>Conclusions</h4>SES is a strong predictor of a complicated course of LRTI in primary care, even after adjusting for conventional risk factors. The incorporation of SES into clinical decision tools and guidelines has the potential to enhance risk-stratification of patients with LRTI in daily practice of primary care, thereby supporting more equitable care.

NEGR1 (neuronal growth regulator 1) has been genetically linked to metabolic and neuropsychiatric disorders; however, its cellular function in insulin-responsive tissues remains poorly understood. Here, we investigated the role of NEGR1 in regulating actin cytoskeletal dynamics and insulin-stimulated GLUT4 trafficking in skeletal muscle. We found that loss of <i>Negr1</i> reduced GLUT4 abundance selectively in predominantly glycolytic skeletal muscles in vivo. Despite preserved insulin-induced Akt phosphorylation, insulin-stimulated GLUT4 translocation was markedly impaired in both Negr1-deficient and NEGR1-overexpressing muscle cells. Mechanistically, <i>Negr1</i> deficiency was associated with enhanced PAK-cofilin signaling and excessive intracellular F-actin accumulation that likely impedes GLUT4 vesicle trafficking. In contrast, NEGR1 overexpression did not increase total F-actin content but induced abnormal peripheral actin organization, resulting in constitutive GLUT4 surface localization and elevated basal glucose uptake. Consistent with these findings, both loss and overexpression of NEGR1 disrupted insulin-induced Rac1-dependent actin remodeling without affecting Akt signaling. Collectively, these results identify NEGR1 as a critical modulator of actin homeostasis required for proper insulin-stimulated GLUT4 trafficking and glucose uptake in skeletal muscle, providing mechanistic insight into the metabolic abnormalities associated with NEGR1 dysregulation.<b>NEW & NOTEWORTHY</b> Neuronal growth regulator 1 (NEGR1) regulates actin cytoskeletal homeostasis required for insulin-stimulated GLUT4 trafficking in skeletal muscle. NEGR1 dysregulation alters PAK-cofilin signaling, induces aberrant F-actin organization, and impairs GLUT4 vesicle movement independent of Akt signaling. Because NEGR1 is a major genetic risk factor for major depressive disorder, these findings reveal a shared actin-based mechanism linking metabolic dysfunction and neuropsychiatric disease.

Structural variants (SVs) are increasingly recognized as important contributors to oncogenesis through their effects on 3D genome folding. Recent advances in whole-genome sequencing have enabled large-scale profiling of SVs across diverse tumors, yet experimental characterization of their individual impact on genome folding remains infeasible. Here, we leveraged a convolutional neural network, Akita, to predict disruptions in genome folding caused by somatic SVs identified in 61 tumor types from the Children's Brain Tumor Network dataset. Our analysis reveals significant variability in SV-induced disruptions across tumor types, with the most disruptive SVs coming from lymphomas and sarcomas, metastatic tumors, and germline cell tumors. Dimensionality reduction of disruption scores identified five recurrently disrupted regions enriched for high-impact SVs across multiple tumors. Some of these regions are highly disrupted despite not being highly mutated, and harbor tumor-associated genes and transcriptional regulators. To further interpret the functional relevance of high-scoring SVs, we integrated epigenetic data and developed a modified Activity-by-Contact scoring approach to prioritize SVs with disrupted genome contacts at active enhancers. This method highlighted highly disruptive SVs near key oncogenes, as well as novel candidate loci potentially implicated in tumorigenesis. These findings highlight the utility of machine learning for identifying novel SVs, loci, and genetic mechanisms contributing to pediatric cancers. This framework provides a foundation for future studies linking SV-driven regulatory changes to cancer pathogenesis.

BACKGROUND: Chemoresistance remains a major barrier in hepatocellular carcinoma (HCC) treatment, and research on personalized therapies for resistant patients is still nascent. This study aimed to construct a preliminary gene-drug resistance map for HCC to advance research in personalized treatment. PATIENTS AND METHODS: Tumor tissues from 51 HCC patients who underwent surgical resection at the First Affiliated Hospital of Chongqing Medical University (December 2019 to June 2021) were used to establish in vitro models. High-throughput drug sensitivity screening (HDSS) and whole-exome sequencing (WES) were performed to identify key resistance-associated genes. Integrated analysis with TCGA database data generated a pharmacogenomic atlas of HCC. RESULTS: Mutational frequencies of key oncogenes and tumor suppressor genes in HCC patients were elevated and largely consistent with The Cancer Genome Atlas (TCGA) data. FAT4 mutations were significantly associated with resistance to epirubicin, doxorubicin, 5-fluorouracil, and XELOX regimens (p < 0.05). KMT2D mutations showed significant correlations with resistance to gemcitabine, 5-FU + cisplatin, ADM + L-OHP, and L-OHP + irinotecan (p < 0.05). PABPC1 mutations were linked to resistance in doxorubicin, apatinib, and ADM + L-OHP regimens (p < 0.05). CONCLUSIONS: FAT4, KMT2D, and PABPC1 are potential key drivers of chemoresistance in HCC, demonstrating cross-regimen associations. These findings may inform personalized chemotherapy strategies for advanced HCC, although further clinical validation is warranted. TRIAL REGISTRATION: This study was registered at the Chinese Clinical Trial Registry ( https://www.chictr.org.cn ; registration number: ChiCTR1900022193; registration date: 2019/03/30).Observational Study.

In recent years, two seemingly unrelated topics, protein disorder and phase separation, have become highly significant in biology. In this way, intrinsically disordered proteins and liquid-liquid phase separation have become two of the hottest topics in the field. In this review, we provide a brief description of intrinsic protein disorder and its effects on the main protein constituents of chromatin, from histones to high-mobility group proteins, heterochromatin HP1, the neuronal methyl-CpG binding protein 2, and the highly specialized arginine-rich protamines of sperm. Following this, we provide a general overview of the role of their intrinsically disordered regions (IDRs) in chromatin condensate formation and present the evolving understanding of the amino acid sequence grammar of IDRs that determines their involvement in in the formation of chromatin condensates. After a brief overview of the theoretical aspects beyond this process, we describe how it is connected to the folding of somatic and male germ cell chromatin. Somatic cell chromatin is organized into compartments. We discuss the characteristics of the phase-separated transcription condensate in compartment A (euchromatin), including the specialized biomolecular condensate in which replication-dependent histone genes are transcribed, and the differential structure and role of HP1 alpha in compartment B heterochromatin compaction across different species.

Chronic pain is a globally prevalent and complex condition, encompassing three primary subtypes, that is, nociceptive, neuropathic, and nociplastic, each with distinct biopsychosocial mechanisms. Chronic pain was historically viewed as a monolithic symptom and managed with opioid-centric models, causing widespread therapeutic failure. While recognition of its heterogeneity has driven a paradigm shift toward precision medicine, tailoring multimodal strategies to the dominant pain mechanism, critical challenges persist. These include difficulty in identifying treatable root causes, limited long-term efficacy of therapies, and significant side-effect burdens. To address these gaps, this review systematically synthesized contemporary knowledge, predominantly from the last decade, on the molecular mechanisms, risk factors, diagnostic frameworks, and therapeutic modalities for chronic pain, framed by its pathophysiological subtypes. Furthermore, it explored two novel frontiers aimed at advancing personalized pain medicine. First, it proposed cold atmospheric plasma as an innovative therapeutic intervention capable of modulating key molecular pathways underlying diverse pain manifestations. Second, it introduced an original neurobiological model positing the hippocampus as a putative sensor for nociplastic pain, interfacing with higher-dimensional information fields. These insights may offer transformative potential for refining diagnostic and therapeutic strategies, potentially revolutionizing the management of chronic pain.

We have investigated the tumoricidal potential of derivatives of protocatechuic acid (PCA) against tumor cells of hematological malignancies (lymphoma and myeloma). PCA (3,4-dihydroxy benzoic acid) belongs to the class of phenolic acids that naturally occur in many plant-derived foods, including olives and white grapes. PCA is a major metabolite of anthocyanin and shows strong <i>in vitro</i> and <i>in vivo</i> antioxidant activity. Novel derivatives of PCA were synthesized by adopting several different strategies, including the formation of amide derivatives, conjugation of carbohydrate moieties employing click reaction, and esterification of PCA with the inclusion of chlorine to form additional derivatives. PCA and its derivatives significantly reduced the viability and long-term growth of the myeloma cells in a dose-dependent manner. Furthermore, the PCA and its derivatives demonstrated dose-dependent tumor cell death via the significant release of lactate dehydrogenase following exposure. The PCA chloride derivative was significantly more effective compared to the other derivatives tested. PCA and its derivatives induce apoptosis of the lymphoma cells and downregulate the expression of PD-L1, suggesting its possible role in disrupting the signaling of the checkpoint pathway, thus regulating the growth of the tumor cells of hematological malignancies. Together, our data demonstrated that PCA and its novel derivatives have therapeutic applications against hematological malignancies.

Alzheimer's (AD) and Parkinson's disease (PD) are prominent neurodegenerative disorders characterized by early synaptic loss, which correlates more closely with clinical symptoms than neuronal death. This synaptic impairment is primarily driven by disruptions in synaptic vesicle (SV) trafficking, a critical process for maintaining synaptic integrity through a tightly regulated cycle involving clustering, docking-priming, Ca²⁺-triggered fusion, and endocytosis. In AD, amyloid-β (Aβ) oligomers interfere with SNARE-mediated fusion and endocytosis, while hyperphosphorylated tau obstructs vesicle mobility and docking, resulting in cumulative toxicity that aggravates SV defects. Conversely, in PD, α-synuclein (α-syn) aggregation alters vesicle clustering, membrane fusion, and recycling, and these effects are further influenced by Leucine-rich repeat kinase 2 (LRRK2)-Rab-related trafficking defects and the selective vulnerability of dopaminergic terminals. Different from previous reviews that address synaptic dysfunction in a broader manner, the present review is specifically organized around the SV trafficking cycle and compares both shared presynaptic endpoints and disease-specific upstream mechanisms in AD and PD. In addition, recent mechanism-oriented therapeutic strategies are summarized. This vesicle-cycle-centered perspective may provide a clearer framework for understanding presynaptic pathology and for guiding the development of earlier and more targeted interventions.

Bipolar disorder (BD) is characterized by mood swings between mania and depression, sharing overlapping symptomatic and genetic risk factors with other mood disorders. Long non-coding RNAs (lncRNAs) show specific spatiotemporal precision in distinct cell types in the human brain, and understanding the precise mechanisms of lncRNAs in mood switching in BD is fundamental to deciphering the key molecular networks underlying BD diagnosis and therapy. In this review, we summarize the classification of BD subtypes, the differences between BD and multiple mood disorders, and the functional potential of lncRNAs in BD. Future studies of these lncRNAs will facilitate the development of RNA-based diagnosis for BD.

Huntington's disease (HD) is caused by expansion of the polyglutamine stretch in the widely expressed huntingtin (HTT) protein. Patients with HD have motor, psychiatric and cognitive changes due to changes in a variety of neural circuits. Somatostatin-expressing interneurons (SST-INs) can regulate neural circuits largely by inhibiting their target cells. Behaviorally, brain-wide inhibition of SST-INs increased anxiety in mice. Silencing striatal SST-INs caused a decrease in movement in the open field. Mutant HTT (mHTT)-expressing mice exhibited abnormal motor, cognitive and psychiatric-like changes, as well as electrophysiological changes in a variety of neurons, including striatal SST-INs. However, it is unknown whether cell-autonomous expression of mHTT in SST-INs contributes to HD-associated behavioral phenotypes or causes abnormal electrophysiological changes in striatal SST-INs. To address these questions, we reduced mHTT expression in SST-INs throughout the brain of BACHD mice. Our findings show that brain-wide reduction of mHTT in SST-INs rescues anxiety-like behavior in male BACHD mice in the light-dark box, without improving performance in the open field or on the rotarod. Additionally, expression of mHTT in striatal SST-INs cell autonomously drives their increased excitability.

Aspartyl-tRNA synthetase (DARS) is implicated in several cancers, but its role in BCR::ABL-negative JAK2V617F-positive myeloproliferative neoplasms (MPNs) is unclear. This study evaluated DARS expression in MPN subtypes and its associations with clinical parameters and survival. Diagnostic bone marrow biopsies from 121 JAK2V617F-positive MPN patients (PV, n = 34; ET, n = 25; PMF, n = 54; MPN-U, n = 8) were stained on a Ventana BenchMark XT immunostainer using anti-DARS antibody (ABclonal A6574). DARS immunoreactive score (IRS) was determined by multiplying intensity (0–3) and percentage of positive cells (0–4). Inter-observer agreement was excellent (κ = 0.89, 95% CI: 0.83–0.95). Patients were stratified using the cohort’s median cutoff (IRS = 9) for survival analysis. Statistical tests included Kruskal-Wallis with effect sizes and multivariate Cox regression. DARS IRS differed across subtypes (H = 14.19, p = 0.003, η²=0.10): PV median 9 with IQR (9–12), ET 9 (8–12), PMF 8 (6–9), and MPN-U 10.5 (6–12). PV vs. PMF differences: intensity p = 0.001 (r = 0.35), IRS p < 0.001 (r = 0.38). DARS IRS correlated inversely with spleen size (ρ=–0.266, p = 0.003, 95% CI: − 0.43 to − 0.09), LDH (ρ=–0.194, p = 0.033), and fibrosis grade (ρ=–0.280, p = 0.002), and positively with hemoglobin (ρ = 0.308, p = 0.001, 95% CI: 0.13–0.47). High DARS expression independently predicted improved leukemia-free survival (LFS) (HR = 0.42, p = 0.007, 95% CI: 0.22–0.80) after adjusting for age, subtype, and fibrosis. DARS is differentially expressed in MPN subtypes. Its association with enhanced LFS identifies DARS as a potential prognostic biomarker warranting validation in larger cohorts.

BACKGROUND: Prematurity is a leading cause of neonatal and childhood mortality, with infections driving early deaths. Innate immunity provides frontline defense after birth and is shaped in part by myeloid-derived suppressor cells (MDSCs). The role of MDSCs in regulation of neonatal immunity, especially in the context of prematurity, remains elusive. We sought to understand the transcriptional landscape of neonatal immune myeloid regulators, specifically differences between preterm and full-term neonates. Insight into specific cellular networks could help understand how to skew preterm MDSCs’ development towards classical immunotolerant and anti-microbial mechanisms. METHODS: This cross-sectional study used single-cell RNA sequencing to characterize the neonatal MDSC transcriptional landscape, developmental trajectories, and predicted signaling networks within 48 h after birth. Peripheral blood mononuclear cells were isolated from 7 preterm neonates (< 36 weeks gestational age), 6 full-term neonates (> 37 weeks), and 6 healthy adult (21–45 years old, control). Primary exposure was premature birth and neonatal intensive care hospitalization from a single-center, academic tertiary care hospital between 2023 – 2024. RESULTS: Among ~ 339,000 cells, preterm neonates exhibited enrichment of polymorphonuclear MDSCs (10.6% ± 5.3%) vs full-term (2.6% ± 1.3%) and adults (0.4% ± 1.3%). Trajectory analysis identified a prematurity-associated differentiation branch characterized by inflammatory signaling, mitochondrial stress, and heightened protein-translation programs, distinct from a conserved, tolerogenic MDSC trajectory present across ages. Cell-communication modeling showed intensified outgoing and incoming signaling via ADGRE, RESISTIN, TGF-β, ANNEXIN, and ICAM networks. Antigen presentation signatures suggested preserved MHC-I output and diminished MHC-II interactions in neonates, with increased MHC-I input to preterm polymorphonuclear MDSCs. CONCLUSIONS: Prematurity is associated with early divergence of MDSC maturation toward an inflammatory and metabolically stressed PMN-MDSC state with altered immune communication. These findings identify cellular mechanisms that may contribute to the heightened tissue damage and infection susceptibility of preterm neonates and highlight MDSC signaling as a potential target for early-life immunomodulatory interventions.

Huntington's disease (HD) is characterized by progressive striatal degeneration associated with mutant huntingtin (mHTT)-related proteostatic disruption and chronic neuroinflammation. Although mHTT-lowering approaches hold therapeutic promise, their capacity to restore the degenerating neural microenvironment remains limited. Here, we evaluated the therapeutic potential of human induced pluripotent stem cell (iPSC)-derived neural precursor cells (s513-NPCs) in two complementary HD models, the acute R6/2 transgenic fragment model and the protracted, full-length YAC128 genomic model. Intrastriatal transplantation of s513-NPCs resulted in sustained functional improvement, including stabilization of motor coordination and attenuation of neuromuscular decline, across both disease contexts. These neuroprotective effects were accompanied by efficient donor cell engraftment and integration within the host striatum. At the molecular level, transplantation was associated with coordinated changes in proteostasis-related pathways, reflected by reduced mHTT aggregate burden and modulation of proteasomal and autophagic markers. In parallel, enhanced local BDNF-TrkB signaling was observed in grafted regions, consistent with improved neuronal support. Notably, transplanted NPCs exhibited context-dependent immunological responses, characterized by attenuation of pro-inflammatory signatures in aggressive disease stages and features of a reparative microenvironment in more protracted settings. Collectively, these findings demonstrate that iPSC-derived neural precursor transplantation confers robust neuroprotective effects in HD models, supporting its potential as a stem cell-based strategy to mitigate striatal pathology and functional decline.

<h4>Background and aim</h4>Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease characterized by aberrant fibroblast activation and excessive extracellular matrix (ECM) accumulation, resulting in irreversible lung remodeling. Although fibroblasts are central to IPF pathogenesis, comprehensive transcriptomic analyses of IPF patient-derived lung fibroblasts remain limited. This study aimed to identify novel molecular contributors to IPF through transcriptomic profiling of lung fibroblasts.<h4>Methods</h4>We performed total RNA sequencing on primary lung fibroblasts isolated from 33 IPF patients and 10 controls, the latter derived from histologically normal lung tissue adjacent to resected tumors. Differentially expressed genes (DEGs) were identified using trimmed mean of M-values normalization and the Exact test with Bonferroni correction. Gene ontology (GO) enrichment was conducted using DAVID, and network analyses were conducted via STRING and GeneClip3.<h4>Results</h4>GO analysis of 475 DEGs (402 upregulated, 73 downregulated) revealed strong enrichment in ECM-related terms consistently observed across three prior IPF fibroblast datasets. Pathway analysis further implicated cytokine-cytokine receptor interaction, complement and coagulation cascades, and immune-related processes. Network analysis identified SERPING1, NR4A1, and C3 as shared hub genes across both STRING and GeneClip3 platforms. Of the top 20 DEGs, 15 had prior associations with fibrosis, while, six (HSD17B2, HMGCLL1, RASL12, HBG1, LOC105375566 and RIPOR3-AS1) were novel. OAS2, an interferon-stimulated gene, emerged as a novel immune-fibrotic axis component in IPF.<h4>Conclusions</h4>This transcriptomic analysis confirms ECM dysregulation as a core feature of IPF pathogenesis and highlights novel DEGs and hub genes with potential roles in fibrosis, providing a foundation for future functional and translational studies.

Cerebral cavernous malformations (CCMs) are vascular lesions in the central nervous system that can cause strokes and seizures. Aggressive CCM growth follows an endothelial cell two-hit mechanism in which enhanced MEKK3-KLF2/4 signaling stimulates PI3K signaling, but how these pathways are linked has been undefined. Here, we use human CCM specimens, two mouse models of CCM disease, and primary human endothelial cells to examine the roles of the major endothelial growth factor receptors, VEGFR2 and TIE2. We find no evidence of augmented VEGFR2 signaling in CCM lesions, and neither genetic nor pharmacologic blockade of VEGFR2 reduced CCM formation in mouse models. Instead, we observe markedly increased phospho-TIE2 levels in human and mouse CCM lesions, MEKK3-KLF2/4-driven induction of TIE2 receptor expression, and almost complete rescue of CCM formation following genetic or pharmacologic TIE2 blockade in mouse models. Our studies identify TIE2 as the molecular link between the MEKK3-KLF2/4 and PI3K signaling pathways during CCM formation and suggest that targeting TIE2 may be an effective means to treat human CCM disease.

Despite the fact that amide and 1,2,3-triazole moieties on their own are widely utilized in designing chemoreceptor structures, their direct connection─amidetriazole is rarely studied. In this work, we present a new class of polyaromatic AIE-active amidetriazoles, their mechanochemical synthesis, and their application as dual-mode anion receptors. Mechanochemical synthesis not only enabled high yields but also shortened the reaction time and reduced the use of solvents and harmful chemicals. The optimization of 1,3-dipolar cycloaddition reactions carried out in solution, as well as in sono- and mechanochemical versions, was mainly based on modifying the copper-ion source, where a nonclassical approach (copper metal mesh) enabled the highest yields to be achieved. The compounds obtained were characterized by spectroscopic methods to fully study their properties derived from the AIE effect. The relationship between their structure and receptor properties was outlined. Ultimately, both AIE-active compounds were successfully employed as anion receptors. Optical and potentiometric methods demonstrated a strong correlation between the receptor structure and its detection properties, as well as the selective interaction of one receptor with SO₄^2- anions.

Breast cancer (BCa) is the most frequently diagnosed malignancy in women worldwide, with approximately 70% of cases driven by oestrogen receptor alpha (ERα). Endocrine therapies aim to suppress ERα signalling activity and form the foundation of current therapeutic strategies. However, a substantial proportion of patients either fail to respond due to intrinsic resistance or acquire resistance over the course of the treatment. This resistance arises through a complex interplay of factors including crosstalk with other signalling pathways such as Notch. Notch signalling, essential for mammary gland development, is aberrantly activated in breast tumours, where it contributes to cancer stem cell maintenance, epithelial-mesenchymal transition, angiogenesis, and metastasis. Notch receptors exert context- and subtype-specific roles: Notch1 and 4 promote tumour aggressiveness, whereas Notch2 often exhibits tumour-suppressive roles. In ERα-positive BCa, ERα and Notch signalling cooperate to drive resistance, whereas in ERα-negative disease, Notch promotes stemness and angiogenesis. While anti-oestrogen therapies effectively inhibit tumour growth, they can paradoxically activate Notch signalling and promote therapeutic resistance. Co-targeting Notch alongside endocrine therapy has been proposed as a strategy to delay the onset of therapeutic resistance. However, clinical development of Notch inhibitors has been limited by toxicity associated with pan-Notch blockade. More selective approaches, such as paralogue-specific antibodies, transcription-complex disruption, rational drug combinations, and advanced delivery platforms, are under active development to overcome these limitations. This review outlines the ERα-Notch crosstalk in BCa and examines current and emerging strategies for targeting Notch to overcome endocrine resistance and improve clinical outcomes.

Phosphorus(P) is essential yet limited, and Norway, like the rest of Europe, remains dependent on imported phosphate rock. Biochar from pyrolysis organic residues offers a potential alternative P source, although quantifying plant-available P is complicated by diverse P speciation. In this study, biochar produced at 400 °C from raw manure, manure digestate, and biologically treated sewage sludge digestate was evaluated using multiple P extraction methods (Olsen, Mehlich-3, ammonium lactate, citric acid, water, oxalate, dithionite, nitric acid). Pot experiments with wheat, spinach, and faba bean were conducted to assess the relationship between extracted P and plant uptake. Results showed that manure-derived biochars had higher agronomic value than sewage sludge biochar, which was limited by Fe/Al-bound P and heavy-metal contamination. Concentrated nitric acid digestion best predicted wheat and faba bean P uptake, while the more common extractants for agronomic evaluation (Mehlich-3, Ammonium Lactate, Olsen, DI-water extracts correlated most strongly with spinach uptake. Digested manure biochar met regulatory requirements for trace metals, but raw manure and sewage sludge biochar exceeded Ni and Zn thresholds, respectively. These findings highlight the need for a tiered approach to biochar P testing and for regulatory frameworks that balance environmental safety with the potential for nutrient recycling.

Mitochondrial dysfunction and dysregulated proteolysis drive Huntington's disease (HD), tauopathy, and related neurodegenerative disorders. Calpain-2, a Ca²⁺-activated protease restrained by calpastatin (CAST), is pathologically overactivated, yet no therapies directly target this axis. We identify A36, a brain-penetrant small molecule derived from CHIR99021 that selectively stabilizes the CAST-calpain-2 complex without inhibiting GSK3. A36 acts as a protein-protein interaction stabilizer, enhancing CAST-calpain-2 binding, preventing CAST degradation, and thereby limiting calpain-2 activation and mitochondrial damage. In patients with HD induced pluripotent stem cell-derived neurons and mutant mouse striatal neurons, A36 normalized mitochondrial morphology and membrane potential, reduced oxidative stress, and improved survival. In vivo, A36 displayed favorable pharmacokinetics and central nervous system exposure; treatment reduced striatal neurodegeneration, mutant huntingtin aggregation, and motor deficits in HD R6/2 mice, and lowered phosphorylated tau, neuroinflammation, and cognitive decline in tauopathy PS19 mice. These findings establish pharmacological stabilization of CAST-calpain-2 as a therapeutic strategy and position A36 as a mechanism-selective modulator with broad neurodegenerative disease potential.

Tandem duplications (TDs) are a common form of genomic rearrangements with both adaptive and pathogenic consequences. While prevalent in genomically unstable cancer genomes, TDs are rarely detected in normal tissues, suggesting the existence of robust protective mechanisms. Here, we identify the histone chaperone TONSL/TONSOKU (tnsl-1 in C. elegans) as a critical suppressor of TD formation. Loss of tnsl-1 results in the accumulation of TDs in two distinct size classes (~25 kb and ~300 kb), arising from different developmental contexts: small TDs emerge in rapidly dividing embryonic cells, whereas large TDs form in slower-dividing germline progenitors. Both classes depend on polymerase theta-mediated end joining (TMEJ), implicating DNA double-strand breaks in their genesis. Inhibition of break-induced replication (BIR) via Pif1 helicase loss reduces TD size, revealing a role for BIR in TD expansion. Remarkably, TONSL-deficient Arabidopsis thaliana exhibit an identical TD signature, highlighting the evolutionary conservation of this genome surveillance mechanism. These findings position TONSL as a cross-kingdom guardian of genome integrity through suppression of TD formation.

Copy number variation (CNV) plays a fundamental role in modulating plant agronomic traits and tumorigenesis in animals. While frequently linked to replication stress, the mechanisms giving rise to CNVs are not fully elucidated. Here we characterize the mutational consequences associated with losing the conserved TONSOKU (TSK/TONSL) pathway (CAF-1-H3.1-TSK), which is required to resolve impaired DNA replication forks. Using Arabidopsis thaliana, we demonstrate that tsk mutants rapidly accumulate large, heritable tandem duplications within their genomes that are consistent with DNA Polymerase θ (Pol θ) activity. These duplications are associated with late replicating heterochromatin enriched in sources of replication stress. We also show that stochastic developmental phenotypes in tsk plants are the result of the DNA Damage Response (DDR), with phenotype suppression occurring when ATR-WEE1 checkpoint signaling is removed. We thus describe a previously uncharacterized source of large tandem duplications that are relevant to understanding genome stability in diverse eukaryotes, and in disease contexts.

Huntington's Disease (HD) is characterized by progressive motor and cognitive decline, largely driven by cortico-striatal synaptic dysfunction. Central to these processes is huntingtin (HTT) protein, which is abundantly present at the synapse. HTT regulates the synaptic vesicle cycle at presynaptic terminals and serves as a scaffold at the postsynaptic density where it modulates receptor dynamics. An expanding network of HTT-interacting proteins (HIPs), crucial for maintaining synaptic structure and function, underscores the role of HTT as a core component of synaptic integrity. This review examines the 30-year research journey that has unveiled HTT pre- and postsynaptic partners, with focus on experimentally validated interactors and their involvement in HD cortico-striatal synaptic dysfunction.

Osteoclasts are essential for bone resorption and interact with osteoblasts during bone remodeling. Ion channels and transporters located in the ruffled border or intracellular vesicles coordinate the transport of various ions and substrates, which is fundamental to the primary functions of osteoclasts. Numerous channels and transporters are implicated in bone metabolic disorders and genetic diseases. Among these, the voltage-gated chloride channel 7 (ClC-7) and vacuolar proton ATPases (V-ATPase) represent the most well-characterized examples in osteoclasts. Using the classification system of the Transporter Classification Database, we reviewed nearly 90 osteoclastic ion channels and transporters, categorizing them into six groups: ATPases, cation channels, anion channels, complex transporters, organic substance transporters, and ATP-binding cassette transporters. We summarized recent advances in their subcellular localization, transported substrates, associated diseases, and physiological roles in relevant biological functions and signaling pathways. Notably, transporters for hydrogen, chloride, phosphate, and calcium are particularly critical for osteoclast function. We also reviewed therapeutic candidates targeting these ion channels and discussed strategies for their future development. As transcriptome and other advanced techniques have identified more channels and transporters in osteoclasts, the diversity and unexplored functions of these molecules may exceed previous understanding. Increased attention to their widespread distributions and interactions could reveal new therapeutic targets for osteoclast-related and other bone disorders.

APOE4 is the leading genetic risk factor for Alzheimer's disease, and chronic stress is a leading environmental risk factor. Studies suggest that APOE4 confers vulnerability to the behavioral and neuropathological effects of chronic stress, representing a potential mechanism by which this genetic variant accelerates Alzheimer's onset and progression. Whether and how APOE4-mediated stress vulnerability manifests in neurons of the hippocampus, a brain region particularly susceptible to stress and Alzheimer's pathology, remains unexplored. Using a combination of in vivo and in vitro experiments in humanized APOE4 and APOE3 knockin mice and primary hippocampal neurons from these animals, we investigated whether and how APOE4 confers sensitivity to glucocorticoids (GCs), the main stress hormones. We found that a hallmark of stress/GC-induced brain damage, tau pathology (i.e., tau accumulation, hyperphosphorylation, and spreading) is exacerbated in APOE4 versus APOE3 mice. Moreover, APOE4 animals exhibit underlying mitochondrial dysfunction and enhanced glucocorticoid receptor activation in the hippocampus, factors that likely contribute to tau pathogenesis in both the presence and absence of stress/GCs. Supporting this concept, opening of the mitochondrial permeability transition pore (mPTP) drives mitochondrial dysfunction and tau pathology in APOE4 mice, while pharmacological inhibition of the mPTP is protective against ApoE4-mediated mitochondrial damage, tau phosphorylation and spreading, and downstream hippocampal synapse loss. These findings shed light on the mechanisms of stress vulnerability in APOE4 carriers and identify the mPTP as a potential therapeutic target for ameliorating Alzheimer's pathogenesis in this population.

Essential amino acids (EAA) may alter bovine mammary epithelial cell (BMEC) activities through regulation of gene transcription. For this study, we used pathway analysis of RNA-sequencing data to identify upstream transcription factors (TF) that potentially mediate the effects of single EAA deficiency on BMEC. Differentiated BMEC were cultured in 1 of 4 treatment media representing normal physiological concentrations of all amino acids (CTL), or ¼ the normal concentration of histidine (LH), lysine (LK), or methionine (LM). After 48 h, rates of DNA synthesis were reduced 30 to 40% by each of the deficiencies. Protein synthesis rate was 30 to 50% lower in LK and LM but was unaffected by LH. Up-regulated upstream TF included those related to nutrient stress, interferon signaling, and suppression of cell population growth. Down-regulated TF were primarily related to cell cycle progression. An array of classic ATF4 targets increased in response to EAA deficiencies including amino acid transporters, branched-chain aminotransferase, aminoacyl-tRNA synthetases, sestrin2 (a leucine sensor), and activators of apoptosis. Interferon signaling emerged as a novel pathway activated by EAA deficiencies, possibly contributing to reduced cell proliferation and protein synthesis. In contrast, the MAPK signaling pathway was unaffected. Expression of hypoxia-inducible factor 1α (HIF1A) increased, consistent with an anti-proliferative role. Downregulation of Forkhead box protein M1 (FOXM1) likely contributed to slower cell proliferation. These findings reveal a complex network of transcriptional regulators and signaling pathways influenced by EAA availability and shed light on the nutritional regulation of milk protein production by dairy cows.

BACKGROUND: Attention-Deficit/Hyperactivity Disorder (ADHD) is a prevalent neurodevelopmental condition with high societal impact. While psychostimulants like methylphenidate (MPH) are first-line treatments, 10–20% of patients show insufficient efficacy, and many experience side effects such as sleep and appetite disturbances. Emerging evidence suggests that neuroinflammation and oxidative stress contribute to ADHD pathophysiology. Animal studies indicate that MPH may induce pro-inflammatory processes, but this has yet to be characterized in humans. METHODS: The ANIME study (ADHD Nutritional, Inflammatory, and Metabolic Endophenotypes) employs a combined case-control and prospective cohort design. A case-control study will compare the immune, inflammatory, and nutritional profiles of 40 children/adolescents with ADHD against 40 age- (± 2 years) and sex-matched typically developing controls. The ADHD group will have a follow-up visit to assess the impact of psychostimulant treatment). The primary outcome is the comparison of the immune and inflammatory markers’ levels, particularly IL-6 levels, at baseline. Secondary outcomes include deep immunophenotyping via spectral flow cytometry (Cytek Aurora) and high-sensitivity multiplex proteomics (NULISA), alongside assessments of polyphenol bioavailability, diet quality (KIDMED), and clinical symptoms. DISCUSSION: ANIME will provide an in-depth characterization of the “inflammatory biotype” in ADHD, specifically examining how psychostimulants interact with pre-existing immune states. By integrating metabolic, nutritional, and clinical data, the study aims to identify biomarkers for treatment response and tolerability, ultimately informing personalized care and future trials of antioxidant-rich adjunct therapies like polyphenols. TRIAL REGISTRATION: Clinicaltrials.gov. Number: NCT07417878 ( https://clinicaltrials.gov/study/NCT07417878 ). Registered on the 13th of Febraury, 2026.

<h4>Background</h4>The left inferior frontal gyrus (IFG) is implicated in both language and motor processes. Its functional reorganization post-stroke, particularly in motor dysfunction without aphasia, remains poorly understood. We investigated effective connectivity (EC) alterations of the left IFG and their neurochemical basis, and responsiveness to neuromodulation in subcortical stroke.<h4>Methods</h4>Cross-sectional analysis included 32 left (LSS) and 27 right (RSS) stroke patients and 40 healthy controls (HCs). Seed-based EC of the left IFG was derived from resting-state fMRI and was compared between groups. Relationships between EC, neurotransmitter density, lesion-derived neurotransmitter indices, and Fugl-Meyer Assessment (FMA) scores were examined. Longitudinally, 30 patients received 14 sessions of cathodal-contralesional sensorimotor cortex (SMC), anodal-ipsilesional M1, or sham transcranial direct current stimulation (tDCS), synchronized with upper limb training. Pre-post FMA and EC changes were compared.<h4>Results</h4>Both LSS and RSS groups showed increased EC from the left IFG to the right cerebellum posterior lobe (CPL) and superior frontal gyrus (SFG), whereas only LSS group showed decreased EC from the left CPL to the left IFG. EC from the left IFG to the right CPL positively correlated with FMA, while EC from the left CPL to the left IFG negatively correlated with FMA in LSS. These EC alterations were significantly associated with serotonergic, dopaminergic, and GABAergic neurotransmitter densities. Crucially, IFG-to-SFG connectivity mediated the relationship between lesion-derived neurotransmitter network damage and lower-limb motor deficits. Longitudinal intervention revealed that different tDCS protocols distinctively modulated the left IFG's EC, with the most robust changes induced by cathodal stimulation of the contralesional SMC.<h4>Conclusions</h4>Collectively, motor-related reorganization occurs in the left IFG post-stroke, characterized by altered EC patterns that are (1) correlated with motor performance, (2) underpinned by specific neurochemical systems, and (3) mediate post-stroke motor impairment. These reorganizations are plastic and respond specifically to targeted neuromodulation, highlighting the left IFG as a potential novel therapeutic target for motor recovery and informing personalized rehabilitation strategies.<h4>Trial registration</h4>All data used in the present study were obtained from the research trials registered on ClinicalTrials.gov (NCT05648552; submitted 5 December 2022) and www.chictr.org.cn (ChiCTR2100044970; submitted 3 April 2021).

<h4>Background & aims</h4>Colorectal cancer (CRC) remains a leading cause of cancer‑related morbidity and mortality worldwide. Although the adenoma-carcinoma sequence and its genetic drivers are well described, the earliest cellular and molecular events initiating tumorigenesis within histologically normal colonic epithelium remain poorly defined. This study aims to identify tumor‑initiating cells (TICs), distinguish them from normal stem‑like cells (nSTMs), and delineate early transcriptional and signaling programs using single‑cell RNA sequencing (scRNA‑seq) from paired normal‑appearing and transformed human colonic tissues.<h4>Methods</h4>Fresh biopsies from histologically normal mucosa and matched polyps, including tubular adenomas, sessile serrated adenomas, and adenocarcinomas, were collected from seven subjects. Single‑cell transcriptomes were generated using the 10x Genomics platform and analyzed with Seurat, Monocle2, CytoTRACE, GSEA/GSVA, RNA velocity, InferCNV, CellChat, and NicheNet. Spatial validation was performed using RNA‑FISH.<h4>Results</h4>We resolved 51,054 high‑quality single‑cell transcriptomes into 33 clusters. Tumor-specific stem-like (tSTM) and deep crypt secretory (tDCS) populations were enriched in adenomas. Subclustering of tSTM identified TIC-like subsets predominantly derived from histologically normal mucosa that localized to the root of lineage trajectories leading to polyp-enriched tSTM states. Compared to nSTMs, TICs exhibited enhanced stemness potential, early epithelial-mesenchymal transition (EMT) and interferon signaling, suppression of oxidative phosphorylation, and distinct genomic and signaling features, indicating early neoplastic reprogramming. ETS2, SLC12A2, and LEFTY1 were identified as TIC‑specific markers; SOD3 and GPRC5A increased along the TIC‑to‑tSTM trajectory. RNA‑FISH confirmed candidate marker localization. Independent validation using the COLONMAP dataset (30 polyps, 35 normal samples) demonstrated that TIC-like cells were predominantly enriched in tubular adenomas but were scarce in serrated lesions. Across this independent cohort, TIC marker genes showed reproducible upregulation in TIC-like populations, supporting the robustness of these observations across cohorts.<h4>Conclusions</h4>Our results identify TICs as the origin of neoplastic stem‑like states in the conventional tubular adenoma pathway and define early transcriptional, metabolic, and microenvironmental reprogramming events that distinguish TICs from nSTMs. In contrast to serrated pathways described in other atlases, our data support a stem‑like expansion model for tubular adenomas and nominate biomarkers with translational potential for early CRC detection and intervention.

Early life stress (ELS) exerts lasting effects on prefrontal cortex (PFC) development and function, contributing to cognitive and emotional vulnerabilities later in life. This review synthesizes findings from human studies and animal models showing that ELS disrupts PFC-dependent cognitive functions, including executive control, attention, working memory, and decision-making. We examine how ELS alters functional connectivity, neurotransmitter systems such as serotonin, GABA, and Dopamine, and hormonal signaling, particularly glucocorticoid regulation. Finally, we explore evidence for epigenetic mechanisms within the PFC that mediate the long-term impact of ELS on cognition and mental health.

<h4>Introduction</h4>Histone demethylation mediated by Jumonji C (JmjC) domain-containing proteins is a pivotal epigenetic mechanism governing plant development and stress adaptation. However, a comprehensive characterization of this family in foxtail millet (<i>Setaria italica</i>) and its wild progenitor, green foxtail (<i>Setaria viridis</i>), remains lacking.<h4>Methods</h4>In this study, we performed a genome-wide identification of 24 <i>SiJMJ</i> and 23 <i>SvJMJ</i> genes, classifying them into five conserved subfamilies. Comparative genomics, promoter cis-element analysis, transcription factor binding site prediction, and expression profiling under various abiotic stresses were conducted to characterize their evolutionary and functional divergence.<h4>Results</h4>Comparative genomics revealed extensive collinearity and structural conservation between the two species, yet significant divergence in promoter cis-elements and transcription factor binding sites suggested potential regulatory remodeling during domestication. Expression profiling indicated distinct tissue-specific patterns, particularly in apical meristems and germination. Under abiotic stresses, <i>SiJMJ17</i> was significantly upregulated under saline-alkali and cold stresses, whereas <i>SiJMJ1</i> exhibited specific upregulation under waterlogging and herbicide treatments.<h4>Discussion</h4>These findings elucidate the evolutionary history of the Setaria JMJ family and highlight candidate demethylases for enhancing stress resilience in C4 cereal crops through epigenetic engineering.

<h4>Background</h4>Neonatal acute liver failure (ALF) is a rare, life-threatening condition often accompanied by complex coagulopathy. Managing this coagulopathy is challenging due to the limitations of conventional coagulation tests and the lack of pediatric-specific transfusion guidelines, often leading to empiric blood product use.<h4>Methods</h4>We present a detailed case report of a neonate with ALF of indeterminate etiology. Clinical and laboratory monitoring included serial conventional coagulation tests (PT, APTT, INR, fibrinogen, D-dimer), coagulation factor assays, and thromboelastography (TEG). The therapeutic management and response to blood product transfusions (fresh frozen plasma, cryoprecipitate, platelets) were documented over a one-month hospitalization.<h4>Results</h4>The patient exhibited severe and persistent abnormalities in standard coagulation parameters (prolonged PT/APTT, elevated INR, hypofibrinogenemia, and deficiency of factors II, V, VII, IX-XII, ATIII, PC, PS) alongside thrombocytopenia. In contrast, TEG revealed a normal reaction (R) time, but a reduced alpha-angle and maximum amplitude (MA). Despite 14 transfusions of fresh frozen plasma, conventional coagulation parameters showed minimal improvement.<h4>Conclusion</h4>This case illustrates the complexity of coagulation monitoring in neonatal liver failure. It underscores the significance of comprehensive global hemostasis assays, such as TEG, which may provide superior guidance for transfusion therapy and help prevent the unnecessary administration of blood products in non-bleeding neonates with ALF.

<b>Background/Objectives</b>: Hepatitis E virus (HEV) infection is an increasingly recognized cause of acute hepatitis in Europe, but short-term in-hospital laboratory dynamics remain insufficiently described in hospitalized cohorts. We aimed to characterize admission biochemical abnormalities and paired admission-to-discharge laboratory changes in hospitalized patients with acute hepatitis E from Craiova, Romania, with exploratory sex- and age-stratified analyses. <b>Methods</b>: We conducted a single-center retrospective observational study including 40 consecutive hospitalized patients with acute hepatitis E during 2024-2025. Admission and discharge laboratory values were compared at the within-patient level, and exploratory subgroup analyses by sex and age class were performed. Given the limited sample size, multivariable analyses were restricted to parsimonious age-adjusted models for selected endpoints. <b>Results</b>: The cohort comprised 22 females (55%) and 18 males (45%), with a mean age of 53.05 ± 21.44 years; two in-hospital deaths occurred. At admission, marked transaminase elevation and frequent hyperbilirubinemia were observed, with 70% of patients having total bilirubin ≥ 2 mg/dL and 40% ≥ 10 mg/dL. During hospitalization, ALT and AST declined markedly, whereas total and direct bilirubin improved more modestly, indicating slower resolution of jaundice/cholestatic abnormalities. Platelets increased, while prothrombin index changes were heterogeneous. Male patients had higher bilirubin values at admission and discharge and more frequent clinically relevant hyperbilirubinemia thresholds; however, these findings should be interpreted cautiously given the small sample size, the retrospective design, and the absence of standardized clinical confounders and mechanistic data. Exploratory age-stratified analyses did not identify robust differences after multiplicity control. <b>Conclusions</b>: In hospitalized hepatitis E, hepatocellular injury markers improved rapidly during hospitalization, whereas cholestatic abnormalities resolved more slowly and often remained clinically relevant at discharge. The observed sex-related cholestatic pattern should be considered exploratory and requires confirmation in larger studies with standardized clinical covariates and longer follow-up. These findings support closer monitoring of bilirubin trajectories at discharge, particularly in male patients, and highlight the need for integrating laboratory dynamics into short-term clinical assessment of hospitalized HEV cases.

Temozolomide and dacarbazine are untargeted anticancer prodrugs that have been widely employed in the treatment of melanoma and glioblastoma. These agents decompose into a short-lived monomethyl triazene intermediate, culminating in the release of a methyl diazonium cation that serves as the DNA-alkylating species responsible for tumour destruction. However, due to their high chemical lability, these agents have been associated with chemotherapy resistance, mutagenicity, tumour relapse, and significant off-target toxicity. One promising strategy towards the resolution of these limitations involves the design of arylmethyl triazene prodrugs, which enable targeted tumour-specific drug delivery. This review explores the various approaches used to selectively deliver alkyl aryl triazenes as alternatives to current therapies. It highlights early chemical strategies such as <i>N</i>-acylation and etherification of monomethyl triazenes, along with associated kinetic studies. The selective activation of novel triazenes in murine and human melanoma cells through a tyrosinase-responsive promoiety is discussed. Recent progress in nitroaromatic-based prodrugs designed to exploit the hypoxic microenvironment of glioblastoma is also examined. Additionally, we summarise the development of combi-triazenes and their underlying chemistries, which enable the simultaneous release of two active therapeutic agents.

Increased intestinal permeability has been implicated in metabolic dysfunction-associated steatotic liver disease (MASLD), but its relationship with liver fibrosis independent of metabolic risk factors remains unclear. The aim of this study was to investigate the relationship between markers of gut-derived immune activation and liver fibrosis in individuals with metabolic disease. We enrolled 139 adults (48.8 ± 11 years; BMI 33.7 ± 9.5 kg/m²; 50% type 2 diabetes); liver steatosis and fibrosis were estimated using the Hepatic Steatosis Index (HSI) and Fibrotic NASH Index (FNI); liver biopsies were available in a bariatric subgroup. Plasma soluble CD14 (sCD14) and lipopolysaccharide-binding protein (LBP) levels were measured by ELISA kits, and the LBP/sCD14 ratio was calculated. MASLD was present in 78% of participants; in these individuals, sCD14 levels correlated with HSI and FNI (both <i>p</i> < 0.01). In multivariable analysis adjusting for age, sex, BMI, waist circumference, and type 2 diabetes, sCD14 was independently associated with advanced fibrosis (OR: 3.16, 95% CI 1.32-7.55; <i>p</i> = 0.010). This association was confirmed by histology (<i>p</i> = 0.02). Overall, these findings point to a link between gut-derived immune activation and fibrotic burden in MASLD and provide insight into the pathophysiological relevance of the gut-liver axis in metabolic disease.

Dental tissue regeneration, particularly alveolar bone and gingival repair, remains a major challenge in regenerative medicine. 3D bioprinting offers patient-specific and anatomically precise constructs, representing an advanced alternative to conventional grafting. In this study, nanohydroxyapatite (nHA), chitosan (CS), and collagen (CoL) were combined to fabricate and characterize 3D bioprinted dental grafts. SEM revealed a highly porous, interconnected architecture favorable for cell infiltration and nutrient exchange. EDS confirmed Ca/P ratios of 2.06 for nHA/CoL and 1.83 for nHA/CS/CoL, both of which are above the stoichiometric 1.67, indicating the presence of additional mineral phases and ion substitutions. FTIR and XRD verified characteristic functional groups and crystalline phases, including B-type HA with carbonate substitution. Mechanical testing showed that pure nHA exhibited the lowest compressive strength, whereas CoL incorporation improved stiffness. The nHA/CS/CoL composite achieved the highest compressive strength, elastic modulus, and toughness, demonstrating superior mechanical resilience. DSC analysis indicated endothermic peaks at 106.49 °C and 351.91 °C, with enthalpy values (264.91 J/g and 15.09 J/g) surpassing those of nHA alone. TGA revealed ~28.8% weight loss across three degradation stages, confirming enhanced thermal stability. In vitro cytocompatibility testing using L929 fibroblasts validated the biocompatibility of the composites. Collectively, the synergy between bioceramics and biopolymers markedly improved both mechanical and thermal performance. These findings position the nHA/CS/CoL scaffold as a promising candidate for clinical applications in dental tissue regeneration. Unlike conventional grafting materials, this study introduces a synergistically optimized nHA/CS/CoL bio-ink formulation specifically designed for extrusion-based 3D bioprinting of patient-specific dental constructs. The core innovation lies in the precise integration of nHA within a dual-polymer matrix (CS/CoL), which bridges the gap between mechanical resilience and biological signaling, achieving a compressive strength that mimics native alveolar bone while maintaining high cytocompatibility.

When using traditional approaches, such as pharmacokinetics and pharmacodynamics, the entire cellular or molecular response to drugs in the body cannot be fully ascertained or established. The oral medication process involves pharmacokinetics, followed by oral microbiomics and then gut microbiomics and pharmacodynamics. Recently, there has been increasing interest in the role of genetics (pharmacogenetics and pharmacogenomics) in both humans and microbiomes, as well as omics alterations (e.g., epigenetic, transcriptomic, proteomic, and metabolomic alterations as a consequence of drug exposure), which can help to ascertain the cellular responses to medications. Both the efficacy and toxicity of a drug are influenced by these factors. To assess these at an individual level, an integrative Personalized Medicine Model may be needed to help with medication management. Two example application cases for SSRIs and statins demonstrate the clinical usefulness of such a model, which can guide clinicians during drug selection and dosing to reduce reliance on trial-and-error, thus potentially improving patient outcomes and safety. Integrating this framework into practical clinical workflows requires the capture, analysis, and translation of multi-omics data in order to realize decision support protocols and actionable drug recommendations. This review also discusses IT requirements and different stakeholder roles. Although the proposed model can guide the treatment of diseases at the individual patient level, further research is still needed before it can be implemented as part of drug development research, clinical care, and healthcare delivery systems.

Hair follicles are highly specialized mini-organs within the skin that drive the production of wool and cashmere, traits of major biological and economic importance in sheep and goats. Despite their microscopic size, hair follicles exhibit extraordinary regulatory complexity, integrating genetic programs with seasonal, endocrine, environmental, and epigenetic cues. Although transcriptional networks and signaling pathways underlying follicle morphogenesis and cycling have been extensively investigated, the post-transcriptional mechanisms that fine-tune these processes remain insufficiently understood. MicroRNAs (miRNAs) have emerged as pivotal post-transcriptional regulators that coordinate cell fate determination, lineage commitment, and tissue homeostasis. Growing evidence indicates that miRNAs play essential roles in hair follicle stem cell maintenance, proliferation, differentiation, apoptosis, and organ-level development, functioning through interconnected regulatory networks rather than isolated linear pathways. By modulating the expression of key follicle-determining genes and signaling components, miRNA-mediated regulation shapes follicle formation, cyclic regeneration, and fiber traits. In this review, we synthesize recent advances in miRNA research related to hair follicle biology, with a particular focus on wool- and cashmere-bearing mammals. We integrate findings across species to propose a systems-level framework in which miRNA networks interface with canonical signaling pathways and epigenetic mechanisms to orchestrate follicle development and regeneration. Conserved and species-specific regulatory principles are discussed to bridge fundamental follicle biology with practical applications in fiber production. Overall, this review highlights miRNAs as a critical yet previously underappreciated regulatory layer in hair follicle biology. A deeper understanding of miRNA-mediated control provides new conceptual insights into wool and cashmere development and offers a foundation for future molecular breeding and precision regulation strategies in livestock.

<h4>Introduction</h4>Androgen deprivation therapy (ADT), widely used in men with advanced prostate cancer, has been implicated in cognitive decline. Evidence from Western cohorts remains inconsistent, with limited long-term and India-specific data.<h4>Methods</h4>We conducted a prospective, observational case-control study between July 2021 and December 2024 with a minimum 36-month follow-up. Men initiating ADT (luteinizing hormone-releasing hormone analogues or bilateral orchidectomy) were compared with a control cohort (benign prostatic hyperplasia or postradical prostatectomy without ADT). Groups were matched for age, baseline Addenbrooke's cognitive examination III (ACE-III) score, G8 frailty score, and gait speed. Cognitive assessments were performed every 6 months using the culturally validated Hindi ACE-III tool. The primary outcome was the change in total ACEII score over 36 months; secondary outcomes included domain-wise changes and correlations with frailty and gait speed.<h4>Results</h4>Sixty men were enrolled (30 per group); 21 men received ADT, and 23 served as controls, completing follow-up. Over 36 months, both groups showed modest declines in cognition, with a mean decline of -2.30 ± 1.13 for the ADT group and -1.87 ± 1.62 for the control group (<i>P</i> = 0.638). Domain-wise changes (attention, memory, fluency, language, and visuospatial) were small and statistically similar. Stratified analyses by frailty (G8 ≤ 14 vs. >14) and gait speed (≤0.6 m/s vs. >0.6 m/s) revealed parallel trajectories in both groups, with no effect of ADT.<h4>Conclusions</h4>In this 36-month prospective study using a culturally validated tool, ADT was not associated with accelerated global or domain-specific cognitive decline compared to a matched cohort without ADT. These findings suggest that in appropriately selected patients, ADT can be administered without undue concern for disproportionate cognitive deterioration.

<title>Abstract</title> <p>Background Alport syndrome is an X-linked inherited kidney disease caused by mutations in type IV collagen genes (COL4A3, COL4A4, and COL4A5) that affect the glomerular basement membrane and can lead to sensorineural hearing loss and eye abnormalities. This disease, which can cause hematuria, proteinuria, and in more severe cases chronic kidney disease, is characterized by bilateral and high-frequency sensorineural hearing loss, eye dysfunction, and in rare cases, aortic aneurysm; however, abnormalities in iron metabolism are not commonly described. Case Presentation: We report a case of persistent and marked hyperferritinemia in a patient with genetically confirmed Alport syndrome, without blood transfusion, significant inflammation, or hereditary hemochromatosis. In 2016, a patient whose etiology of proteinuria and hematuria was investigated showed no abnormalities in iron parameters in laboratory tests performed at that time, while as of 2021, serum ferritin levels were found to be > 1000 µg/L. Abdominal ultrasound and MRI scans revealed severe hepatosplenomegaly. Minimal increases in infection and inflammation markers did not explain this severely elevated ferritin level. Hemochromatosis gene mutations were tested in several different laboratories, and no mutations were detected. Conclusion This case suggests a possible dysregulation of iron metabolism in a patient diagnosed with Alport syndrome, beyond the inflammation associated with conventional chronic kidney disease. The interaction between collagen IV mutations and systemic inflammation and iron regulatory pathways may require more detailed investigation.</p>

Ebola virus (EBOV) causes re-emerging outbreaks of viral hemorrhagic fever with high case-fatality rates. As a negative-sense single stranded RNA virus, EBOV depends on its RNA-dependent RNA polymerase (L protein) to transcribe and replicate the viral genome. This takes place in cytoplasmic inclusion bodies that form following host cell remodeling involving yet unknown signaling pathways. Here, using mass spectrometry-based (phospho-)proteomics, we profiled global protein abundance and site-specific phosphorylation in HEK293T cells expressing an EBOV minigenome system. Our data reveal EBOV-induced rewiring of the host proteome and phosphorylation signaling landscape, including perturbations in cell cycle control, cytoskeletal organization, innate immune regulation, and DNA damage response. Kinase network analysis revealed that Hippo pathway kinases and especially cyclin dependent kinases like CDK2 are central drivers of EBOV replication and transcription. The functional necessity of these signaling pathways is demonstrated via inhibition of CDK family kinases with small molecule inhibitors, which halted EBOV minigenome replication and transcription when administered in the low micromolar range, demonstrating that these pathways represent putative antiviral targets.

Adenosine-to-inosine (A-to-I) RNA editing, catalyzed by adenosine deaminases acting on RNA (ADARs), is a widespread modification in metazoans. Cumulative evidence has revealed the altered A-to-I editing profiles in cancers, but the underlying mechanism remains unclear. Here, we discover the well-known histone lysine methyltransferase enhancer of zeste homologue 2 (EZH2) as an unexplored ADAR interactor and editing regulator in prostate cancer (PCa). Through competing with interleukin enhancer binding factor 2 (ILF2) for ADAR1 binding, EZH2 reshapes the substrate selectivity of ADAR1 and thus exhibits a bidirectional role in editing regulation. Moreover, EZH2 depletion induces the translational repression of transportin-1 (TRN1), which further results in the accumulation of cytoplasmic ADAR1p110 isoform to protect many oncogenic transcripts from degradation. Consistently, depletion of ADAR1 dramatically enhances the sensitivity of cancer cells and tumors to EZH2 selective degraders. Collectively, our study sheds new light on a link between two layers of epigenetic regulations at histone modification and RNA editing levels, demonstrates a previously uncharacterized role of EZH2 in RNA editing and mRNA stability independently of its lysine methyltransferase activity, and reveals the significance of EZH2-ADAR1 cascade in governing RNA editing and mRNA stability, which may provide additional perspectives for the advancement of EZH2-targeting cancer therapies.

BACKGROUND: Lung adenocarcinoma (LUAD) is a prevalent malignancy with poor clinical prognosis, and the specific role of PCDHGA3 in LUAD pathogenesis remains to be fully clarified and understood. METHODS: TCGA, UCSC XENA, and Human Protein Atlas databases were comprehensively utilized to systematically analyze PCDHGA3 expression levels, clinical prognostic significance, DNA methylation status, and mutation patterns. GO, KEGG, and GSEA enrichment analyses were performed to identify related biological pathways and functions. Western blotting, wound healing assays, cell adhesion assays, CCK8 proliferation assays, and transwell migration assays evaluated PCDHGA3 biological functions. Animal models were employed to validate in vivo effects and mechanisms. RESULTS: PCDHGA3 demonstrated low expression in LUAD tissues. High mutation rates and altered methylation patterns indicated potential involvement in LUAD development. PCDHGA3 overexpression notably inhibited cancer cell proliferation and migration abilities in both in vitro and in vivo models. Moreover, PCDHGA3 overexpression markedly elevated E-cadherin protein levels while substantially reducing N-cadherin, phospho-SMAD2, phospho-SMAD3, and TGFβ protein levels. CONCLUSION: PCDHGA3 inhibits LUAD progression and metastasis by suppressing the TGFβ signaling pathway, providing novel therapeutic insights.

BACKGROUND: Saxagliptin is a DPP-4 inhibitor widely used to manage type 2 diabetes, though genetic contributors to variability in response remain unclear. This study aimed to identify genetic variants associated with the efficacy and safety of saxagliptin using genome-wide association studies (GWAS) with data from randomized controlled trials. METHODS: GWAS were conducted on glycated hemoglobin (HbA1c) levels and hypoglycemic events in saxagliptin-treated individuals from 12 phase II or III clinical trials. Additional exploratory traits included fasting glucose, weight, body mass index, systolic and diastolic blood pressure, heartbeat, and renal function. Primary analyses focused on participants of European ancestry, followed by analyses in the overall multi-ancestry cohort. Linear mixed models for continuous traits and Cox proportional hazards models for time-to-event analyses were used, including gene-by-sex and gene-by-treatment interactions. We also performed a transcriptome-wide association study (TWAS) to identify genes whose predicted expression is associated with the response to saxagliptin using the S-PrediXcan framework. RESULTS: Among 1 016 European and 1 826 multi-ancestry saxagliptin-treated participants, no genome-wide significant associations were observed. Suggestive associations included rs2168426:T > C near UBE2E1 for HbA1c (p = 1.0 × 10⁻⁷) and rs138558907:C > T near CCSER2 for hypoglycemia (p = 6.1 × 10⁻⁸). Treatment interaction analyses showed a stronger rs2168426 effect on HbA1c among saxagliptin users compared with non-users (pint = 9.2 × 10⁻³). The TWAS revealed three genes that significantly associated with HbA1c in human tissues: OGFOD3 in aorta (p = 8.3 × 10⁻⁷), TBC1D3D in coronary artery (p = 3.3 × 10⁻⁶), and TYSND1 in amygdala (p = 3.9 × 10⁻⁶). In the multi-ancestry cohort, KDM7A in whole blood was significantly associated with HbA1c (p = 3.6 × 10⁻⁶). CONCLUSION: Larger, diverse studies are needed to identify pharmacogenomic determinants of responses to saxagliptin.

Risk of fractures may be increased in individuals with iron deficiency, iron overload, and/or HFE hemochromatosis. To test this hypothesis, we followed 142,146 Danish general population individuals for a median of 11 years (range:0-41) after study enrolment for hospital and emergency room admissions with fractures. All individuals had blood samples drawn at study enrolment. We measured iron, transferrin saturation, and ferritin in 136,611, 136,555, and 37,990 individuals, respectively, while 132,499 individuals were genotyped for the HFE C282Y and H63D variants. We found a U-shaped relationship between fracture risk and concentrations of plasma iron and transferrin saturation when studying all individuals irrespective of genotype. When studied according to plasma ferritin, fracture risk was increased in individuals with low ferritin concentrations, while risk was not increased in individuals with high concentrations. When compared to non-carriers, HFE C282Y homozygotes had increased risk of any fracture (hazard ratio[HR]:1.38;95%CI:1.09-1.75;p=0.008), and risk was increased even in C282Y homozygotes with normal ferritin concentrations (HR:2.89;95%CI:1.50-5.56), which is important as these individuals would not usually be recommended for HFE genotyping according to clinical guidelines. When compared to non-carriers, risk of fracture of the hip and femur was increased in C282Y homozygotes (HR:1.78;95%CI:1.17-2.70;p=0.007) but surprisingly also in H63D homozygotes (HR:1.21;95%CI:1.00-1.47;p=0.04), C282Y heterozygotes (HR:1.10;95%CI:1.00-1.21;p=0.04), and C282Y/H63D compound heterozygotes (HR:1.23;95%CI:1.00-1.51;p=0.05). The markedly increased fracture risk in C282Y homozygotes with normal ferritin may challenge the presumption that systemic iron accumulation is the primary mechanism causing their increased fracture risk. Further studies are needed to examine whether phlebotomy reduces fracture risk.

Early detection of cancer is essential for effective treatment. However, current prostate cancer screening methods lack sufficient sensitivity and specificity, leading to overdiagnosis and unnecessary treatment. There is also an unmet need to distinguish clinically significant from insignificant prostate cancer. To identify complementary biomarkers for improved screening and diagnosis, we performed transcriptional profiling of cancer-associated transcripts in circulating extracellular vesicles (EVs) isolated from peripheral blood of patients with suspected prostate cancer prior to biopsy and healthy donors. Expression data for 2549 mRNAs were obtained from 28 men. CAPN5 expression was significantly lower, whereas BIRC2, CASP3, CD63, FMO5, IRF6, PFDN1, PRDX6, PSMD2, RIT1, S100A2, THBS1, and XRCC2 were significantly elevated in EVs from patients with significant prostate cancer (n = 14) compared with cancer-free individuals and patients with insignificant disease (n = 14). Candidate biomarkers were subsequently evaluated by in silico validation using the The Cancer Genome Atlas (TCGA) prostate adenocarcinoma dataset and the GEO dataset GSE70768 containing benign and malignant prostate tissues. This analysis identified CASP3, XRCC2, and RIT1 transcripts in circulating EVs as promising biomarkers for the early detection of significant prostate cancer.

<h4>Key points</h4>Lipocalin-2 upregulation paralleled intracellular uromodulin aggregates in mouse and cellular models of autosomal dominant tubulointerstitial kidney disease because of pathogenic uromodulin. The induction of lipocalin-2 was triggered by mutant uromodulin accumulation and endoplasmic reticulum stress in tubular cells. Genetic loss of lipocalin-2 reduced iron deposits in the kidney but did not affect kidney damage, indicating that lipocalin-2 was not driving disease progression.<h4>Background</h4>Autosomal dominant tubulointerstitial kidney disease due to pathogenic UMOD variants (ADTKD- UMOD ) is a toxic proteinopathy caused by intracellular accumulation of mutant uromodulin (UMOD) and endoplasmic reticulum (ER) stress. Lipocalin-2 (LCN2) is an acute phase protein induced by ER stress with context-dependent roles in kidney injury.<h4>Methods</h4>To examine the role of LCN2 in ADTKD- UMOD , we used Umod knock-in mouse models (C171Y, R186S, C125R), urine samples from affected patients, and mIMCD3 cells expressing wild-type or mutant UMOD. LCN2 expression was assessed by immunoblotting, immunostaining, and ELISA. Autophagy was stimulated with Torin1 to evaluate effects on LCN2 induction. UmodR186S/+ mice were crossed with Lcn2-/- mice to determine the impact of LCN2 deficiency on disease progression.<h4>Results</h4>Robust LCN2 induction was observed in kidneys and urine of UmodR186S/+ , UmodC125R/+ , and UmodC171Y/+ mice, correlating with UMOD aggregates and ER stress severity in thick ascending limb cells. In patients, specific UMOD variants were associated with elevated urinary LCN2. In mIMCD3 cells expressing mutant UMOD (C170Y, R185S), treatment with Torin1 reduced aggregates and attenuated LCN2 induction. Genetic deletion of Lcn2 in UmodR186S/+ mice decreased interstitial iron deposition but did not alter UMOD accumulation, interstitial inflammation, or fibrosis.<h4>Conclusions</h4>LCN2 was induced by intracellular UMOD aggregates and ER stress in various models of ADTKD- UMOD . Although it influenced iron handling, LCN2 did not drive fibrosis or inflammation, supporting a role as a biomarker of toxic proteinopathy rather than a therapeutic target.

BACKGROUND Non-cirrhotic non-malignant portal vein thrombosis (PVT) is considered a rare disorder in patients without a risk of thrombophilia. Acute viral infection has been reported to be associated with an increased risk of venous thromboembolism more often in immunocompromised patients. CASE REPORT We hereby present a case of a young man who presented to the hospital with fever, fatigue, and vague abdominal pain. An initial investigation showed mild elevation in liver enzymes, and an ultrasound of the abdomen revealed mild hepatosplenomegaly. The serological test for cytomegalovirus (CMV) and Epstein-Barr virus (EBV) revealed high titers of immunoglobulin M (IgM). The IgG titers for both viruses were negative, suggesting a recent acute infection. A computed tomography (CT) scan of the abdomen demonstrated acute PVT. The patient was shifted to the ICU, and anticoagulation was initiated immediately with unfractionated heparin (intravenous bolus of 80 u/kg followed by 18 u/kg/hr, target aPTT of 1.5-2.3 control). A thorough workup was performed to exclude primary and secondary causes of PVT; all tests were negative. A diagnosis of acute PVT secondary to CMV and EBV infection was made. The patient's symptoms gradually improved, and he was discharged home on oral anticoagulation. CONCLUSIONS Acute CMV and/or EBV infection is associated with the risk of thromboembolism even in immunocompetent patients. Treating it as a provoked risk for thromboembolism will aid in its early detection and management. Additionally, by treating it as a triggered risk, unnecessary lifelong anticoagulation can be avoided.

No abstract available.

The neural processing of subsecond durations recruits a wide network of areas. Although unimodal tuning has been shown in many of these regions, its role and link to perception remain unclear. Here, we used 7T functional MRI while participants performed a visual duration categorization task to characterize unimodal responses along the cortical hierarchy. We found topographically organized neuronal populations tuned to all presented durations in parietal and premotor cortices, and in the caudal supplementary motor area (SMA). In contrast, rostral SMA, inferior frontal cortex, and anterior insula showed neuronal preferences centered around the mean duration, which correlated with the boundary duration participants employed in the task. These differences suggest specialized roles of duration tuning across cortical regions -from discrete to categorical and subjective duration representations. Finally, correlations of neuronal preferences across areas highlighted a hierarchical organization of duration tuning. Together, our findings provide a mechanistic framework for duration perception in vision.

BACKGROUND: Inherited antithrombin deficiency (ATD), a rare autosomal dominant disorder due to SERPINC1 gene mutations, is the most severe inherited thrombophilia. Limited literature exists that focuses on ATD and its mutations in the Chinese population. This study aimed to characterize SERPINC1 gene mutations in a Chinese cohort and to explore their relationship with thrombophilia. METHODS: Coagulation screening results and clinical data were meticulously collected from 23 unrelated probands with ATD and their family members. Genomic DNA was extracted and subjected to PCR amplification and direct sequencing. Putative mutations were analyzed using in silico bioinformatic tools. Mutant antithrombin (AT) proteins were expressed in HEK293 cells, and ELISA was used to detect wild-type and mutant AT. RT-qPCR was used to measure AT mRNA expression in transfected cells. RESULTS: Among the 23 probands, 15 (65.2%) exhibited concurrent reductions in both AT: A and AT: Ag (type I defects), while the remaining 8 (34.8%) had normal AT: Ag levels (type II defects). Genetic analysis revealed a spectrum of 21 distinct mutations across 87.0% (20/23) of the probands. Most were point mutations predicted to be deleterious and were primarily located in exons 5 and 3. Among the 20 mutation carriers, 15 (75%) were heterozygous and most of them experienced thrombosis with identifiable triggers. The other 5 (25%) were compound heterozygous and primarily presented with spontaneous thrombosis. Notably, the missense mutations c.1346T > A and c.442T > C were recurrent. These mutations exhibited high heterogeneity, with no ethnic-specific mutations observed. In vitro expression confirmed that synthesis and/or secretion defects in the mutant proteins are the primary mechanism underlying the antithrombin deficiency. CONCLUSIONS: SERPINC1 gene analysis benefits asymptomatic family members, especially child-bearing women, by informing venous thromboembolism prevention strategies and guiding anticoagulant choice in cases involving heparin-binding site mutations. This underscores the essential role of genetic diagnosis in ATD management.

BACKGROUND: Neuronal Growth Regulator 1 (NEGR1) is a cell adhesion molecule involved in hippocampal circuit development and function. Human genetic studies have identified NEGR1 variants as risk factors for a broad spectrum of neuropsychiatric disorders. These disorders often display sex-specific differences in prevalence, progression, and behavioral impairment, reflecting underlying maladaptive changes in neural circuitry. Findings from preclinical studies using Negr1−/− mice show several hippocampal-based behavioral and anatomical endophenotypes relevant to neuropsychiatric disorders. The hippocampus, a key region implicated in these disorders, exhibits sex-dependent anatomical features that may shape the functional impact of Negr1. However, the mechanisms driving these sex-specific characteristics have not yet been elucidated. Here, we uncover sex-specific molecular signatures and pathways associated with Negr1, using Negr1−/− mice, a genetically relevant animal model for neuropsychiatric risk. METHODS: We performed label-free quantitative proteomic analysis using eight replicates of hippocampi dissected from male and female wild-type, and Negr1−/− mice. Differentially abundant proteins were subjected to functional annotation for Gene Ontology and Protein-Protein interaction using STRING analysis. NEGR1 cellular localization was examined by immunofluorescent in rat brain and human hippocampal sections. RESULTS: Differential expression analysis identified 232 proteins in males and 172 in females. STRING analysis revealed sex-specific regulation of proteins. In males, proteins linked to neurofilament organization, myelin integrity, and postsynaptic structure were downregulated, with parvalbumin (Pvalb, PV) around the central node. In contrast, proteins related to mitochondrial and stress-response pathways were upregulated. Female Negr1−/− hippocampus showed downregulation of proteins involved in translation and amide biosynthetic processes. Colocalization of NEGR1 with PV interneurons in the rat brain and the human hippocampus was observed. CONCLUSIONS: We demonstrate, for the first time, distinct sex differences in the hippocampal proteome and identify molecular networks in Negr1−/− mice. Co-localization of NEGR1 and PV in human brain tissue provides anatomical and translational validation of a proteomic target. These findings provide new insight, offering a valuable resource for understanding NEGR1-related sex-specific mechanisms in neuropsychiatric disorders.

OBJECTIVE: Nutritional deficiencies have been increasingly recognized as potential contributors to adverse cardiovascular outcomes. However, their relationship with clinical outcomes in patients with heart failure (HF) remains unclear. This study aimed to evaluate the association between nutritional deficiency anemia (NDA) and clinical outcomes in patients with HF. METHODS: We conducted a retrospective cohort study, identifying 1,558,577 adult patients diagnosed with HF. Among these, 86,186 patients were classified as having NDA. A 1:1 propensity score matching (PSM) was performed, resulting in 86,181 matched pairs. The primary outcome was a composite of all-cause mortality, HF exacerbation (HFE), and major adverse cardiovascular events (MACE). Secondary outcomes included individual components of the composite outcome. Hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated using Cox proportional hazards models. RESULTS: The composite outcome occurred more frequently in the NDA group than in the control group (HR: 1.414, 95% CI: 1.38–1.449, p < 0.0001). Secondary outcomes further demonstrated an increased risk of all-cause mortality (HR: 1.484, 95% CI: 1.432–1.539), HFE (HR: 1.440, 95% CI: 1.396–1.486), and MACE (HR: 1.278, 95% CI: 1.22–1.338), all with p < 0.0001. CONCLUSION: NDA was associated with worse clinical outcomes in patients with HF, including increased risks of all-cause mortality, HFE, and MACE. These findings suggest that coded nutritional deficiency anemia may represent a clinically relevant prognostic marker in HF populations. Further studies are warranted to clarify the underlying mechanisms and clinical implications of this association.

<h4>Aims</h4>The burden of chronic liver disease (CLD) is increasing. This study aims to identify protein markers for CLD and its progression, and develop a protein-based risk prediction model.<h4>Materials and methods</h4>We used proteome-wide Mendelian randomization (MR), Bayesian colocalization and summary-data-based MR with proteomic data from deCODE Genetics to identify CLD-related proteins. Multivariable MR was used to assess independent protein effects. Protein-protein interaction, druggability and mediation analyses were conducted to prioritise therapeutic targets. We further constructed a protein risk score to predict CLD and composite hepatic event outcomes, comparing its performance with existing clinical predictors.<h4>Results</h4>Genetically predicted levels of 16, 5 and 4 plasma proteins were associated with metabolic dysfunction-associated steatotic liver disease (MASLD), alcoholic liver disease (ALD) and cirrhosis, respectively. IGSF3, FTCD, DCXR, ADH1B and ACY1 were associated with CLD progression. Genetically predicted five modifiable factors (body mass index, waist-hip ratio, glycated haemoglobin, type 2 diabetes, leisure television watching) were associated with CLD-related proteins. Proteomic-based models showed high predictive performance for ALD (C-index = 0.89), liver cancer (C-index = 0.84) and liver failure (C-index = 0.84) in a healthy population without liver diseases at baseline.<h4>Conclusions</h4>This study identified key circulating protein markers for CLD. Protein-based profiling demonstrated strong predictive potential for CLD and related outcomes.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is a prevalent and aggressive subtype of lung cancer, with a 5-year survival rate below 20% due to late-stage diagnosis and drug resistance. Endoplasmic reticulum stress (ERS) and butyrate metabolism (BM) play critical roles in tumor progression, but their co-regulatory features in LUAD remain unclear.<h4>Methods</h4>This study integrated single-cell transcriptome analysis and Mendelian randomization (MR) to identify prognostic genes associated with ERS and BM in LUAD. Public datasets were analyzed using weighted gene co-expression network analysis, differential expression analysis, and MR. A risk model and nomogram were constructed, and immune microenvironment, gene set enrichment, and single-cell analyses were performed to validate findings. Moreover, the expression of prognostic genes was validated in different Non-small cell lung cancer (NSCLC) cell lines through reverse transcription quantitative polymerase chain reaction (RT-qPCR).<h4>Results</h4>Seven prognostic genes (<i>VDAC1</i>, <i>TXNRD1</i>, <i>GDF15</i>, <i>TRIB3</i>, <i>LPL</i>, <i>KCNQ1</i>, <i>PKP2</i>) were identified, RT-qPCR assays confirmed that these genes exhibited significant expression differences in different NSCLC cell lines. The risk model demonstrated that low-risk patients had significantly better survival outcomes. The nomogram exhibited strong predictive accuracy for 1-, 3-, and 5-year survival. Enriched pathways in high-risk patients included olfactory transduction, while low-risk patients showed enrichment in ribosome and complement-coagulation cascades. Immune profiling revealed 13 differentially abundant immune cell types, including M1 macrophages. Single-cell analysis identified macrophages as key players in LUAD. Notably, <i>VDAC1</i>, <i>TXNRD1</i>, and <i>LPL</i> were highly expressed during early macrophage differentiation.<h4>Conclusion</h4>This study identifies seven ERS- and BM-related prognostic genes and highlights macrophages as pivotal in LUAD progression, the expression differences of candidate genes were verified by RT-qPCR assay. These findings provide novel insights into LUAD diagnosis, prognosis, and potential therapeutic targets, offering a foundation for precision medicine strategies. Further validation in clinical cohorts and functional studies is warranted to translate these discoveries into clinical applications.

In response to the growing demand for novel approaches in bone repair, scaffolds that mimic natural bone microstructure and mineralogical composition were developed using a ceramic vat photopolymerization (VPP) method. Due to varying reported results regarding appropriate microstructural characteristics, this study aimed to clarify the best pore size distribution and porosity among the tested scaffolds for an efficient osteogenic response. Scaffolds based on hydroxyapatite both support new bone formation by osteoblasts and can be resorbed by osteoclasts. An average pore size of ∼400 μm and porosity of 45.61% showed the best mechanical properties and osteogenic response, allowing cell penetration, and supporting cell-cell interactions and the differentiation process. When Sr,Mg,Zn-substituted hydroxyapatite is used for scaffold fabrication, the required high sintering temperatures lead to the transformation of hydroxyapatite into <i>β</i>-tricalcium phosphate, a common calcium phosphate used in bone tissue engineering. However, the new mineralogical phase results in different surface properties that do not support appropriate cell attachment on scaffolds with higher negative surface charge and lower wettability. This work emphasizes the potential of ceramic VPP in the development of biomimetic scaffolds that mimic natural bone tissue and provides guidelines on which microstructural characteristics are appropriate for efficient bone regeneration. This study also raises new questions regarding cell attachment on <i>β</i>-tricalcium phosphate-based scaffolds, which are currently under exploration.

<h4>Background</h4>Type 2 diabetes (T2D) is a challenge for the healthcare system. It is a metabolic disease with increased blood sugar with severe complications when it becomes uncontrolled. These complications include diabetic nephropathy (DN), neuropathy, retinopathy neuropathy, and cardiovascular diseases (CVDs). T2D is induced by genetic, environmental, and lifestyle risk factors. Therefore, it is vital to distinguish between genetic risk loci for T2D and those that specifically predispose patients to DN, which may eventually facilitate personalized risk assessment and informed genetic counseling once these markers are clinically validated. The aim of this pilot study was to examine the candidate genes associated with DN using whole-exome sequencing (WES).<h4>Method</h4>We examined the relationship between specific genetic variations and DN in 26 (eight female and 18 male patients) clinically diagnosed DN patients in Tabuk. We utilized WES which was performed on the Illumina NovaSeq 6000 platform. Data were analyzed using an array of bioinformatic tools including FASTQC, Trimmomatic, Ensemble, Bowtie2, GATK, SAMtools, Python Comut, T2Diacod, StringDB, Gene Ontology, and KEGG.<h4>Results</h4>The genes <i>THADA</i>, <i>NOTCH4</i>, and <i>TNXB</i> met the genome-wide threshold and showed minimal T2D association. The DN-specific genes including <i>SP2</i>, <i>CDH3</i>, and <i>ARFGEF2</i> met the suggestive DN threshold, however falling below the T2D significance. The shared genes, <i>INSR, HLA-DQB1</i>, and <i>CRHR1</i>, exhibited possible associations with T2D and DN, positioning them as key candidates for DN. Pathways and biological processes potentially affected by these gene variations include inflammatory pathway, lipid and glucose metabolism, PI3K-Akt signaling, insulin signaling, AMPK signaling pathways, and others. Moreover, we identified putative novel missense variations in the genes <i>SRCAP</i>, <i>PHKG2</i>, <i>TNFRSF6B</i>, and <i>PBX2</i>.<h4>Conclusion</h4>Our exploratory study identified candidate novel variations in the <i>SRCAP</i>, <i>PHKG2</i>, <i>TNFRSF6B</i>, and <i>PBX2</i> genes that may be linked to DN. These results indicate that these genes are potentially involved in DN development. Nevertheless, further verifications through large-scale cohorts and functional protein studies are required to determine the clinical utility of these variants as potential biomarkers.

Sperm cryopreservation is pivotal for conserving fish germplasm, yet cryodamage-induced quality decline limits its application. This study focused on Sichuan bream (<i>Sinibrama taeniatus</i>), an endemic and economically important fish species in the upper Yangtze River. Based on an established cryopreservation protocol, we evaluated sperm quality using computer-assisted sperm analysis (CASA) and fertility assays, followed by a systematic assessment of structural and functional damage via flow cytometry (membrane integrity, mitochondrial potential, reactive oxygen species, and DNA fragmentation), enzymatic assays (energy metabolism and antioxidant enzymes), Western blotting, and ultrastructural observation. Finally, integrated proteomic and metabolomic analyses were employed to elucidate the underlying physiological mechanisms. The results demonstrated that freeze-thawing significantly impaired sperm motility, fertility, and ultrastructure, concurrently disrupting energy metabolism and the antioxidant system. Crucially, multi-omics revealed that these functional declines were linked to dysregulation in key pathways involving cytoskeleton organization, lipid metabolism, energy homeostasis, and oxidative stress, forming a coherent network from initial molecular perturbation to phenotypic dysfunction. This study provides a comprehensive characterization of sperm cryodamage in Sichuan bream, advancing the understanding of fish sperm cryobiology and informing targeted cryoprotection strategy development.

The self-assembly of a novel synthesized chiral dipeptide, Boc-p-nitro-L-phenylalanyl-tyrosine, into supramolecular structures is investigated by optical absorption and photoluminescence spectroscopy as well as single crystal X-ray diffraction. The compound is a diphenylalanine derivative belonging to a family of aromatic dipeptides that spontaneously self-organize into nanostructures through molecular recognition. The dipeptide exhibits several step-like peaks in its absorption band, indicative of self-assembly into quantum-confined nanostructures. In contrast, the parent Boc-p-nitro-L-phenylalanine amino acid lacks these features, indicating that the tyrosine residue favors quantum-confined self-assembly. Crystal structure determination reveals distinct packing styles: Boc-p-nitro-L-phenylalanine forms two-dimensional hydrogen-bonded layers, while the related p-nitro-free Boc-L-phenylalanyl-tyrosine dipeptide organizes into a 3D helical columnar architecture, driven by the additional hydrogen-bonding capacity of the peptide bond and tyrosine hydroxyl group, which favors the formation of a channel-type tetragonal architecture network over the planar sheets of the monomer. Furthermore, the introduction of a tyrosine residue into the Boc-p-nitro-L-phenylalanine molecule alters its supramolecular assembly, as the dipeptide Boc-p-nitro-L-phenylalanyl-tyrosine crystallizes as a monohydrate. The water molecule present in the structure acts as a bridge, participating in a hydrogen-bonding network between the tyrosine hydroxyl groups of neighboring columns through intermolecular interactions.

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy marked by substantial molecular heterogeneity and variable response to gemcitabine-based therapy. While KRAS mutations are nearly universal, the broader RTK-RAS and MAPK signaling architecture and its relationship to treatment response remain incompletely defined. We conducted an integrative clinical-genomic analysis of 184 PDAC tumors stratified by age at diagnosis and gemcitabine exposure, interrogating somatic alterations across curated RTK-RAS/MAPK gene sets. Conversational artificial intelligence agents (AI-HOPE-RTK-RAS and AI-HOPE-MAPK) enabled dynamic cohort construction and pathway-level analyses, with findings validated using standard statistical methods. In late-onset PDAC, ERBB2 and RET mutations were significantly enriched in gemcitabine-treated tumors. Early-onset cases demonstrated differential enrichment of CACNA2D family alterations in non-treated tumors and higher frequencies of FLNB and TP53 mutations in treated disease. Importantly, late-onset patients not treated with gemcitabine who lacked RTK-RAS or MAPK alterations exhibited significantly improved overall survival. These findings reveal age- and treatment-dependent pathway dependencies beyond canonical KRAS status and support a precision oncology framework in PDAC. Conversational AI facilitated rapid, multidimensional clinical-genomic integration to uncover clinically relevant signaling substructures.

Semen parameters, including sperm counts, have rapidly declined in men across the globe over the last five decades. Although this decline remains unexplained, lifestyle factors may affect male fertility. Recently, several studies highlighted a potential link between non-steroidal anti-inflammatory drug (NSAID) usage, such as naproxen and ibuprofen, and declining male fertility. However, the mechanisms by which these common analgesics affect male fertility, including their effects on the blood-testis barrier (BTB), remain poorly characterized. Utilizing an in vitro rhesus macaque non-human primate (NHP) BTB model, we demonstrate that serum levels of naproxen and ibuprofen alter the function of BTB. Following short-term naproxen and ibuprofen treatment of NHP primary Sertoli cells, we show that these NSAIDs increase the transepithelial electrical resistance, indicating an overall strengthening of the Sertoli cell junctions. Furthermore, naproxen and ibuprofen treatment alter the expression of genes involved in maintaining the BTB. Specifically, the genes that were significantly expressed in response to ibuprofen exposure were enriched for human phenotypic abnormalities linked to male factor infertility. Together, these results suggest that short-term naproxen and ibuprofen treatment disrupt the function of the BTB by altering the integrity of the Sertoli cell junctions, proposing a potential role of NSAIDs in male factor infertility.

The 2025 approval of the selective NaV1.8 blocker suzetrigine for acute pain marked a pivotal advance in analgesic drug development. Yet the subsequent failure of Vertex's next-generation NaV1.8 inhibitor VX993 to demonstrate clinical analgesia underscores enduring challenges in translating mechanistic promise into patient benefit. This review examines why promising targets and compounds, spanning NaV and TRP channels, often falter and outlines a path toward more reliable target selection and validation. I first summarize the pain pathway, from nociceptor transduction through spinal processing to cortical perception, emphasizing how inflammation and peripheral sensitization reshape excitability. Historically serendipitous, pain drug discovery now prioritizes molecular precision. Most approved chronic pain therapies act in the CNS and are limited by modest efficacy and adverse effects. Nociceptor-enriched targets (NaV1.7/1.8/1.9; TRP channels) remain attractive, yet redundancy among NaV subtypes and the necessity of blocking targets at the correct anatomical sites complicate translation. Human genetics and multi-omics provide a powerful, unbiased engine for target discovery. Rare high-impact variants offer strong causal hypotheses, while common polygenic contributions illuminate broader susceptibility. Large biobanks increasingly reveal a mismatch between legacy pain targets and genetically supported candidates across neuronal and non-neuronal cells. Human DRG transcriptomics highlight NaV channel redundancy. Human in vitro electrophysiology and PK/PD analyses show suzetrigine achieves ~90-95% NaV1.8 engagement, yet neurons can still fire unless additional channels are blocked. Species differences and drug distribution (including BBB/PNS penetration and P-gp efflux) critically influence efficacy; centrally accessible blockade (e.g., for NaV1.7 or TRPA1) may be necessary to achieve robust analgesia, challenging peripherally restricted strategies. Osteoarthritis illustrates how obesity-driven metabolic inflammation, synovial immune activation, subchondral bone remodeling, and specific nociceptor subtypes converge to drive mechanical pain. Multi-omic integration across diseased human tissues can pinpoint causal processes and cell types, enabling more selective and safer target choices. I propose a practical framework for target validation that integrates: (i) rigorous human genetic support; (ii) cell-type and site-of-action mapping; (iii) human-relevant electrophysiology and PK/PD with verified target engagement; (iv) species-appropriate models; (v) consideration of modality (small molecule, biologic, RNA, targeted protein degradation). Advancing genetically and anatomically aligned targets, tested at the right sites and exposures, offers the best path to genuinely effective, better-tolerated pain therapeutics.

Glutaminyl-peptide cyclotransferase (QPCT, QC) and its isoenzyme glutaminyl-peptide cyclotransferase-like protein (QPCTL, isoQC) are zinc-dependent enzymes that post-translationally catalyze the conversion of N-terminal glutamine or glutamate residues into pyroglutamate (pGlu). The pGlu modification impacts protein-protein interactions, enhances protein stability, and protects proteins from proteolytic degradation. QPCTL and QPCT differ in their subcellular localization, with QPCTL being retained in the Golgi apparatus and QPCT being active in secretory vesicles. Current research focuses on the impact of QPCTL-mediated pGlu formation in cancer and neurodegenerative disorders such as Alzheimer's disease. In cancer, QPCTL is a promising immunotherapy target since QPCTL-mediated CD47 pyroglutamylation prevents macrophages from phagocytosing tumor cells. Moreover, QPCTL shapes the tumor microenvironment by modulating macrophage recruitment and polarization through modification of CCL2. However, QPCTL modulates Butyrophilins on tumor cells and thereby promote their detection and killing by γδ T cells. Hence, QPCTL significantly affects cancer progression, inflammatory processes, and immune regulation. These insights highlight QPCTL's potential as a therapeutic target in oncology, metabolic diseases, and immune-mediated disorders. In this review, we highlight the role of QPCTL in tumor evasion and immune modulation. Moreover, we provide a comprehensive overview about predicted and validated substrates of QPCT/L and about the relevance of QPCT/L in various diseases.

Syntaxin1A (STX1A) is a presynaptic membrane protein that is abundantly expressed in the central nervous system. It is a key member of the soluble N-ethylmaleimide sensitive factor attachment protein receptor protein family. Notably, STX1A acts as a 'molecular hub' in neural networks by regulating presynaptic membrane fusion with synaptic vesicles and the subsequent release of neurotransmitters. In addition to this function, STX1A is crucial for neuronal development, synaptic plasticity, and ion channel regulation. The deficiency or variation of <i>STX1A</i> not only directly disrupts neurotransmitter transmission but also contributes to pathological processes in neurological disorders such as Alzheimer's disease, epilepsy, autism spectrum disorder, and ischemic stroke by interfering with excitatory-inhibitory balance, inducing neuroinflammation, and triggering neuronal apoptosis. The present review summarizes the structure and physiological functions of STX1A, highlights its mechanisms in the pathogenesis of various neurological diseases, and examines its potential as a diagnostic biomarker and therapeutic target for these diseases.

Bone mineral forms both inside and between collagen fibrils in the extracellular matrix. While the morphology of intrafibrillar bone mineral has been hypothesized to be primarily controlled by the size and shape of the restricted spaces inside collagen fibrils within which the mineral forms, what controls the architecture of the extrafibrillar mineral is still an open question. While bone mineral is primarily apatitic in composition, it also contains significant quantities of cell respiration metabolites, in particular, carbonate, citrate, and lactate. An as-yet unanswered question is what, if any, role do these metabolites collectively play in determining the 3D architecture of bone mineral. Here, we propose a composite model of bone mineral that accounts for both intra- and extrafibrillar mineral environments, and to that end, we develop apatitic materials containing citrate and lactate or carbonate that mimic the densely packed ionic environments within which bone mineral forms in vivo. We find that incorporating citrate and lactate leads to complex mineral architectures reminiscent of those in extrafibrillar bone mineral, including mineral crystal curvature. Our results suggest that metabolic acids may play an important role in building the 3D architecture of extrafibrillar bone mineral.

<h4>Background</h4>Neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, multiple sclerosis, and amyotrophic lateral sclerosis, represent a major global health burden and share convergent pathogenic mechanisms, such as mitochondrial dysfunction, oxidative stress, neuroinflammation, calcium imbalance, and neuronal loss. Despite advances in symptomatic management, effective disease-modifying therapies remain limited.<h4>Objectives</h4>This review aims to critically synthesize mechanistic, preclinical, and clinical evidence on α-lipoic acid and biotin as candidate neuroprotective agents in neurodegenerative diseases, with emphasis on shared signaling pathways, therapeutic potential, generally favorable safety profiles, and translational limitations.<h4>Methods</h4>A narrative and integrative review was conducted, encompassing mechanistic studies, preclinical experimental models, and clinical trials and observational studies evaluating ALA and biotin in neurodegenerative diseases. The evidence was qualitatively analyzed with attention to biological plausibility, consistency across models, and clinical relevance.<h4>Results</h4>ALA and biotin modulate key cellular pathways implicated in neurodegeneration, including mitochondrial metabolism, redox homeostasis, inflammatory signaling, and neurovascular function. Preclinical studies consistently report beneficial effects on mitochondrial efficiency, oxidative stress, and neuroinflammatory markers. In contrast, clinical evidence remains heterogeneous, with more extensive evaluation of biotin in progressive multiple sclerosis and more limited or exploratory findings for ALA across neurodegenerative disorders.<h4>Conclusions</h4>ALA and biotin exhibit mechanistic convergence across pathways relevant to neurodegeneration and generally favorable safety profiles. Although current evidence supports their biological plausibility as adjunctive or exploratory therapeutic strategies, clinical outcomes remain inconsistent and appear to be influenced by dosing regimens, disease stage at intervention, and endpoint selection. Well-designed clinical studies are required to define their efficacy, optimal dosing, and disease-specific applicability.

The ongoing panzootic of the highly pathogenic avian influenza (HPAI) H5N1 virus, dominated by clade 2.3.4.4b, constitutes a significant global threat to wildlife, animal health, and public health. Once characterized by sporadic outbreaks, H5N1 has evolved into a sustained, year-round infection with an expanded host range that now includes numerous mammalian species. Its high pathogenicity is primarily driven by the acquisition of a polybasic haemagglutinin cleavage site, enabling systemic viral spread, alongside emerging endothelial and neurotropic properties that contribute to severe disease and high mortality in mammals. Although zoonotic transmission remains limited, H5N1 continues to accumulate mutations associated with mammalian adaptation, particularly within the haemagglutinin and polymerase complex. Notably, recent outbreaks in U.S. dairy cattle highlight the emergence of novel mammalian reservoirs with increased human exposure risk. Concurrently, vaccination strategies are advancing beyond traditional adjuvanted inactivated vaccines toward next-generation platforms, including mRNA and virus-like particle vaccines, designed for rapid deployment and broader immune protection. However, ongoing viral evolution, constrained vaccine availability, and gaps in coordinated surveillance underscore the urgent need for an integrated One Health approach to reduce panzootic risk.

The PRDM (PR domain-containing) family consists of transcriptional regulators characterized by a PR (PRDI-BF1 and RIZ homology) domain, a subtype of the SET domain, and a variable number of zinc finger motifs. Nineteen PRDM family members have been identified in both mice and humans, and increasing evidence supports their roles as epigenetic regulators in development and disease. PRDM proteins share a conserved structure, comprising an N-terminal PR domain with potential histone methyltransferase activity and C-terminal C2H2-type zinc fingers involved in protein-protein, protein-RNA, and protein-DNA interactions. Recent studies indicate that multiple PRDM family members are involved in the regulation of the neuro-motor system, including neural lineage specification, neuronal differentiation, motor function maintenance, and neuromuscular-related pathological processes. This review summarizes current evidence on the functions and regulatory mechanisms of PRDM family members in the neuro-motor system. Overall, PRDM family members act as important epigenetic regulators in the neuro-motor system. Clarifying their molecular mechanisms may contribute to a better understanding of neuro-motor regulation and provide a theoretical basis for future research in exercise and movement science.

We devised a quantitative scoring function to assess the cumulative effects of somatic nonsynonymous single-nucleotide variants (SNVs) on protein-coding genes in patients with ovarian cancer (OvCa) and thyroid cancer (ThCa). The goal is to find novel candidate cancer-related genes for downstream bioinformatics analyses and wet-lab studies. With the Genomic Data Commons as primary data resource, SNV information was extracted from whole-exome sequencing data from patients with these cancers. A cumulative variant scoring function, <i>Q</i>(<i>G</i>), was developed to sum up the deleterious effects of the individual SNVs on gene <i>G</i>. While <i>Q</i>(<i>G</i>) can be computed using any popular functional effect analyzers such as FATHMM-XF, SIFT, PolyPhen, and CADD, we have also established an integrative scoring function <i>iQ</i>(<i>G</i>) that combines the deleterious assessments from different analyzers and demonstrated that <i>iQ</i>(<i>G</i>) is a more effective method for identifying likely cancer-related genes. Based on the <i>iQ</i>(<i>G</i>) rankings, the top three novel genes for OvCa are <i>AHNAK2</i>, <i>UNC13A</i>, and <i>PCDHB4;</i> and those for ThCa are <i>PLEC</i>, <i>HECTD4</i>, and <i>CES1</i>. Furthermore, the top 1% genes with highest <i>iQ</i>(<i>G</i>) scores for each cancer were submitted for KEGG pathway analysis. The results revealed that several genes of the <i>CACNA1</i> family within the type II diabetes mellitus pathway are likely related to both OvCa and ThCa and suggested other molecular interactions that should be further studied in connection with OvCa prognosis and ThCa treatment.

<i>Toxoplasma gondii</i> (<i>T. gondii</i>) is an intracellular parasite known to modulate host immunity and cellular signaling, raising interest in its potential influence on cancer biology. A systematic review was conducted to evaluate experimental evidence on the antitumor or pro-tumor effects of <i>T. gondii</i> infection and parasite-derived antigens and to categorize the underlying mechanisms. PubMed was searched through 9 September 2024, and 54 eligible experimental studies were included (41 in vivo, 10 in vitro, and three combined). Forty-six studies reported antitumor effects, two pro-tumor effects, one stage-dependent divergent effects (acute infection/antitumor vs. chronic infection/pro-tumor), and five highlighted <i>T. gondii</i>-associated cancer-pertinent signaling pathways. Antitumor effects were observed following acute infection and exposure to parasite antigens, certain recombinant proteins, and exosomal microRNA miR-155-5p. Dominant mechanistic categories included activation of innate and adaptive immunity and reversal of tumor microenvironment immunosuppression (notably Th1-driven IL-12/IFN-γ responses, antitumor M1 macrophage polarization), induction of apoptosis, anti-angiogenesis, molecular mimicry and modulation of cancer-pertinent pathways. Conversely, pro-tumor effects were seen with chronic infection and exposure to ROP18 effector protein and miR-21. Future translational research should focus on rigorous evaluation of the safety and efficacy of attenuated non-replicating <i>T. gondii</i> strains and/or select recombinant antigens for potential cancer <i>T. gondii</i>-based immunotherapy.

Dental enamel, the hardest mineralised tissue in the human body, has proven to be an excellent source of ancient proteins, which have been found to survive within dental enamel for at least twenty million years. In archaeological and palaeontological contexts, the enamel proteome is generally considered to be rather small, consisting of about twelve proteins, most of which are unique to enamel. During amelogenesis these proteins undergo in vivo digestion by matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4) as well as serine phosphorylation by ‘family with sequence similarity member 20-C’ (FAM20C) that alter their characteristics. Gaining knowledge of the previously understudied influence of amelogenesis on the archaeological human dental enamel proteome could benefit various palaeoproteomic analysis, especially in an human evolutionary context. Here we present archaeological dental enamel proteomes and explore protein cleavage patterns and sequence coverage to estimate the effects of in vivo digestion, as well as explore phosphorylation patterns. Additionally, we present a new marker based on phosphorylation to estimate genetic sex.

Circadian clocks govern the 24-hour rhythmic activity of organs across the human body and these nycthemeral rhythms are critical for human physical and mental health. Here, we analysed 14,886 samples across 45 human tissues and inferred tissue-specific local phases using CHIRAL to define the rhythmic transcriptome. Local phase inference identified a median of 5,747 rhythmic transcripts across tissues, including 5,531 across brain tissues, indicating that prior donor-level approaches have substantially underestimated the extent of rhythmic transcription in the human brain. In the brain, rhythmic genes were enriched for neurotransmitter pathways, the synaptic vesicle cycle, circadian entrainment and major neurodegenerative disorders. Strikingly, more than 200 genes in Alzheimer’s, Parkinson’s, Huntington’s and prion disease pathways displayed significant 24-hour rhythmicity across cortical, striatal and hippocampal regions, segregating into distinct diurnal and nocturnal phase clusters. These included Parkinson’s disease genes SNCA and PRKN , Alzheimer’s disease genes PSEN1 and APP , prion disease genes PRNP and EIF2AK3 ( PERK ), and Huntington’s disease genes HTT and GPR52 . Among these disease-associated rhythmic transcripts were several targets already under therapeutic investigation for which administration at specific times of day may improve therapeutic efficacy or tolerability. Here, we observe that human tissues, including the brain, exhibit distinct local rhythmic organisation. These findings provide novel insights into disease mechanisms and highlight opportunities for target discovery, drug development and chronotherapeutic intervention.

<i>Helicobacter pylori</i> infection is the main risk factor for gastric cancer. <i>H. pylori</i> easily develop antibiotic resistance and evade host defenses. In-depth knowledge of the first barrier that <i>H. pylori</i> encounter, the gastric surface mucus-producing epithelial cells (SMCs), may enable improved treatment and prevention. This study aimed to characterize SMC gene expression, mucus glycosylation, and identify how <i>H. pylori</i> colonization affects these parameters. The glycosylation of eight <i>H. pylori</i>-infected and eight sham control mice was characterized by mass spectrometry. SMCs from five infected and five sham control mice were extracted with laser microdissection (LCM) and sequenced with RNA sequencing (RNA-Seq). SMCs were characterized by high gene expression for proteins secreted into mucus (<i>Tff1</i>, <i>Gkn1</i>, <i>Gkn2</i>, <i>Psca</i>, and <i>Muc5ac)</i>, mitoribosome RNA, and cytoskeleton proteins. Mucin glycans were large, complex, heavily fucosylated, and dense with H-antigen motifs. Two main glycosylation pathways ending in H-antigen glycans were identified and corroborated with glycosyltransferase expression. Glycosylation was consistent between <i>H. pylori</i>-infected and sham control mice. RNA-Seq data was analysed for differential gene expression, gene set enrichment analysis, and network analysis of functionally-related genes. The analyses revealed that genes required for protein synthesis and oxidative phosphorylation were down-regulated in infected mice. Most up-regulated genes were either interferon-stimulated genes or able to induce interferon production themselves. Depletion of Nkx6-3 occurred in the infected mice, indicating initiation of a pre-cancerous cascade. LCM RNA-Seq of SMCs was thus feasible and enabled characterization of the SMC and definition of a gene set showing how <i>H. pylori</i> infection affects SMCs.

The NOD-like receptor protein 3 (NLRP3) inflammasome is among the most extensively studied multiprotein complexes, driving the maturation of pro-interleukin-1β (pro-IL-1β) and pro-IL-18-into their active forms, IL-1β and IL-18, respectively. The activation of the NLRP3 inflammasome is a multifaceted process triggered by a diverse array of stimuli, including pathogens, environmental particles and endogenous stress signals. Previously, NLRP3 inflammasome activation was considered a straightforward two-step process: signal 1, which induces the expression of NLRP3 and proinflammatory cytokines, and signal 2, which promotes the assembly of the inflammasome complex through mechanisms such as ionic fluxes, mitochondrial dysfunction and lysosomal damage. However, more intricate mechanisms have now been elucidated, particularly regarding the 'priming' step, involving the regulation of its post-translational modifications. Recent studies have comprehensively identified the core components of the NLRP3 inflammasome complex, its interacting complex partners, and regulatory mechanisms. Here we delve into the current understanding of the NLRP3 inflammasome activation mechanisms and explore its regulatory networks. Enhanced insights into the molecular and signaling pathways controlling this specialized inflammasome activation may pave the way for novel applications of NLRP3 inflammasome regulation to advance human health and prevent numerous diseases linked to the NLRP3 inflammasome.

<h4>Background</h4>Ischemic stroke is a leading cause of death and long-term disability worldwide. Recanalization therapies, including thrombolysis and mechanical thrombectomy, restore blood flow, yet many patients experience poor outcomes, a phenomenon known as futile recanalization. Given the short therapeutic window for ischemic stroke, identifying early biomarkers to guide targeted interventions and improve outcomes is critical.<h4>Methods</h4>Using a murine middle cerebral occlusion model that mimics a large vessel occlusion with recanalization, a comprehensive microarray analysis from blood samples collected immediately and 3 hours after recanalization (N=44) was performed. Differentially expressed genes, enrichment pathways, immune cell proportions, enriched cell markers, predicted micro-RNAs, and transcription factors were identified using RStudio. Findings in mice were validated with rat middle cerebral artery occlusion (GSE21136) and patients with stroke (GSE16561) data sets to confirm transcriptional changes in peripheral blood postrecanalization.<h4>Results</h4><i>Il1r2</i>, <i>Cd55</i>, <i>Mmp8</i>, <i>Cd14</i>, and <i>Cd69</i> were early biomarkers poststroke and postrecanalization. Cross-validation revealed <i>Vcan</i> as a differentially expressed gene conserved across species, making it a novel ischemic marker detected as early as 3 hours postrecanalization (4 hours after middle cerebral artery occlusion) in mice, 24 hours after recanalization in rats (middle cerebral artery occlusion-thrombectomy), and within 24 hours from onset in humans receiving recombinant tissue plasminogen activator-thrombolysis. CIBERSORTx and ImmuCellAI-mouse deconvolution showed neutrophil elevation postrecanalization. Leukocyte and neutrophil activation pathways were enriched early after stroke in mice and humans, with stronger upregulation in the female sex. Several regulatory micro-RNAs were identified, and Nuclear Factor Erythroid 4 (NFE4) and Metal Regulatory Transcription Factor 1 (MTF1) emerged as key transcription factors. A coregulatory network underlying neutrophil activity was constructed, highlighting its central role in early responses to ischemia and recanalization, which was enriched in the female sex.<h4>Conclusions</h4>We identified novel early genomic markers for ischemia and recanalization, including the conserved marker <i>Vcan</i>, and highlighted age- and sex-specific immune responses. Mapping a neutrophil-centered coregulatory network provides mechanistic insight into futile recanalization and supports the development of targeted therapies to improve clinical outcomes.

<h4>Background</h4>Ex vivo oxygenated perfusion systems are a promising approach to extend cardiac allograft preservation beyond the typical 4 to 6 hours limit allowed by static cold storage (SCS). Hypothermic oxygenated perfusion (HOPE) has been proven to safely preserve donor hearts, yet its underlying molecular mechanisms have not been extensively evaluated. The aim of the study is to characterize cardiomyocyte viability, transcriptomic and metabolomic responses, and functional recovery of porcine hearts preserved with HOPE for up to 48 hours, including evaluating their ability to regain sinus rhythm following bench-top normothermic reperfusion.<h4>Methods</h4>Seventeen Yorkshire pigs underwent donor cardiectomy. In the first arm, 10 hearts were preserved for up to 48 hours using either SCS (n=5) or HOPE (n=5). Endomyocardial biopsies were collected at 0, 12, 24, and 48 hours for histology, RNA sequencing, flow cytometry, and metabolomics. In the second arm, 6 HOPE-preserved hearts (3, 24, 48 hours) and 2 SCS-preserved heart (3 and 24 hours) underwent 2 hours of normothermic reperfusion to simulate transplantation and assess reanimation.<h4>Results</h4>HOPE preserved cardiomyocyte viability and structural integrity for 48 hours, in contrast to SCS in both arms of the study. RNA sequencing and untargeted metabolomics revealed conserved energy-substrate profiles in HOPE and progressive ischemic metabolite accumulation in SCS. All HOPE hearts regained stable sinus rhythm.<h4>Conclusions</h4>HOPE enables 48 hours ex vivo heart preservation while maintaining cardiomyocyte integrity, normal gross and microscopic architecture, and rapid functional recovery on bench-top reperfusion in a preclinical model. These findings establish a foundation for redefining clinical preservation times and widening geographic donor access.

<h4>Introduction</h4>The objective of this study is to determine the prevalence of hypomagnesemia in patients with calcium pyrophosphate (CPP) arthritis and the role of diuretics on inducing hypomagnesemia.<h4>Methods</h4>This was an observational, cross-sectional, prospectively recruited, case-control study. Cases were confirmed by both chondrocalcinosis in plain X-ray and CPP crystals in synovial fluid. Serum creatinine, magnesium, calcium, phosphate, 25-OH-vitamin D, parathyroid hormone, ferritin and transferrin saturation were measured. Diuretic use, class, and dosage were also recorded.<h4>Results</h4>A total of 568 patients were included, 298 cases and 270 controls. Lower serum magnesium levels were found more frequently among cases, 16% with hypomagnesemia, than in controls [6.4%, (p = 0.01)]. Cases received almost twice as many diuretics as controls (p < 0.001). Comparing cases and controls on diuretic treatment, there were no differences in either magnesium or prevalence of hypomagnesemia. To evaluate the impact of diuretics on serum magnesium, we performed an overall analysis using the entire population. Patients on diuretics showed lower serum magnesium levels compared to patients not on diuretics (1.97 vs 2.05 mg/dl, p < 0.001). Thiazides were associated with lower serum magnesium levels (1.86 vs 2.05 mg/dl; p < 0.001) and hypomagnesemia than loop diuretics (32.1% vs 14.9%; p < 0.004), respectively. Thiazide doses over 12.5 mg/day were associated with higher rate of hypomagnesemia (70.0% vs 11.1%, p = 0.035).<h4>Conclusion</h4>Patients with CPP crystal deposition disease (CPPD) showed lower magnesium levels and higher rate of hypomagnesemia than controls, and were prescribed almost twice as many diuretics. Both classes of diuretics were associated with lower magnesium levels and clinical significant hypomagnesemia. Thiazides induced hypomagnesemia more frequently than loop diuretics, showing dose-dependent association.

The autophagy core machinery carries out the fundamental reactions of autophagosome biogenesis across all forms of bulk and selective macroautophagy. In humans, the core complexes consist of the ULK1 complex (ULK1C), the class III phosphatidylinositol 3-kinase complex I (PI3KC3-C1), the ATG8 proteins and the ATG8ylation machinery, the phosphatidylinositol 3-phosphate (PI3P)-sensing WIPI proteins, the lipid transporter ATG2, and the lipid scramblase and initiation scaffold ATG9. These complexes form a web of interactions that can be initiated by clustering of the FIP200 subunit of ULK1C but also by PI3KC3-C1 or WIPI2. Upon autophagy induction, these interactions are intensified by feed-forward signaling loops. These loops are amplified by WIPI-PI3P interactions and the conjugation of ATG8 proteins to the membrane by the ATG12-ATG5-ATG16L1 complex. Autophagosomes are seeded by ATG9 vesicles, which accrue initiation machinery on their surface and dock onto a PI3P-positive domain of the endoplasmic reticulum known as the omegasome. The omegasome contact site is the focal point for autophagosome growth, which is fed by lipid transport through the ATG2 bridge-like lipid transporter. The core complexes function in a dynamic manner, which makes autophagy vulnerable to stalling when dynamism fails. Disassembly and dissociation of the machinery, which is promoted at least in part by ULK1, is likely to be as important as assembly.

As an intravenous thrombolytic agent, tissue plasminogen activator (tPA) is limited by hemorrhagic transformation (HT) and a narrow therapeutic time window. Here we develop ROS-triggered polymersomes conjugated with fibrin-targeting CREKA peptide (CP) for tPA encapsulation. CP@tPA achieves efficient thrombosis targeting and enhanced thrombolytic efficacy, however, it has not been able to avert the occurrence of hemorrhage subsequent to thrombolysis. Building from our clinical discovery that stroke patients suffer from tPA-induced HT featured strong expression of CD177, recombinant CD177 protein (rCD177) was loaded into CP polymersomes (CP@rCD177) as nanomedicine and administrated prior to CP@tPA thrombolysis. rCD177 can be released in response to elevated ROS within the obstructed vessels, which binds to endothelial cells through interacting with CD31 and efficiently prevents the migration of CD177⁺ neutrophils into the brain parenchyma. The reduced CD177⁺ neutrophils suppress the generation of neutrophil extracellular traps (NETs), thereby dampening the inflammatory activation of microglia and ultimately improving the prognosis of HT. This innovative strategy presents a promising avenue for attenuating hemorrhagic complications post-thrombolysis.

Glioblastomas are the most prevalent and aggressive malignant brain tumors, characterized by hypertranscription and dependence on neurodevelopmental transcription factors. The transcriptional cycle is regulated by phosphorylation of the C-terminal domain (CTD) of RNA polymerase II (RNAPII) by transcriptional cyclin-dependent kinases (tCDKs), including CDK7, CDK9, CDK12, and CDK13. Here we find that glioblastoma stem cells (GSCs) are selectively sensitive to CDK12/CDK13 inhibition, whereas CDK7 and CDK9 inhibition cause non-specific cytotoxicity. This selective targeting halts GSC and organoid proliferation, curtails GSC invasion and suppresses tumor growth in a xenograft mouse model. In GSCs, CDK12/CDK13 inhibition leads to a rapid and genome-wide loss of serine-2 phosphorylation (pSer2) of the RNAPII CTD, abolishing transcriptional elongation and a transcriptional program sustained by key neurodevelopmental transcription factors. CDK12/CDK13 inhibition unexpectedly arrests DNA replication and replication fork progression in a manner distinct from the effect of inhibiting other tCDKs. This dramatic arrest precedes DNA damage response activation and cell cycle arrest, directly linking RNAPII elongation to replication fork dynamics and revealing a previously unrecognized dependence of DNA replication on CDK12/CDK13-RNAPII regulation.

Haemochromatosis is a genetic disorder of iron homeostasis. It can be caused by mutations in genes encoding the iron-regulatory hormone hepcidin (HAMP), and/or genes that regulate hepcidin expression (HFE, HJV, TFR2), or a gain-of-function mutation in the gene encoding hepcidin receptor ferroportin (FPN1/SLC40A1). HFE-related haemochromatosis is prevalent predominantly in individuals of northern European descent. These mutations result in dysregulated levels or activity of hepcidin, leading to high iron-saturation of transferrin followed by progressive liver iron accumulation in the absence of anaemia. To enable and enhance the understanding of haemochromatosis in both researchers and prospective medics, this review collates and discusses the genetic basis and consequent pathophysiology of the different types of haemochromatosis within a single, comparative review. The discussion is supported by figures and a summary table that compares the haemochromatosis types for prevalence, clinical manifestations, primary organs affected, iron-related biochemical parameters and mechanisms of iron loading. Also, gain-of-function ferroportin mutation is compared to ferroportin disease, which is a loss-of-function ferroportin mutation, and shows a tendency to anaemia. Essentially, HFE-related haemochromatosis (common type) and TFR2-related haemochromatosis (rare type) show late-onset, milder and gradual iron loading, and often involve liver and joint damage. In contrast, HJV- and HAMP-related haemochromatosis (rare types) show severe and rapid iron loading in the first three decades of life, with notable cardiac and endocrine complications. Hepcidin levels are more markedly decreased in HJV-related haemochromatosis compared to HFE and TFR2 types. There are minimal to absent levels of hepcidin in HAMP-related haemochromatosis.

With over half of people with HIV (PWH) in the U.S. entering older adulthood, identifying markers to distinguish Alzheimer’s disease (AD) and its precursor, amnestic mild cognitive impairment (aMCI), from other forms of neurocognitive impairment among PWH is urgent. We examined how HIV and aMCI status relate to AD CSF biomarkers in adult PWH and people without HIV (PWoH) characterized for aMCI. We included 80 PWH from the National NeuroHIV Tissue Consortium and 80 PWoH from the Wisconsin Registry for Alzheimer’s Prevention. Binary logistic regressions of AD-related CSF biomarker positivity (Aβ42, Aβ42/Aβ40, t-tau, p-tau181, Aβ42/t-tau) were conducted with HIV serostatus, aMCI status, HIV x aMCI interaction, and demographic covariates. Among PWH, we examined how HIV-disease characteristics relate to AD biomarker positivity. No HIV x aMCI interactions were detected. Regardless of aMCI status, having HIV was associated with higher odds of CSF Aβ42 (OR = 7.97) and Aβ42/Aβ40 (OR = 6.06) positivity, suggesting increased cerebral Aβ plaque burden. Regardless of HIV serostatus, having aMCI was associated with higher odds of CSF p-tau181 positivity (OR = 3.64). Neither HIV nor aMCI status related to Aβ42/t-tau positivity. No HIV-disease characteristics related to AD biomarker positivity. Higher CSF Aβ positivity in PWH versus PWoH, regardless of aMCI status, suggests that HIV disease promotes amyloidosis; however, whether positive CSF biomarkers in PWH raises risk for future cognitive impairment and AD remains to be explored longitudinally. Like PWoH, elevated CSF p-tau181 among PWH may indicate increased risk for AD-related memory impairment.

<h4>Objective</h4>Amnioinfusions in anhydramnios aim to promote fetal lung development, but currently used fluids (Normal Saline [NS], Lactated Ringer's [LR]) fail to mimic the intrauterine environment and increase reactive oxygen species (ROS). We developed a synthetic amniotic fluid (Amnio-well, AW) designed to reduce intrauterine ROS. This study evaluated the pulmonary and gastrointestinal effects of 2 formulations of AW compared with those of NS and LR in a pre-clinical model.<h4>Method</h4>At gestational age E17.5, pregnant rats underwent amniotic fluid replacement with NS, LR, AW, AW plus epidermal growth factor and transforming growth factor-β (AW++), or sham control. Fetal lungs were harvested at E20.5 for histology, fractional airspace, and blinded pathological evaluation. Surfactant protein (SP-A, SP-B, SP-C) expression and inflammatory gene panels were assessed in lungs and gastrointestinal (GI) tissue.<h4>Results</h4>NS and LR lungs demonstrated edema, macrophage infiltration, and reduced airspace (p < 0.001). AW improved SP-B and SP-C relative to control, whereas AW++ suppressed SP-B and SP-C (p < 0.05). Lung gene profiling showed NS/LR induced alterations in histamines, annexins, and immune recruitment, while AW closely resembled control. GI histology was similar across groups, though NS/LR altered TNF, prostaglandin, and adhesion pathways (p < 0.05).<h4>Conclusion</h4>AW reduced lung inflammation and enhanced surfactant expression compared with NS or LR, with minimal GI effects.

BACKGROUND: Dengue shock syndrome (DSS) is characterised by vascular leakage, although myocardial impairment may contribute to haemodynamic instability. The dynamic interplay between volume depletion and cardiac function during resuscitation remains incompletely understood. METHODS: We conducted a prospective cohort study of children aged 6–15 years with DSS admitted to Hospital for Tropical Diseases in Ho Chi Minh City (2018–2020). Serial bedside echocardiography was performed at shock presentation and at 1, 3, 6, 12, and 24 h post-resuscitation, daily until hospital discharge, at follow-up 1–2 weeks later, and during recurrent shock. RESULTS: 90 patients were enrolled; 16 (18%) developed recurrent shock at a median of 14 (IQR 11–17) hours after initial resuscitation. During shock, reduced SVI and CI during shock occurred in the presence of preserved LVEF and small IVC diameters, consistent with a preload-limited state. Transient myocardial impairment (MPI > 0.45) occurred in 40 patients (44%), and improved with fluid replacement. At 6 hours after resuscitation, patients who subsequently developed recurrent shock showed distinct haemodynamic profiles, including higher heart rate, lower SVI, lower lateral e′ velocity, higher E/e’, and lower TAPSE. Reduced lateral e’ velocity was also associated with respiratory distress. CONCLUSIONS: In DSS, preload reduction due to vascular leakage appears to be a major contributor to recurrent shock, although transient myocardial involvement may coexist. Focused echocardiographic assessment 6 h after resuscitation may help identify children at risk of recurrent shock before overt deterioration, supporting earlier risk stratification and more targeted fluid management.

Leptospirosis is a bacterial zoonotic disease caused by Leptospira species, commonly transmitted through direct or indirect contact with contaminated water, soil, or animal urine. Although typically a self-limiting illness, severe forms of leptospirosis (Weil’s disease) can lead to multi-organ dysfunction, including hepatic and renal failure, hemorrhagic manifestations, and altered mental status. This case report describes a 52-year-old male who presented with severe jaundice, nausea, vomiting, diarrhea, and multi-organ dysfunction, including acute kidney injury, liver failure, and thrombocytopenia, following a recent history of fever and self-medication. Initial investigations revealed elevated inflammatory markers, liver enzymes, renal dysfunction, and thrombocytopenia, raising suspicion for leptospirosis. Empiric treatment with doxycycline was initiated, and the diagnosis was confirmed via positive Leptospira IgM (Enzyme-Linked Immunosorbent Assay - ELISA) testing. Despite the severe nature of the illness, the patient showed gradual recovery following supportive care, including hemodialysis and liver protection strategies. This case underscores the importance of early recognition and empirical treatment of leptospirosis in severe cases, as timely intervention can significantly improve outcomes and highlights the need for public health strategies targeting reservoir control and community awareness to avoid its outbreak in the community. Early diagnosis and intensive supportive care, including organ-specific management, are critical for mitigating the high morbidity and mortality associated with multi-organ dysfunction in leptospirosis.

BACKGROUND: Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by pathological expansion of CAG repeats in the HTT gene. A higher CAG repeat count (≥ 55) is generally associated with an earlier age at onset, more rapid disease progression, and a less favorable prognosis. In recent years, deep brain stimulation (DBS) targeting the globus pallidus internus (GPi) has been explored as a therapeutic intervention for medically refractory chorea; however, the available clinical evidence is largely limited to small case series and case reports, and optimal programming often requires balancing therapeutic efficacy against stimulation-related adverse effects. Here, we report an Asian patient with adult-onset HD and a high CAG repeat burden, in whom GPi-DBS using interleaved stimulation (ILS) improved motor symptoms while minimizing stimulation-induced side effects. CASE PRESENTATION: A 28-year-old Asian female with genetically confirmed HD (56 CAG repeats) presented with rapid progression from focal myoclonus of the right thumb to generalized, disabling chorea refractory to pharmacological treatment. Bilateral GPi-DBS was performed. Initial monopolar stimulation yielded limited therapeutic benefit, with only a 12.0% improvement in the Unified Huntington’s Disease Rating Scale–Total Motor Score (UHDRS-TMS), and was associated with blurred vision and gait freezing. Bipolar stimulation increased motor improvement to 30.9%, but induced dysarthria and bradykinesia. Subsequent interleaved stimulation resulted in a 58.2% improvement in motor function without persistent adverse effects. This therapeutic benefit was sustained over a 12-month follow-up period. CONCLUSION: This case provides clinical evidence from an Asian patient with adult-onset Huntington’s disease and a high CAG repeat burden, who underwent bilateral GPi-DBS with ILS. It supports ILS-DBS as an advanced programming strategy capable of optimizing clinical outcomes in HD when conventional programming approaches are limited by stimulation-induced adverse effects, and highlights the importance of individualized postoperative management. Further studies in larger cohorts are warranted to validate the efficacy and safety of ILS-DBS in HD.

<h4>Background</h4>The burden of urinary incontinence (UI) on the American public is great. The impact of this condition will only continue to rise as the population ages, yet the quality of care provided at the primary care level has been inadequate to date. The Optimizing Primary Care Tools for Incontinence MAnagement (OPTIMA) study has been designed as a four-pronged, practice-based incontinence intervention aimed at improving the management of UI by primary care providers (PCP).<h4>Methods</h4>In this pragmatic cluster randomized controlled trial, providers across four Southern California healthcare systems are randomized at the office level to receive incontinence intervention (n = 24 offices; 72 providers) vs. a non-intervention routine primary care cohort (n = 24 offices; 72 providers). The intervention includes (i) academic detailing with physician education and individual performance feedback; (ii) clinical decision support with note templates, order sets, and pop-up EHR alerts; (iii) access to co-management with a dedicated advanced practice provider; and (iv) implementation of an electronic referral service in which a specialist screens referrals for appropriateness. To achieve adequate power, the study will require 720 patients (360 patients per arm, average of 15 patients per office). The primary provider outcome is the quality of UI care, as measured by adherence to a set of 13 quality indicators (QIs) on a 6-month chart review. Secondary provider outcomes include specialty referral rates (overall specialist referrals and presence of appropriate delay of referral) and disease-specific knowledge measured by the Pelvic Floor Awareness and Knowledge Survey (PFAKS). Patient self-reported outcomes include disease-specific knowledge measured by PFAKS, UI severity measured by the International Consultation on Incontinence Questionnaire (ICIQ-SF) and Urinary Distress Inventory (UDI-6), UI response to therapy measured by Patient Global Impression of Improvement (PGI-I), and patient-perceived shared decision making via the 9-item Shared Decision Making Questionnaire (SDM-Q-9). Patient outcomes are measured at baseline, 3-, and 6-month timepoints post-appointment with the study provider.<h4>Discussion</h4>We expect this study to determine the efficacy and impact of comprehensive practice-based interventions on provider quality for UI care.<h4>Trial registration</h4>The study is registered at ClinicalTrials.gov (NCT05534412). Registered on August 1, 2022.

Multiple myeloma (MM), a hematological malignancy characterized by abnormal plasma cell proliferation, remains incurable because of its pervasive resistance to therapy. Consequently, promising therapeutic strategies are urgently needed to improve treatment responses and long-term outcomes in patients with MM. Emerging preclinical evidence indicates that ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation and iron dysregulation, may suppress the progression of MM. Several compounds have demonstrated efficacy in eliminating MM cells through ferroptosis induction. In this review, we summarize the core molecular mechanisms underlying ferroptosis, focusing on its role in MM pathogenesis and the regulatory networks modulating it within the MM microenvironment. We systematically review various ferroptosis-inducing compounds and their documented therapeutic effects in experimental models of MM. This study underscores the translational potential of pharmacological ferroptosis induction as a promising therapeutic strategy for MM.

Methylation is a crucial biochemical reaction involved in a wide range of processes including gene regulation, signal transduction, epigenetics, metabolism and detoxification. A number of methyltransferases (MTases) catalyze transfer of methyl groups from S-adenosyl-L-methionine (AdoMet or SAM) to nucleic acids, proteins, and small molecules, affecting chromatin structure, RNA function, and metabolic pathways. MTase dysregulation is associated with maladies such as cancer, neurodevelopmental disorders, and metabolic syndromes. Advancements in bioinformatics and high-throughput genomics have resulted in identification of ~ 200 human MTase genes, and most of the encoded proteins have now been characterized biochemically. Here, we have classified the human MTases into nine structural homology groups, including a distinct category of methyltransferases with unique structures. Major groups include the versatile seven-β-strand (7BS) MTases, the SET domain MTases which mainly mediate protein lysine methylation, and the SPOUT MTases involved in RNA modification. In addition, we categorized the MTases based on substrate specificity (e.g., nucleic acids, protein, and small-molecule MTases). This article provides a comprehensive classification and structural overview of human MTases, integrating recent nomenclature updates from the HUGO Gene Nomenclature Committee (HGNC). The evolutionary relationships and diversification of methyltransferases are also discussed in the context of structural classification and functional specialization. Emphasis is placed on biological functions, disease associations, and emerging therapeutic potential of the human MTases, particularly in oncology and neurodegenerative research. Despite significant progress, the biological function of many MTases remainselusive, necessitating further research to elucidate their enzymatic mechanisms and potential as drug targets. Understanding the MTase landscape is crucial for advancing biomedical research and developing targeted therapies for methylation-related disorders.

The fovea is a specialized retinal region that supports high-acuity vision through extreme photoreceptor and ganglion cell densities and near one-to-one circuit connectivity. Although present only in a subset of vertebrates, foveae share core functional principles while differing widely in morphology, cellular composition, number, and retinal location, indicating multiple independent evolutionary origins. Here, I review comparative, developmental, and genomic evidence that the fovea represents a case of convergent re-evolution built on conserved central nervous system patterning mechanisms. I highlight retinoic acid signaling as the earliest known molecular pathway demarcating foveal progenitors across distantly related species and discuss how evolutionary modulation of this positional system may repeatedly generate high-acuity retinal specializations.

<h4>Background</h4>Colorectal cancer (CRC) ranks among the most prevalent gastrointestinal malignancies with liver metastasis being the primary cause of CRC-related death. Although surgical and chemotherapeutic interventions continue to improve, patients with hepatic metastases frequently experience recurrence and limited treatment benefits. Liver metastasis is driven by tumor heterogeneity and immune evasion. Therefore, defining the cellular composition of CRC liver metastases may help identify new therapeutic targets.<h4>Methods</h4>Primary CRC and liver metastasis samples were analyzed by single-cell RNA sequencing (scRNA-seq). Seurat was used for quality control, dimensionality reduction, clustering, and cell annotation, and Harmony corrected batch effects. Differential expression with GO, KEGG, and GSEA was performed for enrichment. Copy number variation analysis using inferCNV (v1.1) distinguished malignant from non-malignant cells, with smooth muscle cells (SMCs) and epithelial cells (EPCs) as reference populations. CytoTRACE and Slingshot characterized tumor differentiation trajectories, while CellChat and pySCENIC constructed cell communication and transcriptional regulatory networks. The key factor <i>CEBPG</i> was validated by <i>in vitro</i> functional experiments. Statistical analyses were conducted in R and Python.<h4>Results</h4>scRNA-seq identified five CRC tumor cell subtypes, among which the C4 <i>BCL2L1⁺</i> tumor cells (TCs)subtype was predominantly enriched in liver metastases and displayed enhanced proliferation, metabolic reprogramming, and anti-apoptotic activity. Furthermore, transcription factor analysis suggested that <i>CEBPG</i> might regulate <i>BCL2L1</i> expression to promote tumor survival and migration. Subsequent <i>CEBPG</i> silencing markedly suppressed CRC cell proliferation and invasion. In addition, a BTRS model derived from the C4 <i>BCL2L1⁺</i> TCs subtype effectively stratified patient prognosis, as the high-risk group exhibited elevated expression of immune escape-related genes and impaired immune function.<h4>Conclusion</h4>This study revealed that the C4 <i>BCL2L1⁺</i> TCs subtype might drive CRC progression by promoting metabolic adaptation and immune evasion. In addition, <i>CEBPG</i> functioned as a key regulatory factor that increased tumor malignancy through <i>BCL2L1</i>-mediated survival pathways to some extent. The BTRS model reflected the molecular and immune heterogeneity of CRC and provided a framework for clinical risk stratification and personalized therapy. In summary, this work provides a comprehensive mechanistic framework linking metabolic adaptation, immune escape, and the progression and metastasis of CRC, and identifies potential therapeutic targets.

Cystic fibrosis (CF) is characterized by chronic airway inflammation, yet clinical observations have revealed more favorable COVID-19 outcomes than originally predicted. Several studies demonstrated a significant decrease of SARS-CoV-2 replication in CF-mutated bronchial cells suggesting that CFTR dysfunction may interfere with viral replication, though the underlying mechanisms remain unclear. To elucidate these mechanisms we performed transcriptomic profiling of SARS-CoV-2-infected bronchial epithelial cells with wild-type (WT) or mutated CFTR, using both immortalized and primary airway models. RNA-seq was performed on WT and CF cellular models before and at 24, 48, and 72-hours post-infection. The differentially expressed genes (DEGs) were defined as genes with a log2 fold change>1 between groups (p<0.05) and significant DEGs were subjected to Gene Ontology and KEGG enrichment analysis (p<0.05). Our results reveal that CFTR deficiency impairs SARS-CoV-2 replication not by altering receptor availability (e.g., <i>ACE2, TMPRSS2</i>), but through widespread intracellular remodeling defects. CF cells failed to activate key antiviral and inflammatory responses, including interferon signaling, AP-1 transcriptional complex, and IL-6-mediated pathways. Furthermore, they exhibited defective unfolded protein response, altered calcium signaling, and disrupted ER-mitochondrial communication. Crucially, pH dysregulation and impaired expression of V-ATPase subunits and autophagy-related genes hindered vesicle acidification, double-membrane vesicle formation, and viral assembly. These intrinsic alterations also blunted virus-induced senescence programs. Collectively, our findings indicate that CF cellular environment is intrinsically unfavorable to SARS-CoV-2, limiting its replication and propagation. This study provides a mechanistic basis for the reduced viral burden observed in CF and highlights intracellular pH regulation and organelle homeostasis as potential therapeutic targets against SARS-CoV-2 infection.

<h4>Introduction</h4>Age-related macular degeneration (AMD) represents a multifactorial disease that is influenced by age, genetic, and environmental factors. AMD is characterized by dysfunction of the retinal pigment epithelium (RPE) resulting from oxidative stress, inflammation, and complement activation. As the disease progresses, the loss of the RPE and photoreceptors leads to geographic atrophy, which is a hallmark of dry AMD. Although research is ongoing, there is currently no established effective treatment for dry AMD. Notably, circular RNAs (circRNAs) have been studied in various diseases; however, the role of circRNAs in eye diseases remains poorly understood. To fill this gap, this study aimed to investigate circRNAs as potential therapeutic targets for dry AMD.<h4>Methods</h4>We identified candidate circRNAs using a laser-induced choroidal neovascularization (CNV) model. The function of circAFF3 was investigated through knockdown experiments in ARPE-19 cells, followed by transcriptomic analysis, pathway enrichment, and functional assays, including qPCR, western blotting, immunofluorescence, monocyte adhesion, and measurements of ROS, iron, lipid peroxidation, and apoptosis. Interaction between circAFF3 and p53 was explored using binding prediction, RNA-binding protein immunoprecipitation, and cycloheximide chase assay.<h4>Results</h4>We found that circAff3 levels were downregulated in RPE samples at day 3 after laser injury. Moreover, silencing circAFF3 induced an inflammatory response in ARPE-19 cells. Based on these results, a subsequent transcriptomic analysis of circAFF3 knockdown in ARPE-19 cells was conducted to further elucidate its function. These analyses showed that genes downregulated by circAFF3 knockdown in ARPE-19 cells were significantly associated with retinal degeneration. Additionally, reduced circAFF3 expression promoted a decrease in ID2 levels, resulting in increased oxidative stress and cell death. Our study further demonstrated that circAFF3 directly interacts with p53, thereby regulating ID2 expression in ARPE-19 cells.<h4>Conclusion</h4>Collectively, our study reveals that circAFF3 plays a crucial role in RPE dysfunction by modulating a circAFF3/p53/ID2 pathway, suggesting that circAFF3 can serve as a key regulator with therapeutic potential in dry AMD.

Regular physical activity is a powerful non-pharmacological strategy for preventing and managing cardiovascular diseases (CVDs), including heart failure, by promoting cardioprotective adaptations through molecular mechanisms that remain incompletely elucidated. This review explores the central role of non-coding RNAs (ncRNAs), particularly microRNAs (miRNAs), in exercise-induced cardioprotection, highlighting their interactions within miRNA-lncRNA and miRNA-circRNA axes, as well as the function of exosomal miRNAs as key exerkines facilitating inter-organ crosstalk. Synthesizing current literature, we examine ncRNA biogenesis, canonical functions, and exercise-responsive profiles, focusing on pivotal miRNAs such as miR-1, miR-133, miR-21, miR-126, miR-29, miR-208a, and miR-499; lncRNA-miRNA networks including MALAT1/miR-150-5p, H19/miR-139, and GAS5/miR-217; circRNA-miRNA interactions like circUtrn/miR-132/212; and exosomal miRNAs derived from skeletal muscle (e.g., miR-130a, miR-1), brown adipose tissue (e.g., miR-17-3p), endothelium (e.g., miR-126), and cardiomyocytes (e.g., miR-21-3p). These elements are evaluated in models of physiological cardiac remodeling, myocardial infarction, ischemia-reperfusion injury, diabetic cardiomyopathy, and heart failure, with consideration of influencing factors such as sex, age, and training modality. Exercise-modulated miRNAs differentiate benign "athlete's heart" from pathological hypertrophy by governing angiogenesis, fibrosis, metabolic shifts, and arrhythmia risk, while lncRNA-miRNA and circRNA-miRNA axes regulate apoptosis, inflammation, mitochondrial dynamics, and extracellular matrix remodeling in CVD contexts. Exosomal miRNAs enable remote protection by activating survival, angiogenic, and anti-fibrotic pathways via signaling cascades like PI3K/AKT and NF-κB. Responses exhibit variability based on demographic and exercise variables, underscoring ncRNAs' promise as diagnostic biomarkers, therapeutic targets, or mimics of exercise benefits for heart failure management.

Sepsis is a clinical syndrome defined as life-threatening organ dysfunction caused by a dysregulation in immune response to infection. Dysregulated neutrophil activity plays a critical role in sepsis-induced organ failure through interactions with the vascular endothelial cells during forward and reverse migration, resulting in vascular barrier disruption and increased neutrophil trafficking into vital organs. Therapeutic approaches for treating sepsis are mainly supportive. Due to limited clinical translation from rodent models, complexity of the pathophysiology, and most importantly, the heterogenous nature of sepsis, no significant therapeutics have been successfully developed to address the underlying immune dysregulation. In this review, we will discuss the important gap in knowledge on the fundamental mechanisms of neutrophil-endothelial interaction, the role that neutrophil forward and reverse migration plays in organ damage in sepsis, and how neutrophil and endothelial cell heterogeneity impact cell-cell communication. We will explore emerging methodologies, including novel omic and microphysiological systems, to study the underlying mechanism of neutrophil-endothelial interaction and neutrophil forward migration/reverse migration. Finally, we will review potential therapeutic targets modulating neutrophil-endothelial interaction and the challenges of translating them from bench to bedside.

Beef flavor is a trait difficult to evaluate since different senses (taste, touch, and smell) are involved in its perception. In the last 20 years, 102 Quantitative Trait Loci (QTLs), associated with the variability of different beef flavor notes, have been reported. These QTLs are spread on all chromosomes, including BTA X. In these QTL regions, 2509 genes are located and, among them, 594 are involved in the metabolic processes of lipids, proteins, and carbohydrates, the main meat components for the production of volatile substances responsible for flavor. Only 19 of these genes (<i>ACSM2B</i>, <i>ACSM3</i>, <i>ACSM4</i>, <i>ACSM5</i>, <i>CHID1</i>, <i>DHCR7</i>, <i>EDEM3</i>, <i>GDE1</i>, <i>HEXB</i>, <i>IGF2</i>, <i>INS</i>, <i>NDUFAB1</i>, <i>PIGC</i>, <i>PNPLA2</i>, <i>PRDX6</i>, <i>SCNN1B</i>, <i>SIAE</i>, <i>SMG1</i>, and <i>UMOD</i>) are also present in the QTL regions affecting pork flavor. The applied approach allowed us to strongly restrict the number of candidate genes to affect the variability of both beef and pork flavor.

Neurodegeneration and cancer are fundamentally distinct disorders: one signifies gradual neuronal loss while the latter signifies uncontrolled cell growth and survival. However, emerging evidence explores an inverse association between these conditions, suggesting that they do not arise from independent biological processes. Understanding the context-dependent behaviour of major pathways (for example, p53, PI3K/AKT/mTOR, Wnt, and immune-stress signaling) remains pivotal in elucidating the relationship between these two diseases. Pathways promoting early-life fitness, tissue repair, and tumor suppression in dividing cells can become detrimental later in life for post-mitotic neurons. Cross-species genomics studies reveal how evolution has repeatedly adapted these shared networks to balance cancer resistance with survival. Research on species exhibiting exceptional longevity and disease resistance, including naked mole rats and bowhead whales, shows that cancer resistance and longevity are not fixed traits but rather are controlled by precise regulatory mechanisms. In this review, we integrate insights from broad species genomics and multi-omic and single-cell studies to understand how evolutionarily conserved molecular crosstalks diverge at the interface of cancer and neurodegeneration.

The guinea fowl (<i>Numida meleagris</i>), a thermo-tolerant and disease-resilient poultry species, holds great potential for sustainable poultry production in climate-vulnerable regions. The genomic aspects of this species remain largely understudied. The present study aims to delineate the patterns of domestication and understand the evolutionary dynamics of guinea fowl populations (wild and domestic) across three continents, utilizing whole-genome sequencing data from 122 genomes. The population structure analyses (ADMIXTURE, PCA, phylogeny, F_ST, LD, and MAF) revealed that Indian guinea fowl (CARI) shared close ancestry with Iranian (IRAD) and Chinese (CHID) domesticated populations while remaining distinct from wild lineages. The runs of homozygosity (ROH) identified 49,088 segments, with short fragments (ROHs) preponderant in Indian and domestic populations, reflecting historical inbreeding and effects of domestication cum selection. Copy number variation (CNV) analysis revealed 105,178 CNVs concatenated into 40,067 CNV regions (CNVRs) across 11 populations, establishing the first CNV atlas for guinea fowl at the global level. Gene annotation of overlapping ROH and CNVRs revealed 1080 common candidates across Asian guinea fowl populations, i.e., the Indian guinea fowl (CARI), IRAD, and CHID, including <i>FOS</i>, <i>EPAS1</i>, <i>CD74</i>, and <i>CSF1R</i>. These genes have earlier been associated with immune regulation, stress response, and thermal adaptation. Selection signature scans, integrating intra-population (iHS) and inter-population (XP-EHH) approaches, uncovered genes under positive selection linked to immune response (like <i>BCL11B</i>, <i>IL18</i>, and <i>GPC3</i>), thermo-tolerance (like <i>TRPV4</i> and <i>BAG3</i>), lipid metabolism (like <i>AACS</i> and <i>ELOVL4</i>), and pigmentation (<i>BCO2</i>). These signatures highlight the molecular basis of resilience in guinea fowl and their potential to withstand climate-induced stresses. This study presents the first global CNV atlas for guinea fowl and provides the first comprehensive genomic characterization of the Indian domestic population, integrating ROH, CNV, and selection signature analyses. It offers a comprehensive assessment of guinea fowl genomes (wild and domesticated) across three continents, offering insights into domestication, evolutionary dynamics, and the genetic basis of their adaptation and resilience.

Multiple myeloma (MM) is an incurable plasma cell malignancy characterized by high metabolic activity, chronic endoplasmic reticulum stress, and persistent redox imbalance. Excessive immunoglobulin synthesis and adaptation to the hypoxic bone marrow microenvironment lead to sustained production of reactive oxygen species (ROS). Their excessive accumulation promotes genomic instability, disease progression, osteolytic bone disease, and resistance to therapy. Paradoxically, MM cells adapt to oxidative stress by activating antioxidant and metabolic defense mechanisms, including Nuclear factor erythroid 2-related factor 2 (NRF2)- and Heme Oxygenase 1 (HMOX1)-dependent pathways, metabolic reprogramming, and overexpression of ROS-scavenging enzymes such as peroxiredoxin 6 (PRDX6), allowing survival at the threshold of oxidative toxicity. Evidence indicates that biomarkers of oxidative stress-such as lipid and protein oxidation products, antioxidant enzyme activity, and the Oxidative Stress Score-correlate with disease stage, prognosis, and treatment response. Redox-modulating therapeutic strategies, including pharmacological ROS induction, inhibition of antioxidant defenses, and the use of natural pro-oxidant compounds, are emerging as promising adjuncts to standard MM therapies. Recent studies also highlight the gut microbiota as an indirect regulator of oxidative balance, immune modulation, and metabolic homeostasis in MM. This review summarizes current knowledge on oxidative stress in multiple myeloma, emphasizing its role in pathogenesis, drug resistance, biomarker development, and emerging therapeutic and supportive strategies.

Although tobacco smoking is the leading cause of lung cancer (LC), excessive sugar intake has also emerged as a potential risk factor, yet its mechanistic contribution remains poorly defined. In this study, we investigated how high-fructose intake modulates the tobacco carcinogen-induced LC progression. Co-exposure to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and benzo[a]pyrene (NNK and BaP; collectively referred to as NB) combined with a high-fructose diet markedly accelerated tumor progression in multiple mouse models, including Kras^G12D/+-driven LC and LKB1-deficient (LKB1^KO) lung tumors. NB-induced LC progression was suppressed by restricting glucose metabolism, indicating a metabolic dependency. Mechanistically, NB exposure stimulated transcriptional programs that promote monocyte/macrophage recruitment within the tumor microenvironment and enhanced fructose uptake through both transcriptional and post-transcriptional upregulation of fructose transporters, including glucose transporter 8 (GLUT8). This metabolic reprogramming increased acetylation of histones and signal transducer and activator of transcription 3 (STAT3), leading to transcriptional upregulation of genes governing macrophage differentiation and M2 polarization. Analysis of human LC samples revealed enrichment of pro-metastatic IL-10⁺ and VEGFA⁺ M2 macrophages, which correlated with poor clinical outcomes. Collectively, these findings demonstrate that NB-driven fructose metabolism induces epigenetic reprogramming of macrophages to promote LC progression and identify pro-metastatic M2 macrophages as potential prognostic biomarkers and therapeutic targets.

Hereditary hemochromatosis is a disorder of iron metabolism characterized by increased intestinal iron absorption and progressive parenchymal iron deposition, most commonly associated with homozygous mutations in the HFE gene. In contrast, heterozygous carriers of HFE mutations are generally considered to have low clinical penetrance and are not expected to develop clinically significant iron overload or advanced liver disease. However, iron homeostasis may be substantially altered in the presence of chronic liver inflammation and immune-mediated hepatic injury. We present the case of a woman in her early 60s with a heterozygous HFE C282Y mutation who developed marked hyperferritinemia, biopsy-confirmed hepatic iron overload, and established cirrhosis in the setting of autoimmune hepatitis treated with mycophenolate mofetil. Her clinical course was notable for wide fluctuations in ferritin levels over several years, with a peak exceeding 3,800 ng/mL, and subsequent sustained reduction following therapeutic phlebotomy. Management was complicated by anemia, requiring individualized phlebotomy thresholds and transition to a maintenance strategy. This case highlights the diagnostic challenges of interpreting iron indices in patients with inflammatory liver disease and demonstrates that clinically significant iron overload may occur in HFE heterozygotes when additional disease modifiers are present. It underscores the importance of histologic confirmation of iron overload in cases of genotype-phenotype discordance and reinforces the need for individualized management strategies in complex iron-loading conditions.

<b>Objective</b>: To evaluate the utility of synovial iron quantification using Magnetic resonance imaging (MRI) in assessing structural joint damage in the knee of patients with rheumatoid arthritis (RA). <b>Methods</b>: This cross-sectional study employed a two-stage design. In the initial comparative stage, 6 patients with RA and 5 patients with osteoarthritis (OA) were recruited to compare synovial R2* values, a metric derived from iterative decomposition of water and fat with echo asymmetry and least-squares estimation quantitation (IDEAL-IQ) MRI sequences representing synovial iron content. Following this, the RA cohort was expanded to a total of 51 patients to investigate the association between R2* values and clinical parameters, including disease activity and bone erosion. Synovial fluid iron levels were measured with an Iron Assay Kit and synovial iron deposits were semi-quantified via Prussian blue staining. Associations between R2* and clinical and laboratory parameters, including inflammatory factors and joint damage indices, were analyzed using Spearman's rank correlation. Univariate and multivariate ordered logistic regression models were employed to identify factors associated with bone erosion severity. An R2*-based nomogram was developed and validated using receiver operating characteristic (ROC) analysis and calibration curves. <b>Results</b>: Synovial R2* values were significantly higher in RA patients than those with osteoarthritis (53.66 S^-1 vs. 31.38 S^-1, <i>p</i> < 0.05), consistent with Prussian blue staining results. While synovial R2* values showed no significant correlation with systemic iron metabolic markers, inflammatory indicators, or the Disease Activity Score 28, they were positively correlated with bone erosion severity (ρ = 0.500, <i>p</i> < 0.001) and negatively associated with the joint space width (ρ = -0.307, <i>p</i> < 0.05). Multivariate analysis identified R2* as an independent indicator linked to bone erosion extent (OR = 2358.336, <i>p</i> < 0.001). The R2*-based nomogram demonstrated good discriminative performance. (AUC = 0.83). <b>Conclusions</b>: The R2* value derived from IDEAL-IQ MRI is a reliable tool for quantifying synovial iron and may represent a promising non-invasive imaging biomarker reflecting bone erosion in RA patients.

Neurodegenerative diseases feature diverse pathological protein aggregates, including Lewy bodies in Alzheimer's disease (AD) and skein-like filaments in amyotrophic lateral sclerosis (ALS). The physical mechanisms underlying this morphological diversity remain unclear. Here, we demonstrate that aggregation of the prion-like domain of hnRNPA1 (A1PrD), implicated in AD and ALS, is driven by solution composition and phase transition dynamics. Utilizing 3D timelapse and fluorescence lifetime imaging microscopy, we show that solution conditions modulate phase separation, gelation, and fibrillation, resulting in distinct structures such as fibril, gel, and starburst morphologies. Homotypic and heterotypic interactions between A1PrD and RNA were observed to shift the balance between pathological and physiological condensates. Importantly, amyloid-rich starbursts displayed prion-like infection capabilities toward amyloid-poor condensates. Our findings highlight how the interplay between solution composition and kinetic balances of liquid-liquid phase separation, gelation, and fibrillation shapes the diverse pathological aggregate morphologies characteristic of neurodegenerative diseases.

<h4>Background/objectives</h4>Comparative sociogenomics combines multiple scientific fields to investigate the genetic basis of social behavior across species. Our aim was to uncover the genetic roots of human sociability with possible implications for autism, a neurodevelopmental disorder characterized by social and communication deficits.<h4>Methods</h4>We conducted molecular network analysis on 659 sociability-related genes from different animal species, including humans.<h4>Results</h4>We identified a network of 240 genes strongly associated with autism (<i>p</i> < 10^-15), with 194 inferred. These genes were grouped into 23 functional communities related to cell-cell junctions and communication, inflammatory and synaptic signaling, neurotransmitter receptors and semaphorin signaling among the more enriched meta-pathways. Some network genes were clustered in nine chromosomal bands (FDR < 0.25), indicating genes' functional cooperation, shared evolutionary history, and coordinated regulation, and few genes are physically in linkage with ASD genes (within 0.5 cM) or controlled by human-accelerated regions.<h4>Conclusions</h4>The most compelling inferred autism risk genes are <i>MED12, FZD9</i>, and <i>DMD</i> since they are differentially expressed in autistic brains, physically linked to key autism genes, controlled by human-accelerated regions, or mapped to chromosomal regions enriched in network genes. If validated, they could represent novel biomarkers, advancing the understanding of autism's genetic makeup.

<h4>Background</h4>Bladder urothelial carcinoma (BLCA) is a prevalent malignant tumor within the urinary system. Aspartyl-tRNA Synthetase 2, Mitochondrial (DARS2), a key enzyme involved in mitochondrial protein synthesis, is frequently overexpressed in various cancers and is associated with clinical stage. This study aims to explore the potential of DARS2 as a prognostic biomarker and therapeutic target in BLCA, with the ultimate goal of constructing a prognostic model.<h4>Methods</h4>Expression profiles of BLCA were obtained from The Cancer Genome Atlas (TCGA) database and the Genotype-Tissue Expression (GTEx) database, encompassing a total of 431 samples. Differential gene expression between tumor and normal tissues was first analyzed using the "limma" R package. To identify independent prognostic genes, we integrated various statistical and machine learning approaches, including univariate and multivariate Cox proportional hazards analyses, with variable selection performed via Cox proportional hazards regression with least absolute shrinkage and selection operator (LASSO). For the key gene DARS2, its expression differences were further analyzed using the edgeR method, followed by gene set enrichment analysis (GSEA) to explore associated functional pathways. Furthermore, immune cell infiltration was evaluated using multiple algorithms. The role of DARS2 in BLCA progression was systematically investigated by integrating analyses of gene mutations, tumor mutational burden (TMB), and survival outcomes. Finally, a prognostic model was constructed based on DARS2 expression levels and validated using the GSE13507 dataset from the Gene Expression Omnibus (GEO) database.<h4>Results</h4>Based on the analytical methods described above, we observed that the expression of DARS2 messenger ribonucleic acid (mRNA) was significantly upregulated in BLCA tissues, and its elevated expression was independently associated with adverse patient prognosis. Immune infiltration analysis revealed that DARS2 expression exhibited a significant negative correlation with major immunologically active cells, such as CD4⁺ T cells and CD8⁺ T cells. In terms of genetic mutations, the somatic mutation rate of DARS2 in BLCA was 2.21%, whereas the receptor tyrosine kinase (RTK)/RAS GTPase signaling pathway (RTK-RAS) represented the most frequently mutated oncogenic pathway (72.97%). DARS2 expression showed a moderate positive correlation with TMB, with factors including advanced age, male sex, and higher tumor grade potentially contributing to elevated TMB. Survival analysis further demonstrated that patients with high DARS2 expression had significantly poorer outcomes in overall survival (OS), disease-specific survival (DSS) and progression-free interval (PFI). The risk-scoring model constructed based on DARS2 expression indicated that age, metastasis (M) stage, and risk score all served as independent prognostic factors for BLCA.<h4>Conclusions</h4>Comprehensive analysis suggests that DARS2 holds promise as a potential independent prognostic biomarker for patient stratification in BLCA and may serve as a viable target for the development of immunotherapeutic strategies.

<h4>Background</h4>Zinc finger E-box binding homeobox 1 antisense 1 (<i>ZEB1-AS1</i>) is a long non-coding RNA (lncRNA) related to the progression of several malignancies and a regulator of oncogenesis in several cancers. Therefore, the aim of our study was to explore the prognostic value and immune relevance of <i>ZEB1-AS1</i> in hepatocellular carcinoma (HCC).<h4>Methods</h4>In this study, a comprehensive investigation integrating bioinformatic analysis and <i>in vitro</i> experiments was conducted to evaluate the role of <i>ZEB1-AS1</i> in HCC. HCC-related datasets were retrieved from The Cancer Genome Atlas (TCGA) database to analyze the prognostic value of <i>ZEB1-AS1</i> via univariate and multivariate Cox regression. The potential biological functions and immune-related correlations of <i>ZEB1-AS1</i> were explored through co-expression analysis and functional enrichment using R software (v4.0.5). Furthermore, <i>in vitro</i> assays were performed, including quantitative real-time polymerase chain reaction (qRT-PCR) to quantify <i>ZEB1-AS1</i> expression in HCC and normal liver cells, as well as functional experiments to assess its impact on cell proliferation and motility.<h4>Results</h4><i>ZEB1-AS1</i> overexpression was associated with poor prognosis in HCC patients. <i>ZEB1-AS1</i> expression and tumor stage were independent prognostic factors in patients with HCC. The expression of <i>ZEB1-AS1</i> was significantly increased in HCC cell lines. Functional assays revealed that knockdown of <i>ZEB1-AS1</i> markedly inhibited Huh-7 cell proliferation and suppressed their migratory and invasive capacities. A significant negative association of <i>ZEB1-AS1</i> expression with stromal score and immune score was observed. <i>ZEB1-AS1</i> expression was correlated with multiple immune cells, immune checkpoint inhibitor, immunotherapy response and drug sensitivity. ZEB1-AS1 was involved in the regulation of tumor-associated signaling pathways.<h4>Conclusions</h4>In conclusion, our integrated bioinformatic analysis and <i>in vitro</i> validation suggest that <i>ZEB1-AS1</i> may serve as a potential theoretical prognostic indicator and a target for immune checkpoint reactivity in HCC. These findings provide a novel direction and a theoretical framework for future research involving large-scale clinical cohorts to confirm its clinical application in the long term.

Short tandem repeat expansions in exon 1 of the HTT gene drive Huntington’s disease (HD) pathogenesis, with disease onset and progression heavily influenced by somatic mosaicism and sequence interruptions. While sequencing technologies enable repeat sizing, many computational tools lack the resolution to capture subtle interruption motifs and allele-specific somatic variation. We present STRmie-HD, an alignment-free, de novo framework for interruption-aware genotyping and quantitative profiling of somatic mosaicism at single-read resolution. The tool parses individual reads to quantify uninterrupted CAG tract length, CCG repeat content, and critical interruption variants, including Loss of Interruption (LOI) and Duplication of Interruption (DOI). Validated across Illumina, PacBio SMRT, and Oxford Nanopore platforms, STRmie-HD demonstrates high concordance with reference genotypes and superior sensitivity in identifying rare interruption patterns that conventional tools often overlook. Furthermore, it implements somatic mosaicism metrics to characterize repeat dynamics, successfully distinguishing the higher somatic expansion burden in brain tissues compared to peripheral blood. STRmie-HD offers a comprehensive and extensible solution for high-resolution molecular characterization of HTT variation, providing a robust framework for patient stratification and genetic research in HD. <h4>Graphical Abstract</h4> <h4>Graphical Abstract:</h4> STRmie-HD flowchart. STRmie-HD is a comprehensive analytical framework that processes sequencing reads to analyze CAG/CCG trinucleotide repeats, interruption variants, and somatic mosaicism in the HTT gene. The workflow begins with sequencing reads (FASTA/FASTQ) that can undergo optional custom processing eq]based on the sequencing design. These reads are then fed into a regular expression-based engine (STRmie-HD) to identify CAG and CCG motifs. The identified motifs lead to the estimation of CAG/CCG alleles, visualized as distinct peaks representing different allele sizes, interruption variant assessment, and somatic mosaicism quantification. STRmie-HD produces an HTML output that wraps this information into a report.

Iron-sulfur clusters (ISCs) are redox active cofactors for essential proteins with diverse functions. We demonstrate that Tempol, a redox cycling nitroxide, limits iron bioavailability in a manner distinct from iron chelators, mainly via its effect on ascorbate and iron redox balance. This non-canonical iron limitation triggers upregulation of IRP2 and HIF1α, proteins whose degradation is ferrous iron-dependent, while disrupting ISC synthesis only in a subset of cell lines. Suppression of ISC synthesis inhibits ISC-dependent enzymes, destabilizes ISC proteins, and reduces cell viability, particularly in cells dependent on ISC protein ACO2. These effects are reversed by the reducing agent ascorbate, a cofactor required for multiple enzymes, such as the HIF1α prolyl hydroxylases. Tempol treatment also inhibits ferroptosis, an oxidative form of cell death catalyzed by reduced iron. These results demonstrate ascorbate and cellular iron redox state are essential in iron homeostasis, which is proposed to underlie pathological conditions from neurodegeneration to cancer.

<h4>Background</h4>Focal segmental glomerulosclerosis (FSGS) is a major cause of nephrotic syndrome and chronic kidney disease, characterized by podocyte injury and glomerular scarring. Its heterogeneity complicates early diagnosis and treatment.<h4>Methods</h4>We integrated bulk and single-cell RNA-seq datasets (GSE200818, GSE200828, and GSE176465) to investigate epithelial cell-related molecular alterations in FSGS. Differentially expressed genes (DEGs) were identified and analyzed for functional enrichment. Machine learning algorithms (random forest and support vector machine) were applied to construct a diagnostic gene signature. Immune cell infiltration was assessed using ssGSEA, and molecular subtypes were defined via consensus clustering. Independent datasets (GSE133288 and GSE108109) were used for validation.<h4>Results</h4>A total of 133 epithelial cell-associated DEGs were identified. Enrichment analysis indicated transcriptional regulation and viral infection pathways. Three diagnostic genes-B4GALT5, CSRNP1, and CYP3A5-were selected and shown to have excellent diagnostic accuracy (AUC > 0.97) in both discovery and validation cohorts, including podocyte-enriched samples. Immune profiling revealed increased infiltration of activated CD8⁺ T cells, NK cells, and dendritic cells in FSGS, with B4GALT5 positively correlated with multiple immune subsets. Consensus clustering stratified patients into two molecular subtypes with distinct immune features.<h4>Conclusions</h4>This integrative analysis identifies epithelial cell-related biomarkers and immune-associated subtypes in FSGS, providing promising tools for diagnosis and patient stratification. While robust across multiple datasets, further experimental validation is needed to confirm mechanistic relevance.

Dendritic cells (DCs) are crucial antigen-presenting cells that mediate the interplay between innate and adaptive immunity during lethal infections. Here, we report the key role of reticulophagy regulator 1 (RETREG1), a selective autophagy receptor, in maintaining DC maturation and function in the early stage of sepsis. Mechanistically, activating transcription factor 6 (ATF6) acts as a direct transcription factor regulating RETREG1 expression in response to bacterial lipopolysaccharide-induced endoplasmic reticulum (ER) stress. RETREG1-mediated reticulophagy reduces excessive ER stress via the eukaryotic translation initiation factor 2 alpha kinase 3 (EIF2AK3) signaling pathway and inhibits membrane-associated RING-CH-type finger 8 (MARCH8)-dependent major histocompatibility complex class II (MHC-II) ubiquitination to maintain antigen presentation in DCs. Consequently, Cd11c^creRetreg1^fl/fl, Retreg1^-/-, and Atf6^-/- mice exhibit impaired DC function, leading to immunosuppression and multiple organ failure in experimental sepsis. Exploration of samples from septic patients, combined with single-cell bioinformatics analysis, further suggests that a deficit in reticulophagy in DCs is associated with the development of human sepsis.

Standard preclinical human tumor models lack a human tumor stroma. However, as stroma contributes to therapeutic resistance, the lack of human stroma may make current models less stringent for testing new therapies. To address this, using patient-derived tumor cells, patient-derived cancer-associated mesenchymal stem/progenitor cells, and human endothelial cells, we created a human stroma-patient-derived xenograft (HS-PDX) tumor model. HS-PDX, compared to the standard PDX model, demonstrates greater resistance to targeted therapy and chemotherapy and better reflect patient response to therapy. Furthermore, HS-PDX can be grown in mice with humanized bone marrow to create humanized immune stroma patient-derived xenograft (HIS-PDX) models. The HIS-PDX model contains human connective tissues, vascular and immune cell infiltrates. RNA sequencing analysis demonstrated a 94-96% correlation with primary human tumor. Using this model, we demonstrate the impact of human tumor stroma on recruitment of TAMs and tumor immune exclusion to impact to response to immunologic therapy. We show an immunosuppressive role for human tumor stroma and that this model can be used to identify immunotherapeutic combinations to overcome stroma-mediated immunosuppression. Combined, our data confirm a critical role for human stroma in therapeutic response and indicate that HIS-PDX can be an important tool for preclinical drug testing.

This study aims to investigate the effect of exosomes derived from olfactory mucosa mesenchymal stem cells (OM-MSCs-Exo) on microglial polarization and its potential therapeutic role in Alzheimer's disease (AD). OM-MSCs-Exo were isolated and purified from the mice olfactory mucosa, followed by phenotypic characterization. Proteins transferred by OM-MSCs-Exo were screened using proteomic analysis. The AD model was established in microglial cells and mice with Aβ_1-42. Immunofluorescence and biochemical assays were employed to assess the impact of OM-MSCs-Exo and its secreted protein FGFR1 on microglial polarization. Protein-protein interactions and immunoprecipitation were used to identify the target proteins of FGFR1 in microglial cells. Additionally, the effects of OM-MSCs-Exo-induced microglial polarization on neuronal inflammation and cognitive function in mice were evaluated. OM-MSCs-Exo were successfully isolated and purified. FGFR1 was significantly upregulated in OM-MSCs-Exo compared to OM-MSCs. Aβ_1-42 induced M1 polarization and suppressed M2 polarization of microglia, which was reversed by OM-MSCs-Exo. FGFR1 overexpression in OM-MSCs-Exo further enhanced M2 polarization in microglial cells. Phospholipase C gamma 1 (PLCγ1) was identified as the target of FGFR1, and knocking down PLCγ1 reversed the effects of FGFR1-overexpressing OM-MSCs-Exo. OM-MSCs-Exo alleviated cognitive decline and neuroinflammation in AD mice, with FGFR1 overexpression further enhancing these effects. OM-MSCs-Exo promote M2 polarization of microglia in AD mice through the FGFR1/PLCγ1 pathway, alleviating neuronal inflammation and cognitive dysfunction.

<h4>Objectives</h4>To explore the pharmacotherapeutic efficacy of heparin in the management of meconium-induced acute lung injury (ALI) in near-term newborn rabbits subjected to mechanical ventilation (MV) and ancillary respiratory medications.<h4>Methods</h4>Newborn rabbits at 30-day gestation (term 31 days) were anesthetized, intratracheally intubated and received human meconium-saline suspension, followed by parallel MV with individually adjusted tidal volume in a multi-plethysmograph-ventilator system. When ALI was induced after initial 3-h MV, therapeutic effects of single or combined subcutaneous heparin (100 U/kg), surfactant (200 mg/kg), and inhaled nitric oxide (iNO, 10 ppm), were compared for lung protective ventilation and survival as outcome, analyzed with linear regression models.<h4>Results</h4>Significantly reduced respiratory compliance by meconium was reinstalled during ensuing 7-h MV, with improved survival, among the treatment groups. The impact was verified by lung injury severity, surfactant phospholipid pools, and multiple mRNA expressions of surfactant proteins, lung fluid clearance-related factors, inflammatory mediators, growth factors, endothelial cell injury and coagulation-related factors as subphenotyping biomarkers. The overall benefits of heparin alone, or exerted with the dual and triple regimens, were discernible by both generalized linear model and Cox proportional hazard ratio regression for survival and other major variables as outcome. Its adverse effects were intangible.<h4>Conclusion</h4>The comparable efficacy of heparin, alongside the PS and NO, was corroborated in attenuating meconium-mediated, ventilator-induced ALI, which should warrant clinical investigation to validate.

<h4>Background and objectives</h4>Paediatric patients with haematological disorders frequently require multiple blood transfusions, increasing their risk of developing irregular antibodies. Because of the immunohaematological differences from adults, patterns of alloimmunization in children may vary. This study aimed to determine the prevalence and characteristics of irregular antibodies in paediatric patients with haematological disorders at the National Institute of Hematology and Blood Transfusion (NIHBT).<h4>Materials and methods</h4>A total of 2663 paediatric patients with haematological disorders were screened for irregular antibodies using column agglutination technology. Positive samples underwent antibody identification. Associations between alloimmunization and clinical or transfusion-related factors were assessed using univariate analyses and multivariable logistic regression.<h4>Results</h4>The prevalence of irregular antibodies was 8.04% (95% confidence interval [CI]: 7.0-9.1). Among positive cases, 66.82% had a single antibody, most commonly anti-E (35.51%) and anti-Mi^a (14.02%). The most frequent antibody combination was anti-c and anti-E (13.08%). Factors significantly associated with antibody formation after adjustment included a higher number of transfused red cell units (57.65 vs. 44.53 units, p = 0.000), absence of phenotype-matched transfusions (p = 0.017), non-Kinh ethnicity (p = 0.0003) and a diagnosis of thalassaemia (p = 0.0013).<h4>Conclusion</h4>A relatively high prevalence of irregular red cell alloantibodies was observed in paediatric patients, predominantly involving antibodies of the Rh blood group system and anti-Miᵃ. These findings support the relevance of targeted antigen matching in settings where fully phenotype-matched red blood cells are not consistently available.

Obesity arises from disrupted energy homeostasis, yet the neural mechanisms linking transcriptional regulation to energy expenditure remain unclear. Here, we identify plant homeodomain finger protein 6 (Phf6), a gene mutated in Börjeson-Forssman-Lehmann syndrome (BFLS), as a pivotal regulator of energy balance. Phf6 is enriched in a subset of estrogen receptor 1 (Esr1)-expressing neurons within the hypothalamic medial preoptic area (MPOA). Knockout of Phf6 in the MPOA leads to obesity in a sex-dependent manner by reducing physical activity and energy expenditure, independent of food intake. In female mice, ^MPOAPhf6 neurons respond to physical activity. Activation and inhibition of ^MPOAPhf6 neurons increases and decreases physical activity and energy expenditure, respectively. Phf6 sustains the intrinsic excitability of ^MPOAPhf6 neurons and their responsiveness to estrogen. Circuit mapping identified an ^MPOAPhf6-^VMHvlEsr1 pathway mediating Phf6's effect on metabolism. These findings reveal a neurobiological basis for BFLS-associated obesity and highlight potential therapeutic targets.

Cell lines are indispensable tools in prostate cancer research yet their suitability as models for distance metastasis remains incompletely characterized. Here, we conduct a systematic evaluation study using large-scale public multi-omics data. We reveal substantial genomic differences between cell lines and metastatic patient samples, and meanwhile pinpoint cell lines which more closely resemble metastatic prostate cancer. Notably, hypermutation significantly influences the tumor microenvironment, underscoring the importance of considering mutational burden in model selection. Surprisingly, the widely used PC3 cell line exhibits poor transcriptomic and epigenomic similarity to any prostate cancer subtype, revealing a previously unrecognized limitation. Furthermore, we find existing engineered stem-like cell lines fail to faithfully recapitulate the transcriptomic profiles of mesenchymal stem-like prostate cancer, whereas selected organoids exhibit higher fidelity. Our study provides guidance for cell line selection in metastatic prostate cancer research and highlights the urgent need to develop improved cell lines for the mesenchymal stem-like subtype.

Major histocompatibility complex (MHC) class I and class II molecules present antigens to CD8⁺ and CD4⁺ T cells respectively. Here we uncover a previously unrecognized role for MHC class I in modulating CD4⁺ T cell-mediated immunity. In allogeneic graft-versus-host disease and tumor models, we demonstrate that the absence of MHC class I on target cells significantly increases their susceptibility to CD4⁺ T cell cytotoxicity. Transcriptomic and functional studies suggest that this was because of heightened sensitivity to enhanced ferroptosis of the target cells. In large human transcriptomic and sequencing datasets, a role for CD4⁺ T cells in enhancing immune checkpoint blocker-mediated responses in persons with melanoma and mismatch-repair-deficient colon cancers that have downregulated MHC class I was suggested. These findings revise and expand the known role of MHC class I in CD8⁺ T cell and natural killer cell immunity and demonstrate a previously unrecognized role in CD4⁺ T cell-mediated cancer and alloimmunity.

Air particulate matter (PM2.5 and PM10), can cross the placental barrier, triggering oxidative stress and inflammation that compromise fetal development. These insults lead to placental dysfunction and complications including preterm birth, low birth weight, and preeclampsia. In cell line and placental explant models, urban particulate matter (UPM) increased pro-inflammatory cytokines and oxidative stress pathways, impairing trophoblast invasion, angiogenesis, and nutrient transport, while also altering epigenetic modifications and endoplasmic reticulum function. Rodent studies revealed reduced litter size, placental abnormalities, and fetal growth arrest along with postnatal neurodevelopmental alterations. Human cohorts from high-exposure regions showed elevated low birth weight rates. Proteomic and transcriptomic analyses of rat placenta revealed an inflammatory signature and altered metabolic networks, while gut microbiome dysbiosis suggested links to metabolic disturbances. Importantly, transcriptomic analysis identified IGFBP3 as a major downregulated gene following UPM exposure. IGFBP3, a key regulator of IGF bioavailability, was suppressed by IL1β, establishing inflammation-driven repression as the mechanism. These findings underscore UPM's multidimensional impact on maternal-fetal health and highlight preventive strategies as urgent priorities.

BACKGROUND: Monitoring of unfractionated heparin (UFH) in critically ill patients is challenging due to disease-related alterations of the coagulation system, directly affecting conventional laboratory tests, including activated partial thromboplastin time (aPTT). Measurement of anti-factor Xa activity (Anti-Xa) as an alternative to aPTT often lacks 24/7 availability. Whole blood viscoelastic testing (VET) may provide a feasible point-of-care alternative to guide UFH dosing in this setting. Additionally, VET is readily available in the environment found in our clinic, The aim of this study was to evaluate the diagnostic performance of viscoelastic testing compared with activated partial thromboplastin time for monitoring unfractionated heparin in critically ill patients and to define clinically applicable viscoelastic thresholds using anti-factor Xa activity as the reference standard. METHODS: In this prospective observational study for UFH monitoring Anti-Xa, aPTT, and the VET clotting time IN/HI ratio (ClotPro®; Haemonetics, Boston, MA, USA) were determined in simultaneously taken blood samples. Anti-Xa results were used as the reference standard to categorize ongoing UFH anticoagulation intensity into four categories (prophylactic, sub-therapeutic, therapeutic, and overdosed). Diagnostic performance of aPTT and CT IN/HI ratio were evaluated against Anti-Xa using ROC analysis and Cohen’s Kappa. Test performance was evaluated using ROC analysis, multiclass classification, and weighted Cohen’s kappa. RESULTS: Between September 2020 and July 2022, 466 datasets from 75 ICU patients receiving intravenous UFH were analyzed. Compared to aPTT, AUC values for CT IN/HI were higher for detecting prophylactic (0.817 vs. 0.726), therapeutic (0.690 vs. 0.494) and overdosed (0.845 vs. 0.708) UFH levels. Thresholds of > 1.4, > 1.9, and > 2.3 for the CT IN/HI ratio corresponded to sub-therapeutic, therapeutic, and overdosed ranges, respectively. The CT IN/HI ratio showed moderate agreement with Anti-Xa classification (κ = 0.450), while aPTT agreement was insufficient (κ = 0.220). CONCLUSIONS: The CT IN/HI ratio offers a reliable point-of-care alternative to standard UFH monitoring, in the case it is readily available. It provides better alignment with Anti-Xa activity than aPTT and may improve anticoagulation management in critically ill patients, especially when Anti-Xa testing is delayed or unavailable. TRIAL REGISTRATION: DRKS-ID DRKS00028689 ( https://drks.de/search/de ), PI: Lars Heubner, retrospectively registered 30.03.2022.

<h4>Background</h4>Metabolic dysfunction-associated steatohepatitis (MASH) is a major precursor of hepatocellular carcinoma (HCC), yet the molecular mechanisms linking steatohepatitis to malignancy remain poorly defined. The STE20-type kinase MST3 associates with hepatocellular lipid droplets and regulates metabolic homeostasis and stress responses in the liver. Here, we investigated whether pharmacologic inhibition of MST3 could attenuate the initiation and progression of MASH-associated HCC in vivo.<h4>Methods</h4>The therapeutic potential of MST3 inhibition was evaluated in a mouse model in which MASH-HCC was induced by a single diethylnitrosamine injection followed by 30 weeks of Western-style diet feeding. Liver tumor burden was assessed after 12, 21, or 30 weeks of treatment with Mst3-targeting antisense oligonucleotide (ASO) or a non-targeting control ASO, and histological, biochemical, and mechanistic analyses were performed in the 30-week cohort. In parallel, proteomic profiling of CRISPR/Cas9-generated MST3 knockout and wild-type Huh7 cells was conducted to gain molecular insight into MST3-regulated pathways.<h4>Results</h4>Mst3 ASO therapy had no impact on the onset or aggravation of experimentally induced MASH-associated HCC in mice, despite markedly improving the whole-body metabolic profile and suppressing all key features of MASH. Proteomic profiling of MST3-deficient hepatocytes revealed coordinated activation of mitochondrial and lysosomal pathways, consistent with enhanced fatty acid degradation and catabolic clearance.<h4>Conclusions</h4>Our findings in the selected mouse model of MASH-HCC suggest that MST3 is dispensable for hepatocarcinogenesis, yet its antagonism dampens diet-induced metabolic dysfunction and effectively attenuates MASH severity, challenging the prevailing assumption that targeting MASH driver genes alone is sufficient to prevent HCC development in the context of obesity.

Axons are unique structural and functional features of nerve cells, which play a critical role in regulating neuronal homeostasis. Dysfunction and degeneration of axons (axonopathy) has been established as an early and prominent contributing mechanism to the pathogenesis of neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. In this review, we briefly summarize the structure and function of axons, and highlight recent advances in the understanding of the role of axons in health and disease. We argue that axons are a potential target for developing novel therapies for neurodegenerative diseases.

Liver metastasis remains a major challenge in cancer treatment, yet its cellular and molecular landscape remains poorly defined at the pan-cancer level. Here, we construct a single-cell transcriptomic atlas of liver metastases across multiple cancer types by analyzing 100 single-cell RNA sequencing samples, profiling over 460,000 cells, and identifying 121 distinct cellular subtypes. We define 4 representative cellular programs (CPs) associated with liver metastasis, revealing how cellular composition and intercellular interactions within the tumor microenvironment drive metastatic progression and immune modulation. These CPs recapitulate a dynamic transition from immunoactive states, marked by natural-killer-cell-mediated immune surveillance and macrophage-driven angiogenesis, to immunosuppressive environments dominated by regulatory T cell infiltration and immune exclusion. The shift is marked by progressive alterations in immune infiltration, stromal remodeling, and tumor-intrinsic adaptations, elucidating key mechanisms of immune evasion and metastatic niche formation. Our study provides a high-resolution framework for understanding the heterogeneity and evolution of liver metastasis and highlights the potential of CP-based stratification to inform therapeutic strategies targeting the metastatic tumor microenvironment.

Diffuse mild traumatic brain injury (mTBI) often leads to persistent post-concussive symptoms (PCS) such as fragmented sleep, yet the brain loci and cellular mechanisms that link injury to disability remain obscure. We tested the hypothesis that repeated blast-induced mTBI provokes a chronic myelinopathy with accompanied microglial response in the pontine reticular formation, a brainstem region that modulates arousal and sleep, and that this pathology statistically mediates persistent PCS burden. Using spatially resolved single cell phenotyping in a mouse model of blast-mTBI, we found that only repeated mTBI established persistent activation of disease-associated microglia and phagocytosis of myelin in the pontine reticular formation. Parallel studies in veterans with repeated blast-mTBI revealed identical microglial nodules on neuropathological exam up to two decades after documented blast injury, while diffusion tensor imaging confirmed a dose-dependent pontine myelin disruption that statistically mediated sleep disturbance and broad PCS. Together, these data identify pontine white matter pathology as both a biomarker and mechanistic driver of chronic PCS after repeated diffuse mTBI, highlighting brainstem microglia and oligodendrocytes as rational therapeutic targets.

Heightened and persistent inflammation during the periparturient period impedes health and performance in early lactation. Identifying mitigation strategies capable of attenuating inflammatory cascades without interfering with the inflammation necessary to support normal physiological processes is of significant interest. Dietary supplementation of Scutellaria baicalensis extract (SBE; Dairy Relieve, Elanco Animal Health, Greenfield, IN) has been shown to improve milk performance, an effect hypothesized to be mediated by its anti-inflammatory and antioxidant properties. Thus, study objectives were to evaluate the effect of SBE supplementation on performance and inflammation in primiparous and multiparous cows and to determine the optimal feeding duration. Holstein cows (n = 404; 80 primiparous and 324 multiparous) were used in a randomized complete block design and randomly assigned within block (parity and expected calving date) to 1 of 4 dietary treatments: (1) control diet (CON), (2) 10 g/d of SBE from 255 ± 3 d carried calf (DCC) to 21 DIM (SBE42), (3) 10 g/d SBE from 255 ± 3 DCC to 90 DIM (SBE111), and (4) 10 g/d SBE from 1 to 90 DIM (SBE90). Cows were housed in a freestall barn and received the same base ration. Dietary treatments were administered individually by restraining cows in headlocks for a minimum of 10 min/d during which the top-dress pellet (113.5 g/d) containing 10 g of SBE was administered. Cows were milked 3 times daily for the first 6 wk of lactation and twice daily for the remainder of the data collection period (i.e., 40 wk). Milk yield was recorded from wk 1 to 40, and milk samples for composition analysis were obtained every 2 wk from wk 1 to 15 relative to calving. Blood samples were obtained via coccygeal venipuncture at 7, 21, and 90 DIM from a random subset of cows (24 cows/treatment). Data were analyzed using mixed effects repeated measures ANOVA. A preplanned contrast was used to estimate differences in CON versus cows supplemented with SBE to 90 DIM (SBE111 + SBE90). Over the 40-wk lactation, milk yield increased 1.5 kg/d in SBE111 relative to CON cows. Milk yield was similar in SBE42 and CON cows. Feeding SBE to 90 DIM (SBE111 and SBE90) tended to increase energy-corrected milk yield and milk fat and protein yields, whereas it tended to decrease MUN over the first 15 weeks of lactation relative to CON. Regardless of prepartum feeding, SBE supplementation to 90 DIM tended to increase circulating sirtuin-1 concentrations. Relative to CON, circulating α-1-acid glycoprotein (AGP) increased in SBE-fed cows at 7 DIM, whereas concentrations decreased at 21 DIM. In summary, the optimal feeding window of SBE for improving postpartum performance for cows enrolled in the current study was from 255 DCC to 90 DIM. Changes in circulating sirtuin-1 and AGP concentrations with SBE supplementation support the hypothesis that improved performance with SBE supplementation is partially mediated by the anti-inflammatory properties of SBE.

Despite adjuvant imatinib therapy, a substantial proportion of patients with high-risk gastrointestinal stromal tumor (GIST) experience disease recurrence. This study aimed to identify prognostic factors associated with poor recurrence-free survival (RFS) and develop a predictive nomogram for personalized clinical decision-making. In the present study, a retrospective analysis was conducted of 269 patients with modified Armed Forces Institute of Pathology-classified high-risk GIST who underwent macroscopically complete resection and received adjuvant imatinib therapy at 11 Taiwanese centers between 2013 and 2023. All patients were intended to receive 3 years of adjuvant imatinib and were categorized into disease-free (n=204) and recurrence (n=65) cohorts. Independent predictors of poor RFS were identified through multivariate Cox regression analysis and incorporated into a nomogram. Model performance was assessed using concordance index (C-index) and calibration plots. The cohort comprised 149 males and 120 females with a median age of 60.9 years (range, 29-88 years). At a median follow-up of 62.6 months, the 1-, 3- and 5-year disease-free survival rates were 97.0, 91.7 and 76.4%, respectively. Multivariate analysis identified male sex [hazard ratio (HR): 1.76, P=0.039], non-gastric tumor origin with duodenum showing highest risk (HR: 6.15, P<0.0001) and shorter imatinib duration with <1 year treatment showing HR: 3.91 (P=0.002) as independent predictors of poor RFS. The resulting nomogram demonstrated robust predictive performance with good calibration and C-index of 0.72. Nomogram-based prognostic stratification identified three distinct groups with 5-year survival rates of 99.2% (favorable prognosis), 68.1% (moderate prognosis) and 16.7% (poor prognosis). In the present study, a clinically applicable nomogram was developed that accurately predicts poor RFS in patients with high-risk GIST following surgery and adjuvant imatinib therapy. While external validation is needed before clinical implementation, this prognostic tool may facilitate individualized treatment strategies and patient counseling.

<h4>Background</h4>Poorly differentiated endometrial carcinoma in Black African women is under-characterized at the transcriptomic level, although it is known for aggressive subtypes. We conducted the first RNA-seq analysis of formalin-fixed, paraffin-embedded (FFPE) tumors from Black South African women to explore population-specific gene expression, alternative splicing, and novel isoforms.<h4>Methods</h4>Seventy-six FFPE tumor samples passed RNA quality thresholds (DV200 ≥ 30%) and were sequenced on the PacBio Onso™ short-read platform. StringTie assembled transcripts and estimated transcripts per million (TPMs); differential expression was quantified by mean log₂ fold-change (|log₂FC| ≥ 0.5). SUPPA2 computed percent spliced-in (PSI) values for five splicing event classes. Reactome over-representation analysis identified enriched pathways, and novel isoforms were validated by read coverage and expression support.<h4>Results</h4>Cohort-level analysis (n = 76) revealed the coordinated activation of signaling and broad repression of transcriptional programs. We detected 17,990 dysregulated transcripts (4,483 upregulated and 13,507 downregulated; range -4.81 to +2.99). Prominent upregulated transcripts included GJA1 (+2.99), GRN (+2.60), ZC3H3 (+2.58), PCSK4 (+2.55), HSPB7 (+2.55), BICRA (+2.49), LINC02692 (+2.23), MKNK2 (+2.03), and RARA (+0.76). Strong downregulation was observed for MRPL15 (-4.81), ZSCAN23 (-4.46), NUF2 (-4.37), RTN1 (-4.33), EIF3I (-4.31), CDC73 (-3.36), MYC (-2.96), FRS2 (-2.90), and zinc-finger factors including ZNF569 (-3.36), ZNF573 (-2.92), ZNF793 (-2.33), and ZNF382 (-1.87). Reactome enrichment linked the upregulated set to MAPK family/GPCR signaling, while the downregulated categories were associated with gene expression (transcription), RNA polymerase II transcription, cell cycle, and metabolism. Alternative splicing was pervasive, with recurrent high-magnitude events at cancer-relevant loci including GJA1 (SE), NF2 (A3/A5), LIN37 (A5), ECT2 (A3/A5), BCL2L11 (A3), UQCRH (A5), CSE1L (A3), BROX (A3), and multiple ZNFs. StringTie identified previously unannotated isoforms at LINC02692 (+2.23), LINC01605 (-0.62), ZNF793 (-2.33), ZNF382 (-1.87), and retained-intron transcripts at RAD51 (+1.30), RPS24 (-0.97), and RPS3A (-0.62).<h4>Conclusions</h4>Poorly differentiated endometrial carcinomas in Black South African women show a distinct MAPK-linked activation pattern, along with transcriptional repression and extensive splicing changes. Aligning with findings in African Americans, this cohort highlights the unique aspects of isoform-resolved splicing and zinc-finger repression, suggesting that translational control, retinoid signaling, and RNA processing may be targets for biomarkers and therapy.

Alzheimer's disease (AD) is a prevalent neurodegenerative disorder for which effective treatments remain elusive. This review aims to explore the roles, mechanisms, and therapeutic potential of three principal ginseng components, including ginseng polysaccharides (GPS), ginseng proteins (GP), and ginseng glycoproteins (GGP), in the prevention and management of AD. We systematically reviewed recent literature related to these components in AD research. By analyzing evidence from cellular experiments, animal models, and preliminary clinical studies, we evaluated their effects on core pathological processes. These ginseng-derived compounds exert neuroprotective effects via multiple pathways. Specifically, they inhibit the aggregation of amyloid-β (Aβ) and reduce the hyperphosphorylation of tau protein. Furthermore, they demonstrate significant anti-neuroinflammatory and antioxidant activities, which protect neurons from damage and enhance cognitive functions, including memory and learning. The efficacy of these components has been consistently demonstrated across various AD experimental models. In conclusion, GPS, GP, and GGP exhibit promise as multitarget therapeutic agents against AD, underscoring a potential pathway for developing novel natural product-based treatments. Although current preclinical results are promising, further rigorous clinical trials are necessary to validate their efficacy and safety in humans. Therapeutic strategies targeting these components may therefore offer new hope for AD patients.

<h4>Background</h4>The placebo effect in early-phase metabolic dysfunction-associated steatotic liver disease (MASLD) trials is poorly defined but may substantially influence endpoint interpretation and proof-of-concept decisions. A ≥30% relative reduction in magnetic resonance imaging-proton density fat fraction (MRI-PDFF) is an established noninvasive surrogate of histologic improvement. This study aimed to evaluate short-term placebo responses and 1-year longitudinal changes in hepatic fat content and metabolic parameters in phase Ib/IIa MASLD trials.<h4>Methods</h4>A total of 41 MASLD participants from the placebo arms of five phase Ib/IIa clinical trials (2020-2024) were prospectively enrolled, with 18 completed the 1-year follow-up. Hepatic fat content (MRI-PDFF), body weight, liver enzymes, and lipid parameters were assessed from baseline to week 56.<h4>Results</h4>After 4-5 weeks of placebo treatment, 20% of participants achieved ≥30% hepatic fat reduction, with mean absolute and relative decreases of 2.48% (SD: 2.88) and 16.47% (SD: 23.46), respectively. Body weight decreased by 2.5 kg (SD: 1.8), corresponding to a relative reduction of 2.73% (SD: 1.97). Additionally, liver enzymes and most lipid parameters declined, whereas triglycerides showed a modest increase. During 1-year follow-up, hepatic fat content and body weight rebounded from end-of-treatment levels but remained below baseline. Liver enzymes partially rebounded but stayed below baseline, while lipid parameters exceeded baseline values.<h4>Conclusion</h4>Early-phase MASLD trials show a measurable, heterogeneous placebo effect with partial rebound after treatment cessation. These findings underscore the importance of explicitly accounting for short-term placebo-associated changes when designing early-phase trials, selecting surrogate endpoints and interpreting preliminary efficacy signals.

Vascular surgery is an important procedure that is carried out to treat diseases of the entire vascular system. Intraoperative hemostasis, precise vision of intricate vascular anatomy, restenosis, implant-associated infection, and inadequate endothelium regeneration are among the enduring difficulties that continue to limit vascular surgery. Advanced biomaterial techniques are required since conventional materials and imaging technologies often fail to meet these multifactorial limitations. By enabling synthetic nanomaterials with programmable physicochemical qualities that directly interact with biological systems at the molecular and cellular levels, recent developments in nanotechnology have sparked a paradigm shift in vascular surgery. The limitations of conventional contrast media are addressed by nanostructured contrast agents, such as superparamagnetic iron oxide nanoparticles, which significantly improve vascular imaging resolution and diagnostic sensitivity across magnetic resonance modalities. Biodegradable polymers and surface-engineered coatings are examples of controlled-release nanocarriers that enable targeted administration of antiproliferative and antithrombotic drugs, lowering systemic toxicity and minimizing restenosis. In addition to providing long-lasting antibacterial action to avoid surgical site infections, nano-engineered surface topographies and coatings on grafts and stents accelerate endothelialization and reduce thrombogenicity. Additionally, some nanomaterials' inherent catalytic and redox modulation properties aid in better wound healing and inflammation resolution. With a focus on mechanistic insights into hemostasis, imaging enhancement, drug transport, implant integration, and regenerative processes, this review thoroughly summarizes recent preclinical and translational studies on the use of nanomaterials in vascular surgery. Furthermore, we outline the present obstacles to clinical translation, such as biocompatibility, long-term safety, and manufacturing scalability, and suggest future paths for incorporating nano-enabled techniques into evidence-based surgical practice.

<h4>Background</h4>Folate metabolism plays a pivotal role in tumor proliferation. However, the relationship between folate metabolism-related genes (FMGs) and the tumor immune microenvironment (TIME) in pancreatic cancer (PC) remains unclear. This study aimed to identify key FMGs and investigate whether FMGs are related to TIME in PC.<h4>Methods</h4>Transcriptomic data from 732 PC patients were obtained from public databases, and 78 FMGs were gathered from the Molecular Signature Database (MSigDB). Pan-cancer analysis of genetic alterations in FMGs was performed. Patients with PC were stratified into two subtypes using non-negative matrix factorization (NMF). Clinical data and pathological sections from 150 PC patients were retrospectively collected as a validation cohort. The levels of dihydrofolate reductase (DHFR), FAP⁺ and α-SMA⁺ in cancer-associated fibroblasts (CAFs), CD8⁺ tumor-infiltrating lymphocytes (TILs), Foxp3⁺ TILs, CD206⁺ tumor-associated macrophages (TAMs) were analyzed using immunohistochemistry.<h4>Results</h4>Copy number variation (CNV), single nucleotide variation (SNV), methylation, and risk levels of FMG in pan-cancer were confirmed. Compared to Cluster 1, Cluster 2 demonstrated significantly poorer overall survival (OS) (P<0.05), increased sensitivity to chemotherapy drugs, lower immune cell counts, and more immunosuppressive cells in TIME. DHFR was identified as the folate metabolism-driving gene in PC. DHFR was an independent predictor of poor prognosis (P=0.001). DHFR expression was strongly associated with CD206⁺ TAMs (P<0.001) and Foxp3⁺ T cells (P<0.05).<h4>Conclusions</h4>FMG expression heterogeneity significantly impacts PC prognosis and TIME. DHFR, a central folate metabolism enzyme, demonstrates critical associations with TIME modulation and clinical outcomes in PC.

<h4>Background</h4>Robotic-assisted thoracic surgery (RATS) and video-assisted thoracic surgery (VATS) have been utilized in the context of complex pulmonary segmentectomy. However, the extant literature offers a paucity of information with regard to the early outcomes and inflammatory impacts of these approaches inadequately. We analyzed perioperative outcomes, inflammatory markers, and complications in 31 RATS and 66 VATS cases.<h4>Methods</h4>A retrospective study was conducted on 97 patients undergoing complex pulmonary segmentectomy for pulmonary nodules at The Affiliated Hospital of Guangdong Medical University between July 2022 and October 2024. Short-term surgical outcomes and inflammatory markers were compared.<h4>Results</h4>Among 97 patients with comparable baseline characteristics, no significant differences were observed in intraoperative blood loss, extubation time, or overall complication rates between RATS (n=31) and VATS (n=66). However, RATS demonstrated shorter operative time (174.00±50.67 <i>vs.</i> 224.24±61.65 min, P<0.001), reduced inflammatory responses [postoperative white blood cell counts: (9.90±2.10)×10⁹/L <i>vs.</i> (13.31±4.03)×10⁹/L, P<0.001], and fewer dissected lymph nodes (7.60±4.79 <i>vs.</i> 12.48±7.80, P=0.002). RATS also exhibited superior ergonomic design and three-dimensional (3D) visualization.<h4>Conclusions</h4>For patients diagnosed with early-stage pulmonary nodules, RATS significantly shortens operative time, mitigates inflammatory responses, and enhances ergonomic efficiency compared to VATS.

The link between the intestinal microflora and cancer outcomes has been recognized for over a decade. Several recent studies have demonstrated that the gut microbiome is associated with the efficiency of T-cell checkpoint blockade therapy for cancer, raising interest in strategies to harness this effect via consumption of live microorganisms (probiotics). The probiotic <i>Clostridium butyricum</i> strain MIYAIRI 588 (CBM588) improves response rates and overall survival in patients receiving immune checkpoint inhibitor (ICI) therapy for non-small cell lung cancer and metastatic renal cell carcinoma but the mechanism underlying this benefit remains unclear. Here, we show that CBM588 spores induce a population of Vγ9Vδ2 T-cells from the peripheral blood of healthy donors and lung cancer patients. A subset of these T-cells responded to, and directly lysed, cancer cell lines via a butyrophilin 3A-dependent mechanism. In patients taking CBM588 alongside checkpoint blockade, using samples from a cohort of 38 patients, peripheral blood Vδ2⁺ T-cells expressed the activation marker CD69 more frequently than in those receiving checkpoint blockade alone and the frequency of Vδ2⁺CD69⁺ cells increased following initiation of CBM588 treatment (p = 0.0041). Pleural effusions from patients receiving ICI with CBM588, although available from only three individuals, also showed a notable shift in the local γδ T-cell compartment from the expected Vδ1 dominance towards Vδ2 cells, suggesting altered recruitment or retention of Vδ2 cells at the tumour site. Across the patient cohort, higher post-treatment frequencies of CD69⁺ Vδ2 T-cells were associated with improved survival and more favourable clinical outcomes. These findings provide a potential mechanism by which manipulation of the intestinal microflora might contribute to cancer prognosis through effects on immune effector cells with intrinsic anticancer properties.

<h4>Background and aims</h4>HFE hemochromatosis (HH) may cause arthritis, cirrhosis, and hepatocellular carcinoma (HCC). Cirrhosis is the main risk factor for HCC in HH. Arthritis is common in HH, strongly associated with cirrhosis and readily diagnosed. Thus, we evaluated whether arthritis might be of clinical utility in predicting the future risk of HCC in HH.<h4>Methods</h4>Two hundred four clinically well-characterized patients with HH were recruited between 1974 and 2013 and attended for regular follow-up at a single center over a median duration of 15.2 years. Clinical, biochemical, and cirrhosis status were recorded at diagnosis. The occurrence of HCC was defined as a documented diagnosis in the medical record.<h4>Results</h4>Ten of the 204 patients with HH developed HCC during a follow-up; 9 of these had recorded arthritis status. Seven of 88 patients with arthritis at diagnosis of HH subsequently developed HCC compared with 2 of 115 patients without arthritis (relative risk = 4.57, 95% confidence interval (CI) 1.11-19.07, <i>P</i> = .042). The sensitivity, specificity, positive predictive value, and negative predictive values were 78% (95% CI 45%-96%), 58% (95% CI 51%-64%), 8% (95% CI 4%-15%), and 98% (95% CI 93%-100%). Patients with arthritis and cirrhosis at diagnosis who went on to develop HCC had significantly higher alcohol consumption at baseline compared with those individuals who had arthritis and cirrhosis but did not progress to HCC (median 100 g/d vs 45 g/d, <i>P</i> = .0312).<h4>Conclusion</h4>Arthritis combined with higher alcohol consumption at diagnosis of HH is associated with an increased risk of future development of HCC.

<h4>Background</h4>Non-alcoholic fatty liver disease (NAFLD), recently termed metabolic dysfunction-associated steatotic liver disease (MASLD), is increasingly recognized as a multisystem metabolic disorder associated with significantly elevated risk of cardiovascular disease (CVD). Atherogenic Index of Plasma (AIP) and carotid intima-media thickness (CIMT) are considered as surrogate markers of atherosclerotic cardiovascular disease (ASCVD). As CVD is the common link between NAFLD and parameters, such as AIP and CIMT, the present study was conducted to determine any association and discriminatory ability of AIP and CIMT in relation to NAFLD severity.<h4>Methods</h4>This was a hospital-based cross-sectional study, conducted in the Department of General Medicine in collaboration with the Department of Radiodiagnosis at Kalinga Institute of Medical Sciences (KIMS), KIIT, Deemed to be University, Bhubaneswar, Odisha, India, from March 2023 to February 2025. A total of 151 participants (75 NAFLD cases and 76 controls) were enrolled during the study. NAFLD was graded by ultrasonography. AIP was calculated as log₁₀(triglyceride (TG)/high-density lipoprotein cholesterol (HDL)-C), Fatty Liver Index (FLI) was derived using the Bedogni formula, and CIMT was measured bilaterally using B-mode ultrasonography. In addition, correlation, multivariate regression, and receiver operating characteristic (ROC) curve analyses were performed.<h4>Results</h4>Among 151 participants (75 NAFLD cases and 76 controls), patients with NAFLD demonstrated significantly higher TG and lower HDL-C levels (p<0.001). The AIP was markedly elevated in patients with NAFLD compared to controls (0.82±0.24 vs. 0.42±0.17, p<0.001). Bilateral CIMT values were significantly greater in patients with NAFLD than in controls (right: 1.01±0.36 mm vs. 0.63±0.07 mm; left: 1.02±0.40 mm vs. 0.62±0.10 mm; p<0.001). Across ultrasound-defined NAFLD grades, TGs, AIP, and FLI increased progressively, whereas HDL-C declined significantly (p<0.001 for trend). FLI showed strong positive correlations with CIMT (right: ρ=0.607; left: ρ=0.663; p<0.001). Multivariate regression identified body mass index (BMI), TGs, HDL-C, AIP, and CIMT as independent determinants of NAFLD severity. The ROC analysis demonstrated excellent diagnostic performance for AIP (area under the curve (AUC) = 0.917) and combined AIP+CIMT (AUC = 0.951), indicating high discriminatory accuracy for identifying NAFLD.<h4>Conclusions</h4>NAFLD is strongly associated with increased atherogenic burden and subclinical carotid atherosclerosis. AIP and CIMT correlate with disease severity and demonstrate good diagnostic utility in identifying advanced NAFLD. These findings reinforce the systemic cardiometabolic nature of NAFLD and support the potential role of simple lipid indices and vascular imaging in early CVD risk stratification.

<b>Background/Objectives</b>: In the area of pharmacology and clinical research, it is necessary to use versatile technologies able to quantify last-line antibiotic molecules with high specificity and sensitivity. This article describes the development of two types of immunosensors based on amperometric and surface plasmon resonance (SPR) measurements and their applicability in the measurement/assessment of therapeutic drug monitoring (TDM) of four last-line antibiotics such as vancomycin, colistin, daptomycin and meropenem in human plasma. In this study, ligand immobilization by preconcentration assays, sensor surface regeneration, determination of sensitivity and correlation of plasma sample quantification results by HPLC were considered. <b>Results</b>: In the case of the electrochemical biosensor the IC50 values obtained were 3.49 μg/L for vancomycin (VAN), 5.44 μg/L for colistin (COL), 0.82 μg/L for meropenem (MER) and 5.10 μg/L for daptomycin (DAP). For the SPRi biosensor the LODs achieved were 19 ng/mL for VAN, 9 μg/L for COL, 12 μg/L for MER and 12.3 μg/L for DAP. Finally, both electrochemical biosensor and the SPRi optical biosensor showed that for the four antibiotics the standard deviations were less than 10% with respect to the HPLC results, with ranges for VAN between ~5-6 µg/mL, for COL ~0.2-0.7 µg/mL, for MER ~4.5-5.5 µg/mL and for DAP ~0.09-0.65 µg/mL. <b>Conclusions</b>: These kinds of biosensors provide a precise and sensitive strategy, together with real-time determination, to quantify last-line antibiotics, with working ranges like those shown by robust techniques such as HPLC and great potential for the clinic.

<title>Abstract</title> <p>Background Inter-patient expression heterogeneity is not merely statistical dispersion but a structured biological signal encoding distinct therapeutic antigen states. Current antigen discovery frameworks over-prioritize mean expression and systematically underinterpret this variance structure, potentially obscuring the most clinically actionable targets. We asked whether inter-patient transcriptomic heterogeneity could be operationalized as a decomposition framework to reveal recurrent, biologically interpretable modes of therapeutic antigen biology. Methods We developed TANK (Tumor Antigen prioritization by variance-based raNKing) as a heterogeneity decomposition framework - not primarily a ranking method, but an approach to resolving recurrent antigen modes from patient-level transcriptomic distributions. TANK was applied across 33 TCGA cancer types (n > 11,000 patients, 60,656 genes). A 10-gene reference panel was predefined based on independent clinical development status. Non-randomness was confirmed against 1,000 random gene set controls (empirical p < 0.0001). External validation was performed in two independent gastric cancer cohorts (GEO GSE26942, n = 217; ACRG GSE66229, n = 400). Single-cell validation was conducted across three cancer types totaling 28,617 annotated tumor epithelial cells. Mode 3 candidates were characterized by survival analysis, immune correlation, and DepMap CRISPR dependency. Beyond the reference panel, CLDN6 was examined as a showcase candidate across the same dimensions. Results TANK resolved four recurrent modes of therapeutic antigen heterogeneity - tumor-restricted rare activation (Mode 1: PRAME), lineage-dependent expression (Mode 2: CLDN18), tumor-enriched heterogeneous expression with functional stratification (Mode 3: MSLN/OLFM4/VSIG1/MUC16), and mean-dominant baseline (Mode 4: ERBB2/EGFR) - each with distinct implications for patient stratification, immune context, and translational modality. Concordance between TANK and MAD confirms the signal reflects a robust variance-associated structure rather than any single metric artifact. Beyond the reference panel, CLDN6 ranked top 0.22% (comparable to FDA-approved CLDN18 at top 0.16%), with significant adverse survival association (p = 0.0049), immune-cold correlates (CD274 r=-0.204, p < 0.0001), and directional CRISPR dependency across gastric (mean=-0.227), colorectal (mean=-0.242), and lung cancer cell lines (mean=-0.288). Conclusions Inter-patient transcriptomic variance defines a previously underutilized axis of antigen biology encoding recurrent therapeutic modes that are systematically inaccessible to mean-based approaches — CV-based ranking recovers 0/9 reference targets and DESeq2 fails to prioritize CLDN18 and CLDN6 within the top 20%, while variance decomposition places both in the top 0.22%. The four-mode framework enables prospective mapping of novel candidates to distinct therapeutic strategies from heterogeneity structure alone, without prior biological knowledge of the candidate. CLDN6 exemplifies this capacity: nominated solely from variance structure, its convergent multi-dimensional evidence positions it as a high-priority oncofetal antigen candidate for patients with limited therapeutic options.</p>

Copy number variants (CNVs) can alter disease susceptibility by gene deletion, duplication, and other mechanisms, and have been implicated in neuropsychiatric diseases. However, their rarity or de novo nature impedes linkage analysis. Therefore, we identified recurrent CNVs (rCNVs) in Native Americans with low genetic admixture and high prevalence of alcohol use disorder (AUD) and other psychiatric disorders. Large (> 200 kb) rCNVs were abundant in Plains Indians (PI) and Southwest American Indians (SWI), almost all carrying at least one rCNV, with some CNVs found in both geographically and linguistically distinct tribes. In patients carrying rCNVs, gene deletions led to haploinsufficiency, and duplications led to increased gene dosage. Haplotype analysis revealed a common chromosome 6p21.33 recurrent CNV (rCNV) that persisted in Native Americans for at least 750 generations, leading to haploinsufficiency of at least two genes. Gene-based CNV burden and CNV count did not predict AUD or other psychiatric disorders. However, an rCNV, found in PI and duplicating three genes within the 22q11.2 velocardiofacial syndrome region, showed nominally elevated odds ratios in generalized linear mixed models accounting for kinship as a random effect and age and sex as fixed covariates. For AUD, the odds ratio was 3.19 (95% CI 1.12-9.07, p = 0.03), and for psychiatric diagnosis, the odds ratio was 4.80 (95% CI 1.37-16.82, p = 0.014). These associations, calculated with adjustment for relatedness, did not remain statistically significant after correction for multiple testing, illustrating the challenge of linking CNVs to behavior even if they are recurrent and potentially of large effect. The effects of most CNVs are undetectable via genome-wide association studies with single SNPs, and in cosmopolitan populations, most CNVs are nonrecurrent, sometimes affecting similar genomic regions but differing in their properties, and origins. Recurrent CNVs having ancient origins are prevalent in Native American populations providing an opportunity to examine their relationships to disease risk. However, we observed that neither gene-based CNV burden nor individual rCNVs did not predict AUD or other psychiatric outcomes, after adjustment for the numbers of rCNVs tested.

Heat stroke (HS) is the most severe heat-related emergency, and its pathophysiology remains largely unknown, especially for exertional HS (EHS), which affects younger populations, athletes, and manual workers. Herein, we performed single-cell-transcriptomics, T cell receptor sequencing, and flow cytometry of PBMCs from 9 healthy control participants, 9 patients with heat exhaustion, and 9 patients with EHS to explore complex immunological responses associated with HS pathobiology. We showcased that granzyme-positive T cells and CD56dim NK cells with high cytotoxicity features and IL-1B+NLRP3+ monocytes with high inflammation and pyroptosis scores were enriched in HS, while the CD161+ T cells with innate immune-like, low cytotoxicity, and clonal expansion features were reduced in HS. Importantly, elevated granzyme-positive T and NK cells might interact with monocytes to induce pyroptosis of hepatic and renal cells and target organ injuries, and blocking the NLRP3 inflammasome pathway prior to the induction could alleviate organ injury in HS. This study offers deeper insights into the pathogenesis of HS, supporting the development of optimal treatment strategies.

Maternal obesity alters breast milk composition in ways that may predispose infants to excess adiposity. Although maternal exercise during lactation has been associated with favorable shifts in milk metabolites in humans, the mechanisms by which exercise remodels the mammary gland and milk lipid profile to influence offspring metabolism remain unclear. We developed a mouse model incorporating daily moderate treadmill exercise only during lactation, using lean (LN) and diet-induced obese (OB) dams, and leveraged indirect calorimetry, stable isotope tracer respirometry, and mammary epithelial cell (MEC) proteomics assays. Maternal obesity broadly remodeled the MEC proteome, decreasing enzymes of de novo fatty acid synthesis and altering lipid transport and oxidative pathways. These molecular adaptations in OB dams corresponded to higher milk triglyceride content and shifts in fatty acid composition, including suppressed medium-chain fatty acids (MCFAs). The exercise (EX) intervention during lactation reset MEC protein networks, enhancing protein translation and vesicle transport pathways, whereas decreasing fatty acid desaturation, relative to the sedentary (SED) group. In OB dams, the exercise intervention increased milk MCFA levels and partially corrected the proinflammatory omega-6 fatty acid bias. Offspring suckling OB-EX dams exhibited enhanced in vivo fatty acid oxidation, partially rescuing obesity-associated impairments in metabolic fuel preference. Together, maternal exercise during lactation remodels mammary metabolism and milk fatty acid composition in obese dams, which in turn, enhances postnatal lipid oxidation. These findings highlight lactation as a modifiable window, wherein maternal activity influences milk composition and early life metabolism.<b>NEW & NOTEWORTHY</b> Maternal obesity alters milk fatty acid composition, with consequences for postnatal metabolism. Maternal exercise during lactation in obese dams remodeled the mammary epithelial cell proteome, increasing medium-chain fatty acids in milk and enhancing offspring lipid oxidation.

<h4>Background</h4>Fat Mass and Obesity-Associated (FTO) is linked to multiple myeloma (MM) progression, but its action mechanisms are poorly understood.<h4>Methods</h4>Machine learning and correlation analysis identified genes with consistent expression patterns as FTO-associated genes. Additionally, the expression levels of FTO and its associated genes in MM tissues were validated through immunohistochemistry. The impact of FTO-associated genes expression on the immune microenvironment of MM was assessed through immune infiltration analysis. Single-cell RNA sequencing (scRNA-seq) elucidated cellular expression patterns.<h4>Results</h4>FTO was found to be significantly upregulated in MM and associated with poor prognosis. Four potential FTO-associated genes (CHRM3, GINS3, RRM2, and SHCBP1) were identified. Correlation and immunohistochemical analyses further focused the FTO-CHRM3 axis. Functional enrichment analysis unveiled that FTO and CHRM3 were involved in "DNA replication" and "cell cycle". The infiltration of M2 macrophages and eosinophils were altered in high FTO/CHRM3 expression groups. ScRNA-seq highlighted the monocyte-specific expression of SLC8A1 in the calcium signaling pathway in MM.<h4>Conclusion</h4>The FTO-CHRM3 axis contributes to MM progression via calcium signaling pathway dysregulation, and the exploration of its underlying mechanisms provides insights for targeted MM interventions.

<h4>Introduction</h4>Bladder cancer (BLCA) is a malignant tumour that occurs on the mucosa of the bladder. It accounts for the first place in the incidence of genitourinary tumours in China. BLCA is characterized by high recurrence rate and poor survival rate. There is still a research gap regarding super-enhancer-related genes (SERGs) in BLCA.<h4>Methods</h4>The The Cancer Genome Atlas Bladder Urothelial Carcinoma (TCGA-BLCA) and GSE31684 were subjected into this study. In addition, the Super-Enhancer Archive database was used to identify SERGs. Differential expression analysis was used to analyse the differentially expressed genes (DEGs) between the BLCA and control groups. The DEGs were overlapped with SERGs to get candidate genes in TCGA-BLCA, which were analyzed for Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Univariate Cox, Least Absolute Shrinkage and Selection Operator (Lasso) regression and multivariate Cox regression analyses were used to build the risk model for BLCA. Survival analyses and validation of the model were performed by Kaplan-Meier (K-M) curves and Receiver Operating Characteristic (ROC) curve, respectively. In addition, using the estimating relative subsets of RNA transcripts (CIBERSORT) algorithm, 22 immune cell proportions were calculated. The drug sensitivity was also analyzed in this study.<h4>Results</h4>First of all, based on the TCGA-BLCA, 70 DE-SERGs were yielded. A prognosis model based on MXRA7, PLEKHG4B and ATP2B4 was finally constructed. ROC curves revealed that the prognosis model was a good predictor of BLCA outcomes. Immune infiltration analysis revealed that risk score was positively associated with T cells CD4 memory resting, Mast cells resting and Macrophages M2 and negatively associated with Dendritic cells activated and T cells CD8. Besides, AZD8186, BMS-754,807, JQ1, KRAS (G12C) Inhibitor and NU7441 were the top five sensitivity drugs for BLCA.<h4>Conclusion</h4>Three genes (MXRA7, PLEKHG4B and ATP2B4) were identified to construct a SERG-related model in BLCA, which provides a basis for understanding BLCA pathogenesis and new insights into BLCA treatment.

γδ T cells are unconventional lymphocytes that bridge innate and adaptive immunity by combining recognition of stress-induced ligands independently of classical major histocompatibility complex molecules with the capacity to undergo clonal expansion and long-term adaptation. Their unusual ability to detect malignant transformation using semi-invariant T-cell receptors, butyrophilin recognition and natural killer-like receptors positions them as powerful effector cells in tumors that evade classical immune escape mechanisms. Furthermore, distinct γδ subsets have distinct phenotyping and specific tissue-residencies, which could be leveraged to modulate immunological responses. We evaluate engineered therapies and different experimental platforms for studying γδ T cell biology. We conclude that next-generation cancer treatments should strategically integrate γδ T cells into synthetic immunology, individualized modeling, and combinatorial regimes.

Epigenetic dysregulation is a fundamental cancer hallmark, and lysine demethylase 1 (LSD1) is a central target for cancer intervention. Developing novel LSD1 inhibitors with high selectivity, favorable bioavailability, and safety for acute myeloid leukemia (AML) remains challenging. We developed DC551040, a highly potent, selective irreversible LSD1 inhibitor with good tolerability in Phase I AML clinical trial (CTR20222026). DC551040-LSD1 complex crystal structure uncovered a new binding pocket, providing molecular insights for subsequent LSD1 inhibitor design. Given the significant role of LSD1 in epigenetic regulation, we performed comprehensive transcriptomic and proteomic analyses to investigate gene and protein expression dynamics following DC551040 treatment in an MV-4-11 xenograft model. These analyses revealed that multiple immune and inflammation related pathways are activated upon DC551040 treatment, including the key members STAT5, NF-κB, and AKT, suggesting the potential for adaptive resistance. Through a search of the Connectivity Map (CMAP) database, we identify homoharringtonine (HHT), an approved anti-leukemia drug, which mimics the anti-transcriptional activation of inflammatory pathways. Subsequent in vitro and in vivo experiments validated the efficacy of combining HHT with DC551040, demonstrating a synergistic antitumor effect and extended survival in MV-4-11 disseminated xenograft model mice. Together, this study not only introduces a novel LSD1 inhibitor but also delves into the molecular mechanisms underlying LSD1 inhibitors, while proposing a promising combination therapy for AML individuals in clinical trials.

Calcium signaling is essential for neuronal development and function, yet its role in the differentiation of immortalized neuronal cell lines remains poorly understood. In this study, we investigated the involvement of voltage-gated calcium channels (VGCCs) in the differentiation of the F11 cell line, a widely used translational model derived from the fusion of mouse neuroblastoma cells with rat dorsal root ganglion neurons. Differentiation was associated with greater KCl-evoked increases in intracellular Ca2+ and with upregulated expression of VGCC-encoding genes, particularly those for L-type channels. Pharmacological inhibition of L-type VGCCs reduced the KCl-evoked intracellular Ca2+ increase and impaired neurite outgrowth, underscoring their role in differentiation. Additionally, overexpression of CaV1.3 in non-differentiated F11 cells enhanced neuronal features, including greater KCl-evoked increases in intracellular Ca2+ and neurite outgrowth. However, under differentiation conditions, CaV1.3 overexpression disrupted the acquisition of neuronal features and coincided with increased intracellular oxidative stress. Notably, this effect was more pronounced for CaV1.3 than for the higher activation-threshold CaV1.2 channels. These findings highlight the dual role of L-type VGCCs in neuronal differentiation and oxidative stress regulation, demonstrating that their precise modulation is critical for proper neuronal development. Moreover, the shared molecular pathways between F11 cell differentiation and neurogenesis reinforce the translational value of this model for drug discovery efforts targeting oxidative stress-mediated neuronal dysfunction, such as in neurodegenerative diseases and neuropathic pain.

Myostatin (MSTN) is a well-established negative regulator of muscle growth and development in mammals. Genetic variations within MSTN are linked to differences in sheep musculature, particularly the double-muscle phenotype. This study represents the first comprehensive polymorphism analysis within intron 1 of the MSTN in the Ukrainian Carpathian Mountain (UCM) sheep breed combining molecular and bioinformatic approaches. Sequencing data of samples from UCM sheep revealed eight previously reported single-nucleotide polymorphisms (c.373+241T>C, c.373+243G>A, c.373+246T>C, c.373+249T>C, c.373+259G>T, c.373+323C>T, c.373+563G>A, c.373+607G>A) and one novel polymorphism (c.373+283T>C). Bioinformatic analysis evaluated potential functional effects of these intronic polymorphisms, including changes in pre-mRNA stability, proximity to transcription factor binding sites, and possible pre-miRNA formation, using in silico approaches, including molecular dynamics simulations of predicted pre-miRNA structures. Predictions identified c.373+607G>A and the novel c.373+283T>C polymorphisms as potential candidates for further functional investigation and association studies. This study demonstrates the value of combining molecular genetic and bioinformatic approaches for characterizing intronic polymorphisms and supporting a deeper understanding of functional genetic variation in livestock.

Traceability is a key requirement for compliance in food supply chains, ensuring transparency, safety, and product authenticity across all stages of production and distribution. Although advanced technologies have been introduced to support compliance efforts, blockchain stands out for its potential to provide secure, tamper-proof, and verifiable traceability. However, adoption of blockchain remains limited due to technical complexity, high implementation costs, and fragmented integration between supply chains. To address these challenges, this study proposes a unified blockchain-enabled services framework for the implementation of streamlined, standardized, and accessible traceability. The framework includes a network architecture, traceability-driven operational rules, a UML sequence diagram defining system interactions, and algorithms to record and verify data. Demonstrated through an Australian seafood traceability scenario, the framework was validated by deploying a prototype digital platform and smart contracts on the blockchain. The results show that the framework enables reliable traceability with minimal integration effort while supporting interoperability and data integrity. Although not required, the study highlights the potential role of regulatory bodies as service providers to strengthen compliance alignment. In general, this approach presents a scalable, cost-effective, and practical solution to improve traceability in supply chains through blockchain technology.

<h4>Background</h4>Obesity is a recognized risk factor for numerous cancers. Although several biological mechanisms have been proposed to explain obesity-associated carcinogenesis, the extent to which excess adiposity influences tumor genomic profiles remains incompletely understood. In particular, whether obesity-related selective pressures shape cancer-specific mutational landscapes is still underexplored.<h4>Methods</h4>A pan-cancer analysis of non-synonymous somatic mutations across 14 tumor types using data from The Cancer Genome Atlas (TCGA) has been conducted. Body mass index (BMI) at diagnosis was analyzed as a continuous variable. Associations between gene mutations and BMI were assessed using logistic regression models adjusted for age, sex, and tumor mutational burden, with false discovery rate correction. Genes were prioritized using a two-step ranking strategy based on mutation frequency and regression strength. Functional inactivation, exon-level mutation distribution, and Gene Ontology enrichment analyses were performed for significantly BMI-associated genes.<h4>Results</h4>In particular, bladder urothelial cancer (BLCA) resulted as the most frequently mutated neoplasia in association with higher body mass index. Among Eighty-six genes significantly associated with BMI in BLCA, a prioritized set of ten genes (BRCA2, DNAH9, GRIA4, PLXNA4, UNC13C, FCGBP, SF3B1, ELP1, NES, TRERF1) has been selected for further analyses. Overweight and obese patients exhibited distinct BMI-specific exon-level mutational patterns and concurrent deleterious mutations across multiple candidate genes. Functional inactivation analysis suggested loss-of-function mechanisms in most top-ranked genes, while Gene Ontology (GO) analysis highlighted deregulation of extracellular matrix-related pathways.<h4>Discussion</h4>These findings support a role for obesity in shaping the genomic landscape of tumors, highlighting the importance of integrating clinical parameters such as BMI into genomic studies to determine the potential impact of obesity on tumor evolution, heterogeneity, and treatment response.

Endometriosis is a common gynecological disorder in which inflammatory and immune responses play a crucial role in its development and progression. This study aimed to identify potential inflammation-related biomarkers for the diagnosis and therapeutic monitoring of endometriosis. Differentially expressed genes (DEGs) between endometriosis and control groups were identified using the limma R package. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed with the clusterProfiler R package to explore functional categories and biological processes associated with the DEGs. Immune cell proportions were estimated using CIBERSORT and xCell, followed by correlation analysis between gene expression and immune cell ratios. Data were obtained from the GEO datasets GSE104948 and GSE116626. Under the criteria |fold-change (FC)| > 1 and p-value < 0.05, a total of 357 DEGs were identified, including 136 down-regulated and 221 up-regulated genes. GO analysis revealed enriched biological processes, such as regulation of cell-cell adhesion mediated by cadherin, cell-cell adhesion mediated by cadherin, and response to interleukin-6. Functional enrichment included extracellular matrix structural constituents and protein-binding activities. KEGG analysis highlighted pathways related to protein digestion and absorption. Three inflammation-related genes, PGI2 synthase (PTGIS), E26 transformation-specific homologous factor (EHF), and collagen type X alpha 1 (COL10A1), were identified as potential biomarkers for endometriosis. In 12Z endometriotic epithelial cells, PTGIS knockdown reduced viability, enhanced apoptosis, and impaired migration and invasion, whereas PTGIS overexpression had the opposite effects. Collectively, this study suggests that PTGIS, EHF, and COL10A1 may serve as valuable predictors for the progression of endometriosis.

The skin is the largest human organ and a site of substantial disease burden, yet its cellular and molecular organization across the body is largely undefined. Here we construct an organ-wide single-cell spatial atlas of ~1.2 million cells from normal adult human skin, resolving the location of 45 cell types across 114 samples encompassing 15 anatomic sites. We uncover site-specific stereotypic cell-type composition and their organization into ten multicellular neighborhoods, most notably a perivascular neighborhood reminiscent of skin-associated lymphoid tissue. Within this neighborhood, ligand-receptor (L-R) analyses identify a central role for tumor necrosis factor in maintaining CCL19⁺ perivascular fibroblasts, highlighting homeostatic immune-stromal crosstalk. Finally, comparing neighborhood dynamics in spatial transcriptomics of skin disease, we find pan-disease immune alterations in this perivascular neighborhood, suggesting spatial compartmentalization of pathogenic activity. Thus, multicellular neighborhoods underlie the skin's multiscale molecular to macroanatomic organization, orchestrate cell-cell interactions and anatomic site specialization and exhibit architectural disruption in disease.

No abstract available.

Social competition exerts a diverse set of influences on neural development and behavior during adolescence and yet the precise underlying molecular mechanisms remain poorly understood. Here, we demonstrate that individual rodents that rank lower in social hierarchies are more vulnerable to drug abuse. Proteomic analysis revealed a crucial role of the mesocorticolimbic netrin-1/DCC/UNC5 pathway within the nucleus accumbens (NAc) in mediating the impact of social competition. We found that mice with a conditional knockout of the deleted in colorectal cancer (DCC) gene in dopamine neurons were more likely to achieve higher social rank but exhibited increased drug-seeking behaviors. Following dopamine fiber immunostaining, these outcomes were attributed to ectopic mesolimbic dopamine fibers, which enhanced risk-taking behavior in winners that had DCC knockout. Collectively, our work elucidates a molecular mechanism through which social competition influences adolescent brain development and behavior, particularly in relation to drug susceptibility.

OBJECTIVE: This study aimed to investigate the effect of miRNAs differentially expressed in serum exosomes on endothelial cells after myocardial infarction. DESIGN & METHODS: Specific pathogen-free male Sprague–Dawley rats with acute myocardial infarction (AMI) were established by ligating the anterior descending branch of the left coronary artery. Hematoxylin–eosin staining was used to detect myocardial histopathological changes during model evaluation. The differentially expressed miRNAs carried by the serum exosomes were detected using the Agilent Rat miRNA Gene Chip on the Illumina platform. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed using the Database for Annotation, Visualization, and Integrated Discovery. The culture supernatant of hypoxic H9c2 cells was collected and centrifuged to obtain exosomes. The expression of miR-484 was verified using real-time fluorescence quantitative PCR (qRT‒PCR). miR-484 mimics were transfected into human umbilical vein endothelial cells (HUVECs) to construct cell models of miR-484 overexpression. Cellular proliferation, scratch wound, and tube formation assays were conducted with HUVECs. A dual-luciferase reporter assay was used to validate the target gene of miR-484. RESULTS: Serum exosomes were sequenced on an Illumina sequencing platform, and 22 differentially expressed miRNAs between the model and sham groups were detected (P < 0.05). Among them, miR-484 was prioritized for further evaluation, and its downregulation in MI serum exosomes was confirmed through both gene chip analysis and animal experiments. miR-484 expression increased in HUVECs after culture with exosomes from hypoxia-exposed H9c2 cells. miR-484 expression upregulation suppressed proliferation, migration, and vascular formation in HUVECs, indicating that miR-484 plays an antiangiogenic role. Bioinformatics analyses showed that rno-miR-484 and the VEGFA 3’UTR possess base binding sites. The dual-luciferase reporter assay showed that rno-miR-484 could significantly downregulate the expression of luciferase in r-Vegfa-3UTR-WT (P < 0.01). VEGFA expression was downregulated after miR-484 mimic transfection in HUVECs. CONCLUSIONS: Our study revealed that miR-484 may inhibit angiogenesis after myocardial infarction by targeting VEGFA, providing a novel focal point for the diagnosis and treatment of this disease.

Evidence for an association between insulin-like growth factors (IGF) and multiple myeloma (MM) is inconsistent. We examined total IGF-I concentrations and risk of MM by combining baseline serological data among UK Biobank participants (n = 444 187; 732 incident MM) with a two-sample Mendelian randomisation (MR) analysis using identified genetic variants associated with circulating total IGF-I and IGF-binding protein 3 (IGFBP-3) in the InterLymph consortium (2434 MM and their 2567 controls). Finally, additional lymphoid neoplasm (LN) subtypes were included for comparison with the main hypothesis. Circulating IGF-I level was positively associated with MM risk Hazard ratio-HR-per one standard deviation-SD-increase (HR_1-SD = 1.11, 95% confidence interval [CI]: 1.01-1.22; p-value = 0.03), especially closer to diagnosis. Genetically inferred IGF-I levels were associated with increased MM risk (odds ratio [OR] = 1.27, 95% CI: 1.05-1.54) but not with any other LNs. Genetically inferred IGFBP-3 levels showed no associations with any LN evaluated. Corroborating previous findings, in a secondary analysis, IGF-I levels were associated with the risk of chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL) in males with higher body mass index (HR_{1-SD in obese male} = 1.36, 95% CI: 1.14-1.61). Our serological and MR analyses suggest a contributing role of IGF-I in the susceptibility of MM; the CLL/SLL findings warrant further investigation considering sex-specific adiposity.

This study explores the inflammatory response observed in the pancreas and pancreatic lymph nodes (pLNs) during the natural history of type 1 diabetes (T1D). Using multicell-resolution spatial transcriptomics (ST), we profile individuals without diabetes (ND), at-risk autoantibody-positive (AAb+) individuals, and T1D donors. In the T1D pancreas, we observed global upregulation of inflammation-associated transcripts, including REG family genes, C3, SOD2, and OLFM4. In the T1D pLN, LTB was significantly upregulated within the lymphoid follicles. Using an orthogonal subcellular-resolution ST platform on an independent donor set, we identified follicular B cells as the primary source of LTB in the pLN and observed increased LTB expression in lymphocytes in insulitic lesions proximal to CCL19/CCL21-expressing endothelium. Collectively, these findings highlight lymphotoxin-β and downstream chemokine signatures in the pancreatic lymphatics as well as within the insulitic lesion, which can inform future therapeutic interventions.

Osteosarcoma (OS), the most common primary malignant bone tumor in children and young adults, is characterized by aggressive behavior, frequent metastasis, and resistance to chemotherapy, resulting in poor clinical outcomes. Increasing evidence indicates that OS progression is not solely driven by tumor-intrinsic factors but is strongly influenced by dynamic interactions within the tumor microenvironment (TME). This literature review synthesizes current research on the roles of endothelial cells, fibroblasts, mesenchymal stromal cells, immune populations, and osteoclasts in OS pathogenesis, with emphasis on cell-cell interactions mediated by direct contact, soluble factors, and extracellular vesicles. The studies demonstrate that these interactions promote tumor proliferation, immune evasion, extracellular matrix remodeling, metastatic dissemination, and therapeutic resistance. Adaptive responses of both tumor and stromal cells to environmental stressors contribute to chemoresistance and disease progression. Collectively, our findings highlight the multifactorial nature of OS driven by complex cellular crosstalk within the TME. Understanding these mechanisms highlights the limitations of conventional chemotherapy and encourages the development of combined therapeutic approaches, including targeted therapies, immunomodulation, and microenvironmental interventions. Continued investigation into tumor-microenvironment interactions may facilitate the identification of actionable targets and improve personalized treatment approaches for OS.

Yamakagashi (<i>Rhabdophis tigrinus</i>) is a widely distributed snake species in Japan. Yamakagashi causes venom-induced consumption coagulopathy (VICC) when the amount of infused venom is high, and bites can be fatal if antivenom treatment is delayed. However, yamakagashi antivenom is an unapproved treatment, and its storage capacity is limited, preventing its prompt administration. Therefore, we investigated the application of commercially available drugs, namely tranexamic acid and antithrombin III, in the treatment of VICC caused by yamakagashi venom in a rat model. Furthermore, we investigated the combination of each drug with recombinant thrombomodulin α. Administration of tranexamic acid or antithrombin III alone failed to extend rat survival or correct changes in blood coagulation markers, such as prothrombin time, fibrinogen concentrations, and D-dimer levels, in yamakagashi venom-treated rats. However, combined administration of recombinant thrombomodulin α and tranexamic acid extended rat survival and partially restored blood coagulation markers. Therefore, the combination of recombinant thrombomodulin α and tranexamic acid might represent a useful therapeutic regimen for yamakagashi venom exposure.

<b>Background and Clinical Significance</b>: Mixed-phenotype acute leukemia (MPAL) is a rare hematologic malignancy characterized by the co-expression of myeloid and lymphoid markers and is associated with poor prognosis. Myeloid sarcoma (MS), particularly in the mediastinum, is an uncommon extramedullary manifestation and is rarely reported in association with MPAL. <b>Case Presentation</b>: We report a rare case of mediastinal MS with biphenotypic features and pericardial extension occurring concurrently with MPAL, highlighting diagnostic challenges, therapeutic strategies, and long-term outcomes. We describe the clinical course, diagnostic workup, treatment, and follow-up of a 21-year-old woman who presented with cardiac tamponade secondary to a mediastinal mass. Histopathology and immunophenotyping established the diagnosis of mediastinal MS associated with MPAL (B/myeloid, NOS). Management included surgical cytoreduction, intensive induction chemotherapy, and consolidation with allogeneic hematopoietic stem cell transplantation (allo-HSCT) from an unrelated donor. Fertility preservation with oocyte retrieval, in vitro fertilization (IVF), and embryo cryopreservation was performed prior to conditioning. A focused literature review of MPAL cases with extramedullary involvement was conducted. The patient achieved complete remission following induction therapy and underwent allo-HSCT. Despite the historically poor prognosis of mediastinal MS and MPAL, she remains in sustained complete remission 13 years after diagnosis. A literature review identified only eight reported cases of MPAL with extramedullary disease, with mediastinal involvement described in a single case and allo-HSCT performed in only two patients. <b>Conclusions</b>: This case illustrates a rare presentation of MPAL with mediastinal myeloid sarcoma and cardiac tamponade, demonstrating that aggressive multimodal therapy including allo-HSCT may achieve durable remission even in high-risk presentations. Early multidisciplinary management and consideration of fertility preservation are essential in young patients.

Alterations in tight junction (TJ) organization and dysregulation of cancer stem cell (CSC)-associated markers are increasingly recognized as molecular features linked to colorectal cancer (CRC) progression, heterogeneity and clinical outcome. Bioactive dietary compounds such as spermidine (SPD) and eugenol (EUG) have been proposed as modulators of cancer-related molecular pathways; however, their combined effects on CRC spheroid models relevant to molecular characterization remain insufficiently defined. In the present study, the molecular impact of SPD and EUG, administered individually or in combination, was evaluated in primary and metastatic CRC spheroids. First-generation spheroids derived from Caco-2 and SW620 cells were exposed to SPD, EUG, or SPD+EUG at the time of seeding, and spheroid growth and self-renewal capacity were monitored across successive generations. The expression of TJ- and CSC-associated markers was assessed at both the transcript and protein levels using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blotting and immunohistochemistry. The combined SPD+EUG treatment was associated with a marked reduction in spheroid area and self-renewal capacity in both CRC models. Baseline molecular profiling revealed higher TJ marker expression in Caco-2 spheroids and enrichment of CSC-associated markers in SW620 spheroids. Treatment-induced modulation of CSC- and TJ-related transcripts was observed; however, transcript-level changes were not consistently mirrored at the protein level, indicating the involvement of post-transcriptional regulatory mechanisms. In particular, Occludin (OCLN), Zonula occludens-1 (ZO-1), CD133, ALDH1A1, SOX2 and VE-cadherin exhibited divergent RNA and protein expression patterns depending on cell type and treatment condition. Collectively, these findings underscore the relevance of three-dimensional CRC spheroid models for molecular profiling studies and highlight the importance of integrating transcript- and protein-level analyses when evaluating bioactive compounds with potential diagnostic and translational relevance in colorectal cancer.

Progesterone receptor (PR) regulates gene expression through recruiting coregulators and general transcription factors by activation functions AF1 and AF2. AF1 localizes to the non-conserved and disordered N-terminal domain and is believed to facilitate tissue- and gene-specific activity. Our previous proteomic analysis identified three key residues (K464, K481 and R492) in AF1 that are monomethylated. Methylation mimic mutations KKR → FFF created hypoactive PR, whereas the KKR → QQQ mutation generated hyperactive PR in gene reporter assays. The current study used these mutants to determine the roles of AF1 in PR regulation of cellular activities and global gene regulation in breast cancer cells MCF-7. AF1-FFF mutation attenuated PR regulation of cell proliferation and apoptosis in response to progestin, whereas AF1-QQQ mutation enhanced these effects. AF1-FFF mutation attenuated gene regulation by progestin in ~60% of PR target genes, including genes involved in cell proliferation, hypoxia and TNFα signaling. However, the AF1-FFF mutation had little effect on ligand-independent gene regulation, suggesting distinct mechanisms of gene regulation by liganded and unliganded PR. Intriguingly, impaired activity of methylation mimic mutant PRB-FFF is associated with greater chromatin binding in ChIP-Seq analysis, corresponding to a stronger association between PRB-FFF and Steroid Receptor Coactivator-1 (SRC-1), a member of the p160 family of nuclear receptor coactivators, as was previously reported. In conclusion, PR AF1 is important for the core activities of liganded PR in regulating ~half of target genes and cell proliferation. AF1 monomethylation may modulate PR-chromatin interactions through stronger association with coregulators, thereby decelerating chromatin binding kinetics. This is supported by PRODIGY's prediction of higher binding affinities of monomethylated AF1 and methylation mimic mutant with SRC-1.

<h4>Background</h4>Chromophobe renal cell carcinoma (ChRCC) is characterized by the accumulation of abnormal mitochondria, a high rate of mitochondrial DNA (mtDNA) mutations, and altered oxidative metabolism. There are no existing circulating biomarkers to distinguish metastatic ChRCC from clear cell renal cell carcinoma (ccRCC).<h4>Methods</h4>High-throughput plasma proteomic profiling using the SomaScan platform was performed in 18 ChRCC (including 16 metastatic ChRCC) and 197 metastatic ccRCC patients. Data were harmonized to generate a unified 7K-protein matrix.<h4>Results</h4>Differential expression analysis was performed using limma (version 3.62.2). Of 7272 quantified human plasma proteins, 209 were differentially expressed between ChRCC and ccRCC. Upregulated proteins in ChRCC included essential β-oxidation enzymes such as ECH1 (enoyl-CoA hydratase 1) and ECI1 (enoyl-CoA delta-isomerase 1), suggesting increased long-chain fatty acid degradation. Creatine and energy-buffering pathways were also represented, with increased CKMT1A (Creatine Kinase, Mitochondrial 1A) in ChRCC. KIM-1 (Kidney Injury Molecule-1) and leptin were lower in ChRCC, consistent with the known upregulation of these proteins in ccRCC. Pathway enrichment analyses revealed an overrepresentation of mitochondrial protein degradation, fatty acid β-oxidation, and respiratory electron transport in ChRCC, suggesting that ChRCC sheds a unique mitochondrial signature into the peripheral circulation. A bootstrap-based LASSO logistic regression restricted to upregulated mitochondrial proteins in ChRCC vs. ccRCC consistently selected ECI1 and CKMT1A. The LASSO model achieved an AUROC of 0.964.<h4>Conclusions</h4>Compared to ccRCC, the plasma proteome of metastatic ChRCC is dominated by mitochondrial metabolic enzymes, revealing a systemic metabolic phenotype strikingly aligned with the known histologic accumulation of abnormal mitochondria in ChRCC cells.

Curvularins, a class of macrocyclic lactones, have cytotoxic, antimicrobial, and anti-inflammatory properties. Curvularin, a 12-membered macrolactone, was used as a scaffold to design and synthesize structurally modified analogues to investigate structure-activity relationships and improve biological efficacy. Three series of curvularin-based analogues, Cur-5H-OMe, Cur-4P-OMe, and Cur-OMe, were synthesized with the same core structure but different substituent sizes and positions. Nine representative derivatives were evaluated for anti-inflammatory, anticancer, antibacterial, and antifungal activities. In LPS-stimulated RAW 264.7 macrophages, most compounds inhibited nitric oxide (NO) production in a concentration-dependent manner but exhibited cytotoxicity at high concentrations. Cytotoxicity assays against HaCaT cells and human cancer cell lines (HCT116, HeLa, and A375) revealed limited selectivity toward cancer cells. Antimicrobial evaluation indicated selective activity against the Gram-positive bacteria, <i>Staphylococcus aureus</i>. Compound <b>23</b> exhibited superior antibacterial potency compared with kanamycin and notable antifungal activity against <i>Candida albicans</i>. This study provides a versatile synthetic platform and identifies key structural features of curvularin derivatives, demonstrating their potential as anti-inflammatory and antimicrobial lead compounds.