Gene Literature Dashboard

Tumor dormancy represents a clinically significant but poorly understood state in which disseminated cancer cells persist in a quiescent, non-proliferative state, evading conventional therapies and driving late relapse. This review summarizes recent advancements in experimental models-both <i>in vitro</i> and <i>in vivo</i>-that recapitulate the full spectrum of dormancy, including its induction, maintenance, and reactivation. Crucial intrinsic pathways such as ERK/p38 signaling shifts, epigenetic remodeling, and metabolic adaptations and microenvironmental and immune-mediated cues that regulate dormant cell fate are discussed. Therapeutic strategies aimed at maintaining dormancy, reactivating dormant cells for elimination, or directly targeting their survival pathways have been highlighted. By integrating insights from model systems, molecular regulation, and therapy, this review aims to provide a comprehensive framework that informs future efforts to target dormant cancer cells and ultimately reduce recurrence and improve patient outcomes.

Hydroxyapatite (Hap) is a prominent biomaterial used as an effective implant material in bone tissue engineering, but its use presents some points of weakness in bone regeneration efficiency. Different biofunctionalization strategies have been utilized to increase the regenerative Hap capacities. Carnosine (Car) or β-alanyl-l-histidine dipeptide has received much attention due to its beneficial effects in osteoarticular diseases and bone tissue healing. Hap functionalized in noncovalent mode with Car at a nominal Ca:Car molar ratio (10:1, 2:1, and 1:1) was synthesized. The Hap-Car composites were characterized by using X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopies. The structural and morphological feature comparisons indicate a similarity between Hap-Car10:1 and Hap. The Hap-Car composites and Hap bind copper present at submicromolar concentration in the complete culture medium, determined by inductively coupled plasma-optical emission spectroscopy. Hap-Car composites enhance the biological properties of Hap in in vitro assays and promote the mineralization process and the expression of alkaline phosphatase, osteocalcin, vascular endothelial growth factor, brain derived neurotrophic factor, and bone morphogenetic protein-2 in hFOB1.19 cells. The protective and regenerative activities of the metal ion are also related to the intracellular chaperone copper chaperones for superoxide dismutase.

<h4>Background</h4>Lower respiratory tract infection (LRTI) remains the leading infectious cause of morbidity and mortality globally. Key bacterial pathogens include Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Streptococcus pneumoniae. This study examined the prevalence and antimicrobial resistance patterns of major bacterial pathogens from community- and hospital-acquired LRTIs across six major hospitals in Vietnam.<h4>Methods</h4>Between January 2022 and May 2023, 1000 bacterial isolates were collected through an isolate-based surveillance. Species identification and antimicrobial susceptibility testing were performed by VITEK-2/Phoenix M50, with MICs determined by E-test or broth microdilution. Multiplex PCRs were used to detect common AMR genes.<h4>Results</h4>A. baumannii (49.6%), P. aeruginosa (21%), K. pneumoniae (18.6%) were predominant, followed by S. aureus (6.7%), E. coli (3.9%) and S. pneumoniae (0.2%). Most isolates (94.4%) were identified from hospital-acquired cases. High prevalence of MDR and carbapenem resistance were identified in A. baumannii (96% and 95%), P. aeruginosa (56.7% and 57.1%), and K. pneumoniae (78% and 69.2%), respectively. Notably, resistance to ceftazidime-avibactam was detected in K. pneumoniae (34.3%), P. aeruginosa (29%), and E. coli (7.7%), while colistin resistance was found in K. pneumoniae (18.2%) and A. baumannii (2.8%). MRSA prevalence was 79.1%, though S. aureus remained susceptible to vancomycin, linezolid and ceftaroline. Most bla_NDM-positive K. pneumoniae (62/71, 87.3%), E. coli (2/2, 100%), and P. aeruginosa (23/25, 85.2%) showed resistance to ceftazidime-avibactam. Whole genome sequencing revealed that the bla_NDM-positive but ceftazidime-avibactam susceptible isolates (9 K. pneumoniae and 2 P. aeruginosa) carried truncated bla_NDM. Overall, ceftazidime-avibactam was effective against K. pneumoniae, E. coli, and P. aeruginosa isolates carrying ESBL, ESBL and bla_OXA-48, or ESBL and bla_KPC. Alternatively, no detectable AMR genes were found in 35 ceftazidime-avibactam resistant P. aeruginosa isolates.<h4>Conclusions</h4>Carbapenem-resistant Gram-negative pathogens were predominant among hospital-acquired LRTIs in Vietnam, with notable resistance to ceftazidime-avibactam and colistin. The lack of effective treatment for A. baumannii remains a major concern. We found a strong correlation between AMR phenotype and genotype among K. pneumoniae and E. coli, supporting gene-based therapy to guide ceftazidime-avibactam use. However, the presence of disrupted bla_NDM underscores the need to re-evaluate commercial PCR assays for carbapenemase detection.

<h4>Background</h4>Epidemiological studies have suggested that dried fruit intake may be associated with a lower risk of breast cancer (BC), yet the genetic mechanisms underlying this association remain unclear. This study aimed to explore the potential genetic relationship between dried fruit intake and BC susceptibility.<h4>Methods</h4>We conducted a comprehensive genetic analysis using genome-wide association study (GWAS) data for dried fruit intake and BC, focusing on individuals of European ancestry. We assessed genome-wide and region-specific genetic correlations using several complementary methods, including linkage disequilibrium score regression and regional genetic mapping. To identify shared genetic regions, we applied statistical approaches that integrate information across traits and improve the detection of common genetic signals.<h4>Results</h4>Modest but statistically significant negative genetic correlations were observed between dried fruit intake and BC, including its subtypes. Regional analysis revealed shared signals across multiple chromosomal regions. Five candidate loci were consistently identified as shared between the traits, including BCL11A, MAD1L1, MLLT10, JMJD1C, and RP11-795H16.2.<h4>Conclusion</h4>This study provides evidence for a genome-wide genetic link between dried fruit intake and BC risk, identifying several loci that may be shared between the traits. These findings may help improve our understanding of BC development and offer preliminary leads for future dietary prevention and personalized interventions, pending further experimental validation.

<h4>Purpose</h4>To investigate the effect of school education on the prevalence of high myopia.<h4>Patients and methods</h4>This prospective cross-sectional study, conducted in schools across Hubei Province, included 1,017,622 students from 103 county-level administrative regions between October 2021 and November 2023. Refractive measurements and basic demographic data were collected for all participants. The prevalence of high myopia and the differences in prevalence across grade levels were calculated. Regression discontinuity was used to assess the effects of age and education on high myopia prevalence.<h4>Results</h4>A total of 1,017,622 students participated, with 540,860 (53.15%) boys, and an average age of 11.93 ± 3.06 years. Over three years, the average prevalence of high myopia was 4.75%. Prevalence increased with grade level, with the largest difference observed between grades 9 and 10. Age showed no significant effect on high myopia prevalence. Each additional year of education led to an increase in prevalence by 1.26% (95% CI: -1.87, -0.65, P=0.000) in 2021, 1.20% (95% CI: -1.88, -0.52, P=0.001) in 2022, and 1.26% (95% CI: -2.49, -0.04, P=0.04) in 2023.<h4>Conclusion</h4>Increased grade, rather than age, is the key factor driving the rise in high myopia prevalence. Interventions targeting myopia risk factors during schooling could potentially reduce the growing prevalence of high myopia.

Dental mesenchymal stem cells (MSCs) play an essential role in the development of immature permanent teeth. Bacterial infection of the pulp and periapical tissues of immature permanent teeth, the associated oral pathogens, and their virulence factors affect the viability, proliferation, differentiation, and cytokine secretion of MSCs. Bacteria and virulence factors can also trigger an inflammatory response that induces pro-inflammatory cytokine secretion and destroys odontogenic MSCs in the pulp and periapical region, negatively affecting the development of immature permanent teeth. The present study explored the role and mechanisms of oral pathogens associated with pulpitis and apical periodontitis and their virulence factors concerning odontogenic MSCs. The findings can contribute to the clinical treatment of pulpitis and apical periodontitis of immature permanent teeth, providing a theoretical basis for improving its clinical efficacy.

Antihypertensive drugs are widely used for the treatment of hypertension, and the choice of drug and dosage is based on trial and error. The variability in drug response and adverse reactions leads to the poor adherence to treatment. Epigenetic modulation is one of the major mechanisms that may contribute to the variability in drug responses, and microRNAs (miRNAs) can serve as crucial epigenetic regulators and have also been reported to be associated with hypertension pathogenesis. The objective of this study is to investigate the regulatory effects of commonly used antihypertensive drugs on the endothelial miRNome in human aortic endothelial cells. We aim to integrate miRNA expression data with proteomic analyses to elucidate drug-induced molecular mechanisms relevant to hypertension treatment. Whole genome small RNA sequencing was performed, followed by whole proteome analysis using LC-MS/MS comparing between control and treated samples. The treatments induced significant differential regulation of several miRNAs and proteins; among these, a few reflected reverse relationships with miRNA regulation and protein expression. Certain miRNAs and their corresponding target proteins seem to distinguish between good therapeutic outcomes and potential side effects. This study unravels the potential role of drug-induced miRNAs in inducing post-transcriptional modifications to cause the differential expression of certain proteins that may induce not only therapeutic effects or drug side effects but can also indicate the potential for drug-repurposing in other diseases.

A novel structure-functional-integrated particle featuring dual micromechanical interlocking property with resin matrix was constructed through surface modification of urchin-like serried hydroxyapatite (UHA) in this work, and the effect of this modification strategy on physicochemical and biological properties of dental resin composite was also investigated. A porous silica coating layer was anchored onto UHA surface via a simple template method in an oil-water biphase reaction system, and the coating time had a prominent effect on the coating thickness and morphology-structure of the particle. When these particles with different porous silica coating thickness were used as fillers for dental resin composite, results showed that UHA/PS5 (porous silica coating reaction time: 5 h) exhibited the optimal 3D urchin-like structure and a desirable porous silica coating thickness. Additionally, UHA/PS5 formed the best dual physical micromechanical interlocking structure when mixing with resin matrix, making the dental resin composites presented the desirable matrix/filler interfacial bonding, and the excellent physicochemical-biological properties, especially for flexural strength and water sorption-solubility. In vitro remineralization and cellular biological properties confirmed that the coating layer did not compromise their remineralization activity. The use of UHA/PSx provides a promising approach to develop strong, durable, and biocompatible DRCs.

<h4>Background</h4>Respiratory infections in poultry, particularly those caused by <i>Escherichia coli</i>, are a major concern in veterinary medicine due to their economic impact and increasing antibiotic resistance. Nanotechnology-based antimicrobial delivery systems have emerged as promising alternatives to conventional treatments.<h4>Aim</h4>This study aimed to evaluate the cytotoxicity and antibacterial activity of various nano-antibiotic formulations-comprising antibiotics, herbal extracts, and silver nanoparticles-against <i>E. coli</i> strains isolated from chickens with respiratory disease.<h4>Method</h4>The nanoantibiotic formulations were developed by the Department of Biomedical Nanomaterials, Institute of Materials Science, Vietnam, and included combinations such as polymer-garlic, polymer-<i>Phyllanthus urinaria</i>, polymer-garlic-silver, polymer-<i>P. urinaria</i>-silver, polymer-doxycycline + florfenicol, polymer-doxycycline + oxytetracycline, and their respective silver-conjugated forms, along with chitosan. Cytotoxicity was assessed using Vero cells, and antibacterial activity was determined <i>in vitro</i> using the dilution method in 96-well flat-bottom plates.<h4>Results</h4>The study found that cytotoxic effects were observed at dilutions of 1:32 and 1:64 when the formulations combined antibiotics, herbal extracts, and nanosilver. Importantly, this combination significantly enhanced bactericidal activity compared with formulations containing only antibiotics or herbal extracts.<h4>Conclusion</h4>The integration of nanosilver with antibiotics and herbal extracts in nanoparticle systems shows promising antibacterial efficacy against <i>E. coli</i>, with tolerable cytotoxicity at appropriate dilutions. These findings suggest the potential of such nanoformulations as effective alternatives in managing bacterial respiratory infections in poultry.

<h4>Background</h4>Swine influenza viruses (SIVs) is a highly infectious viral disease in pig population. This disease is caused by the influenza A virus. According to the World Organization for Animal Health (WOAH), three subtypes of SIVs have been officially reported, including H1N1, H1N2, and H3N2. However, information about this pathogen circulated in Vietnam is limited, especially during the last 5 years.<h4>Aim</h4>This research aimed to examine the molecular characteristics of SIVs circulating in Vietnam. A total of 213 swab samples from 18 slaughterhouses in Thanh Hoa province, North Central Vietnam, were collected between 2024 and early 2025, and SIVs were detected by realtime RT-PCR. The characteristics of these viruses were further determined based on the phylogenetic analysis of the Hemagglutination (HA) and Neuraminidase (NA) genes.<h4>Methods</h4>SIV subtypes were determined by realtime RT-PCR. The full-length HA and NA genes were amplified by PCR. Phylogenetic and characterization analysis of HA and NA nucleotides and proteins were performed using bioinformatics. Virus isolation and titration were performed in the Madin-Darby canine kidney (MDCK) cell line.<h4>Results</h4>Of 213 samples, 24 were positive for SIVs, representing an 11.27% prevalence at the slaughterhouses (SIVs were confirmed in 5 out of 18 slaughterhouses). These samples included 17 H1N1, 1 H1N2, and 6 H3N2 subtype samples. Phylogenetic analysis of the HA genes showed that four H1N1 subtypes belonged to the H1N1pdm09 lineage, three to the Eurasian "avian-like" (EA) H1N1 lineage, and one to the TR-H1N2 lineage. Additionally, four H3N2 subtypes were classified into clade IV of the recent human-line (RH) H1N1. Based on the NA gene sequences, seven viruses belonged to the EA H1N1 lineage, three to the RH-H3N2 lineage, and one to the TR-H1N2 lineage. Receptor-binding and antigenic site analysis revealed that all isolates could bind to both human- and avian-like receptors. Furthermore, <i>in vitro</i> analysis of the growth kinetics of SIVs showed that 12 SIV isolates could efficiently replicate in MDCK cell lines.<h4>Conclusion</h4>Our study demonstrated that all three WOAH subtypes have been circulating in Vietnam during 2024 and early 2025. H1N1 and H3N2 subtypes are predominant, suggesting a potential threat to human health. This highlights the importance of active surveillance as a critical step for swine influenza prevention and control in Vietnam.

Intestinal epithelium is derived from Lgr5⁺ intestinal stem cells (ISCs) located at the crypt base. However, how small nucleolar RNAs (snoRNAs) regulate ISC stemness remains elusive. Here, a conserved snoRNA, Snord15b, that is highly expressed in ISCs is identified. Snord15b knockout abolishes the self-renewal capacity of ISCs and impairs epithelial regeneration. Mechanistically, Snord15b interacts with interleukin enhancer-binding factor 2 (Ilf2) to recruit splicing factors, which mediates alternative splicing of Btrc to form a short isoform of Btrc, resulting in abrogation of E3 ligase complex formation for ubiquitination of β-catenin. Subsequently, stable β-catenin translocates into the nucleus of ISCs for activation of the Wnt/β-catenin signaling pathway, leading to ISC stemness maintenance and intestinal regeneration. Of note, knockout of Snord15b or Ilf2 increases β-catenin ubiquitination to suppress activation of Wnt signaling. Furthermore, Btrc knockout blocks β-catenin ubiquitination to enhance the stemness of ISCs and intestinal regeneration. These findings reveal that Snord15b-Ilf2 association mediates alternative splicing of Btrc short isoform to inhibit β-catenin ubiquitination for ISC stemness maintenance.

<h4>Opinion statement</h4>Pancreatic adenocarcinoma remains a leading cause of cancer-related mortality worldwide. Although surgery can be curable for a subset of patients, the five-year overall survival remains less than 15%. Despite extensive molecular characterization of pancreatic cancer, cytotoxic chemotherapy has served as the major component in therapeutic management. A major driver of pancreatic adenocarcinoma is mutations in KRAS, present in over 90% of cases. However, attempts to inhibit KRAS through upstream and downstream targets through the mitogen-activated protein kinase pathway have not been successful in the past. Despite this, multiple KRAS inhibitors have recently entered clinical trials and have shown promising results. These inhibitors have the potential to dramatically alter the landscape of treatment. In parallel, immunological approaches utilizing vaccines and bispecific antibodies are also in clinical development. Given these rapid new developments, the future of pancreatic cancer treatment will likely be determined by discovering the appropriate combinations of targeted and immune-based treatments.

The blood-brain barrier (BBB) maintains brain homeostasis through specialized functions including tight junction formation and selective transport of brain endothelial cells (ECs). While ECs are generally thought to rely primarily on glycolysis for energy production, the transcriptional mechanisms underlying their metabolic specialization in the brain endothelium remain poorly understood, especially considering the brain's extraordinary energy demands. Through comparative transcriptomic analysis, it is demonstrated that brain endothelial cells are enriched for mitochondrial function genes, with forkhead box protein 1 (FOXQ1) being selectively expressed in cerebral vasculature. Conditional knockout of Foxq1 in endothelial cells results in severe mitochondrial dysfunction, including disrupted cristae morphology, reduced oxygen consumption, and impaired adenosine triphosphate (ATP) production. Mechanistically, FOXQ1 directly regulates two key pathways: calcium signaling through huntingtin-associated protein (HAP1)-mediated endoplasmic reticulum (ER)-mitochondrial calcium transfer, and mitochondrial structural integrity via AarF domain-containing protein kinase 1 (ADCK1)-dependent cristae organization. These findings reveal that brain endothelial cells rely on oxidative phosphorylation rather than glycolysis alone, challenging the prevailing metabolic paradigm for endothelial cells. This work establishes FOXQ1 as an important regulator of brain endothelial metabolism and provides new insights into the molecular basis of cerebrovascular specialization, with implications for understanding vascular dysfunction in neurological diseases.

The total synthesis of compound 1, a chiral diglyceride metabolite present in both humans and fungi, was achieved via a seven-step route, affording the target molecule in 2.33% overall yield. The synthetic strategy involved: (1) selective protection of the terminal hydroxyl group of chiral ketal 2 with a sterically hindered benzyl group, followed by ketal deprotection to yield benzyl ether 4; (2) protection of the terminal hydroxyl group of benzyl ether 4 with a bulky silyl protecting group, and subsequent esterification of the remaining hydroxyl with erucic acid to generate ester 6; (3) removal of the silyl protecting group from ester 6, followed by esterification of the liberated hydroxyl group with pentadecanoic acid to afford ester 8; and (4) selective deprotection of the benzyl group of ester 8 to furnish compound 1. Network pharmacology and molecular docking studies identified 196 potential targets of compound 1, with AKT1, ALB, CASP3, EGFR, HSP90AA1, IGF1, and SRC highlighted as potential hub targets. In silico analysis suggested potential therapeutic applications in diabetes, neuro-systemic diseases, thyroid hormone disorders, lipid disorders, tumors, and gynaecological diseases. Furthermore, in silico screening using three databases identified 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) as a potential biological target. However, in vitro HMGCR inhibition assays failed to demonstrate a significant reduction in cholesterol levels in human blood upon treatment with compound 1. These findings contribute to the understanding of the chemical synthesis and biological activity of chiral diglycerides and provide a basis for future research in this area.

Leucine zipper like transcription regulator 1 (LZTR1) is amplified in acral melanomas, is required for melanocytes and melanoma cell proliferation, and it induces anchorage-independent growth, by yet unknown mechanisms. We therefore performed comprehensive studies to identify its activity in melanomas employing proximity biotinylation and co-immunoprecipitation combined with LC-MS/MS proteomics and molecular characterization. The results show that LZTR1 regulates the ubiquitin proteasome system in melanoma cells and also associates with actin-related proteins and actin cytoskeleton organization. Its downregulation suppresses the protective effect of the autophagy-initiating ULK1 and AMBRA1, regulators of normal cell survival and proliferation, and upregulates the sequestosome 1 (SQSTM1/p62), an autophagic cargo adapter which mediates selective degradation of ubiquitinated proteins. In contrast, overexpression of LZTR1 provides growth advantage under environmental stress, enhancing cell invasion, by activating ERBB3 receptor and its downstream targets PYK2 and SRC tyrosine kinases that regulate the cytoskeleton, actin organization, cell spreading, cell migration and adhesion. LZTR1 is a "safeguard" for melanoma cells under stress and its downregulation can be exploited for melanoma therapy.

<h4>Background</h4>Before Zika virus (ZIKV) infections were observed in the Americas, an association between ZIKV and microcephaly or other congenital malformations was not well documented. Initial reports suggested strong associations between ZIKV and congenital malformations, but plausible estimates of causal effects from prospective studies with adequate sample size and covariate data were few.<h4>Methods</h4>From 2016-2018, the Zika in Infants and Pregnancy (ZIP) study enrolled pregnant people before 18 weeks gestation or with confirmed symptomatic ZIKV in a prospective cohort across 10 sites in South and Central America, and in Puerto Rico. Pregnancies were followed monthly through delivery and 6 weeks postpartum. Infants were followed quarterly to age 12 months. Prespecified co-primary analyses evaluated the associations between a composite endpoint of adverse fetal, neonatal, and infant outcomes with intrauterine ZIKV exposure overall and with symptomatic intrauterine ZIKV exposure. Secondary analyses separately evaluated the association of intrauterine ZIKV exposure with individual components of the primary endpoint.<h4>Results</h4>Six thousand one hundred pregnant participants were included in the primary analysis, including 61 with ZIKV infection during pregnancy confirmed by a ZIKV-specific RNA test. For the primary analyses, the relative risk (RR) for the composite endpoint associated with any ZIKV exposure was 1.64 (95% CI: 0.65, 4.13) and with symptomatic ZIKV exposure 1.08 (95% CI: 0.15, 7.64). Sensitivity analyses provided similar results. Secondary analyses showed significant adjusted RRs [95% CI] for stillbirth (4.28 [1.39, 13.21]), infant death within six weeks (6.20 [1.08, 35.60], and fetal loss before 20 weeks (3.72 [1.82, 7.59]).<h4>Conclusions</h4>The ZIP study identified an elevated but not statistically significant risk of the primary composite outcome with intrauterine ZIKV exposure, and a significantly increased risk of some adverse fetal and infant outcomes with intrauterine ZIKV exposure in secondary analyses. Fewer than expected infections observed during pregnancy, coincident with a waning epidemic, limited study power to evaluate risk. Combining data from multiple cohorts for future meta-analysis may better define the risks of intrauterine ZIKV exposure.<h4>Trial registration</h4>NCT02856984. Registered August 5, 2016. Retrospectively registered.

High-altitude environments, characterized by hypoxia, low temperatures, and intense ultraviolet radiation, pose significant challenges to human physiology and health. DNA methylation, as a key epigenetic regulatory mechanism, plays a central role in human adaptation to high-altitude environments and in disease pathogenesis. Current research indicates that high-altitude native populations (such as Tibetans and Andeans) modulate the methylation of hypoxia-responsive genes like EPAS1 and EGLN1 to enhance oxygen transport efficiency and energy metabolism patterns, while simultaneously suppressing excessive erythropoiesis and oxidative stress damage. This epigenetic regulation not only compensates for the lag in genetic adaptation over time but also forms synergistic networks with genetic variations. For instance, the functional SNPs of the EPAS1 gene are co-localized with its differentially methylated regions, revealing a delicate balance between genetic and epigenetic interactions under environmental stress. On the other hand, aberrant methylation patterns may disrupt the homeostasis of the HIF pathway, leading to acute and chronic high-altitude illnesses. This article provides a review of the recent research progress in plateau medicine and DNA methylation (up to 2025), including human clinical studies and animal model research. This includes research on high-altitude adaptation/acclimatization, as well as studies on inadequate adaptation to high altitude in relation to acute and chronic high-altitude-related diseases, cognitive decline, and pregnancy risks. By elucidating the core mechanisms underlying the "environmen - epigenetics - phenotype" axis, this work aims to provide a theoretical foundation for precision health interventions in high-altitude regions.

<b>Background:</b> Osteoporosis is a prevalent metabolic bone disorder with complex molecular underpinnings. Emerging evidence implicates endoplasmic reticulum stress (ERS) in its pathogenesis; however, systematic exploration of ERS-related genes (ERSRGs) remains limited. This study aimed to identify ERS-related differentially expressed genes (ERSRDEGs) in osteoporosis, construct a diagnostic model, and elucidate associated molecular mechanisms. <b>Methods:</b> Three osteoporosis datasets (GSE56815, GSE230665, and GSE7429) were integrated after batch effect correction and normalization. ERSRGs were curated from GeneCards, and ERSRDEGs were identified by intersecting co-differentially expressed genes (Co-DEGs) across datasets. Functional enrichment (gene set enrichment analysis [GSEA], gene set variation analysis [GSVA], Gene Ontology [GO], and Kyoto Encyclopedia of Genes and Genomes [KEGG]) and immune infiltration analyses were performed. Diagnostic models were developed using support vector machine (SVM) and least absolute shrinkage and selection operator (LASSO) regression, validated via receiver operating characteristic (ROC) curves, nomograms, and decision curve analysis. Experimental validation included immunohistochemistry and quantitative reverse transcription polymerase chain reaction (qRT-PCR) in ovariectomized (OVX) mice. Regulatory networks (TF-miRNA-RBP-drug) and protein structure predictions were generated using bioinformatic tools. <b>Results:</b> Fifty six ERSRDEGs were identified, enriched in apoptosis, autophagy, and cytokine signaling pathways. A diagnostic model comprising seven genes (CYB5R4, RAB1B, UFSP2, RNF13, SERP1, CES2, and C1QBP) demonstrated high accuracy (area under the curve (AUC) > 0.9) in both training and validation datasets. Immune infiltration analysis revealed distinct patterns of activated B cells, CD8⁺ T cells, and macrophages between high- and low-risk groups. Regulatory networks highlighted interactions with 52 transcription factors (TFs), 42 miRNAs, and 27 therapeutic compounds. Experimental validation in OVX mice confirmed upregulated expression of C1QBP, CYB5R4, RAB1B, and UFSP2 at protein/mRNA levels, aligning with bioinformatic predictions. <b>Conclusions:</b> This study establishes ERSRDEGs as critical players in osteoporosis pathogenesis and provides a clinically translatable seven-gene diagnostic model for early osteoporosis detection. The integration of multiomics analyses uncovered key pathways, immune dynamics, and regulatory networks, while experimental validation reinforced the role of specific ERSRGs. These findings provide novel insights into ERS-mediated mechanisms and therapeutic targets for osteoporosis management.

Molecularly imprinted polymers (MIPs) are polymeric receptors for a targeted template molecule that are traditionally formed using a combination of functional monomers and crosslinkers. While investigating novel crosslinkers for MIPs, one of these (2-(methacryloylamino)ethyl-2-methylacrylate (referred to as N,O-bismethacryloyl ethanolamine or "NOBE")) performed better when used alone versus in combination with other monomers. This introduced the concept of one monomer molecularly imprinted polymers, given the acronym OMNiMIPs, and prompted studies provided in this report that clarify OMNiMIPs have fundamental differences compared to traditionally formulated MIPs. Enantioselectivity studies using BOC-L-tyrosine as a standard template showed that NOBE OMNiMIPs afforded higher-performing MIPs compared with traditional MIPs, have significantly higher binding capacities, and have an internal hydrogen-bonded crosslinking structure that contributes to the morphological stability of the binding site structure. Based on the adventitious discovery of NOBE OMNiMIPs, new analogs based on the NOBE structure were developed and evaluated for further enhancement of molecular recognition performance and novel capabilities of OMNiMIPs. While the majority of the new OMNiMIPs exhibited enantiomeric selectivity toward BOC-L-tyr, improvements were not observed compared with NOBE.

Lateral Spreading Tumors (LSTs) are a type of non-polypoid lesion known for their flat morphology, which often leads to them going undetected. However, especially nongranular (NG) LSTs have the potential for malignant transformation. Recent advances in endoscopic technologies have improved the detection of these lesions. Despite growing research interest in their role in colorectal cancer (CRC) development, a comprehensive molecular characterization of LSTs is still lacking. The aim of this review is to highlight the current knowledge of the molecular characteristics of LSTs, that may help in determining whether LSTs can be prognostic indicators and identifying cases where they may rapidly progress to CRC through characteristic molecular pathways. From a mutational point of view, LSTs seem to be more closely associated with inflammatory bowel diseases (IBDs) than with polypoid lesions. Nonetheless, they have peculiar epigenetic and genetic traits, which set them apart from other adenomas or bowel diseases. Elucidating their role in CRC development would provide benefits for their classification and management, by enhancing clinical surveillance strategies for patients diagnosed with these lesions in order to improve the efficient prevention of colorectal cancer.

d-Tagatose is a rare sugar that exhibits intriguing biological properties, such as its role as a low-calorie sweetener and its ability to reduce the glycemic response. Consequently, the synthesis of d-tagatose derivatives is a crucial endeavor for the advancement of their functionalities, as well as elucidation of their biological properties. In this study, we present the α-selective glycosidation of a 1,3,4,6-tetra-<i>O</i>-benzoylated d-tagatofuranosyl donor with various glycosyl acceptors. In contrast to d-allulose, which is the C3,C4-epimer of d-tagatose and does not exhibit the neighboring group effect, the current d-tagatofuranosyl donor demonstrated significant neighboring group participation, achieving high α-selectivity ratios up to α:β = 99:1. This method was also applicable to the synthesis of α-d-tagatofuranosylceramide, which has potential as a novel functional molecule. Meanwhile, the glycosylation of sterically congested glycosyl acceptors, such as 2-hydroxycumene, resulted in poor α-selectivity, which may be attributed to the interaction with the C1-benzoyloxy group of d-tagatofuranosyl donors in the transition state.

<b>Background</b>: Although <i>Presenilin-1</i> (PSEN1) mutations are classically associated with early-onset Alzheimer's disease (AD), spastic paraparesis (SP) may occasionally represent as an initial or even isolated clinical manifestation. <b>Methods</b>: We report the novel association of a PSEN1 mutation (Leu282Arg) with isolated SP at onset in a patient with a family history of early-onset AD. Additionally, we reviewed previously published cases describing similar presentations related to PSEN1 mutations. <b>Results</b>: The age of reported patients ranged from 24 to 60 years. The most common clinical course included the presence of cotton wool plaques and a progressive development of cognitive decline following the onset of SP. A positive family history of either motor or cognitive symptoms was consistently observed. <b>Conclusions</b>: Our findings emphasize the clinical importance of considering PSEN1 mutations in the differential diagnosis of patients presenting with spastic paraparesis, particularly in the presence of cognitive symptoms, cerebral amyloid angiopathy, or a family history of AD.

Heavy metal contamination is a serious concern affecting the safety of tap water sources. Hence, this study evaluated physicochemical quality indices, carcinogenic and non-carcinogenic health hazard derived from the level of toxic metals in tap water in Gondar city, Ethiopia. The results revealed that except dissolved oxygen, salinity and nitrite, all quality attributes were below the allowable quality standards. The average concentrations for iron (Fe), copper (Cu), lead (Pb), chromium (Cr) and cadmium (Cd) were ranged from 0.003 mg/L to 5 mg/L, 0.475 mg/L to 0.752 mg/L, 0.14 mg/L to 0.703 mg/L, 0.261 mg/L to 2.182 mg/L, and 0.035 mg/L to 4.286 mg/L, respectively. The mean levels of metals in different areas decreased in the order: AR > AZ1 > PS1 > AZ3 > PS2 > MR > PS3 > SHD > AZ2 > CL. Except for Cu, the concentration of Fe, Pb, Cr, and Cd exceeded the safe limits described by WHO/FAO. According to principal component analysis and cluster analysis, anthropogenic activities were found to be the major source of metals. Chronic daily intake (CDI), target hazard quotient (THQ), hazard index (HI), and incremental lifetime cancer risk assessment (ILCR) were employed to evaluate human health risks. Except for Pb in AZ1, PS3, and AR, the values of THQ for both ingestion and dermal pathways from the analysed metals for adults were within the safety limits (THQ < 1). However, the distribution pattern of HI values were presented in the decreasing order: PS1 > PS2 > AZ3 > MR > PS3 > AR > AZ2 > AZ1 > SHD > CL. Except, the HI values in CL, all values were greater than one (HI > 1), indicating that tap water in these areas may pose non-carcinogenic health risk. The analysis of carcinogenic health risks indicated that the lifetime cancer risk (ingestion and dermal exposure pathways) of heavy metals were in accordance with the acceptable range for tap water (10^-6 - 10^-4). This finding provides valuable input for the development of precise action plans aimed at elevating water quality standards in the studied areas.

FAM134B, originally characterized as an oncogene in esophageal squamous carcinoma, has also been implicated in the pathogenesis of hereditary sensory and autonomic neuropathy type IIB (HSAN2B). It is recognized as the inaugural endoplasmic reticulum (ER)-phagy receptor in mammals containing an LC3-interacting region, which facilitates its interaction with LC3 and GABARAP proteins in the autophagosome. ER-phagy, a critical process involved in ER quality control, selectively degrades superfluous or damaged ER fragments in lysosomes, thereby maintaining ER and protein homeostasis. This review offers an in-depth analysis of FAM134B's structure, function, and regulation, emphasizing its role in infectious diseases, neuropathies, cancer, metabolic disorders, degenerative conditions, and cardiovascular diseases. The evidence presented highlights the need for further research on FAM134B as a potential therapeutic target in human diseases.

The repeated head impacts experienced by athletes have attracted significant interest from both the public and the scientific community; however, the neurobiological effects following the games are not well understood. For example, a single football match carries the risk of repeated concussive and subconcussive head impacts, which can increase the risk of developing neurodegenerative diseases. Chronic traumatic encephalopathy (CTE) is one of the neurodegenerative conditions athletes often face or are unaware of. However, addressing the disease progression in CTE is difficult to determine due to several reasons, such as the failure to identify risk factors, difficulty in differentiating CTE from other neurodegenerative diseases, and the lack of a specific mechanism by which CTE leads to tau protein accumulation. In addition, CTE symptoms overlap with other neurodegenerative conditions, such as Alzheimer's disease (AD) and Parkinson's disease (PD), which poses a challenge to producing specific targeted therapy. In this case, ultrasound represents a promising non-invasive technique that enables clear visualization of brain structures and may modulate neuronal activity. The term ultrasound encompasses various modalities; for example, high-intensity focused ultrasound (HIFU) employs thermal energy to ablate cells, whereas low-intensity pulsed ultrasound (LIPUS) delivers mechanical energy that activates molecular signaling pathways to impede the progression of CTE. Therefore, the LIPUS application could potentially minimize the risk of damage in the surrounding tissues of the brain and reduce the disease progression in individuals with CTE. Nevertheless, limited studies have been reported in the literature, with a poor mechanistic approach. Hence, this review aims to highlight the molecular signaling pathways, such as AKT, MAPK, and ERK, affected by LIPUS and emphasize the need for additional research to clarify its mechanistic effects in CTE management. Ultimately, this review aims to contribute to a nuanced understanding of LIPUS as a therapeutic strategy in addressing the complexities of CTE and its associated neurodegenerative disorders.

Radiotherapy is one of the most commonly prescribed anticancer modalities in the clinic, which is widely recognized as an effective and safe treatment for a broad spectrum of solid tumor indications. Interestingly, there is increasing evidence that the tumors can dynamically modulate cell fate decisions after ionizing radiation (IR) exposure, which is beneficial for escaping the radiation-induced antitumorigenic cell damaging and immunostimulatory impacts. Consequently, the regulatory network of cell fate determination could be a promising target for enhancing the susceptibility of tumor cells to various radiotherapeutic modalities. In this review, we provide a comprehensive account on the mechanisms of post-radiation cell fate control in tumor cells to escape programmed cell death (PCD) including apoptosis, necrosis, pyroptosis and ferroptosis, while special emphasis is placed on the development of synthetic agents for the therapeutic modulation of post-radiation tumor cell fate decisions to facilitate tumor cell eradication, focusing on their therapeutic utility for amplifying the RT-induced direct tumor cell damage as well as promoting the post-IR antitumor immunity. We envision that these synthetic cell fate regulatory technologies could provide new avenues for improving radiotherapeutic efficacy.

<b>Rationale:</b> Abdominal aortic aneurysm (AAA) is a life-threatening cardiovascular disease lacking clinical predictors and effective pharmacologic therapies. The cellular heterogeneity and molecular changes of different cell types during AAA have been revealed in human and mouse aortas by single-cell RNA sequencing (scRNA-seq) technology. However, the heterogeneity and plasticity of endothelial cells (ECs) in AAA remain poorly characterized. <b>Methods:</b> scRNA-seq was performed on the abdominal aorta from angiotensin II (AngⅡ) and salt-induced AAA mice. Additionally, public scRNA-seq data of human and mouse AAA were analyzed with a focus on ECs. Cellular and animal experiments were conducted to validate EC heterogeneity and to investigate the role of SRY (sex-determining region on the Y chromosome)-box transcription factor 18 (Sox18) in endothelial-to-mesenchymal transition (EndMT) during AAA formation. <b>Results:</b> Unbiased clustering analysis identified 20 clusters encompassing 11 cell types. Four subpopulations of ECs were identified in AngⅡ and salt-induced mouse AAA models: <i>Cd36</i> ⁺ lipid-handling ECs, <i>Fn1</i> ⁺ mesenchymal-like ECs, <i>Lrg1⁺</i> pleiotropically activated ECs, and <i>Mmrn1</i> ⁺ lymphatic-like ECs. Similar results were observed in human AAA scRNA-seq data. Endothelial dysfunction and EndMT were detected at single cell solution and validated experimentally. Sox18 was identified as a potential EndMT regulator. Sox18 downregulation was confirmed in both human and mouse aortic aneurysm. <i>In vitro</i>, Sox18 siRNA transfection induced EndMT and increased EC permeability via PI3K/Akt signaling pathway. <i>In vivo</i>, EC-specific Sox18 overexpression inhibited EndMT and attenuated AAA formation. <b>Conclusion:</b> Our data reveal the heterogeneity and transcriptional signatures of ECs in AAA at single cell solution, and demonstrate the previously unrecognized role of Sox18-mediated EndMT in AAA, providing novel insights and a promising therapeutic target for AAA intervention.

Schizophrenia (SCZ) is highly polygenic, and its biological underpinnings remain unclear. In this study, a cost-effective strategy of including sub-threshold GWAS (subGWAS) loci (i.e., 5 × 10^-8 < P ≤ 10^-6) in analysis is explored to increase the inference power of novel pathways. A total of 180 subGWAS loci are identified from SCZ GWAS studies and are shown to contain substantial true genetic association signals. By jointly modeling GWAS (sigGWAS) and subGWAS loci, 304 high-confidence risk genes (HRGs) are identified, as well as a novel category of biological processes detected only in subGWAS loci, i.e., dendrite development and morphogenesis (DDM). Two candidate DDM genes (CUL7 and DCC), whose risk alleles in GWAS are associated with increased expression, are examined, and it is observed that upregulation of these genes leads to reduced neurite length. It is further revealed that the DDM genes lead to disrupted regulatory programs of the transcription factors CUX1/2 and NEUROD1. Collectively, the study identifies DDM as a novel biological process in SCZ susceptibility, with particular implications for DCC- and CUL7-mediated alterations in neurite development and reveals regulatory programs involved in perturbation of the two candidate genes.

The global annual output of liquid crystal monomers (LCMs) continues to increase, yet associated environmental and health risks remain poorly characterized. Assessing the toxic effects of the LCM mixture and individual components is critical for risk evaluation. Organoid models, characterized by highly biomimetic crypt-villus structures and cellular heterogeneity, serve as the preferred in vitro platforms for risk assessments. Our results reveal that exposure to the LCM mixture inhibits organoid expansion rates and induces abnormal crypt branching. Specifically, four machine learning models predicted that 4-cyano-4'-ethoxybiphenyl (EBCN) significantly contributes to intestinal toxicity. Exposure to EBCN alone reduces the proportion of viable cells, impairs intestinal barrier function, and disrupts stem cell homeostasis. Specifically, upregulated expression of the intestinal stem cell marker olfactomedin 4 (OLFM4) aligns with molecular signatures of inflammatory bowel disease patients in the Gene Expression Omnibus (GEO) data set. Additionally, the application of the Adverse Outcome Pathway framework identifies complement component C3 as the initial event, with downstream immunosuppression mediating EBCN-induced intestinal toxicity. In conclusion, this study highlights potential adverse effects of LCMs on gut physiology and emphasizes the urgent need for comprehensive health risk assessments for both the LCM mixture and individual LCM constituents.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent chronic liver disorder globally, affecting 30% of the population and causing a significant healthcare burden due to its increasing incidence and limited therapeutic options. Arachidonic acid (AA) is a key bioactive lipid precursor that generates eicosanoids, such as prostaglandins, leukotrienes, and epoxyeicosatrienoic acids, via 3 distinct enzymatic pathways: cyclooxygenase, lipoxygenase, and cytochrome P450. Emerging evidence indicates that AA-derived metabolites and pathway factors contribute to the progression and severity of MASLD and liver fibrosis. This review systematically summarizes the pathophysiological roles of AA metabolism in MASLD and liver fibrosis, focusing on mechanisms involving lipid accumulation, liver inflammation, fibrogenesis, and related cellular processes. In addition, we discuss potential therapeutic targets within the AA metabolic pathway in MASLD and liver fibrosis, highlighting emerging clinical advances targeting AA metabolites and pathway factors to improve these pathological conditions.

<h4>Background</h4>Double-negative T (DNT) cells (CD3+CD4-CD8-NK1.1-) demonstrate immunoregulatory functions in maintaining hepatic immune homeostasis. This study investigates how energy metabolism impacts DNT cell survival and immunoregulatory functions, exploring potential therapeutic applications for autoimmune hepatitis.<h4>Methods</h4>We compared DNT cells with conventional CD4+ T cells through lipidomic analysis, fatty acid β-oxidation (FAO) assessment, and single-cell RNA sequencing. Cells were treated with fatty acids (oleic acid and palmitic acid) and the FAO inhibitor Etomoxir (Eto). We evaluated cell survival, proliferation, and function through flow cytometry and reverse transcription-quantitative polymerase chain reaction. Transcriptome sequencing identified key regulatory molecules. The therapeutic potential was assessed in a Concanavalin A (ConA)-induced autoimmune hepatitis mouse model receiving DNT and DNT-Eto cell treatments.<h4>Results</h4>DNT cells showed higher fatty acid content, FAO levels, and related gene expression compared with CD4+ T cells. Fatty acid supplementation enhanced DNT cell proliferation and immunoregulatory function, whereas FAO inhibition significantly impaired cell survival and function. Transcriptome analysis identified OX40 as a key regulator of DNT cell survival and function, regulated by FAO-activated pSTAT4. In the ConA-induced murine model, therapeutic administration of DNT cells significantly ameliorated the severity of autoimmune hepatitis compared with the ConA-treated control group. Meanwhile, DNT-Eto-treated groups showed more severe liver injury and elevated liver enzymes compared with DNT-treated groups. In vivo analyses revealed that DNT cells exhibited superior survival, function, and CD4+ T cell inhibition compared with Eto-treated or OX40 KO-DNT cells.<h4>Conclusions</h4>FAO regulates DNT cell survival and immunoregulatory function through the pSTAT4-OX40 pathway, enhancing their protective effect against autoimmune hepatitis.

<h4>Background</h4>Novel therapeutic approaches are actively explored and evaluated, but applying these innovations to practice depends critically on the recruitment of volunteers for clinical trials. This study aimed to assess the motivations, barriers, and willingness of the general population in Vietnam to participate in clinical trials for novel cancer treatments.<h4>Methods</h4>This analytical cross-sectional research involved the distribution of self-administered online and printed questionnaires to residents across central and southern Vietnam in December 2024. A structured 23-item questionnaire was developed based on a literature review of studies from the UK, US, and Germany. Items were culturally adapted for Vietnam and translated using a WHO-adapted four-step process, including forward translation, expert panel review, cognitive interviews with 25 participants, and final revision. Face and content validity were assessed during this process. Logistic regression analysis was conducted to determine the factors associated with the respondents' willingness to participate in the aforementioned clinical trials.<h4>Results</h4>The questionnaires were completed by 851 participants, with the majority aged 40-50 years (30.3%). The respondents also indicated a preference for treatments with prior clinical use and coverage by health insurance (65.6%). The primary motivation for participation was intensive monitoring of conditions (83.7%). The most frequently cited barrier were concerns regarding the high risk associated with less-tested treatments (74.1%). A total of 33.6% expressed a willingness to participate in clinical trials, but fewer (18.9%) were willing to allow such involvement for their children or other family members. Finally, Kinh ethnicity, good health, and positive attitudes toward novel cancer treatments were significantly associated with an increased willingness to participate in clinical trials (p < 0.05).<h4>Conclusion</h4>This study provides critical insights into the motivation of and barriers to participation in clinical trials for novel cancer treatments. Addressing barriers and promoting motivations can contribute to supporting cancer clinical trial conductions and then improving cancer treatment effectiveness in Vietnam.

Understanding diseases as the result of continuous transitions from a healthy system is more realistic than understanding them as discrete states. Here, we designed the spectrum formation approach (SFA), a machine learning-based method that extracts key features contributing to disease state continuity. We applied the SFA to transcriptomic data from patients with progressive liver disease and neurodegenerative movement disorders to examine its effectiveness in identifying biologically relevant gene sets. The SFA identified transcription factors that potentially regulate liver inflammation and voluntary movement. In neurodegenerative disorders, the SFA also identified genes regulated by ETS-1, with unclear effects on movement. In functional assessment using human iPSC-derived neurons, ETS-1 overexpression disrupted dopamine receptor balance, reduced GABA-producing enzyme levels, and promoted cell death. These findings suggest that the SFA enables the discovery of regulatory factors capable of modifying disease states and provides a framework for the continuity-based interpretation of biological systems.

High-dose copper (Cu) supplementation is used as an alternative to antibiotic growth promoters for improving gut health in broiler chickens. The aim of this study was to evaluate the effects of hydroxychloride Zn (HyZ) and Cu (HyC) at different inclusion levels on productive traits and gut health biomarkers in broiler chickens. A total of 990 Ross 308 broilers were assigned to 55 floor pens (18 birds per pen) and received five dietary treatments as follows: (T1) an inorganic diet (INO) with 15 mg/kg Cu from CuSO₄ and 100 mg/kg of Zn from ZnSO₄; (T2) a hydroxychloride diet with 15 mg/kg of Cu from HyC and 100 mg/kg of Zn from HyZ; (T3 and T4) hydroxychloride diets with 80 mg/kg Zn from HyZ and either 100 (T3) or 150 (T4) mg/kg of Cu from HyC; and (T5) a hydroxychloride diet containing 80 mg/kg Zn from HyZ and Cu from HyC supplied at 200 mg/kg in the starter phase (1-10 days), 100 mg/kg in the grower phase (10-24 days), and 60 mg/kg in the finisher phase (24-35 days). Gut integrity and cecal bacterial populations were assessed on day 21, while carcass composition, liver, tibia and excreta mineral profiles, and tibia strength were evaluated on day 35 of the experiment. Hydroxychloride diets significantly improved body weight by ~ 4% at day 35 compared to INO (P < 0.05). Feed conversion ratio improved by 2.5% with T2 over INO, with further improvements in T3 (3.8%) and T4 (3.2%) (P < 0.05). Birds on T5 performed similarly to T3 throughout the trial (P > 0.05). INO birds had the lowest breast meat yield and the highest abdominal fat percentage (P < 0.05). Hydroxychloride diets reduced serum FITC-d levels and cecal Enterobacteria counts while increasing tibia breaking strength, Zn, and P levels (P < 0.01), and tended (P = 0.06) to increase cecal Bifidobacteria populations. In conclusion, these results suggest that replacing ZnSO₄ and CuSO₄ with HyZ and HyC has the potential to enhance gut health, body weight gain, and feed efficiency in broiler chickens.

Kabuki syndrome type 1 is a congenital multisystem disorder caused by KMT2D mutations. While some studies suggest that KMT2D deficiency may lead to autistic-like behaviors, the role of KMT2D in social behavior remains unconfirmed due to a lack of animal model evidence. In this study, we developed a mouse knockdown model and a zebrafish knockout model to investigate the role of KMT2D in synaptic function and behavioral patterns. We also performed an RNA sequencing analysis to delve into the molecular and cellular mechanisms of KMT2D. Results showed that Kmt2d deficiency in mouse and zebrafish both exhibited autistic-like behaviors including social behaviors defects and repetitive behavior. Additionally, knockdown of Kmt2d in the mouse hippocampus decreases excitatory and increases inhibitory synaptic transmission, disrupting the excitation-inhibition balance-a hallmark of autistic-like behaviors. RNA sequencing analysis revealed that under conditions of low kmt2d expression, differentially expressed genes were associated with glutamatergic and GABAergic synapses, supporting the dysregulation of excitation-inhibition balance in the hippocampus. Taken together, our research elucidates the critical role of KMT2D in modulating animal social behavior, potentially through its impact on synaptic excitation-inhibition balance.

Immune cells play a central yet poorly understood role in metabolic dysfunction-associated steatotic liver disease and metabolic dysfunction-associated steatohepatitis (MASLD/MASH), a global cause of liver disease with limited treatment. Limited access to human livers and lack of studies across MASLD/MASH stages thwart identification of stage-specific immunological targets. Here we provide a unique single-cell RNA sequencing atlas of paired peripheral blood and liver fine-needle aspirates from a full-spectrum MASLD/MASH human cohort. Our findings included heightened immunoregulatory programs with MASH progression, such as enriched hepatic regulatory T cells, monocytic myeloid-derived suppressor cells, TREM2⁺S100A9⁺ macrophages and S100^hiHLA^lo type 2 conventional dendritic cells. Hepatic cytotoxic T cell functions increased with inflammation, but decreased with fibrosis, while acquiring an exhausted signature, whereas natural killer cell-driven toxicity intensified. Our dataset proposes immunological mechanisms for increased fibrogenesis and vulnerability to liver cancer and infections in MASH and provides a basis for a deeper understanding of human immunological dysfunction in chronic liver disease and a roadmap to new targeted therapies.

Hypoxia is a key feature of solid tumors, contributing to therapeutic resistance and poor prognosis. Targeting hypoxic environments presents an opportunity to enhance drug delivery selectivity and improve treatment outcomes. Among nanocarriers, mesoporous silica nanoparticles (MSNs) have demonstrated great potential for drug delivery; however, achieving precise control over drug release remains a challenge. In this work, we develop an enzyme-responsive MSN system for targeted drug delivery in hypoxic tumors. MSNs were loaded with the chemotherapeutic drug doxorubicin (Dox) and capped with an azobenzene (Azo) gatekeeper. The bulky Azo group acts as a responsive molecular gate that remains closed under normoxic conditions but undergoes enzymatic cleavage by azoreductases, which are overexpressed in hypoxic tumor microenvironment. This cleavage triggers the uncapping of the pores, inducing the release of Dox specifically in the hypoxic region, minimizing premature drug leakage and off-target toxicity. In vitro studies with A549 cells (which overexpress azoreductase) and THP-1 cells (with low expression of azoreductases) under normoxic and hypoxic conditions, demonstrated a significant increase in Dox release and cytotoxicity in the A549 cells compared with the THP-1, which was more pronounced under hypoxia. These findings highlight the potential of enzyme-responsive MSNs as a promising strategy for selective drug delivery in hypoxic tumors.

Understanding the genomic basis of human proteomic variability provides powerful tools to probe potential causal relationships of proteins and disease risk, and thus to prioritise candidate drug targets. Here, we investigated 6432 plasma proteins (1533 previously unstudied in large-scale proteomic GWAS) using the SomaLogic (v4.1) aptamer-based technology in a Scottish population from the Viking Genes study. A total of 505 significant independent protein quantitative trait loci (pQTL) were found for 455 proteins in blood plasma: 382 cis- (P < 5×10^-8) and 123 trans- (P < 6.6×10^-12). Of these, 31 cis-pQTL were for proteins with no previous GWAS. We leveraged these pQTL to perform causal inference using bidirectional Mendelian randomisation and colocalisation against complex traits of biomedical importance. We discovered 43 colocalising associations (with a posterior probability > 80% that pQTL and complex traits share a causal variant), pointing to plausible causal roles for the proteins. These findings include hitherto undiscovered causal links of leukocyte receptor tyrosine kinase (LTK) to type-2 diabetes and beta-1,3-glucuronyltransferase (B3GAT1) to prostate cancer. These new connections will help guide the search for new or repurposed therapies. Our findings provide strong support for continuing to increase the number of proteins studied using GWAS.

Exosomes derived from tumors are critical agents in intercellular communication and the tumor microenvironment, offering a rich source of signatures for renal cell carcinoma (RCC) diagnosis. Conventional diagnostic techniques often suffer from limited sensitivity and can be invasive. This study presents an innovative approach using near-infrared (NIR) digital PCR (dPCR) with black phosphorus-embedded gelatin microcarriers for profiling exosomal miRNAs and modulating STAT3 signaling and macrophage polarization. Microcarriers produced via microfluidics, characterized by their phase-change and photothermal properties, are subjected to thermal cycling using a custom NIR source. The study identified a 4.2-fold increase in miR-210 levels in RCC cells (ACHN and A498) compared to normal cells (HK-2), with miR-126 and miR-30c levels decreasing by 7-9 times. Additionally, the method achieved a 20-fold enrichment of miRNA-34 in exosomes, leading to reduced STAT3 expression and decreased M2 macrophage polarization after co-incubation. This pioneering dPCR method provides a robust tool for early RCC detection through exosomal miRNA profiling and opens new avenues for therapeutic exosome engineering. The study underscores the potential of dPCR-based exosome genotyping in identifying cancer biomarkers and developing novel treatment strategies.

<h4>Background</h4>The intestine is one of the most sensitive organs to ionizing radiation (IR), and radiation-induced intestinal injury (RIII) impacts the quality of life of patients undergoing radiotherapy. There are limited early diagnostic biomarkers and specific medicines clinically approved for RIII. Therefore, we sought to identify new theranostic targets to prevent RIII and to facilitate the reestablishment of the intestinal epithelium. Circular RNAs (circRNAs) are widely appreciated as pervasive regulators of many diseases and multiple biological processes, while whether and how specific circRNAs are involved in radiation-induced intestinal injury remains largely unknown.<h4>Methods</h4>Differentially expressed circRNAs were analyzed and verified via RNA sequencing. The function of an intestine-specific circRNA (termed circDmbt1(3,4,5,6)) on cell proliferation, apoptosis, and DNA damage level after radiation was explored in vitro, and the underlying mechanism was further investigated. Ultimately, intestinal organoids and mice model were used to verify the role of circDmbt1(3,4,5,6) on radiation-induced intestinal injury.<h4>Results</h4>Primarily expressed in intestinal stem cells, CircDmbt1(3,4,5,6) was downregulated in mice intestines after 14 Gy abdominal radiation and showed timely relationship with intestinal injury level. CircDmbt1(3,4,5,6) promoted the proliferation and alleviated cell apoptosis and DNA damage level of intestinal epithelial cells and promoted organoids survival after radiation compared with control groups. In vivo experiments showed that compared with control groups, overexpression of circDmbt1(3,4,5,6) could increase intestinal length; enhance epithelial integrity and the percentage of proliferative cells, stem cells, paneth cells, and goblet cells; and promote intestinal adaption after radiation. Mechanistically, circDmbt1(3,4,5,6) protects intestines from IR via circDmbt1(3,4,5,6)/miR-125a-5p/STAT3.<h4>Conclusions</h4>CircDmbt1(3,4,5,6), a novel promising RIII bio-marker, responses rapidly at the early stage after 14 Gy abdominal irradiation, and exogenous expression of circDmbt1(3,4,5,6) could promote intestinal fitness in RIII. We reveal that the circDmbt1(3,4,5,6)/miR-125a-5p/STAT3 axis is important to the regeneration of the intestinal epithelium after radiation-induced damage, providing a potential diagnostic and therapeutic target for RIII.

<h4>Background</h4>Celiac disease is associated with HLA-risk haplotypes, but non-HLA genes and environmental factors are also linked to disease susceptibility. In this study, we explore the molecular pathways involved in celiac disease by analyzing the differential expression of genes in both the gut and peripheral blood across various celiac disease phenotypes.<h4>Methods</h4>Whole genome RNA sequencing was performed on 283 samples from intestinal mucosa and peripheral blood from 72 cases with either active, potential, or treated celiac disease and 73 disease controls. Enrichr pathway analysis of top differentially expressed genes was performed.<h4>Results</h4>Overall, 7565 genes in intestinal biopsies and 542 genes in blood samples were differentially expressed between cases and controls. Compared with controls, immunoglobulin heavy variable 5-51 (IGHV5-51) (p = 1.05 × 10^-14) and tissue transglutaminase (TGM2) (p = 5.29 × 10^-10), encoding for TG2, the main autoantigen in celiac disease, were two of the top up-regulated genes in intestinal biopsies from celiac cases. TGM2 was also slightly upregulated in blood cells from cases with active disease compared with controls (p = 0.05). The topmost differentially expressed genes in peripheral blood were HLA-DQB1, HLA-DQB2, and GSTM1. Among pathways identified containing transcriptionally differentiated genes were antioxidant defense systems (e.g., nuclear factor (erythroid-derived 2)-like 2 (Nrf2), glutathione, ergothioneine, and peroxisome metabolism), as well as MHC class 1 antigen presentation, amino acid transport, mTORC1, bilirubin and lipid metabolism, liver homeostasis, the complement system, and interferon signaling.<h4>Conclusions</h4>Differentially expressed genes in cases and controls indicate crosstalk between molecular pathways involved in antioxidant defense, immune regulation, and nutrient signaling in the pathogenesis of celiac disease.

<h4>Objective</h4>Over the past decade, significant advances in genetic testing for ataxia have improved diagnostic accuracy, informed clinical trial eligibility, guided treatment decisions, and enabled cascade testing of at-risk relatives. While guidance exists for other neurogenetic conditions, there are no standardized guidelines on genetic counseling and testing for individuals with unexplained ataxia.<h4>Methods</h4>We conducted a comprehensive literature review on genetic counseling and testing in ataxia, identifying 7362 articles. After removing 2971 duplicates, 4391 articles were screened by two authors using the Evaluation of Genomic Applications in Practice and Prevention (EGAPP) framework. In areas lacking clear published evidence, we convened a multidisciplinary expert panel with clinical and genetic expertise in ataxia. Following conflict resolution and additional filtering, 68 articles were included in our guidance development.<h4>Results</h4>Based on this evidence and expert consensus, we developed 20 recommendations addressing indications for genetic testing in hereditary ataxia, components of pre- and post-test counseling, testing options, insurance considerations, interpretation of test results, and appropriate referral to genetic counseling services. Major themes include the importance of formal genetic counseling and suggesting whole genome sequencing as first-line testing, with an emphasis on detecting repeat expansions.<h4>Conclusion</h4>These evidence-based, consensus-driven recommendations aim to support clinicians in evaluating patients with unexplained ataxia in order to provide timely evaluation and care, both for patients and their at-risk relatives.

The 2nd MObility for Vesicle research in Europe (MOVE) Symposium, held in Belgrade-Serbia, from October 8 to 11, 2024, showcased the dynamic and interdisciplinary nature of extracellular vesicles (EVs) research in Europe. Organized by eight National EV Societies under the MOVE initiative, the event gathered over 280 attendees from 28 countries, promoting collaboration and scientific exchange. The symposium featured eight keynote lectures, 48 oral and 126 poster presentations, and sessions dedicated to EV-related tools and industry innovations. The scientific program was structured around seven core themes: EV biogenesis and signal transmission, roles of EVs in health and disease, EV-based biomarkers, interspecies communication, novel EV preparation and analysis techniques, therapeutic and regenerative applications, and the manufacturing of native and engineered EV products. Supported by 18 sponsors and the Ministry of science, technological development and innovation of the Republic of Serbia, the symposium also highlighted the MOVE Fellowship Program and offered rich networking opportunities. This landmark event reinforced MOVE's promising mission to promote excellence, mobility, and resource sharing in EV research across Europe.

NUTM1-rearranged (NUTM1-R) infant acute lymphoblastic leukaemia (ALL) is a newly identified subgroup of non-KMT2A-R infant ALL, with ACIN1::NUTM1 the most frequent fusion. KMT2A-R and NUTM1-R infant ALL are characterized by fewer copy number alterations. Moreover, the gene expression profile in NUTM1-R infant ALL characteristically reveals upregulation of the genes that are involved in KMT2A-R infant ALL development, including HOXA9 and HOXA10. However, the direct association of NUTM1-fusion with this gene expression remains unexplored. Clinically, in sharp contrast to KMT2A-R infant ALL, the prognosis of NUTM1-R infant ALL is excellent, although its drug sensitivity profile remains unclear. Here, we newly identified an ACIN1::NUTM1-positive ALL cell line, KOPN32, which was previously established from a relapsed infant-ALL case, as the only cell line with NUTM1-fusion. KOPN32 had fewer copy number alterations, like KMT2A-R ALL cell lines. Both KOPN32 and an ACIN1::NUTM1-inducible ALL model, which was established using the ACIN1::NUTM1 fusion cDNA subcloned from KOPN32, revealed upregulation of HOXA9, HOXA10, SKIDA1 and BMI1, indicating direct involvement of ACIN1::NUTM1 fusion in the upregulation of these genes. In the drug sensitivity to eight standard agents, KOPN32 showed high sensitivity to both daunorubicin and vincristine; both are crucial agents in the induction therapy for infant ALL.

Neuronal growth regulator 1 (NEGR1) has been identified as a critical risk factor for major depressive disorders in humans. Although NEGR1 is predominantly expressed in the brain, its deletion in mice (Negr1^-/-) results in abnormalities in peripheral tissues, suggesting a role beyond the nervous system, particularly in intracellular lipid trafficking. However, the role of NEGR1 in testosterone production has not yet been elucidated. Here, we demonstrate that Negr1^-/- mice exhibit significantly reduced serum and testicular testosterone levels, accompanied by diminished male reproductive behaviors. The expression of key testosterone-synthesizing enzymes was downregulated in Leydig cells, and histological analysis revealed disorganized testicular and epididymal structures with lipid droplet accumulation in testicular cells. Additionally, Negr1^-/- mice displayed a significant increase in abnormal sperm morphology. Notably, testosterone supplementation alleviated their impaired sexual behaviors and mitigated anxiety- and depression-like phenotypes. These findings highlight a crucial role for NEGR1 in testicular function, particularly in testosterone production and spermatogenesis, underscoring the intricate link between hormonal balance and mental health.

Vulvar diseases are a neglected area of women's health, profoundly affecting patients' QOL. Lichen sclerosus is a chronic inflammatory vulvar skin disorder leading to severe itching, pain, scarring, and an increased risk of malignancy. Despite this burden, the molecular pathogenesis of vulvar lichen sclerosus is not well-understood, limiting treatment options. In this study, we analyze lesional, nonlesional, and healthy vulvar skin using technologies including spatial and single-cell transcriptomics. Our findings identify unifying molecular changes across multiple cell types in lesional vulvar lichen sclerosus skin, including keratinocyte stress response, necroptosis, and basal/stem cell depletion. Chronic T-cell activation, enhanced cytotoxicity, aberrant cell-cell communication, and elevated IFN-γ/JAK/signal transducer and activator of transcription signaling were also observed. Functional studies suggest keratinocytes' dual role as both targets of microenvironmental signaling (eg, IFN-γ) and sources of inflammatory alarmins (eg, S100A8/9). This work reveals keratinocytes as central players in vulvar lichen sclerosus pathogenesis and identifies potential biomarkers and therapeutic targets for future research.

<h4>Background</h4>Liver transplantation (LT) in patients aged ≥70 years is feasible with selected donors.<h4>Background</h4>Short-term outcomes were comparable to those in younger recipients.<h4>Background</h4>Elderly patients had higher intensive care unit (ICU) stay and transfusion needs.<h4>Background</h4>Advanced age should not be a contraindication for LT when carefully evaluated.<h4>Background</h4>A retrospective analysis of liver transplants was performed, comparing patients over and under 70 years of age. The elderly group was transplanted with careful donor selection and obtained results comparable to those of the younger group.<h4>Background</h4>This study aims to show that elderly patients over 70 years of age can have good results after liver transplantation, comparable to patients under 70 years of age, with good donor selection and perhaps additional points to favor their position on the waiting list.<h4>Background</h4>Liver transplantation (LT) is increasingly recognized as a treatment option for various diseases affecting a growing elderly population. However, its use in patients over 70 years of age remains controversial in centers with suboptimal outcomes or high waitlist mortality.<h4>Aim</h4>The aim of this study was to evaluate the effectiveness of LT as a treatment option for elderly patients aged 70 years or older, in comparison with younger recipients.<h4>Methods</h4>This retrospective study was conducted based on medical record data from 309 liver transplant recipients treated by the same surgical team across three hospitals - two located in São Paulo, São Paulo state (SP) and one in Rio Branco, Acre state (AC). Patients were divided into two groups for comparison: those aged up to 69 years (Group I) and those aged 70 years or older (Group II).<h4>Results</h4>Donor characteristics were similar between the two groups, except for a higher norepinephrine dose in Group I (p<0.05). Group II showed greater transfusion requirements and longer intensive care unit (ICU) stays (p<0.05), as well as higher rates of malnutrition and comorbidities. Notably, 90-day survival was comparable between the groups.<h4>Conclusions</h4>Patients aged 70 years or older can achieve outcomes comparable to those of younger recipients, provided they receive grafts from carefully selected donors. This population should not be excluded from transplant waitlists, and specific allocation policies or scoring adjustments should be considered to ensure equitable access.

<h4>Introduction</h4>Adrenocortical carcinoma (ACC) is a rare malignancy with poor prognosis, limited treatment options, and high recurrence rates. Surgery and mitotane-based chemotherapy remain the standard of care, and new treatment strategies are needed. Iron oxide nanoparticles (IONPs) offer promise as theranostic agents due to their modifiability for selective uptake and imaging.<h4>Methods</h4>We investigated the uptake, toxicity, and impact on steroidogenesis of dopamine-coated Fe/Fe₃O₄ core-shell IONPs in three ACC cell lines (H295R, HAC-15, and MUC-1). Uptake was assessed using flow cytometry, confocal microscopy, and TEM. A multicellular transwell model including human endothelial cells (HUVEC) and primary monocytes was used to simulate physiological barriers to delivery.<h4>Results</h4>IONP uptake by ACC cells was concentration- and time-dependent, with optimal uptake at 10 µg/mL. Nanoparticles localised primarily to the cytoplasm and vesicular compartments. At this concentration, IONPs did not impair ACC cell viability, proliferation, metabolic activity, or forskolin/angiotensin II-stimulated steroidogenesis. Higher concentrations (≥20 µg/mL) led to aggregation and reduced viability in some cell lines. In the transwell model, primary monocytes and endothelial cells also avidly absorbed IONPs, reducing nanoparticle availability to ACC cells.<h4>Conclusion</h4>ACC cells actively internalise IONPs without significant impairment of viability or steroidogenesis at pharmacologically relevant concentrations. However, non-specific uptake by monocytes and endothelial cells reduces delivery efficiency. These findings highlight the need for strategies to enhance tumour-specific targeting and improve biodistribution in future theranostic applications.

<i>HTT1a</i> was identified in human and mouse Huntington's disease brain as the pathogenic exon 1 mRNA generated from aberrant splicing between exon 1 and 2 of <i>HTT</i> that contributes to aggregate formation and neuronal dysfunction. Detection of the huntingtin exon 1 protein (HTT1a) has been accomplished with Meso Scale Discovery, Homogeneous Time Resolved Fluorescence and immunoprecipitation assays in Huntington's disease knock-in mice, but direct detection in homogenates by gel electrophoresis and western blot assay has been lacking. Subcellular fractions prepared from mouse and human Huntington's disease brain were separated by gel electrophoresis and probed by western blot with neoepitope monoclonal antibodies 1B12 and 11G2 directed to the C-terminal eight residues of HTT1a. In caudate putamen of an allelic series of 6-month-old Huntington's disease knock-in mice (Q50, Q80, Q111, Q140 and Q175), HTT1a migration was inversely correlated with CAG repeat length and appeared as a sodium dodecyl sulphate soluble high molecular mass smear in Q111, Q140 and Q175 mice but weakly in Q80 and not in wild-type mice or Q50 indicating a CAG repeat size threshold for detecting HTT1a. HTT1a immunoreactivity diminished if 1B12 and 11G2 antibodies were preincubated with an eight amino acid peptide containing the C-terminus of HTT1a but not with an unrelated peptide sequence. Migration of HTT1a and its high molecular mass smear changed with age in caudate putamen of Q111, Q175 and YAC128 mice. Reducing levels of MutS Homolog 3 (MSH3) protein >84% in Q111 mice caudate putamen with small interfering RNA to <i>MSH3</i>, a modifier of CAG repeat expansion, significantly reduced levels of the high molecular mass smear suggesting that the effects of curbing CAG repeat expansion on HTT1a were quantifiable. A prominent 56-60 kDa doublet detected by 1B12 and 11G2 antibodies in lysates from human Huntington's disease brain was not blocked by preincubation with C-terminal HTT1a blocking peptide and also appeared in brains of Parkinson's disease patients. 1B12 and 11G2 antibodies did not immunoprecipitate huntingtin (HTT) proteins from either Huntington's disease mouse or human brain lysates using conditions that pulled down full-length HTT with anti-HTT antibody 2B7. Altogether, these data show that 11G2 and 1B12 antibodies can be used in western blot assays to track and quantify immunoreactive HTT1a levels, solubility and subcellular localization in Huntington's disease mouse brain.

Left ventricular noncompaction (LVNC), also called noncompaction cardiomyopathy (NCM), is a myocardial disease that affects children and adults. Morphological features of LVNC include a noncompacted spongiform myocardium due to the presence of excessive trabeculations and deep recesses between prominent trabeculae. Incidence and prevalence rates of this disease remain contentious due to varying clinical phenotypes, ranging from an asymptomatic phenotype to fulminant heart failure, cardiac dysrhythmias, and sudden death. There is a strong genetic component associated with LVNC, and nearly half of pediatric LVNC patients harbor an identifiable genetic mutation. Recent studies have identified LVNC-associated mutations in genes involved in intercellular trafficking and cytoskeletal integrity, in addition to well-known mutations causing abnormal cardiac embryogenesis. Currently, the diagnosis is based on symptoms, as well as various diagnostic criteria, including echocardiography, electrocardiograms, and cardiac magnetic resonance imaging. Meanwhile, clinical management is primarily focused on the prevention of complications, such as heart failure, thromboembolic events, life-threatening arrhythmias, and stroke. Continued research is focusing on the genetic etiology, the development of gold-standard diagnostic criteria, and evidence-based treatment guidelines across all age groups. This review article will highlight the genotype-phenotype relationship within pediatric LVNC patients and assess the latest discoveries in genetic and molecular research aimed at improving their diagnostic and therapeutic management.

Autophagy is a fundamental catabolic pathway critical for maintaining cellular homeostasis in the central nervous system (CNS). While neuronal autophagy has been extensively studied, growing evidence highlights the crucial roles of astrocytic autophagy in CNS physiology and pathology. Astrocytes regulate metabolic support, redox balance, and neuroinflammatory responses. These functions are closely linked to autophagic activity. The disruption of astrocytic autophagy contributes to synaptic dysfunction, chronic inflammation, myelin impairment, and blood-brain barrier instability. Dysregulation of astrocytic autophagy has been implicated in the pathogenesis of multiple neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. This review summarizes the molecular mechanisms of autophagy in astrocytes and delineates its role in intercellular communication with neurons, microglia, oligodendrocytes, and endothelial cells. Furthermore, we will discuss current pharmacological approaches targeting astrocytic autophagy, with particular attention to repurposed agents such as rapamycin, lithium, and caloric restriction mimetics. Although promising in preclinical models, therapeutic translation is challenged by the complexity of autophagy's dual roles and cell-type specificity. A deeper understanding of astrocytic autophagy and its crosstalk with other CNS cell types may facilitate the development of targeted interventions for neurodegenerative diseases.

<h4>Background</h4>Ovarian cancer is an immunologically cold tumor that is treated with surgery and a chemotherapy regimen of platinum agents with taxanes. Paradoxically, elevated levels of several immune markers are effective at predicting prognosis for patients with ovarian cancer, though it is not clear how chemotherapy might influence this. Chemotherapy elicits immunogenic cell death, yet tumor-controlling doses of chemotherapy are also immunotoxic.<h4>Objectives</h4>To evaluate interactions of chemotherapy with the immune system, we studied the impact of chemotherapy in an aggressive mouse model of ovarian cancer developed within our lab.<h4>Methods</h4>Using a single-cell transcriptomics sequencing approach, supported by flow cytometry, we evaluated the influence of a first-line therapy, cisplatin and docetaxel, and a second-line therapy, pegylated liposomal doxorubicin (PLD), on control of tumor growth and on tumor-associated immune populations of cells.<h4>Results</h4>Both chemotherapy approaches were effective at controlling tumor growth and selectively depleted tumor cells from distinct transcriptional clusters. Both chemotherapies also resulted in relative increases in immune populations compared to untreated tumor-bearing mice, but immune populations from PLD-treated mice were more abundant and expressed a greater fraction of maturity-associated transcripts and increased proportions of tumor resident macrophage populations. PLD treatment selectively upregulated MHC class II on tumor cells, and this could be replicated in vitro across ovarian cancer cell lines and in patient tumor cells ex vivo.<h4>Conclusions</h4>Altogether, the results support the notion that PLD has a greater capacity for immunopotentiation, which may be important to consider if immunotherapy approaches are adapted for ovarian tumors in the future.

<b>Background:</b> Irradiation (IR) targets cancer cells, but also the tumor microenvironment, including the tumor's blood vessels. In addition to tumor endothelial cell (TEC) apoptosis, IR can lead to TEC activation, potentially increasing immune cell infiltration. However, the changes underlying the IR-induced activation of endothelial cells (ECs) are poorly understood. This study investigated dose- and time-dependent molecular and functional responses of murine and human EC lines to IR in vitro and TECs in vivo in murine tumor models of colorectal carcinoma. <b>Methods:</b> HUVEC, EA.hy926, and Hulec5a, as well as murine bEND.3, 2H11, and SVEC4-10 EC lines, were irradiated with single doses of 2-10 Gy. EC proliferation and survival after IR were assessed by staining all nuclei (Hoechst 33342) and dead cells (propidium iodide) every 24 h for 5 days using the Cytation 1 Cell Imaging Multi-Mode Reader. RNA sequencing analysis of HUVECs irradiated with 2 Gy and 5 Gy at 24 h and 72 h after IR was conducted, focusing on processes related to EC activation. To validate the RNA sequencing results, immunofluorescence staining for proteins related to EC activation, including Stimulator of Interferon Response cGAMP Interactor 1 (STING), Nuclear factor kappa B (NF-κβ), and Vascular cell adhesion molecule 1 (VCAM-1), was performed. To validate the in vitro results, the response of TEC in vivo was analyzed using publicly available RNA sequencing data of TECs isolated from MC38 colon carcinoma irradiated with a single dose of 15 Gy. Finally, murine CT26 colon carcinoma tumors were immunofluorescently stained for STING and NF-κβ 24 and 48 h after IR with a clinically relevant fractionated regimen of 5 × 5 Gy. <b>Results:</b> Doses of 2, 4, 6, 8, and 10 Gy led to a dose-dependent decrease in proliferation and increased death of ECs. RNA sequencing analysis showed that the effects on the transcriptome of HUVECs were most pronounced 72 h after IR with 5 Gy, with 1014 genes (661 down-regulated and 353 up-regulated) being significantly differentially expressed. Irradiation with 5 Gy resulted in HUVEC activation, with up-regulation of the immune system and extracellular matrix genes, such as <i>STING1</i> (log₂FC = 0.81) and <i>SELE</i> (log₂FC = 1.09), respectively; and down-regulation of cell cycle markers. Furthermore, IR led to the up-regulation of immune response- and extracellular matrix (ECM)-associated signaling pathways, including NF-κβ signaling and ECM-receptor interaction, which was also observed in the transcriptome of irradiated murine TECs in vivo. This was confirmed at the protein level with higher expressions of the EC activation-associated proteins STING, NF-κβ, and VCAM-1 in irradiated HUVECs and irradiated TECs in vivo. <b>Conclusions:</b> IR induces changes in ECs and TECs, supporting their activation in dose- and time-dependent manners, potentially contributing to the anti-tumor immune response, which may potentially increase the infiltration of immune cells into the tumor and thus, improve the overall efficacy of RT, especially in combination with immune checkpoint inhibitors.

Circular RNAs (circRNAs) are increasingly recognized as regulators of gene expression, although their roles in hematopoietic differentiation remain relatively understudied. This study compares circRNA expression profiles between erythroblasts derived from human fetal liver and bone marrow CD34⁺ hematopoietic stem cells using publicly available RNA-seq datasets (GEO: GSE90878). Twelve samples from each developmental source were analyzed. Differential expression analysis was performed, and circAtlas 3.0 was employed to predict interactions between circRNAs, microRNAs (miRNAs), and RNA-binding proteins. Differentially expressed miRNAs were curated from miRNA-seq data (GEO: GSE110936) profiling the same cell types. Principal component analysis of circRNA expression profiles demonstrated clear separation between erythroblasts from fetal liver and bone marrow, which was statistically confirmed by PERMANOVA (<i>p</i> = 0.001); though this effect size is small (R² = 0.065). One circRNA, circALS2(4).1, was significantly upregulated in bone marrow-derived erythroblasts (adjusted <i>p</i> < 0.05), and ten additional circRNAs showed suggestive evidence for differential expression (adjusted <i>p</i> < 0.1). The resulting interaction networks reveal distinct circRNA landscapes and suggest regulatory circuits that may contribute to developmental differences in human erythropoiesis, indicating that the functions of circRNAs in hematopoietic development remain to be further elucidated.

Cyclin-dependent kinase-like 5 (CDKL5) deficiency disorder, which is a developmental and epileptic encephalopathy occurring in 1 in every 40,000 to 60,000 live births, was the subject of this computational investigation. This study provided a comprehensive list of missense variants (156) seen in the human population within the CDKL5 protein. Furthermore, the list of CDKL5 binding partners was updated to include four new entries. Computational modeling resulted in 3D structure models of twenty-four CDKL5-target protein complexes. The CDKL5 stability changes upon the above-mentioned missense mutations that were modeled, and it was shown that the corresponding folding free energy changes (ΔΔG_folding) caused by pathogenic variants are much larger than the ΔΔG_folding caused by benign variants. The same observation was made for the binding free energy change (ΔΔG_binding). This resulted in a protocol that allowed for the reclassification of missense variants with unknown or conflicting significance into pathogenic or benign. It was demonstrated that such reclassification is more reliable than using leading tools for pathogenicity predictions, since the latter failed to correctly predict known pathogenic/benign variants. Furthermore, the study demonstrated that pathogenicity is linked with the disturbance of thermodynamics quantities such as ΔΔG_folding and ΔΔG_binding, paving the way for development of therapeutic solutions.

Artemisinin and its derivatives are widely recognized for their exceptional antimalarial efficacy. Recently, accumulating evidence indicates therapeutic potential beyond malaria. Despite these advances, detailed mechanisms and pharmacological limitations remain incompletely defined. This review summarizes their pharmacological activities and molecular mechanisms associated with oncology, immunoregulation, and metabolic disorders. Mechanistically, these compounds exert potent antitumor effects by inducing oxidative stress, arresting the cell cycle, triggering apoptosis, and inhibiting angiogenesis. They likewise modulate immune responses, re-establishing immune homeostasis and enhancing the effectiveness of immunotherapeutic strategies. Preliminary evidence also suggests involvement in metabolic regulation, pointing to promising avenues for treating metabolic disorders. Given alternative mechanisms of artemisinin and its derivatives, we also discuss the trinity modulation network among antitumor activity, immunoregulation, and metabolic homeostasis. We anticipate that future research will address these knowledge gaps, thereby enhancing the clinical utility of artemisinin and its derivatives and improving patient outcomes across diverse pathologies.

<h4>Background</h4>Lung cancer (LC) is the most common cause of cancer-related death worldwide, while there are limited treatment methods. Resveratrol (RESV), a natural food-derived compound, has attracted attention around the world for its anti-LC effects. However, little is known about the efficacy and safety of RESV for LC.<h4>Purpose</h4>This study aimed to provide preclinical evidence for the efficacy and safety of RESV for LC, and to find the optimal dose and duration.<h4>Methods</h4><i>In vivo</i> studies of RESV against LC, published before 24 July 2024, were retrieved from PubMed, Embase, Web of Science, and Cochrane Library. The CAMARADES checklist was used to assess study quality. Primary outcomes were tumor volume and tumor weight. Secondary outcomes included body weight, lung metastases number, and the apoptotic cell proportion. Statistical analysis was performed using RevMan 5.3 and Stata 16.0. Dose-duration-effect model was conducted to determine the optimal dose and duration, and the toxicology of RESV was predicted through the ProTox 3.0 platform.<h4>Results</h4>A total of 23 studies involving 425 animals were included. The methodological quality of included studies was medium-to-low. RESV significantly reduced tumor volume, tumor weight, and lung metastases number, and increased apoptotic cell proportion, while having no effect on body weight. High heterogeneity was observed, and subgroup analysis suggested that the heterogeneity was partly attributed to the dose of RESV. The optimal dose and duration of RESV were 30-100 mg/kg and 25-28 days, respectively. The median lethal dose of RESV was 1,560 mg/kg.<h4>Conclusion</h4>RESV demonstrated a significant inhibitory effect on LC <i>in vivo</i>. However, the lower research quality and high heterogeneity call for more high-quality preclinical studies to be conducted. Before achieving clinical translational research on RESV, the problem of low bioavailability of RESV needs to be solved.

Pathogenic bacteria of the genus <i>Brucella</i> cause a severe threat for public health and agricultural economics. The World Health Organization considers brucellosis to be one of the most serious and also neglected zoonotic diseases. The use of traditional whole-cell brucellosis vaccines complicates the differentiation between infected and vaccinated animals (DIVA). Moreover, diagnostics based on lipopolysaccharide of <i>Brucella</i> are susceptible to false positive results. Structural features of <i>Brucella</i> O-antigens make synthetic oligosaccharides promising agents for the development of diagnostic tools and vaccines against brucellosis. Here we report the synthesis of spacer-armed di-, tri-, tetra- and penta-4,6-dideoxy-4-formamido-α-(1→2)-d-mannopyranosides which are related to the A-epitope of <i>Brucella</i> O-antigen. The key α-(1→2)-linked disaccharide thioglycoside donor was synthesized by employing the strategy of orthogonal glycosylation of thioglycoside acceptor with trichloroacetimidate donor. Sequential block-wise assembly yielded a series of desired compounds, which were subsequently deprotected and converted into target molecules and then into their fluorescein-labeled conjugates. The obtained conjugates were employed as tracers in a fluorescence polarization assay (FPA) to detect anti-<i>Brucella</i> immunoglobulins. Among the studied compounds, the trisaccharide conjugate showed the greatest difference in median FP signals between <i>Brucella</i>-positive and <i>Brucella</i>-negative sera samples making it a promising candidate for developing FP diagnostic assays. The decreased FP signal in the cases of tetra- and pentasaccharide tracers can be associated with the known "propeller-effect" due to the rotational mobility of the part bearing the fluorescent label and of the fluorescein itself and/or the enlarging of the distance between the fluorescein part and the antibody-oligosaccharide complex. This observation demonstrates the advantages of using synthetic relatively small synthetic tracers with well-defined structure in comparison with heterogeneous fluorescein-labelled O-polysaccharides which are in use today in spite of the fact that they contain poorly characterized amounts of label attached along the polysaccharide chains.

The increasing prevalence of marital infertility and the persistent desire for offspring have become more significant issues over past decades. Considering the potential genetic, hormonal, and anatomical causes, it is evident that the analysis of infertility is complex, necessitating the development of innovative therapies to address various challenges and dilemmas. The interdisciplinary collaboration of multiple fields fosters scientific progress, such as the development of new research models, reproductive mini-organoids, enhancing the chances of successful parenthood even in challenging cases. Since the fifth decade of the 20th centurymarked by the <i>in vitro</i> fertilization of an egg cell, the birth of Louise Brown (the first test-tube baby), the methods of embryo cryopreservation, the discovery of induced pluripotent stem cells (iPSC), and the genetic editing technology CRISPR-Cas9-research has been advancing towards promising directions for studying infertility causes and testing potential therapeutic interventions in controlled conditions. Gene therapy stands as a significant pillar, with 2017 witnessing promising experimental advancements in repairing mutations responsible for hypertrophic cardiomyopathy. Attempts were also made to create Human Immunodeficiency Virus (HIV) immunity by disabling the CCR5 gene, leading to the birth of twins with this variation. Progress in innovative therapies has kept pace with advancements in artificial intelligence, poised to revolutionize reproductive medicine by minimizing human errors. Machine learning (ML) algorithms are being integrated into embryo selection processes, predicting their implantation potential, raising concerns among various nations about eugenics and the interference with human nature. These concerns form a highly debated legal and political pillar. The growing automation is driven by arguments related to the increasing problems of future challenges, such as environmental changes or declining gamete quality. Scenarios under consideration include the development of advanced assisted reproduction technologies and support programs. Theoretical possibilities of alternative methods for organism development are being explored, though they remain constrained by the necessity of rigorous human studies.

Vitamin C (VC), a pleiotropic molecule with context-dependent redox properties, exhibits dual roles in cancer biology through dose-dependent mechanisms. While nutritional VC intake demonstrates chemopreventive effects by scavenging carcinogen-induced reactive oxygen species (ROS) and maintaining genomic stability, high-dose intravenous VC acts as a prooxidant to selectively kill tumor cells via ROS-mediated deoxyribonucleic acid (DNA) damage, adenosine triphosphate (ATP) depletion, and HIF-1α degradation. Preclinical studies reveal VC's ability to reprogram the tumor microenvironment (TME) through collagen hydroxylation-mediated extracellular matrix remodeling, Treg suppression, and enhancement of CD8+ T cell infiltration. Importantly, VC synergizes with conventional therapies by radio-sensitizing hypoxic tumors through oxygen-sparing effects and reversing platinum resistance via glutathione depletion. Early-phase clinical trials corroborate VC's safety profile and potential to ameliorate chemotherapy-induced fatigue and nephrotoxicity. However, translational challenges persist, including the lack of pharmacokinetic standardization between oral and intravenous routes, tumor-type-specific response heterogeneity, and incomplete understanding of VC's immunomodulatory dynamics. Emerging strategies integrating VC with checkpoint inhibitors and TME-targeted nano-delivery systems show promise in preclinical models. This review synthesizes mechanistic insights from redox biology and immunometabolism to clinical trial data, proposing a framework for optimizing VC-based combination therapies while addressing critical gaps in biomarker development and dose scheduling. Deciphering the molecular determinants of VC's context-dependent anticancer effects may accelerate its rational clinical deployment.

Breast cancer brain metastasis (BCBM) remains fatal with elusive mechanisms. Here, we unveil the first circRNA m5C methylation landscape in BCBM through MeRIP-seq (methylated RNA immunoprecipitation next-generation sequencing) identifying 7465 BCBM-specific m5C peaks versus 5929 in primary breast cancer (BC). A total of 48 hypermethylated and 128 hypomethylated m5C sites in BCBM (231-BR) were identified compared to BC. Bioinformatics enrichment analysis revealed hypermethylated circRNAs enriched in ERBB/VEGF signaling pathways. Among 8 validated differentially methylated circRNAs, <i>hsa_circ_0004516</i> was consistently upregulated in BCBM tissues/cells and exhibited NSUN2-dependent m5C modification. Mechanistically, NSUN2-mediated m5C methylation enhanced <i>hsa_circ_0004516</i> stability, evidenced by significantly shortened half-life upon NSUN2 depletion. Crucially, catalytic mutant NSUN2 (C271A/C321A) abolished this effect. Functional assays demonstrated that <i>hsa_circ_0004516</i> knockdown in 231-BR cells suppressed proliferation, migration, and invasion by reducing p-AKT (Ser473) levels. The AKT activator SC79 reversed these phenotypic impairments, definitively linking <i>hsa_circ_0004516</i>-driven metastasis to AKT signaling activation. Our study establishes the NSUN2-m5C-<i>hsa_circ_</i>0004516-AKT axis as a novel therapeutic target and biomarker for BCBM.

<b>Background/Objectives:</b> The use of collagen-based scaffolds in dentition tissue engineering has gained significance and importance in the field as they are structurally equivalent and biologically compatible with the native extracellular matrix (ECM). In this review, collagen-composite scaffolds for pulp, alveolar bone, and periodontal regeneration are analyzed in terms of materials, fabrication techniques, and clinical outcomes. <b>Methods:</b> Recent developments in collagen scaffolds are highlighted in this review, with a focus on type I collagen due to its structural strength and arginine-glycine-aspartic acid (RGD) motifs, which promote cell adhesion and differentiation. Composite materials, freeze-drying, electrospinning, and 3D bioprinting, which are used to improve the functionality of the scaffold, are key developments. <b>Results:</b> This review shows progress in collagen-based scaffolds for restoring dental tissues, such as dentin, gingival tissue, or bone, in humans. Electrospinning and 3D bioprinting are new manufacturing techniques that enhance the functionality of scaffold devices, and incorporating bioactive molecules increases the regenerative capacity; however, stability and long-term efficacy are still problems. <b>Conclusions:</b> Although they have a lot of potential, collagen-composite scaffolds face challenges like rapid degradation and limited mechanical strength. To make long-lasting, tailored dental regeneration therapies feasible, future research needs to improve smart biomaterials, gene delivery, and personalized designs for dental regenerative therapy.

<h4>Background</h4>Deferred cord clamping (DCC) is beneficial for preterm infants, but there are concerns about the safety of DCC during Cesarean deliveries (CD) under general anesthesia (GA). We evaluated maternal and neonatal outcomes in preterm CD under GA vs. regional anesthesia (RA) after implementing 180 s of DCC.<h4>Methods</h4>This retrospective single-center observational study included CD at <33 weeks gestation, delivered between January 2018 and December 2023. The cord was clamped before 180 s for concerns of maternal bleeding or infant apnea after 30-45 s stimulation. Data was collected from reports from electronic medical records, neonatal intensive care unit database, and manually from the medical records of the patient. Multivariable regression analysis was used to assess the effect of anesthesia type and DCC on outcomes, adjusting for confounders.<h4>Results</h4>This study included 170 mothers and 194 infants, and 84.9% of the infants received DCC ≥ 60 s. The GA group had a higher percentage of emergency CD and a lower median duration of DCC (105 s vs. 180 s, <i>p</i> ≤ 0.001) compared to RA. In multivariate regression analysis, GA was associated with lower odds (95% CI) of umbilical artery pH < 7 [0.1, (0.0, 0.6)], base deficit ≥ 16 [0.0, (0.0, 0.5)], and higher odds of necrotizing enterocolitis [28.2, (1.4, 560.0)]. GA was not associated with maternal hemorrhage, delivery room (DR) resuscitation, or other major neonatal morbidities or mortality. DCC ≥ 60 s was associated with lower maternal blood loss [Regression coefficient -698, (-1193, -202)], lower odds of transfusion [0.4, (0.1, 1.0)], DR resuscitation [0.4, (0.2, 0.8)], and chronic lung disease [0.4, (0.2, 0.9)], and higher survival without major morbidities [2.8, (1.2, 6.8)].<h4>Conclusions</h4>DCC was performed in a majority of CD under GA by adhering to protocols to shorten DCC in cases where maternal or fetal safety was threatened. GA with DCC was not associated with increased neonatal resuscitation or major neonatal morbidities and was associated with lower maternal hemorrhage and transfusion.

Exercise addiction is described in the literature as a compulsive behavior associated with adverse health symptoms. Currently, knowledge about the biological and social factors that trigger the development of this behavior is still lacking, and there are no published studies on genetic variants associated with the disorder. Because of this, we genotyped specific polymorphisms in the genes <i>DRD1</i> (rs265981), <i>DRD2</i> (rs1800497), <i>BDNF</i> (rs6265), <i>HFE</i> (rs1799945), <i>ACTN3</i> (rs1815739), <i>PPARA</i> (rs4253778), <i>PPARGC1A</i> (rs8192678), and <i>AMPD1</i> (rs17602729) to investigate whether they were associated with exercise addiction. In total, 469 men and women, comprising athletes and non-athletes between the ages of 18 and 50, were enrolled in the study. Each participant provided an oral swab sample for genetic analysis and completed the Negative Addiction Scale questionnaire that tests for physical exercise addiction. For the <i>DRD2</i> polymorphism, there was a significant association of the <i>GG</i> genotype with asymptomatic participants and of the <i>AA</i> genotype with participants symptomatic for exercise addiction. Additionally, for the <i>BDNF</i> polymorphism, the <i>CC</i> genotype was associated with symptomatic participants, and the <i>T</i> allele was associated with asymptomatic individuals. However, all associations were found by evaluating the SNP individually, and this demonstrates the difficulty in studying variables related to behavioral phenotypes.

Mitochondrial aging plays a central role in the functional decline of the central nervous system (CNS), with profound consequences for neurological health. As the brain is one of the most energy-demanding organs, neurons are particularly susceptible to mitochondrial dysfunction that arises with aging. Key features of mitochondrial aging include impaired mitochondrial dynamics, reduced mitophagy, increased production of reactive oxygen species (ROS), and accumulation of mitochondrial DNA (mtDNA) mutations. These alterations dramatically compromise neuronal bioenergetics, disrupt synaptic integrity, and promote oxidative stress and neuroinflammation, paving the path for the development of neurodegenerative diseases. This review also examines the complex mechanisms driving mitochondrial aging in the central nervous system (CNS), including the disruption of mitochondrial-organelle communication, and explores how mitochondrial dysfunction contributes to neurodegenerative diseases, such as Alzheimer's, Parkinson's, Huntington's, and amyotrophic lateral sclerosis. By synthesizing current evidence and identifying key knowledge gaps, we emphasize the urgent need for targeted strategies to restore mitochondrial function, maintain cognitive health, and delay or prevent age-related neurodegeneration.

Condensin plays a central role in mitotic chromosome organization and segregation by mediating long-range chromatin interactions. However, the extent to which cellular metabolic status influences condensin function remains unclear. To gain insights into the relationship of metal ion homeostasis and the function of condensin, we conducted genome-wide chromatin immunoprecipitation sequencing (ChIP-seq) using Schizosaccharomyces pombe under iron- or zine-deficient conditions. Under iron- or zinc-deficient conditions, ChIP-seq results revealed a selective reduction in condensin binding at high-affinity target loci, particularly genes regulated by Ace2 and Ams2, while cohesin binding remained largely unaffected. Hi-C analysis showed that iron depletion weakened chromatin interactions at these condensin targets and centromeres, without disrupting global genome architecture. DNA fluorescence in situ hybridization (FISH) confirmed that iron deficiency impaired long-range associations between centromeres and Ace2 target loci at the single-cell level. Notably, iron deficiency led to chromosome segregation defects during mitosis, suggesting that diminished condensin occupancy compromised genome stability. These changes occurred without significant alterations in condensin protein levels or global transcription, indicating a direct effect of metal ion availability on condensin activity. Collectively, our findings revealed a previously unrecognized regulatory axis in which cellular metal ion homeostasis modulated condensin-dependent chromatin organization and mitotic chromosome segregation, offering new insights into the integration of metabolic state with genome maintenance.

Atherosclerosis is a major cause of cardiovascular disease, and accurate preclinical models are essential for developing effective therapies. The Ossabaw pig model has high potential due to its physiological similarity to humans, but its molecular characterization remains limited. To address this, we performed single-nucleus RNA sequencing of over 36,000 nuclei from aortas of Ossabaw pigs fed chow or atherogenic diets. Our analysis revealed activation of brain-derived neurotrophic factor (BDNF), transforming growth factor β (TGF-β), SPP1, and interleukin-2 (IL-2) signaling pathways in atherosclerosis, along with smooth muscle cell transitions to synthetic and pro-osteogenic states, endothelial-to-mesenchymal transition, and TREM2+ immune cell phenotypes. Advanced fibrotic, immune cell-infiltrated plaques with calcification sites paired with molecular changes closely resembled those seen in human atherosclerosis. Together, our findings establish the Ossabaw pig as a valuable translational model and provide high-resolution data to support its use in preclinical cardiovascular research.

MAP1B (microtubule-associated protein 1B) encodes a cytoskeletal regulator critical for neuronal migration, axon guidance, and cortical circuit formation. Disease-causing variants (DCVs) in MAP1B have recently emerged as a cause of neurodevelopmental disorders characterized by intellectual disability, epilepsy, and cortical malformations, including periventricular nodular heterotopia (PVNH) and polymicrogyria (PMG). However, the phenotypic and neuroimaging spectrum associated with MAP1B-related disease remains incompletely defined. We describe seven affected individuals from three unrelated families with pathogenic MAP1B variants. Clinical, neuroimaging, and genetic data were analyzed in the context of emerging literature to delineate the pathogenic mechanisms and phenotypic variability associated with MAP1B dysfunction. All individuals carried loss of function MAP1B variants. Clinical features included global developmental delay, intellectual disability, behavioural dysregulation, and focal epilepsy. Neuroimaging revealed anteriorly predominant PVNH in four of five cases with neuroimaging available. These findings reinforce MAP1B's central role in cytoskeletal regulation, neuronal positioning, and synaptic connectivity. Functional data from animal and cell models support a mechanism involving impaired microtubule stabilization, altered growth cone dynamics, and dysregulated axon branching. Our case series expands the clinical and radiological phenotype associated with MAP1B-related disorders and highlights its position as a key cytoskeletal regulator in human corticogenesis. Systematic genotype-phenotype correlation and functional studies are needed to inform diagnostic interpretation and explore therapeutic avenues in MAP1B-associated disease.

Periprosthetic joint infection (PJI) is the most common and difficult to treat form of arthroplasty failure. While treatment with debridement, antibiotics, and implant retention (DAIR) is preferable to one- or two-stage implant exchange based on morbidity and cost, outcomes are not successful in all cases selected for this management strategy. DAIR is currently recommended when infection is perceived to be in an "acute" phase, based on symptom duration; despite this selection strategy, DAIR failure rates are high. A more objective means of predicting DAIR success could improve patient outcomes by limiting DAIR failures. To explore this possibility, advanced proteomic analysis of sonicate fluid samples from 95 subjects with PJI was performed using proximity extension assay (PEA), with 77 samples additionally undergoing complementary proteomics analysis with liquid chromatography with tandem mass spectrometry (LC-MS/MS). Sparse partial least squares differentiation analysis (sPLS-DA) separated samples from patients who underwent DAIR with known outcomes (<i>n</i> = 19) into successful or unsuccessful groups, predicting DAIR surgical outcome (area under the curve [AUC]: 1; mean "leave one out" AUC: 0.6). Proteins predictive of DAIR surgical outcome were significantly enriched for the "protein repair" Gene Ontology (GO) pathway (<i>P</i>-adj = 0.02). Additionally, proteomic signatures associated with symptom duration in PJI categorized samples into short and long symptom duration PJI. In summary, an investigation of 7,261 unique human proteins separated PJI-associated samples based on symptom duration and identified a proteomic profile predictive of DAIR success.IMPORTANCEChronic infections are generally understood to last months to years, with acute infections lasting days to weeks. The transition from acute to chronic infection is, however, poorly understood. Periprosthetic joint infection (PJI) has been considered more challenging to treat when "chronic" than when "acute." A surgery preferred for its recovery time and cost-debridement, antibiotics, and implant retention (DAIR)-is recommended for PJI management when symptom duration is short; yet, even in this select patient group, DAIR is associated with a high treatment failure rate. A means to better identify those predicted to have a successful outcome if they undergo DAIR is needed. Here, over 7,000 proteins from human clinical PJI samples were measured and shown to be able to separate samples based on symptom duration. Importantly, a proteomic profile predictive of DAIR success was identified.

Ovarian cancer (OC) remains one of the deadliest gynecological malignancies. Immune checkpoint blockade (ICB) inhibitors efficacy in OC has been minimal, highlighting the need for a deeper understanding of the immune microenvironment in OC. Recent studies suggest that DNA methylation and transcription factors may influence the response to immunotherapy. This study aims to classify ovarian cancer into distinct immune subtypes by integrating DNA methylation and transcription factor data through comprehensive bioinformatics analysis. Using data from The Cancer Genome Atlas (TCGA), we identified twelve differentially methylated genes (DMGs) associated with transcription factors and categorized OC into two immune subtypes, C1 and C2.The C1 subtype exhibited higher levels of immune infiltration and better prognosis, characteristic of immune "hot" tumors, whereas the C2 subtype was associated with lower immune infiltration and poorer prognosis, indicative of immune "cold" tumors. A prognostic prediction model based on four key genes-KRT81, PAPPA2, FGF10, and FMO2-was developed using the least absolute shrinkage and selection operator (LASSO) and Cox regression analyses. This model effectively stratified the TCGA OC cohort into high- and low-risk groups and was validated by predicting patient survival outcomes. Additionally, drug sensitivity analysis revealed potential therapeutic targets for different risk groups, offering new avenues for precision treatment in ovarian cancer. Immunohistochemical tests confirmed the potential of KRT81 as a prognostic marker for ovarian cancer. Our findings enhance the understanding of the molecular characteristics of the OC immune microenvironment, propose novel biomarkers for prognosis, which may potentially improve the prognosis of OC.

The GGGGCC nucleotide repeat expansion (NRE) mutation in the <i>C9ORF72</i> (C9) gene is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Neuronal activity plays an essential role in shaping biological processes within both healthy and neurodegenerative disease scenarios. Here, we show that at baseline conditions, C9-NRE-induced pluripotent stem cell-cortical neurons display aberrations in several pathways, including synaptic signaling and transcriptional machinery, potentially priming diseased neurons for an altered response to neuronal stimulation. Indeed, exposure to two pathophysiologically relevant stimulation modes, prolonged membrane depolarization or a blockade of K⁺ channels, followed by RNA sequencing, induces a temporally divergent activity-dependent transcriptome of C9-NRE cortical neurons compared with healthy controls. This study provides new insights into how neuronal activity influences the ALS/FTD-associated transcriptome, offering a dataset that enables further exploration of pathways necessary for conferring neuronal resilience or degeneration.<b>NEW & NOTEWORTHY</b> A recent study using iPSC-derived cortical neurons reveals how neuronal activity drives gene dysregulation in <i>C9ORF72</i>-linked ALS/FTD. We uncover synaptic dysfunction, peroxisomal dysregulation, and NPAS4-linked transcriptional shifts, highlighting key disease-modifying pathways. Could these insights pave the way for new therapeutic targets? Explore our research and generate your own discoveries using our interactive dataset included in the link in the article.

Skeletal muscle is essential for life as it enables physical movement, maintains posture, is crucial for breathing, and serves as a major site for energy and carbohydrate metabolism. Pathological conditions that reduce skeletal muscle mass and function-such as muscular dystrophies, motor-neuron diseases, cancer, type-2 diabetes, or aging-have detrimental effects on human health, reducing quality of life and survival. Currently, exercise is the only validated treatment for increasing muscle mass and function, but it is impractical for bedridden patients or the frail elderly. Significant advances in understanding the molecular mechanisms underlying atrophy of slow- or fast-twitch muscle fibers have identified numerous previously unknown key players that may show promise as potential drug targets. Here, we review these recent advances and discuss the potential of these discovered mechanisms as therapeutic targets to combat muscle wasting.

Rieske oxygenases (ROs) are a diverse family of nonheme iron enzymes that catalyze a wide array of oxidative transformations in both catabolic and biosynthetic pathways. Their catalytic repertoire spans dioxygenation, monooxygenation, oxidative <i>N</i>- and <i>O</i>-dealkylation, desaturation, sulfoxidation, C-C bond formation, <i>N</i>-oxygenation, and C-N bond cleavage─reactions that are often challenging to achieve selectively through synthetic methods. These diverse functions highlight the increasing importance of ROs in natural product biosynthesis and establish them as promising candidates for biocatalytic applications. Despite extensive study, our understanding of how ROs orchestrate these diverse reactions at the molecular level remains incomplete. In particular, the transient, dynamic nature of electron transfer events and the limited structural characterization of oxygen-bound intermediates hinder our understanding of how structural features govern electron transfer efficiency, O₂ activation, and the origins of their catalytic diversity. Recent findings challenge traditional views of the RO catalytic cycle and underscore the importance of integrating static structural data with dynamic studies of redox interactions. In this Perspective, we explore emerging insights into the structural and mechanistic basis of RO function. We focus on how the architecture of the oxygenase component shapes reactivity, electron transfer, and redox partner interactions. Finally, we discuss current limitations and future opportunities in harnessing ROs for biocatalysis, emphasizing the potential of engineering approaches─particularly the optimization of redox partner compatibility─to expand their functional utility.

Enamel is a cellular, nonregenerative, highly mineralized tissue essential for the mechanical durability and wear resistance of human teeth. Combating its degradation necessitates effective remineralization strategies, with hydroxyapatite (HAp) playing a central role in both natural and synthetic enamel restoration. Fluoride incorporation enhances HAp stability, forming fluoridated hydroxyapatite (FHAp), which is widely used to prevent or resist dental caries and improve remineralization. However, a mechanistic understanding of demineralization and remineralization remains incomplete due to the limitations of conventional ex situ techniques, which fail to capture real-time crystal dissolution and growth dynamics. In this study, we developed and applied a facile synthesis method for oriented FHAp nanocrystals under ambient pressure and at body temperature. This unlocks the possibility of direct in situ liquid imaging using atomic force microscopy (AFM) that serves as a platform for direct observation of demineralization and remineralization processes at the nanoscale. Investigation of the morphology, spectroscopy, and mechanical properties of nanocrystals grown in different conditions elucidated the effect of the substitution rate of fluorine through both in situ and ex situ studies. The findings presented offer a generic approach for understanding the re/demineralization mechanisms in enamel and demonstrate the potential for charting biomimetic enamel restoration pathways.

<h4>Background</h4>DDX27, a member of the DEAD-box RNA helicase family, plays a pivotal role in RNA metabolism and is essential for diverse cellular processes, including transcription, pre-mRNA splicing, translation, and ribosome biogenesis. Recent findings have implicated DDX27 as a substantial contributor to tumorigenesis and cancer progression across various malignancies, establishing its significance as a molecular hub that interacts with key oncogenic partners such as major vault protein (MVP) and nucleophosmin 1 (NPM1).<h4>Methods</h4>We conducted systematic search in the following comprehensive academic databases: PubMed, MEDLINE or Web of Science. The keywords such as DDX27, DEAD-box protein 27, RNA helicase DDX27 and cancer, tumor or carcinoma were used for searching. This review consolidates the existing literature on DDX27, examining its structural features and biological functions within the context of tumorigenesis. We systematically explore the molecular mechanisms by which DDX27 influences tumor development and progression, focusing particularly on its roles across different cancer types, including colorectal cancer (CRC), gastric cancer (GC), breast cancer (BC), hepatocellular carcinoma (HCC), and oral squamous cell carcinoma (OSCC). Key molecular mechanisms such as NF-κB activation and ERK1/2 phosphorylation involved in DDX27-related pathways are discussed.<h4>Results</h4>Our comprehensive summary elucidates the context-dependent roles of DDX27 across various cancers, highlighting its associations with advanced disease stages, metastasis, and therapeutic resistance. We also assess the potential of DDX27 as a diagnostic and prognostic biomarker, correlating its expression levels with negative clinical outcomes.<h4>Conclusion</h4>Novel therapeutic strategies targeting DDX27 are proposed, including RNA interference techniques (siRNA and shRNA), miRNA-based therapies (miR-617 mimics), pathway modulation, and synthetic lethality approaches. Furthermore, we identify notable limitations in current research surrounding DDX27 and offer potential avenues for future investigation. These innovative strategies present significant promise for the development of precision cancer therapies aimed at improving treatment outcomes for patients.

<h4>Introduction</h4>Diffuse Pleural Mesothelioma (DPM) is a rare and incurable cancer. Immune checkpoint inhibitors (ICIs) marked some advances but only for a limited fraction of patients. Improving response prediction to ICIs is currently a clinical need in DPM. Deletion of CDKN2A gene, in chr9p21.3, is one of the most frequent alterations in DPM. As in other settings, deletion of CDKN2A locus has been associated with an immunosuppressive phenotype. Here we investigated the consequences of CDKN2A deletion (CDKN2Adel) on the tridimensional organization and function of immune infiltrate in DPM.<h4>Methods</h4>A retrospective cohort of 89 DPMs was analyzed and assessed for CDKN2Adel through digital droplet PCR. Immune-profiling was assessed by analyzing 770 immune-related genes by digital profiling. Finally, morphologically resolved, high-dimensional transcriptomic approach was used to reconstruct the spatial architecture of immune-tumor interaction in wild-type and deleted FFPE samples.<h4>Results</h4>CDKN2Adel was detected in 41.5% of DPMs and was associated with reduced survival (p = 0.04). Bulk gene expression identified 373 differentially expressed genes, of which 98.6% were downregulated in CDKN2Adel samples. These genes were enriched in several immune categories, suggesting significant immune deprivation in deleted tumors. Deconvolution analysis confirmed a major depletion of infiltrating immune cells including effector populations. Spatial transcriptomics revealed that this immunosuppressive phenotype was different according to histotype and prominent in the sarcomatoid lesions.<h4>Conclusion</h4>These data demonstrated that CDKN2Adel deeply affects the spatial organization of immune microenvironment by depleting immune-signaling and reducing or preventing immune infiltration, supporting the potential implementation of this alteration as ICIs predictive biomarker in DPM.

Most genetic loci linked to polygenic traits are in non-coding regions, with complex regulation and linkage disequilibrium (LD), complicating causal variant and gene prioritization. We used multiplexed single-cell CRISPR interference and activation perturbations to investigate cis-regulatory element (CRE) and gene expression relationships within tight LD in the endogenous chromatin context. We demonstrated the prevalence of multiple causality in perfect LD (pLD) for independent expression quantitative trait loci (eQTLs) and uncovered fine-grained genetic effects on gene expression within pLD, which are difficult to decipher using traditional eQTL fine-mapping or existing computational methods. We found that over one-third of the causal CREs lack classical epigenetic markers prior to perturbation, and we functionally validated one of these hidden regulatory mechanisms. Leveraging Multiome single-cell epigenetic and sequence perturbations, we highlighted the regulatory plasticity of the human genome. Our study will guide the exploration of missing causal mechanisms underlying molecular trait regulation and disease development.

Cancer is the second leading cause of death globally and in the United States, second only to cardiovascular disease. Unlike many cardiovascular conditions, cancer is often less preventable, manageable, and curable-even with ongoing technological advancements in medicine. The adverse effects of cancer treatments on cancer patients remain profound due to shared cellular characteristics between cancerous and normal cells; one of the primary adverse effects is treatment-induced inflammation. These inflammatory responses aim to eliminate cancerous cells but often damage normal tissues. Notably, inflammatory side effects vary considerably across the growing diversity of therapeutic approaches. This study reviewed studies between 2007 and 2024, comparing the inflammatory profiles associated with five major radiation therapies (RTs): Three-Dimensional Conformal Radiation Therapy (3D-CRT), Intensity-Modulated Radiation Therapy (IMRT), Image-Guided Radiation Therapy (IGRT), Stereotactic Body Radiation Therapy (SBRT), and Proton Beam Therapy (PBT)-each characterized by distinct mechanistic and therapeutic features. In addition to each radiation modality eliciting distinct inflammatory responses, tissue-specific variability further complicates clinical outcomes. Accordingly, this review also undertakes a cross-tissue comparison of radiation-induced inflammation, with a focus on the gastrointestinal (GI) system, central nervous system (CNS), and skin. However, the variation in treatment modalities and organ-specific inflammatory biomarkers greatly hinders direct comparison across studies. Finally, this review highlights potential inflammatory mitigations, including ambroxol, that may be employed synergistically with RTs, minimizing side effects and enhancing patient outcomes. Taken together, while all modalities offer therapeutic value alongside certain limitations, proton-based therapy demonstrates the greatest potential for minimizing toxicity though its broader adoption remains limited by cost-effectiveness concerns.

Despite clear evidence that vitamin C levels are depleted in the brains of Alzheimer's disease (AD) patients, dietary supplementation has consistently failed in clinical trials, suggesting a critical bottleneck not in systemic supply, but in its transport into brain cells. Here, we identify this bottleneck as a progressive downregulation of the ascorbate transporter, Slc23a2, also known as SVCT2, in microglia. Then we hypothesized that bypassing this cellular deficiency via targeted SVCT2 overexpression in microglia could either prevent the onset of pathology or rescue established functional deficits. Indeed, overexpressing SVCT2 in microglia before disease onset in 5xFAD mice triggered a profound redox reprogramming, resulting in a unique "hybrid" neuroprotective microglial phenotype that co-expressed both homeostatic and disease-associated markers. Functionally, this leads to decreased amyloid plaque burden and strengthens the synaptic bioenergetic capacity, which consequently prevents the development of synaptic and memory deficits. Strikingly, when employed after disease establishment, SVCT2 overexpression rescued synaptic plasticity and memory performance despite not affecting the existing amyloid burden. This rescue was driven by changes in the microglial secretory pathways. Collectively, these findings resolve a long-standing clinical paradox by establishing that neuroprotection depends not on systemic vitamin C intake but on the brain's cellular uptake machinery. This offers a mechanistic explanation for the failure of dietary supplementation in AD and identifies SVCT2 as a promising therapeutic target against the neurodegenerative process in AD.

Lipophilic yeasts of the genus <i>Malassezia</i> are commensal fungi that constitute the normal skin microbiota but may become pathogenic. These fungi, especially <i>M. furfur</i>, convert tryptophan into various alkaloid indoles such as malassezione, which may serve as virulence factors. To facilitate testing of malassezione as an aryl hydrocarbon receptor agonist and potential glucokinase activator, we developed a convenient synthetic route from commercially available indole-3-acetic acid. Treatment of the <i>N</i>-Boc-protected indole-3-acetic acid with <i>N</i>-ethyl-<i>N</i>'-(3-dimethylaminopropyl)carbodiimide (EDC) in the presence of DMAP generates the <i>N</i>,<i>N</i>'-Boc-protected malassezione, which upon deprotection yields malassezione in an overall yield of ca. 20%. This is an improvement over the preparation of the isonitrile followed by an Fe hydride initiated isonitrile-olefin intramolecular coupling reaction, which generated malassezione with an overall yield of ca. 5%. Furthermore, the present method may also be used to prepare related compounds.

Schwann cells (SCs) are the primary glial cells of the Peripheral Nervous System (PNS), which insulate and provide protection and nutrients to the axons. Technological and experimental advances in neuroscience, focusing on the biology of SCs, their interactions with other cells, and their role in the pathogenesis of various diseases, have paved the way for exploring new treatment strategies that aim to harness the direct protective or causative properties of SCs in neurological disorders. SCs express cytokines, chemokines, neurotrophic growth factors, matrix metalloproteinases, extracellular matrix proteins, and extracellular vesicles, which promote the inherent potential of the injured neurons to survive and accelerate axonal elongation. The ability of SCs to support the development and functioning of neurons is lost in certain hereditary, autoimmune, metabolic, traumatic, and toxic conditions, suggesting their role in specific neurological diseases. Thus, targeting, modifying, and replacing SC strategies, as well as utilizing SC-derived factors and exosomes, have been considered novel therapeutic opportunities for neuropathological conditions. Preclinical and clinical data have demonstrated that SCs and SC-derived factors can serve as viable cell therapy for reconstructing the local tissue microenvironment and promoting nerve anatomical and functional recovery in both peripheral and central nerve injury repair, as well as in peripheral neuropathies. However, despite the promising successes of genetic engineering of SCs, which are now in preclinical and clinical trials, improving tactics to obtain 'repair' SCs and their products from different sources is the key goal for future clinical success. Finally, further development of innovative therapeutic approaches to target and modify SC survival and function in vivo is also urgently needed.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and the third leading cause of cancer-related death. Hyperactivation of oncogenes and suppression of tumor suppressor genes/proteins drive HCC initiation and progression. MicroRNAs (miRNAs) critically modulate HCC biology by regulating proliferation, apoptosis, and metastasis. Acting either as tumor suppressors or oncomiRs, they shape core signaling pathways, including PI3K/Akt/mTOR, Hippo-YAP/TAZ, Wnt/β-catenin, RAS/MAPK, and p53. Their dysregulation in tissues and body fluids renders them promising diagnostic biomarkers and therapeutic targets. Preclinical studies demonstrate that miRNA-based strategies-either restoring tumor-suppressive miRNAs (e.g., miR-34a, miR-125a-5p) or inhibiting oncogenic miRNAs (e.g., miR-660-5p)-can suppress HCC progression and reduce treatment resistance. Combination approaches, such as pairing miR-122 mimics with miR-221 inhibitors or delivering miR-326 via nanoparticles, further enhance efficacy by simultaneously targeting multiple oncogenic pathways. This review summarizes recent advances in miRNA-mediated regulation of HCC signaling and highlights their clinical potential, including ongoing trials of miRNA-based diagnostics and therapeutics for early detection, prognostication, and personalized treatment.

Phosphorus deficiency significantly limits soybean production across 74% of China's arable land. This study investigated the molecular mechanisms enabling soybean to access insoluble phosphorus through transcriptome sequencing of the Heinong 48 variety across four developmental stages (Trefoil, Flower, Podding, and Post-podding). RNA-Seq analysis identified 2755 differentially expressed genes (DEGs), with 2506 up-regulated and 249 down-regulated genes. Notably, early developmental stages showed the most substantial transcriptional reprogramming, with 3825 DEGs in the Trefoil stage and 10,660 DEGs in the Flower stage, compared to only 523 and 393 DEGs in the Podding and Post-podding stages, respectively. Functional enrichment analysis revealed 44 significantly enriched GO terms in the Trefoil stage and 137 in the Flower stage, with 13 GO terms shared between both stages. KEGG pathway analysis identified 8 significantly enriched pathways in the Trefoil stage and 21 in the Flower stage, including key pathways related to isoflavonoid biosynthesis, alpha-linolenic acid metabolism, and photosynthesis. Among 87 differentially expressed transcription factors from 31 families, <i>bHLH</i> (8.08%), <i>bZIP</i> (7.18%), and <i>WRKY</i> (5.94%) were most prevalent. These findings provide genetic targets for developing soybean varieties with improved phosphorus acquisition capacity, potentially reducing fertilizer requirements and supporting more sustainable agricultural practices.

<h4>Background</h4>Type 1 myocardial infarction (T1MI) is an acute ischemic event triggered by the rupture of a coronary atherosclerotic plaque. The pathogenesis of T1MI is highly complex, involving disturbances in iron metabolism, cell apoptosis, immune activation, and inflammatory responses. In recent years, ferritinophagy, a novel autophagic mechanism regulating iron homeostasis, has attracted increasing attention for its role in cardiovascular diseases. However, its precise involvement in T1MI remains to be fully elucidated. This study aims to systematically analyse the mechanism of ferritinophagy in T1MI and explore its potential connection to immune and inflammatory responses.<h4>Methods</h4>Exosomes were isolated from coronary thrombi of T1MI patients and subjected to comprehensive transcriptomic profiling. Differentially expressed lncRNAs and mRNAs were validated through functional assays, including RIP, FISH, ChIP, and m6A methylation experiments. Cardiomyocyte models and integrated bulk and single-cell RNA sequencing were used to clarify cellular context and regulatory networks, with particular emphasis on YTHDF family proteins. Bioinformatics analyses, including GO and KEGG, were employed for pathway annotation.<h4>Results</h4>Electron microscopy confirmed the presence of exosomes in coronary thrombi. Thrombus-derived exosomes (TEs) induced pronounced ferritinophagy in cardiomyocytes, evidenced by increased autophagosomes, ROS, apoptosis, and iron overload, with these effects ameliorated by the ferroptosis inhibitor Fer-1. Transcriptomic and functional analyses identified lncRNA FENDRR as highly enriched in TEs, with FENDRR and P53 acting in concert to regulate NCOA4 and system Xc-. Mechanistically, FENDRR directly binds P53, and both upregulate m6A modification in cardiomyocytes, specifically through upregulation of YTHDF1 and downregulation of YTHDF3. Inhibition of either FENDRR or P53 reverses these changes. Single-cell RNA-seq analysis revealed significant upregulation of TP53, NCOA4, and YTHDF1, alongside downregulation of YTHDF3 in macrophages from plaque tissue, linking ferritinophagy, autophagy, and immune-inflammatory responses.<h4>Conclusion</h4>This study is the first to reveal the critical role of the "FENDRR-m6A-NCOA4" regulatory axis as a critical mediator of ferritinophagy in T1MI. It also suggests that immune cells may participate in the immune-inflammatory response associated with myocardial injury via ferritinophagy. Our research provides multi-omics evidence of the interaction between iron homeostasis, immunity, and inflammation in T1MI, offering potential therapeutic strategies for targeting ferritinophagy and related RNA modification pathways.

Colorectal cancer (CRC) is a common and aggressive malignancy of the gastrointestinal tract with a severe disease burden. The role of Traditional Chinese Medicine (TCM) and its natural active ingredients in enhancing the therapeutic effects of radiotherapy and chemotherapy and preventing the recurrence and metastasis of CRC has been increasingly recognized. Herba <i>Patriniae</i> has shown significant clinical efficacy for the treatment of CRC. Flavonoids has been found to be one of the main active anticancer components of Herba <i>Patriniae</i>. This review summarizes the latest findings from clinical trials and <i>in vitro</i> studies on anticancer mechanisms of Herba <i>Patriniae</i>, and discusses the role of the flavonoids in combination therapy against CRC. These flavonoids exert anticancer effects through diverse mechanisms. For instance, they prevent the development of precancerous lesions, regulate the cell cycle, modulate CRC cell proliferation, promote tumor cell apoptosis, inhibite epithelial-mesenchymal transition, reverse drug resistance, and modulate gut microbiota by acting on several key signaling pathways, such as PI3K/Akt/mTOR, Wnt/β-catenin, and EGFR/ERK/MAPK. Future research should prioritize clarifying the specific dosage and safety of flavonoids under different pathological conditions, further conducting large-scale, rigorously designed clinical studies to determine the efficacy differences of flavonoids for patients with different pathological types of CRC and simultaneously delving into the mechanisms of their anti-colorectal cancer effects, as well as their interactions with the intestinal microbiota and tumor microenvironment.

<h4>Background</h4>The increasing incidence of asymptomatic pulmonary nodules (PNs) underscores the need for accurate malignancy estimation to guide early-stage lung cancer management. This study aimed to identify risk factors for malignant PNs and develop a risk prediction model.<h4>Methods</h4>This study enrolled 691 patients with PNs (444 training, 247 validation) and 62 healthy controls. Clinical, imaging, and serum data were collected. Transcriptome sequencing was performed to identify circular RNAs (circRNAs) associated with malignant PNs. Additionally, Tumor-associated antigens and tumor-associated autoantibodies (AAbs) were measured. Univariate logistic regression analysis was employed to identify risk factors for malignant PNs, followed by multivariate logistic regression to establish a risk prediction model. Finally, receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of the risk prediction model.<h4>Results</h4>Among five tumor-associated antigens, only neuron-specific enolase (NSE) levels were significantly higher in malignant versus benign PNs (P<0.001). AAb positivity rates and number of positive AAbs were elevated in malignant PNs. Transcriptome sequencing revealed hsa_circCFLAR_008 was upregulated in malignant PNs (P=0.04). The risk prediction model was established [area under the curve (AUC), 0.8241], and was validated with a concordance statistic (C-statistic) of 0.8344 in an independent cohort.<h4>Conclusions</h4>A risk prediction model for malignant PNs was established. Hsa_circCFLAR_008 enhanced the diagnostic performance of the model and served as a novel biomarker for malignant PNs.

<h4>Background</h4>Genetic factors play a significant role in human male infertility, with about 4% of infertile men currently identified with genetic reasons, yet most (60-70%) still lack a definitive diagnosis and remain unexplained. Similar to other medical fields, the advent of next-generation sequencing (NGS) has resulted in the discovery of a growing array of genetic variations in infertility issues affecting both genders. With the rising count of newly discovered genes, precise diagnoses are now possible for cases of male infertility that were once considered idiopathic. Nonetheless, substantial proof supporting the gene-disease relationships (GDR) remains absent in numerous instances.<h4>Objective and rationale</h4>The year 2019 and 2021 saw the release and revision of the standardized clinical validity evaluation for monogenic reasons behind male infertility. In this report, we offer an extensive review to methodically assess all existing data (spanning from 1 Jan, 2020, to 24 Sep, 2024) regarding the singular causes of either isolated or syndromic male infertility, hormonal imbalances, or reproductive irregularities in male reproductive organs.<h4>Search method</h4>The PRISMA protocols were utilized to gather comprehensive data from PubMed and Web of Science regarding the genetics of human male infertility and disorders of sex development (DSD) resulting in infertility, spanning from 1 January 2020 to 24 September 2024. The pathologies examined encompass both isolated infertility and syndromic male infertility, along with disorders of the endocrine and reproductive systems. A standardized scoring system was used to evaluate whether pathogenic variations in a particular gene lead to a recognized phenotype. Each GDR received a conclusive rating, ranging from no evidence to definitive.<h4>Outcomes</h4>Out of 19885 identified and screened publications, 229 were chosen for gene and variant analysis. Our research has pinpointed 191 genes and confirmed 191 GDRs, encompassing all documented single-gene reasons for male infertility and DSD. Additionally, our research pinpointed 100 genes with at least a moderate connection to male infertility or atypical genitourinary development traits. The study did not take into account associated genetic risk factor(s) or oligogenic/polygenic causes of male infertility.<h4>Systematic review registration</h4>http://www.crd.york.ac.uk/PROSPERO, identifier CRD42024593082.

Head and neck squamous cell carcinoma (HNSCC) is an aggressive cancer with a complex molecular landscape. Despite extensive research, our understanding of the molecular mechanisms remains incomplete, hindering the development of effective therapeutic strategies for this disease. Long non-coding RNAs (lncRNAs) have emerged as crucial factors in cancer biology, regulating key networks across various malignancies. These molecules exert their regulatory functions through interactions with nucleic acids or proteins, thereby influencing signaling pathways within tumor cells. Consequently, lncRNAs play a significant role in key processes like cell proliferation, metastasis, immune evasion, and treatment resistance. This review offers a comprehensive overview of current knowledge regarding lncRNA-mediated mechanisms in HNSCC. The first section explores how lncRNAs influence tumor processes through various modulation mechanisms, including transcriptional and post-transcriptional regulation, chromatin remodeling, and epigenetic modifications. We also highlight the impact of lncRNAs on specific signaling pathways that control essential cellular functions (e.g., proliferation, apoptosis, angiogenesis, invasion, metastasis). Ultimately, this underscores the promising potential of lncRNAs as diagnostic biomarkers and therapeutic targets capable of enhancing patient care in oncology. Gaining a deep understanding of how lncRNAs modulate carcinogenic mechanisms may yield innovative approaches for early detection, personalized treatment, and improved clinical outcomes for HNSCC patients.

The convergence of nanotechnology and cancer immunotherapy represents a promising frontier in the fight against cancer, offering new avenues for enhancing therapeutic efficacy and precise personalized cancer treatments. This review presents recent advancements in nanomedicine that strategically target and modulate various immune cells to enhance anti-tumor responses and overcome barriers posed by the immunosuppressive tumor microenvironment for cancer treatment. By exploring advance strategies such as photoimmunotherapy, sonodynamic therapy, therapeutic nanovaccines, and surface-modified immune cells, we highlight how nanoparticles can go beyond drug delivery to actively reprogram immune pathways. Despite significant advances in cancer immunotherapy, current nanomedicine approaches struggle with poor targeting, immune evasion, systemic toxicity, <i>etc.</i> This review highlights recent breakthroughs that address these challenges by focusing on treatment that are induced using effective immune cells, innovative approaches to tumor reduction through targeted drug delivery systems, the development of enhanced combinational immunotherapy techniques, and the modification of immune cells using nanoparticles to enhance their anti-tumor activity through immune cell-specific strategies. These nanotechnological innovations hold the potential to overcome existing limitations in cancer immunotherapy, paving the way for more effective and personalized treatments to reshape cancer immunotherapy through synergistic, targeted, and immune-responsive strategies.

We evaluated the analytical performance of the TriVerity test system, a benchtop system for rapid measurement and interpretation of host messenger RNAs (mRNAs) measured quantitatively from PAXgene Blood RNA specimens using the TriVerity cartridge on the Myrna instrument. In approximately 30 minutes, 3 scores are generated from 29 host mRNAs for the likelihood of a bacterial infection, a viral infection, and illness severity (7-day need for ICU-level care), each falling into 1 of 5 interpretation bands (very low, low, moderate, high, and very high). Reproducibility, precision, limit of detection, linearity, interference, and sample and cartridge stability were determined per Clinical Laboratory Standards Institute standards. An operator survey assessed TriVerity's ease of use. Reproducibility demonstrated SDs meeting the acceptance criteria of <5.5 score units. The lowest concentration at which 100% of replicates showed measurable amplification was 1 × 10⁶ cp/mL of <i>in vitro</i> RNA transcripts; the limit of quantification and detection were equivalent to blood containing 500 leukocytes/µL. Interference was not observed for 17 potential interferents. The cartridge was stable at room temperature (RT) for 9 and 12 months using accelerated testing at 37°C. TriVerity results were equivalent using fresh and frozen PAXgene Blood RNA samples. All operators agreed or strongly agreed that the TriVerity test system was easy to use. In summary, the analytical performance presented here, along with diagnostic accuracy established in the SEPSIS-SHIELD trial, demonstrates that the TriVerity system is reliable and user friendly, assisting clinicians in managing patients with suspected acute infections and sepsis in acute care settings.<h4>Importance</h4>The prompt diagnosis of acute infections and sepsis is critical for better patient outcomes. We introduce a groundbreaking messenger RNA-based test, the first of its kind, designed to diagnose the presence of infection and predict illness severity in adult patients with suspected acute infections or sepsis. Several key findings regarding the accuracy and robustness of the test system are presented, which will be relevant to laboratories or acute care settings implementing the test for patient care. Furthermore, these findings may assist clinical researchers in developing analytical trial protocols aimed at the combined evaluation of RNA-based multi-marker tests with both diagnostic and prognostic test characteristics.

High myopia (HM) is a severe form of refractive error that results in irreversible visual impairment and even blindness. However, the genetic and pathological mechanisms underlying this condition are not yet fully understood. From a cohort of 1015 patients with HM in adolescents, likely pathogenic missense mutations were identified in the <i>ZC3H11A</i> gene in four patients by whole exome sequencing. This gene is a zinc finger and stress-induced protein that plays a significant role in regulating nuclear mRNA export. To better understand the function and molecular pathogenesis of myopia in relation to gene mutations, a <i>Zc3h11a</i> knockout (KO) mouse model was created. The <i>Zc3h11a^+/-</i> mice exhibited significant shifts in refraction toward myopia. Myopia-related factors, including <i>Tgfβ1, Mmp2,</i> and <i>Il6,</i> were found to be upregulated in the retina or sclera, and electroretinography and immunofluorescence staining results showed dysfunction and reduced number of bipolar cells in the retina. Transmission electron microscopy findings suggest ultrastructural abnormalities of the retina and sclera. Retinal transcriptome sequencing showed that 769 genes were differentially expressed, and <i>Zc3h11a</i> was found to have a negative impact on the PI3K-AKT and NF-κB signaling pathways by quantitative PCR and western blotting. In summary, this study characterized a new candidate pathogenic gene associated with HM and indicated that the ZC3H11A protein may serve as a stress-induced nuclear response trigger, and its abnormality causes disturbances in a series of inflammatory and myopic factors. These findings offer potential therapeutic intervention targets for controlling the development of HM.

Perturbations in protein quality control lead to the accumulation of misfolded proteins and protein aggregates, which can compromise health and lifespan. One key mechanism eliminating protein aggregates is aggrephagy, a selective type of autophagy. Here we reveal that fragmentation is required before autophagic clearance of various types of amorphous aggregates. This fragmentation requires both the 19S proteasomal regulatory particle and the DNAJB6-HSP70-HSP110 chaperone module. These two players are also essential for aggregate compaction that leads to the clustering of the selective autophagy receptors, which initiates the autophagic removal of the aggregates. We also found that the same players delay the formation of disease-associated huntingtin inclusions. This study assigns a novel function to the 19S regulatory particle and the DNAJB6-HSP70-HSP110 module, and uncovers that aggrephagy entails a piecemeal process, with relevance for proteinopathies.

Phenotype switching is a form of cellular plasticity in which cancer cells reversibly move between two opposite extremes: proliferative versus invasive states^1,2. Although it has long been hypothesized that such switching is triggered by external cues, the identity of these cues remains unclear. Here we demonstrate that mechanical confinement mediates phenotype switching through chromatin remodelling. Using a zebrafish model of melanoma coupled with human samples, we profiled tumour cells at the interface between the tumour and surrounding microenvironment. Morphological analysis of interface cells showed elliptical nuclei, suggestive of mechanical confinement by the adjacent tissue. Spatial and single-cell transcriptomics demonstrated that interface cells adopted a gene program of neuronal invasion, including the acquisition of an acetylated tubulin cage that protects the nucleus during migration. We identified the DNA-bending protein HMGB2 as a confinement-induced mediator of the neuronal state. HMGB2 is upregulated in confined cells, and quantitative modelling revealed that confinement prolongs the contact time between HMGB2 and chromatin, leading to changes in chromatin configuration that favour the neuronal phenotype. Genetic disruption of HMGB2 showed that it regulates the trade-off between proliferative and invasive states, in which confined HMGB2^high tumour cells are less proliferative but more drug-resistant. Our results implicate the mechanical microenvironment as a mechanism that drives phenotype switching in melanoma.

Clinical studies demonstrate that comorbidity between colorectal cancer (CRC) and depression is common, leading to a higher mortality risk among CRC patients. However, the mechanisms remain largely unexplored. The role of core brain regions in comorbidity of CRC and depression and whether modulating these regions can improve both depression-like symptoms and CRC progression have yet to be clarified. In this study, we establish a mouse (Mus musculus) model of CRC and observe that mice with orthotopic colorectal cancer (CRC mice) display depression-like behaviors. Through c-FOS mapping, network analysis, correlation analysis, and inverse tracing, we identify the anterior cingulate cortex (ACC) as a central node within the depression-related brain network in CRC mice. Notably, inhibiting ACC activity not only alleviates depression-like behaviors but also mitigates CRC-induced neuronal damage and reduces CRC tumor progression. These findings underscore the critical role of the ACC in comorbidity of CRC and depression and suggest that ACC-targeted interventions may hold therapeutic potential for CRC patients with comorbid depression.

Acral melanoma (AM) is the predominant subtype of melanoma in Asians. Early detection and prevention can significantly improve patient outcomes; however, there is a lack of effective early biomarkers for predicting AM metastasis. Here, we employed single-cell and spatial transcriptomics analyses to investigate early microsatellite lesions of AM and identify biomarkers of invasiveness in these lesions. Our results characterize a highly immunosuppressive microenvironment and metabolic process shifts in early AM microsatellite lesions that promote the metastatic potential. The transcription factor SOX6 is overexpressed in microsatellite lesions and marks a population of highly invasive melanoma cells. The pro-invasive role of overexpressed SOX6 was validated in vivo and in vitro, including its ability to enhance tumor invasion by upregulating cellular glycolysis, disrupt fatty acid transport, and increase intracellular phosphatidylcholine content. This study suggests that SOX6-overexpressing melanoma cells are the main driver subpopulation promoting early invasion of AM and establishes SOX6 and fatty acid transport processes as biomarkers and potential therapeutic targets for early melanoma metastasis.

<h4>Background</h4>In recent years, obsessive-compulsive disorder (OCD) has emerged as the fourth most prevalent mental disorder. However, the relationship between OCD and various factors, including demographic characteristics, lifestyles, and dietary habits remains uncertain. This study aims to explore the connection between various factors and OCD.<h4>Methods</h4>A cross-sectional survey was conducted from January 14 to February 15, 2023, targeting the general population through an online platform. Demographic characteristics, lifestyles, dietary habits, and OCD symptoms were assessed using a validated classification scale. To identify potential risk factors for OCD, Lasso and logistic regression analyses were performed. Subsequently, the relative importance of these factors was evaluated through machine learning models, including Support Vector Machine, Random Forest, and Back Propagation Neural Network.<h4>Results</h4>Among the 1572 participants, 30.40% were categorized as high-risk for OCD. Logistic regression analysis revealed that having a medical history (OR = 1.709, 95% CI: 1.064-2.738), sleep disorders (OR = 1.460, 95% CI: 1.005-2.137), gastrointestinal (GI) symptoms (OR = 1.562, 95% CI: 1.057-2.311), and abnormal defecation (OR = 1.431, 95% CI: 1.013-2.018) significantly increased the risk of OCD (all P < 0.05). These findings were consistent with the results from machine learning models, further supporting the critical role of these factors in OCD risk.<h4>Conclusion</h4>This study identified medical history, GI symptoms, sleep disorders and picky eating as significant risk factors for OCD. These findings offer valuable insights into the etiology of OCD and suggest potential avenues for targeted interventions and preventive strategies aimed at reducing OCD risk.

Eukaryotic cells use a multi-layered immune response to combat intracellular pathogens. The ubiquitin ligase ZNFX1 has emerged as a crucial yet little understood player that regulates the immune response while protecting against RNA viruses. Our study unveils the molecular mechanism of ZNFX1, mediated by the joint activity of a helicase serving as a nucleic acid sensor and a non-conventional E3 module featuring a split active site. We demonstrate that single-stranded RNA stimulates E3 activity by fostering dimerization of ZNFX1 subunits that translocate along nucleic acid tracks. Juxtaposed E3 domains complement each other, leading to the ubiquitination of ZNFX1 itself and engaged RNA molecules, while clustering nucleic acids into dense nucleoprotein particles. We show that the E3 ligase activity of ZNFX1 protects cells during an immune response and propose that ubiquitin-coated particles formed by ZNFX1 represent part of an ancient mechanism to regulate both foreign and host RNA in the cell.

<h4>Rationale</h4>People with cystic fibrosis (pwCF) exhibit chronic and hyperactive neutrophilia which results in a progressive loss of lung function. CF neutrophils have elevated store operated Ca²⁺ entry (SOCE) relative to healthy non-CF neutrophils, which contributes to persistent neutrophilia. The vast majority of pwCF now take CFTR modulators such as elexacaftor/tezacaftor/ivacaftor (ETI), which effectively increase CFTR function in multiple organs including the lung. However, ETI's impact on neutrophils is poorly understood. Orai1 is a plasma membrane Ca²⁺ channel that contributes to SOCE. We have developed a novel peptide (ELD607) that specifically inhibits Orai1, which we evaluated in CF neutrophils.<h4>Objectives</h4>To characterize Orai1/SOCE in neutrophils from pwCF taking ETI, and to evaluate the impact of SOCE inhibition by ELD607 on pwCF neutrophil Ca²⁺ signaling/function.<h4>Methods</h4>Peripheral blood neutrophils were isolated by negative selection. SOCE was characterized using fluorescent approaches. Protein expression was characterized by proteomics and confocal microscopy. Neutrophil degranulation was measured using a multiplex assay.<h4>Measurements and main results</h4>Proteomic analysis revealed major global differences between non-CF and pwCF neutrophils, despite use of ETI. Several proteins involved in SOCE, including Orai1, were significantly elevated in pwCF neutrophils. ELD607 dose-dependently inhibited SOCE, leading to reduced neutrophil degranulation. Ca²⁺ homeostasis was significantly elevated in pwCF compared to non-CF neutrophils. ELD607-sensitive SOCE inversely correlated with lung function (FEV1pp).<h4>Conclusions</h4>Our findings highlight SOCE as a novel biomarker of CF lung disease. ELD607 can be used to reduce SOCE and subsequent degranulation in CF neutrophils. We therefore hypothesize that ELD607 may be of benefit in the management of inflammation in pwCF.

Disseminated cysticercosis (DCC) is a rare parasitic infection caused by the systemic spread of <i>Taenia solium</i> larvae. This study presents a retrospective analysis of diverse radiological manifestations in five confirmed cases of DCC. Patients presented with varied symptoms, including localized swelling, pain, tingling sensations, and breakthrough seizures. High-resolution ultrasonography (USG) revealed typical cystic lesions, with or without scolex, surrounded by inflammatory changes or calcifications, while computed tomography (CT) and magnetic resonance imaging (MRI) provided additional confirmation, especially in neurocysticercosis cases. Imaging patterns included myo-cysticercosis, soft tissue cysticercosis, orbital involvement, and nodular-calcified neurocysticercosis. Recognition of these radiologic patterns is vital for prompt diagnosis and appropriate management, particularly in endemic regions. USG proved invaluable as a non-invasive, cost-effective, first-line tool for diagnosing soft tissue involvement, while CT and MRI were essential in evaluating central nervous system (CNS) lesions and disease staging.

Skeletal and dental mineralization relies on a precisely regulated sequence of events culminating in apatite deposition onto collagen fibrils. Matrix vesicles (MVs), extracellular vesicles released by mineralization-competent cells, play a pivotal role in this process through the catalytic activity of alkaline phosphatase (TNAP). The lipid composition of MVs, particularly phosphatidylserine (PS)-calcium complexes, facilitates the nucleation of amorphous calcium phosphate and apatite formation. However, the interplay between the TNAP structure, the lipid membrane environment, and its enzymatic activity remains incompletely understood. Biomimetic models of MVs, as proteoliposomes made with dipalmitoylphosphatidylcholine (DPPC) and various TNAP mutants, were used to investigate the TNAP's activity and mineralization potential. Molecular docking and site-directed mutagenesis revealed that specific cysteine substitutions near TNAP's catalytic and anchoring sites influence structural stability, enzymatic activity, and incorporation into lipid bilayers. Notably, TNAP mutants S221C and P307C exhibited enhanced catalytic efficiency in DPPC liposomes, while A420C showed reduced activity due to steric hindrance near the catalytic site. Solid-state NMR and cryo-TEM analyses confirmed hydroxyapatite formation, with significant contributions from lipid-anchored TNAP to the mineralization process. These findings highlight the critical influence of the lipid environment on TNAP's functional properties and provide insights into the mechanisms governing biomineralization and related pathologies, including hypophosphatasia associated with various TNAP mutations. The study underscores the importance of ATP and pyrophosphate hydrolysis by TNAP in modulating apatite formation and reveals the role of specific TNAP mutations in regulating enzymatic activity, stability, and mineral propagation. Understanding these interactions could lead to alternate therapeutic strategies in treatment and regenerative medicine.

The transcription factor PAX8 is expressed in most ovarian tumors, being associated with increased tumorigenesis. Although recent studies have addressed the gene regulatory functions of PAX8 in ovarian cancer, an integrative analysis of multi-omic and patient data is required to identify the core regulatory network of PAX8 and its prognostic and therapeutic value. Here, we integrate PAX8 chromatin binding and accessibility data in ovarian cancer cells with transcriptomic and patients' data to gain insight into the core gene regulatory network orchestrated by PAX8 in ovarian tumors. Integration of differential chromatin accessibility, transcription factor binding, and gene expression upon PAX8 knockout provides a core regulatory network that explains most of the genes regulated by PAX8. We combine these target genes with patient expression data and find a PAX8 gene signature associated with tumor stemness, a property related to therapy resistance. Indeed, we show that the PAX8 gene signature predicts disease outcome and response to therapy in ovarian cancer patients. Finally, we validated experimentally our results from bioinformatic analyses, thus reassuring their robustness. Our findings uncover a PAX8 core network that represents a promising strategy for targeted antitumor therapies and open new pathways to fight against ovarian cancer resistance.

Remifentanil, a widely used ultra-short-acting μ-opioid receptor agonist in clinical anesthesia, is strongly associated with postoperative hyperalgesia (remifentanil-induced hyperalgesia, RIH), posing significant challenges to postoperative pain management. RIH is characterized by an abnormally heightened pain perception following opioid withdrawal, and its underlying mechanisms are complex and multifactorial. Current research highlights the roles of central sensitization, peripheral sensitization, and multiple interacting molecular pathways. These include NMDA receptor activation, glial cell activation, neuroinflammation, disinhibition of inhibitory neurotransmission, and dysfunction of the descending pain modulation system. Additionally, alterations in ion channel expression, synaptic plasticity enhancement, and peripheral responses to inflammatory mediators contribute critically to RIH development. Individual factors such as age, sex, genetic polymorphisms, and surgical type significantly influence the risk of RIH. Although substantial progress has been made in elucidating the molecular mechanisms of RIH, a unified theoretical framework and effective clinical strategies remain lacking. Future studies should emphasize multi-omics approaches and clinically relevant experimental models to uncover key regulatory targets and provide a theoretical basis for individualized analgesic interventions.

<h4>Background</h4>Cancer stem cells (CSCs) are crucial contributors to the development and progression of esophageal cancer (ESCA). This study utilized single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing (RNA-seq) to identify gene signatures of CSCs in ESCA, aiming to construct a prognostic tumor stem cell marker signature (TSCMS) model.<h4>Methods</h4>We analyzed scRNA-seq and RNA-seq data of ESCA. CytoTRACE was used to quantify the stemness of tumor-derived epithelial cell clusters. The TSCMS model was developed using Lasso-Cox regression, and its prognostic significance was evaluated via Kaplan-Meier survival analysis, Cox regression, and ROC curve analysis. Drug response predictions were conducted using the pRRophetic package. Functional studies of TSPO in ESCA cells included bioinformatics analysis, quantitative reverse transcription PCR (qRT-PCR), Western blotting, immunohistochemistry, and cell proliferation assays.<h4>Results</h4>Distinct cell cluster stemness potentials were identified using CytoTRACE. The TSCMS model consists of 18 tumor stemness-related genes. High-risk patients showed reduced immune and ESTIMATE scores, along with elevated tumor purity. Notable differences in immune infiltration and chemotherapy sensitivity were observed between risk groups. TSPO was found to be positively correlated with RNA expression-based stemness scores in various tumors, including ESCA. Its expression was diminished in ESCA cell lines and clinical tumor tissues, with low expression correlating with poor prognosis. Overexpression of TSPO inhibits the proliferation of ESCA cells and the formation of tumor clones. In a mouse model of esophageal carcinoma <i>in situ</i>, TSPO expression was significantly lower than in normal tissues.<h4>Conclusion</h4>This study underscores the prognostic significance of the TSCMS model in ESCA, elucidates the immune landscape and treatment response, and identifies TSPO as a potential therapeutic target.

Digestive cancers are a significant global public health challenge due to their high incidence and mortality rates. Emerging research highlights the pivotal role of circular RNA (circRNA) and autophagy in the progression of digestive cancers. This review provides a summary of recent findings, showing that circRNA-mediated autophagy could either promote or suppress tumor development in esophageal, gastric, liver, pancreatic, and colorectal cancers. CircRNAs could regulate autophagy in digestive cancers by acting as competitive endogenous RNAs, influencing downstream target genes and signaling pathways. Dysregulated circRNAs contribute to tumor onset, progression, and chemoresistance by altering autophagy levels. Targeting specific circRNAs to modulate autophagy may offer potential as a diagnostic, prognostic biomarker, or therapeutic strategy for digestive cancers.

Corneal nerve integrity is vital for maintaining ocular surface health and visual clarity, but damage from injury or disease can lead to pain, persistent epithelial defects, and even vision loss. A deeper understanding of how corneal nerves regenerate at the molecular level is key to developing therapies that restore both anatomical structure and function. In this review, we bring together current insights into the pathways that drive corneal nerve repair after injury. We outline the major signaling pathways that promote neuronal survival, axon extension, and nerve-epithelial interactions, along with evolving research around novel modulators that could improve repair outcomes. Although advances in imaging and molecular therapies have led to significant progress in promoting nerve regrowth, functional sensory recovery often lags. This gap in recovery emphasizes the need for research approaches that align anatomical restoration with sensory function. In this review, we aim to clarify the mechanisms underlying corneal nerve regeneration (and their intersections) and identify opportunities for improving patient outcomes.

Octacalcium phosphate (OCP) is a calcium phosphate compound with a layered structure in which apatite layers, which have a structure similar to hydroxyapatite, and hydrated layers are stacked alternately. OCP can incorporate various carboxylate ions into its interlayers. OCPs with incorporated carboxylate ions, also known as OCP carboxylates (OCPCs), are organically modified at the molecular level. OCPCs are an attractive research target in a wide range of fields, from basic inorganic chemistry to applied materials chemistry. Therefore, it is expected that a comprehensive overview of recent research on OCPCs will be useful in progressing this field. This review focuses on recent advances in OCPCs, namely their synthesis, the identification of new types of carboxylate ions that can be incorporated into OCP interlayers, the steric structure estimation of the interlayer carboxylate ions, and applications of OCPCs as functional materials. OCPC-based functional materials include fluorescent materials, artificial bones, and adsorbents. Furthermore, based on existing studies, challenges in OCPC research and future research directions are described.

Iron is essential for cellular respiration, oxidative defense, and host immunity, but its dysregulation is increasingly associated with metabolic disorders, such as obesity and type 2 diabetes. In these diseases, regional iron accumulation occurs in adipose tissue, independent of systemic overload. This process disrupts the mitochondrial redox balance, induces ferroptotic stress, and activates the innate immune pathways. Recent studies have highlighted the NLRP3 (nucleotide-binding domain, leucine-rich repeat, pyrin domain-containing protein 3) inflammasome and its effector cytokine interleukin-1β (IL-1β) as important mediators of the interface between iron and inflammation. In both adipocytes and macrophages, labile iron increased reactive oxygen species (ROS) production and promoted inflammasome formation. Simultaneously, metabolic stress factors upregulate hepcidin expression, suppress ferroportin activity and exacerbate intracellular iron retention. These molecular events converge to maintain low-grade inflammation and impair insulin signaling. Despite these compelling associations, direct mechanistic evidence remains limited, particularly with respect to depot-specific responses and cell type resolution. In this review, I examine the current evidence linking iron handling and inflammasome biology in adipose tissue, focusing on ferroptosis, thioredoxin-interacting protein (TXNIP) signaling, and spatial mapping of iron-cytokine networks. I also discuss novel therapeutic strategies targeting iron overload and inflammasome activation, including chelation, hepcidin modulation, and inflammasome inhibition in the context of metabolic diseases.

<h4>Background</h4>Acute leukemia (AL) is one of the most prevalent pediatric malignancies with highly heterogeneous clinical outcomes. Coagulation-related genes (CRGs) play a crucial role in tumours, but their value in combination with clinical factors for prognostic prediction in AL is unclear. This study aims to develop a prognostic model based on the CRGs signature, with the goal of improving prognostic monitoring and identifying potential therapeutic targets for pediatric AL.<h4>Methods</h4>We collected transcriptomic and clinical data of pediatric AL patients from the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) and The Cancer Genome Atlas (TCGA) databases, including age, gender, and white blood cell count (WBC). Molecular subtypes related to CRGs were identified via non-negative matrix factorization (NMF). A CRGs-based gene signature was developed using the least absolute shrinkage and selection operator (LASSO) and regression analyses. The model was built on a training set and validated independently. Time-dependent receiver operating characteristic (ROC) was used to assess the predictive accuracy of the model for 1-, 3-, and 5-year overall survival (OS). Nomograms were constructed combining CRGs characteristics and clinical factors, and their clinical utility was assessed using calibration curves and decision curve analysis (DCA). Immune infiltration was quantified using the single-sample gene set enrichment analysis (ssGSEA) and the microenvironment cell populations-counter (MCPcounter) algorithm. Kaplan-Meier (K-M) survival analysis was performed to assess the correlation between signature gene expression and OS. Moreover, molecular docking was utilized to investigate the potential interactions between signature genes and small-molecule drugs. Expression of key genes was confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR).<h4>Results</h4>A total of 103 AL patients were included as a training set. Risk stratification based on the median risk score of CRGs showed a significant difference in OS between the two groups (P<0.001), with the low-risk group having a better prognosis. The area under the curves (AUCs) of the model for 1-, 3-, and 5-year survival prediction in the training set were 0.711, 0.762, and 0.718, respectively, and the AUC values in the independent validation set also showed good agreement. Analysis integrating risk scores with clinical data indicated that the CRGs signature could serve as an independent prognostic factor. The nomogram constructed based on CRGs features and key clinical variables showed good fit and potential clinical net effect. Molecular docking analysis revealed stable binding interactions between <i>PROS1</i> and the small-molecule drugs, avatrombopag and lusutrombopag.<h4>Conclusions</h4>In this study, a robust prognostic model incorporating CRGs was constructed to effectively predict survival outcomes in paediatric AL patients. The model helps to enable individualised risk stratification and guide targeted therapy. In addition, avatrombopag and lusutrombopag as potential therapeutic agents provide new ideas for precision medicine in paediatric AL.

<h4>Background</h4>BIRC5, also known as survivin, is the smallest but functionally most complex member of the inhibitor of apoptosis protein (IAP) family and plays an important role in tumorigenesis, recurrence, and chemoresistance, this study aimed to investigate its impact on the clinical and tumor microenvironmental features of lung adenocarcinoma (LUAD), together with its prognostic values.<h4>Methods</h4>Clinical and transcriptomic data of 535 LUAD samples, 59 normal lung, and 54 patients with non-small cell lung cancer (NSCLC) received immune checkpoint blockades (ICBs) were analyzed. Immune infiltration analysis was conducted to uncover the relationship between tumor microenvironmental features and BIRC5 expression level. The prognostic values of BIRC5 were also evaluated with log-rank test and Cox regression analysis.<h4>Results</h4>LUAD had a significantly higher BIRC5 expression level than normal lung tissues. The elevated BIRC5 expression was markedly associated with unfavorable clinical outcomes. Transcriptomic and single-cell sequencing data analysis revealed that tumors with high BIRC5 expression was correlated with multiple pathways' enrichment. Immune infiltration analysis indicated a negative correlation between BIRC5 expression and infiltration levels of CD8+ T cells, dendritic cells (DCs), and natural killer (NK) cell in LUAD, but a positive correlation was observed between BIRC5 expression and regulatory T cells (Tregs) infiltrations. Importantly, NSCLC patients received ICB with high BIRC5 expression had dramatically shorter progression-free survival (PFS; 1.2 <i>vs.</i> 4.5 months; P=0.01) and overall survival (OS; 3.1 <i>vs.</i> 12.7 months; P=0.005) than those with low BIRC5 expression.<h4>Conclusions</h4>These findings suggested that high BIRC5 expression was associated with DNA damage/repair, cell invasion and proliferation related pathways enrichment and increased Tregs infiltration, which would result in inferior outcomes in NSCLC received ICB.

Biological-sequence representation methods are pivotal for advancing machine learning in computational biology, transforming nucleotide and protein sequences into formats that enhance predictive modeling and downstream task performance. This review categorizes these methods into three developmental stages: computational-based, word embedding-based, and large language model (LLM)-based, detailing their principles, applications, and limitations. Computational-based methods, such as k-mer counting and position-specific scoring matrices (PSSM), extract statistical and evolutionary patterns to support tasks like motif discovery and protein-protein interaction prediction. Word embedding-based approaches, including Word2Vec and GloVe, capture contextual relationships, enabling robust sequence classification and regulatory element identification. Advanced LLM-based methods, leveraging Transformer architectures like ESM3 and RNAErnie, model long-range dependencies for RNA structure prediction and cross-modal analysis, achieving superior accuracy. However, challenges persist, including computational complexity, sensitivity to data quality, and limited interpretability of high-dimensional embeddings. Future directions prioritize integrating multimodal data (e.g., sequences, structures, and functional annotations), employing sparse attention mechanisms to enhance efficiency, and leveraging explainable AI to bridge embeddings with biological insights. These advancements promise transformative applications in drug discovery, disease prediction, and genomics, empowering computational biology with robust, interpretable tools.

Hepatocellular carcinoma (HCC), the most prevalent primary liver cancer, continues to pose a significant global health burden due to its high mortality rate. In addition to genetic alterations, epigenetic aberrations, including DNA methylation, histone modifications, chromatin remodeling, and noncoding RNA (ncRNA) dysregulation, play critical roles in HCC initiation and progression. Notably, miR-375 and miR-483-5p are among the most dysregulated miRNAs in HCC, with their altered expression levels closely associated with tumor stage and patient survival. These epigenetic modifications offer promising therapeutic avenues due to their reversibility and dynamic nature. Furthermore, specific epigenetic signatures such as CDH1 promoter hypermethylation and HOTAIR overexpression are being explored as potential biomarkers for early detection and treatment response. In this chapter, we review recent advances in the epigenetic landscape of HCC and discuss their diagnostic and therapeutic implications, highlighting their potential to improve patient outcomes through personalized medicine approaches.

<h4>Background</h4>Urinary tract infections (UTIs) are common infectious disorders affecting 50% of healthy women at least once during their lifetime. Antibiotic treatment for UTIs may increase the prevalence of antimicrobial resistance and side effects, highlighting the need for safe and efficacious alternatives. This study investigated the effect of acute supplementation with UClear, a D-mannose product, on UTI symptoms.<h4>Materials and methods</h4>Women with at least 2 uncomplicated UTI symptoms, as assessed using the UTI Symptom Assessment (UTISA) questionnaire and urine nitrite and leukocyte testing, were randomized to receive UClear or a placebo for 3 days. The study outcomes included changes in the UTI symptom severity and bother scores, complete resolution of UTI symptoms, urine culture data, quality of life assessed using the RAND 36-Item Short Form Survey questionnaire, and rescue medication use. Study outcomes were measured from baseline to days 2, 3, and 4.<h4>Results</h4>The total UTISA severity scores were not significantly different between the groups. Participants supplemented with UClear had lower total UTISA bother scores after 3 days than the placebo group (<i>p</i> = 0.027). On day 4, 43% of participants receiving UClear had complete resolution of "frequency of urination," based on the severity scores, compared with 20% of participants receiving the placebo (<i>p</i> = 0.032). A greater proportion of participants supplemented with UClear reported improvements in the severity and bother scores of "incomplete voiding" (<i>p</i> ≥ 0.020) and "urgency of urination" (<i>p</i> ≥ 0.059), compared with the placebo group. The proportion of participants in the UClear group with urinary microbial growth improved from 50% at baseline to 29% by day 4 (<i>p</i> = 0.020). UClear supplementation was safe and well tolerated.<h4>Conclusions</h4>After 3 days of UClear supplementation, urination-related UTI symptoms improved compared with those of the placebo group. These improvements corresponded with significant improvements in the degree of discomfort due to those symptoms. Further investigations in at-risk populations are warranted to understand the efficacy of UClear in resolving UTIs.

<h4>Introduction: </h4> Langerhans cell histiocytosis (LCH) is a rare clonal myeloid disorder characterized by tissue infiltration with pathological antigen-presenting cells morphologically and phenotypically similar to Langerhans cells. Activating mutations in the MAPK/ERK signaling pathway play a key role in the pathogenesis of LCH. The aim of this study was to characterize the morphological and molecular features of the neoplastic process. <h4>Materials and Methods:</h4> This study presents a clinical case of a 34-year-old patient with a rare mixed form of LCH combined with Erdheim-Chester disease (ECD), involving multiple organs including kidneys, skin, bones, ENT organs, eyes, and the central nervous system. The patient also had diabetes insipidus, chronic external otitis, and presumptive hereditary forms of anemia and hemochromatosis. A comprehensive histopathological evaluation was performed, including histochemical, immunohistochemical, and immunofluorescent analyses, alongside molecular genetic testing using whole exome sequencing (WES) of blood and skin samples. The patient demonstrated a partial clinical and laboratory response to targeted therapy with trametinib and desmopressin. <h4>Results:</h4> Comparison of WES data from blood and skin revealed 71,953 shared variants consistent with likely germline changes, as well as 3,081 unique somatic variants in skin and 2,633 unique variants in blood. Low-level mosaic BRAF V600E mutation (VAF = 1%) was detected in the skin, alongside pathogenic germline variants in the STEAP3 (c.523-2A&gt;T, NM_182915.3) and SLC40A1 (c.332T&gt;A, p.Met111Lys, NM_014585.6) genes. Immunofluorescence analysis of the trephine biopsy revealed diverse cellular populations, including sparse tryptase-positive mast cells, which showed minimal contact with each other or other labeled cells. The populations of Vimentin+CD11b–CD34+ and Vimentin+CD11b+CD34– cells (excluding endothelial cells) accounted for median values of 0.68% and 7.78%, respectively, of the total Vimentin+ cells. Regarding CD68 and S100 expression, CD68+ cells predominated (median = 55.61%), S100+ cells constituted a median of 43.02%, whereas CD68+S100+ cells were the rarest (median = 2.174%), reflecting the transition of Langerhans cells to LCH cells, encompassing up to 4.81% of the cellular population. This hybrid microenvironment paralleled the patient’s systemic inflammatory activity (elevated acute-phase reactants), multi-organ involvement, and renal interstitial fibrosis/tubular atrophy with reduced kidney function, without serological or histological evidence of vasculitis. <h4>Conclusion:</h4> This case illustrates the histological heterogeneity and hybrid pathology of mixed histiocytosis, where canonical dendritic LCH cells coexist with macrophage-rich ECD-like stroma. An integrated approach involving a multidisciplinary clinical team, thorough molecular genetic testing, and the application of targeted therapy is essential for improving prognosis and quality of life in young patients.

<h4>Background</h4>Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths worldwide, often diagnosed at advanced stages with limited treatment options. Despite advancements in systemic therapies, including the use of lenvatinib, the survival rate for advanced HCC remains low due to drug resistance and tumor heterogeneity.<h4>Method</h4>This study employed single-cell sequencing and spatial transcriptomics to investigate intra-tumor heterogeneity and identify subpopulations of malignant cells with inherent drug resistance. Tumor samples were obtained from patients undergoing lenvatinib, and differential analysis was performed to identify biomarkers associated with drug resistance.<h4>Results</h4>Analysis revealed significant differences in treatment response between different HCC nodules within the same patient, indicating intra-tumor heterogeneity. Single-cell sequencing and spatial transcriptomics identified distinct cell clusters within tumors, with specific subpopulations showing inherent resistance to lenvatinib. Differential analysis identified 17 common upregulated genes, including PCK1 and ALDH1A1, which were significantly associated with drug resistance. Pseudotime analysis further supported the role of PCK1 and ALDH1A1 as potential biomarkers of inherent drug resistance in HCC. This study highlights the importance of tumor heterogeneity and identifying biomarkers associated with drug resistance in HCC.<h4>Conclusions</h4>The findings suggest that PCK1 and ALDH1A1 may serve as potential biomarkers for predicting treatment response and guiding therapeutic strategies in HCC patients.

<h4>Background</h4>Our previous research revealed that PRDI-BF1 and RIZ homology domain containing protein 14 (PRDM14) accelerated human non-small cell lung cancer (NSCLC) cell migration. However, the internal mechanism remains unclear.<h4>Objective</h4>Herein, we aimed to explore whether PRDM14 modulates NSCLS cell growth and metastasis via H3K27me3-mediated phosphatidylethanolamine-binding protein 1 (PEBP1)/Raf-1/MEK/ERK signaling.<h4>Methods</h4>PRDM14 overexpression plasmid (OE-PRDM14), siRNA targeting PRDM14 (si-PRDM14) or si-jumonji domain-containing protein 3 (si-JMJD3) were transfected to alter PRDM14 or JMJD3 expressions. Vemurafenib (VMF) was utilized to inhibit Raf-1/MEK/ERK signaling. Cell proliferation was tested via Edu staining and colony formation assay. Cell migration and invasion were evaluated through two-chamber transwell assay. The protein levels of polycomb repressive complex 2 (PRC2), tri-methylation at lysine 27 of histone H3 (H3K27me3) and PEBP1/Raf-1/MEK/ERK signaling were measured via western blotting. qPCR was used to test mRNA expressions of PEBP1, c-Fos, c-Jun and c-Myc.<h4>Results</h4>OE-PRDM14 promoted A549 cell proliferation, migration and invasion, elevated PRC2 and H3K27me3 expressions, lowered PEBP1 expression and activated Raf-1/MEK/ERK signaling. si-PRDM14 had contrary influences. VMF weakened the influences of OE-PRDM14 on A549 cell functions and Raf-1/MEK/ERK signaling, but had no significant effects on PEBP1 and H3K27me3 levels. si-JMJD3 accelerated the influences of PRDM14 overexpression on A549 cell functions, PEBP1 and H3K27me3 expressions, as well as Raf-1/MEK/ERK signaling.<h4>Conclusions</h4>PRDM14 promoted NSCLC proliferation, migration and invasion via modulating PRC2/H3K27me3-mediated PEBP1/Raf-1/MEK/ERK signaling activation. PRDM14 might be as a new target in NSCLC therapy.

<i>Streptomyces</i> are naturally endowed with the capacity to produce a wide array of natural products with biomedical and biotechnological value. They have garnered great interest in synthetic biology applications given the abundance of uncharacterized biosynthetic gene clusters (BGCs). However, progress has been hindered by the limited availability of genetic tools for manipulating these bacteria. Several representative CRISPR-Cas systems have been established in <i>Streptomyces</i> to streamline experimental workflows and improve editing efficiency. Nevertheless, their broader applicability has been constrained by issues such as nuclease activity-related cytotoxicity and the large size of effector proteins. To address these challenges, we present <i>Streptomyces</i>-compatible TnpB-assisted genome editing (STAGE), a genetic toolkit based on ISDra2 TnpB, which is approximately one-third the size of Cas9 and enables precise, site-specific gene editing. We demonstrated that STAGE introduces genetic mutations with high efficiency and minimal off-target effects in two industrially important <i>Streptomyces</i> strains. Building on this platform, we developed STAGE-cBEST and STAGE-McBEST, enabling single and multiplexed C·G-to-T·A base editing, respectively, with editing efficiencies exceeding 75%. To further enhance performance, we engineered the ISDra2 TnpB system using an AI-assisted protein engineering framework, resulting in two variants that achieve nearly 100% genome editing efficiency. Additionally, through sequence homology analysis, we identified a TnpB ortholog from the same biological origin of ISDra2 TnpB, which also functions effectively as a gene editing tool. Our study establishes STAGE as a highly precise, programmable, and versatile genome editing platform for <i>Streptomyces</i>, paving the way for advanced genetic manipulation and synthetic biology applications in these industrially important bacteria.

Aspartimide formation remains a persistent challenge during Fmoc chemistry solid phase peptide synthesis. This review outlines various strategies to suppress base-mediated aspartimide formation, including the use of ester β-carboxyl protecting groups, non-ester-based β-carboxyl masking groups, and backbone-protecting groups. In addition, alternatives to the Fmoc group are explored that are cleavable under nonbasic conditions. The work discussed in this review highlights that the continued development of compatible and scalable tactics toward aspartimide prevention is essential for advancing peptide synthesis in both research and industry.

<h4>Introduction</h4>Post-ligation cardiac syndrome (PLCS) represents a state of severe post-intervention cardiopulmonary instability, seen in up to 50% of extremely premature infants after surgical closure of the patent ductus arteriosus (PDA); yet an evidence-based approach to treatment of this condition does not exist. The objective of this study is to determine the efficacy and safety of prophylactic milrinone in reducing incidence of PLCS and/or mortality within the first 7 days following PDA closure. The central hypothesis is that administration of intravenous milrinone will reduce the incidence of PLCS or death within 7 days of PDA closure either by percutaneous device (PCD) closure or surgical ligation (SL).<h4>Methods and analysis</h4>MIDAS (MIlrinone for prevention of post-patent Ductus Arteriosus closure Syndrome) is a multicentre, double-masked, randomised, placebo-controlled trial conducted across 19 neonatal intensive care units (NICUs) in the USA with an anticipated enrolment of 316 infants over 2 years. Prior to PDA closure (PCD or SL), infants will be randomised to either of two interventions that will be started after a 30-min period of stability and within 90 min following PDA closure: Group A (milrinone): milrinone infusion at an initial dose of 0.33 mcg/kg/min accompanied by a slow intravenous bolus of 10 mL/kg of 0.9% NaCl; Group B (placebo): 0.9% saline infusion of equivalent volume. Infants will be eligible if (1) gestational age at birth ≤27 weeks and 6 days and postnatal age ≤90 days at intervention; (2) haemodynamically significant PDA with minimum transductal diameter ≥1.0 mm prior to PDA closure or classified as at least small; (3) decision by clinical team to proceed with PDA closure via SL or PCD based on clinical and echocardiography features of haemodynamic significance; (4) invasive or non-invasive positive pressure respiratory support (does not include low-flow (<2.0 L/min) nasal cannula) for at least 2 days prior to PDA closure. The primary outcome is a composite of PLCS or death within 7 days of PDA closure. An intent to treat analysis will be applied using robust Poisson or logistic regression with generalised estimating equations (GEE) to account for centre effects and randomisation stratification factors.<h4>Ethics and dissemination</h4>The study has been approved by the University of Utah single Research Ethics Board (#00185742). Results will be disseminated through conferences, peer-reviewed publications, contributing to the enhancement of knowledge in post-PDA closure management and safety and efficacy of milrinone in preterm infants.<h4>Trial registration number</h4>NCT06679855.

Understanding the brain's molecular diversity requires spatially resolved maps of transcripts and proteins across regions and compartments. Here, we performed deep spatial molecular profiling of the mouse hippocampus, combining microdissection of 3 subregions and 4 strata with fluorescence-activated synaptosome sorting, transcriptomics, and proteomics. This approach revealed thousands of locally enriched molecules spanning diverse receptor, channel, metabolic, and adhesion families. Integration of transcriptome and proteome data highlighted proteins tightly linked to or decoupled from mRNA availability, in part due to protein half-life differences. Incorporation of translatome data identified roles for protein trafficking versus local translation in establishing compartmental organization of pyramidal neurons, with distal dendrites showing increased reliance on local protein synthesis. Classification of CA1 synapses revealed contributions from kinases, cytoskeletal elements, and adhesion molecules in defining synaptic specificity. Together, this study provides a molecular atlas of the hippocampus and its synapses (syndive.org), and offers insights into spatial transcript-protein relationships.

Lineage switching (LS) is the conversion of cancer cell lineage during the course of a disease. LS in leukemia cell lineage facilitates cancer cells escaping targeting strategy like CD19 targeted immunotherapy. However, the genetic and biological mechanisms underlying immune evasion by LS leukemia cells are not well understood. Here, we conduct a multi-omics analysis of patient samples and find that lineage-switched acute myeloid leukemia (LS AML) cells with KMT2A rearrangement (KMT2A-r) possess monocytic myeloid derived suppressor cell (M-MDSC)-like characteristics. Single-cell mass cytometry analysis reveals an increase in the M-MDSC like LS AML as compared to those of lineage-consistent KMT2A-r AML, and single-cell transcriptomics identify distinct expression patterns of immunoregulatory genes within this population. Furthermore, in vitro assays confirm the immunosuppressive capacity of LS AML cells against T cells, which is analogous to that of MDSCs. These data provide insight into the immunological aspects of the complex pathogenesis of LS AML, as well as development of future treatments.

The immune response against pathogens involves multiple cell state transitions and complex gene expression changes. Here, we establish a single-cell in vivo new RNA labeling sequencing method (scIVNL-seq) and apply it to survey time-resolved RNA dynamics during immune response to acute enteric infection with Salmonella. We show that the detection of new RNA synthesis reflects more realistic information on cell activation and gene transcription than total RNA level. Interplay of RNA synthesis and degradation modulates the dynamics of total RNA. The bone marrow macrophages are first primed at a very early stage upon Salmonella infection. In contrast, the innate immune response of macrophages in intestine is limited. Notably, intestinal CD8⁺ T cells and plasma cells are rapidly and specifically activated at the early stage post infection. Intestinal late enterocytes quickly express MHC-I molecules and present Salmonella antigen to CD8⁺ T cells for their activation, serving as antigen presenting cells for the initiation of adaptive immunity. Our findings reveal the RNA control strategies and the dynamic activation rules of immune cells in response to Salmonella infection, challenging the doctrine boundary between innate immunity and adaptive immunity against bacterial infection.

There are striking sex differences in the prevalence and symptomology of Major Depressive Disorder. Here, we conduct the largest sex-stratified genome wide association and genotype-by-sex interaction meta-analyses of Major Depressive Disorder to date (Females: 130,471 cases, 159,521 controls. Males: 64,805 cases, 132,185 controls). We identify 16 and eight independent genome-wide significant variants in females and males, respectively, including one novel variant on the X chromosome. Major Depressive Disorder in females and males shows substantial genetic overlap with a large proportion of variants displaying similar effect sizes across sexes. However, we also provide evidence for a higher burden of genetic risk in females which could be due to female-specific variants. Additionally, sex-specific pleiotropic effects may contribute to the higher prevalence of metabolic symptoms in females with Major Depressive Disorder. These findings underscore the importance of considering sex-specific genetic architectures in the study of health conditions, including Major Depressive Disorder, paving the way for more targeted treatment strategies.

Lung recurrence following hepatectomy is a common and clinically significant complication in patients with hepatocellular carcinoma (HCC), often leading to poor prognosis. However, the underlying mechanisms and reliable predictive biomarkers remain poorly defined. We performed cDNA microarray analysis to identify genes associated with lung recurrence after hepatectomy in HCC patients and identified lysyl oxidase (LOX) as a candidate. We further evaluated the association between LOX expression, circulating tumor cell (CTC), and microvessel density (MVD). The predictive value of serum LOX levels was assessed in both training and validation cohorts. LOX expression was significantly elevated in HCC patients who developed lung recurrence post-hepatectomy and was associated with worse prognosis. High intratumoral LOX expression correlated with increased CTC counts and elevated MVD. In vitro, LOX overexpression enhanced HCC cell migration and invasion, while LOX knockdown suppressed these phenotypes. Serum LOX levels demonstrated predictive potential for postoperative lung recurrence in both cohorts. LOX overexpression is closely associated with lung recurrence after hepatectomy in HCC patients. LOX may serve as a potential biomarker, and its serum level could be used to predict postoperative lung recurrence.

Valvular heart disease affects 2.5% of the population and is frequently associated with congenital heart disease. Blood flow is critical for valve formation, but the cellular mechanosensors translating flow into the transcriptional regulation of valve development remain undiscovered. Here, we identify that primary cilia and blood flow in mouse embryos regulate early valve development in vivo by regionally controlling endothelial-to-mesenchymal transition (EndoMT) through the modulation of Krüppel-like factor 4 (Klf4) in the endocardial cushions. Endocardial ciliation decreases during cushion development in regions of high shear stress, correlating with KLF4 downregulation and EndoMT progression. Mouse embryos lacking cilia exhibit blood flow-dependent accumulation of KLF4 and impaired cushion cellularization. Single-nucleus RNA sequencing revealed that the cilia-knockout and contractility-knockout endocardium fails to progress through EndoMT pseudostages, retains endothelial markers, and has reduced EndoMT and mesenchymal genes that KLF4 antagonizes. These data indicate that endocardial primary cilia function as mechanosensors in cushion development through the regional regulation of KLF4.

The DNA Damage Response (DDR) is a highly regulated process that safeguards genomic integrity against DNA lesions. Increasing evidence supports a reciprocal relationship between damaged chromatin architecture and the signalling pathways that coordinate the DDR. However, the mechanisms underlying this interplay in response to transcription-blocking DNA lesions remain largely unexplored. Here, we show that stalling of RNA polymerase II (RNAPII) at such lesions induces local chromatin acetylation, mediated primarily by the histone acetyltransferase p300. The resulting chromatin relaxation stimulates the dissociation of mature co-transcriptional spliceosomes from nascent RNA and promotes RNA:DNA hybrid (R-loop) formation, leading to ATM activation. In turn, activated ATM modulates chromatin conformation by phosphorylating histone H2A.X and triggering p38MAPK/MSK1-dependent histone H3S10 phosphorylation. Our findings highlight the cross-regulation between chromatin state and ATM signalling as a key component of the cellular response to transcription stress.

A hallmark of Alzheimer's disease (AD) is the accumulation of extracellular amyloid-β plaques in the brain. Amyloid-β is a 40-42 amino acid peptide generated by proteolytic processing of amyloid precursor protein (APP) via membrane-bound proteases. APP is a transmembrane protein, and its trafficking to sites of proteolysis represents a rate-limiting step in AD progression. Although APP processing has been well-studied, its trafficking itinerary and machinery remain incompletely understood. To address this, we performed an unbiased interaction screen for interactors of the APP cytosolic tail. We identified previously characterised APP binders as well as novel interactors, including RABGAP1. We demonstrated that RABGAP1 partially co-localises with APP and directly interacts with a YENPTY motif in the APP cytosolic tail. Depletion or overexpression of RABGAP1 caused mistrafficking and misprocessing of endogenous APP in human and rodent neurons. This effect is dependent on the GAP activity of RABGAP1, demonstrating that RABGAP1 affects the trafficking of APP by modulating RAB activity on endosomal subdomains. This novel trafficking mechanism has implications for other NPXY cargoes and presents a possible therapeutic avenue to explore.

Toxoplasma gondii is an obligate intracellular parasite that causes severe illness in infants infected during pregnancy and in immunocompromised individuals. This parasite manipulates host cells through effector proteins that promote its survival and replication. While the phosphatases in the PP2C family have been shown to regulate host immune responses and contribute to the virulence and pathogenicity of various pathogens, the specific biological functions of PPM3H in T. gondii and its role in host-pathogen interactions remain unclear. In this study, we demonstrate that knockout of ppm3h significantly reduces the virulence and pathogenicity of T. gondii. In contrast, that high expression of ppm3h in the less virulent PRU induced by replacing the ppm3h gene elements of RH strain can enhance its pathogenicity, indicating a direct contribution of PPM3H to virulence in expression-independent manner. Furthermore, PPM3H significantly influenced host gene expression, with differentially expressed genes predominantly enriched in immune and inflammatory pathways. Weighted gene co-expression network analysis identified host immune genes, including chemokines such as Cx3cl1 and Ccl22, as co-expressed with ppm3h. Also, ppm3h co-expressed with T. gondii rhoptry genes including rop18, a well-known virulence factor, suggesting a role for PPM3H in coordinating host-pathogen interactions. Our findings establish that PPM3H enhances T. gondii virulence by modulating the host immune and inflammatory responses. PPM3H does not impact parasite gene expression, invasion or replication in vitro, supporting its role as an immune modulator rather than a general fitness factor. This suggests that T. gondii's pathogenicity arises not only from immune evasion but also from the active induction of host immune and inflammatory responses mediated by PPM3H.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disease caused by an abnormal expansion of cytosine-adenine-guanosine (CAG) trinucleotidein the huntingtin gene. Mutant huntingtin (mHTT) expression in neurons and glial cells affects neuron and astrocyte functions and leads to the loss of medium spiny neurons of the striatum. Brain cholesterol pathway is severely affected by HTT mutation in neurons and astrocytes, contributing to HD pathogenesis. Decreased cholesterol production and transport by astrocytes impair synapse maturation and neurotransmission. Brain cholesterol metabolism is maintained by cholesterol hydroxylation into 24-hydroxycholesterol by the neuronal enzyme cholesterol 24-hydroxylase (CYP46A1). CYP46A1 is decreased in affected brain regions in HD patients and mice. AAV-CYP46A1 striatal delivery was shown to restore cholesterol metabolism with neuroprotective effects in two mouse models of HD, characterized by mHTT aggregates' reduction, improved transcriptomic profile, and Brain-Derived Neurotrophic Factor (BDNF) signaling, and preservation of striatal neurons. From a therapeutic perspective, we intended to clarify the detailed mechanisms and the specific role of neurons and astrocytes in the therapeutic effects of AAV-CYP46A1 delivery. We first evaluated CYP46A1 expression in astrocytes in HD post-mortem putamen at a late stage of disease progression. To determine the specific contribution of CYP46A1 expression in astrocytes compared to neurons on the HD phenotype, we assessed the effects of AAV-CYP46A1 striatal injection under the control of astrocytic (GFA2) or neuronal (hSYN) promoters in R6/2 mice. Overall, equivalent transgenic CYP46A1 protein levels, both astrocytic and neuronal targeting, mitigate medium ppiny neuron (MSN) atrophy and improve spine density in R6/2 mice. Reduction of mHTT aggregates in neurons is similar when CYP46A1 is overexpressed in neurons or in astrocytes. However, astrocyte targeting reduces mHTT aggregates in neurons and astrocytes, while restricted neuronal targeting reduces mHTT aggregates in neurons only. Altogether, astrocytic targeting of CYP46A1 expression in CYP46A1-tested animals combines cell-autonomous and non-cell-autonomous mechanisms of action, with improved phenotypic correction compared to neuronal-restricted targeting. Allowing expression in both cell types with higher expression levels of CYP46A1 showed overall better efficacy. We demonstrate that astrocyte-neuron combined targeting with AAV-CAG-CYP46A1 delivery increases therapeutic efficacy. This study brings new evidence that CAG-mediated CYP46A1 striatal overexpression significantly modifies the transcriptome in R6/2 mice for pathways involved in synaptogenesis and inflammation, suggesting targeting both astrocytes and neurons provides benefits for HD phenotypic correction.

Periodontitis is a multifactorial inflammatory disease involving pathogenic biofilm formation, amplified oxidative stress, and impaired tissue regeneration. In addition to its complicated pathology, effective treatment of periodontitis is challenged by a dynamic oral microenvironment that prevents drug retention. To overcome these issues, an anti-bacterial, ROS-scavenging, and tissue-regenerative hydrogel system (HQUP@TF127) is developed. In this triple-functional HQUP@TF127, a ROS-responsive gatekeeper on hollow mesoporous silica nanoparticles enabled the spatiotemporally controlled release of quercetin, a naturally occurring anti-inflammatory and osteogenic ingredient. The covalent attachment of the antibacterial, 4-terpineol with thermosensitive Pluronic F127 prolonged retention time, thereby ensuring deep penetration and eradication of subgingival pathogens. HQUP@TF127 restored endoplasmic reticulum homeostasis and, maximized the osteogenic potential of periodontal ligament stem cells. In a rat model of periodontitis, HQUP@TF127 effectively suppressed osteoclast activation by inhibiting inflammatory infiltration and collagen degradation. Micro-computed tomography analysis confirmed an increase in bone mineral density and periodontal tissue regeneration. HQUP@TF127, addressed the multifactorial pathology and obstacles to local drug administration and, requires further translational research by virtue of its triple synergistic mechanisms of action and advantages in local drug delivery.

RNA interference (RNAi, also known as RNA silencing) is one of the most important plant defense responses against viral invasion. Although major components of the RNAi pathway, steps leading to viral small interfering RNA biogenesis, and viral counterdefense strategies via RNAi suppressors have been well studied, the broader roles of RNAi in viral infection and seed transmission remain less thoroughly characterized. In particular, the increasing complexity of RNAi-associated mechanisms and their integration with other biological processes have not been comprehensively summarized. Increasing numbers of studies have identified non-canonical RNAi pathways, novel host factors involved in RNAi, and the possibility of small RNAs acting across kingdoms to modulate plant-virus-vector tritrophic interactions. In this review, we provide an overview of the roles of RNAi in plant viral infections and describe recent advances, with emphasis on the discoveries of novel positive and negative RNAi regulators, potential signaling pathways upstream and downstream of antiviral RNAi, and the prospects and challenges of double-stranded RNA applications, either expressed from transgenes or supplied exogenously via spraying. We also discuss how these findings reshape current views on antiviral RNAi, highlight remaining knowledge gaps, and examine how these advances influence plant-virus co-evolution while informing strategies for managing plant virus diseases and reducing their impact.

DNA probe design plays a critical role in biosensor-based disease diagnostics, gene expression analysis and environmental monitoring. Traditional probe designs primarily target lower-copy genetic sequences, often leading to low detection sensitivity due to limited hybridization events. This study introduces a novel probe design strategy that leverages highly repetitive DNA sequences as target sites to amplify biosensor signals without requiring PCR-based amplification. The computational selection process is conducted using a custom-developed bioinformatics tool to identify repetitive sequences across the entire <i>Mycobacterium tuberculosis</i> genome, independent of gene boundaries. The identified sequences are then cross-referenced against the <i>Homo sapiens</i> genome using BLAST to minimize host cross-reactivity. The analysis revealed that a 23 bp sequence repeated 39 times in <i>M. tuberculosis</i> exhibits only 78% sequence identity with human DNA and is present in just two copies within the human genome. This suggests that the selected probe may yield substantially stronger hybridization signals for <i>M. tuberculosis</i> relative to human cfDNA, thereby enhancing biosensor sensitivity. The computational methodology introduced in this study provides a robust framework for designing high-sensitivity biosensors, enabling more effective infectious disease diagnostics, environmental monitoring and clinical point-of-care testing.

No abstract available.

<b>Objective:</b> Mauriac syndrome (MS) is a rare condition linked to inadequate glycemic control in type 1 diabetes mellitus (T1DM) and has also rarely been reported in patients with neonatal diabetes. MS manifests as growth failure, delayed puberty, cushingoid features, and hepatomegaly. The condition can be associated with complications like dyslipidemia, retinopathy, and nephropathy. The main objective of this study was to describe the magnitude of the condition, clinical features, management, and outcome of Sudanese children and adolescents with MS due to inadequate control of diabetes in our center. <b>Study Design and Methods:</b> This is a cross-sectional hospital-based study. All medical records of patients with MS were reviewed. Data, including demographics, clinical features, investigations, management, and outcome, were obtained. Patients were re-educated and management intensified then followed up. <b>Results:</b> Thirty-seven MS patients were enrolled in this study, with a male predominance of 59.5%. Neonatal diabetes was diagnosed in 5.4% of the patients, while others had T1DM. The median age at diagnosis of MS was 12 years. The diagnosis was based solely on clinical findings, including a history of prolonged unsatisfactory glycemic control, short stature, and hepatomegaly. Regarding the outcome, eight children (21.6%) were lost to follow-up, one patient died (2.7%), seven (18.9%) had a static condition, and those who showed improvement were 21 (56.8%). Signs of improvement were a decrease in liver size with or without an increase in growth velocity. Nephropathy was the most common associated complication; it was seen in 33.3% of our cohort. Some got it at a very young age. <b>Conclusions:</b> Despite many efforts that have been made to achieve better glycemic control in children with T1DM, MS still exists in our setting. Though liver biopsy is the gold standard for diagnosis, being invasive, the diagnosis could be made conservatively, based on clinical features and response to treatment. The condition can be reversed with good metabolic control.

Epithelioid mesothelioma (EM) is a pleural malignancy whose many histopathologic patterns may overlap considerably with those of lung adenocarcinoma (LAC) or poorly differentiated squamous cell carcinoma (SCC). This study aimed to evaluate the diagnostic role of SOX6 immunohistochemical expression in EM, study its differential expression in EM, LAC, and SCC, and evaluate the utility of various combinations of SOX6 with established EM markers calretinin and D2-40. The study included 39 EM, 21 LAC, and 11 SCC cases. SOX6 expression was detected in 71.8 % of EM cases. Conversely, all SCC cases were SOX6-negative, and only two LAC cases were SOX6-positive (P < 0.001). The sensitivity and specificity of SOX6 in identifying EM was 71.8 % and 93.8 %, respectively. Calretinin and D2-40 expression was detected in 100 % and 97.4 % cases of EM, respectively. The diagnostic sensitivity of SOX6 for EM in combination with D2-40 and/or calretinin was higher than SOX6 as a solitary marker. Notably, the sensitivity of calretinin and/or SOX6 positive expression was 100 % higher than that of SOX6 combination with D2-40. Although the sensitivity of SOX6 is lower than that of other established markers for EM, it may be a fairly specific marker for the diagnosis of EM. Therefore, the inclusion of SOX6 into an immunohistochemical panel may have diagnostic utility in distinguishing between EM and lung carcinomas. However, more research is needed on a wider array of tumor types from various organs to truly understand its global specificity.

<i>NDUFV1</i> encodes NADH: ubiquinone oxidoreductase core subunit V1, a key component of mitochondrial Complex 1. Biallelic pathogenic variants in this gene produce a broad and variable phenotypic spectrum in affected individuals, including ophthalmoplegia, developmental delays, brain imaging abnormalities, and recurrent episodes of emesis and lactic acidemia. We report female siblings compound heterozygous for two missense variants (Arg40Gln, Val245Met) in <i>NDUFV1</i> with unusual presentations of this condition. The 6-year-old proband showed normal growth and neurodevelopment until recently when weight loss and recurrent vomiting were noticed and brain imaging abnormalities consistent with Complex 1 deficiency were documented. She developed lactic acidemia without a clear precipitating factor and that, incongruously, was associated with profound alkalosis with blood pH as high as 7.83. We describe management of her acute illness during a hospital admission with aggressive sodium bicarbonate and sodium acetate replacement, and eventual recognition that anxiety-related hyperventilation contributed substantially to her transient profound alkalosis. We review the complex interplay of lactic acidemia due to mitochondrial Complex 1 deficiency, metabolic acidosis from acute loss of bicarbonate, respiratory alkalosis from hyperventilation and hypocapnia, and other concomitant medical issues, as well as her distinctive neuroradiological findings. Her 14-year-old sister was diagnosed retrospectively despite an earlier initial presentation, and manifests greater neurocognitive effects, similar neuroradiological signs, but no history of acute metabolic decompensation. These cases expand the phenotypic spectrum of this rare inherited illness, provide new information about its presentation and intrafamilial variability, and offer insight relevant to management of life-threatening metabolic crises associated with this disorder.

Lipid peroxidation stands as a prominent hallmark and a prerequisite for the onset of ferroptosis. Lipid metabolism holds a pivotal role in regulating this process, forming the metabolic foundation for cellular sensitivity to ferroptosis. Studies in lipid metabolomics reveal that the activation of Polyunsaturated fatty acids (PUFA), specifically arachidonic acid and adrenoic acid (AdA), mediated by acyl-CoA synthetase long-chain family member 4 (ACSL4), represents a critical step in generating lipid peroxidation substrates. The expression level or enzymatic activity of ACSL4 emerges as a potential indicator of cellular susceptibility to ferroptosis. Additionally, other members of the ACSL family can indirectly influence the occurrence of ferroptosis by modifying the fatty acid composition of the cell membrane. Given the high expression of ACSL4 in various human tumors, targeting lipid peroxidation with ACSL4 as the focal point may pave a new path in tumor therapy. This article provides a brief overview of the primary structure and function of ACSL4, its role in lipid peroxidation, and summarizes the current advancements in drug development targeting ACSL4 and lipid peroxidation.

To align sleep-wake behavior and internal physiology with the Earth's 24-h light-dark cycle, organisms rely on circadian clocks-endogenous timekeeping systems that anticipate and adapt to daily environmental changes. These clocks are governed by transcription-translation feedback loops that produce rhythmic oscillations in gene expression, including key regulators such as PERIOD and CRYPTOCHROME. The timing and stability of these proteins are tightly controlled by post-translational mechanisms, including ubiquitin-mediated degradation. The ubiquitin-proteasome system (UPS) ensures that clock proteins are cleared at precise times within the circadian cycle, a process which is essential for resetting the molecular clock and sustaining robust circadian rhythms. Disruption of this process can have profound impacts on human health and contribute to impairments in sleep timing, circadian phase, and rhythm amplitude. In this review, we focus on the mechanistic role of the UPS in circadian clock regulation, summarize key E3 ligases and deubiquitinating enzymes implicated in clock protein turnover, and highlight the essential role of the UPS on sleep timing and overall circadian biological homeostasis.

<i>Mycobacterium tuberculosis</i> (Mtb) is the causative agent of tuberculosis capable of manipulating and circumventing the host's immune system to establish infection. Ubiquitination plays a crucial role in the host's response to pathogens; however, the global alterations in protein ubiquitination during Mtb infection remain poorly understood. To elucidate the regulatory roles of ubiquitination in the immune response to Mtb, we investigated the ubiquitome of human macrophages following Mtb infection. In our study, we identified a total of 1,618 proteins exhibiting altered ubiquitination levels, with 1,182 lysine-ubiquitination sites in 828 proteins showing increased ubiquitination and 1,077 sites in 790 proteins displaying decreased ubiquitination. Bioinformatics analyses revealed that most proteins involved in the immune response were upregulated, including those associated with autophagy, lysosome, the NF-κB signaling pathway, necroptosis, and ferroptosis. Furthermore, the ubiquitination levels of numerous proteins involved in conserved physiological processes, such as ribosome biogenesis, spliceosome function, nucleocytoplasmic transport, and mRNA surveillance, were also altered, suggesting that these pathways may be regulated by ubiquitination during Mtb infection. The extensive pool of ubiquitinated proteins and sites identified in this study will serve as a valuable resource for understanding the regulatory mechanisms of the ubiquitination system in immune responses during Mtb infection.

The giant freshwater prawn (<i>Macrobrachium rosenbergii</i>) has economic significance in the aquatic industry, but production is impacted by infectious diseases induced by various pathogens. Herein, we investigated the impact of adding feed additives to the diet of <i>M</i>. <i>rosenbergii</i> to promote health. Diets were formulated to contain different levels of <i>Centella asiatica</i> (L.) Urb. crude extracts (1, 5, and 10 g kg^-1), with growth performance and innate immune parameters assessed after 28 days of feeding. Real-time quantitative PCR (RT-qPCR) was employed to determine the mRNA levels of <i>serine proteinase inhibitor</i> (<i>SPI</i>) and <i>alpha-2 macroglobulin</i> (<i>Mr-2α2M</i>) from 12 h to 28 days of feeding. Prawns feeding at 5 and 10 g kg^-1 showed statistically significant differences in specific growth rate, lysozyme assay, and phenoloxidase activity. The expression levels of all the immune-related genes studied were significantly upregulated in prawns fed with supplemented diets compared to the control group. Findings revealed that the observed upregulations varied in response to alterations in feeding time and concentration. Furthermore, 16S rRNA analysis showed that the supplemented diets at 10 g kg^-1 supplementation increased beneficial bacteria (<i>Lactococcus</i> sp.) and reduced pathogenic taxa (e.g., <i>Candidatus Hepatoplasma</i>, <i>Flavobacteriaceae</i>, <i>Weeksellaceae</i>, <i>Thiothrix</i> sp.).

Regenerative medicine is the branch of medicine that aims to repair and regenerate damaged tissues and presents promising avenues for addressing a wide range of currently incurable diseases. Regenerative medicine is based on the use of cell therapy with stem cells that can differentiate into differentiated cells of specific tissues. There are various types of stem cells, which are different in potential and derivation. The aim of this review is to summarize the types of stem cells most studied and recently discovered, from adult stem cells to innovative induced pluripotent stem cells (iPSCs), for regenerative medicine purposes. The stem cells involved in the identification of new regenerative therapeutic approaches are analyzed here through a classification based on the tissues' embryonic derivation: stem cells from ectodermal derivation tissues, stem cells from mesodermal derivation tissues, stem cells from endodermal derivation tissues, and iPSCs.

The poultry industry, a globally fast growing agricultural sector, provides affordable animal protein due to high efficiency. <i>Gallus gallus domesticus</i> are the most common domestic birds. Hybrid chicken breeds (crosses) are widely used to achieve high productivity. Maintaining industry competitiveness requires constant genetic selection of parent stock to improve performance traits. Genetic studies, which are essential in modern breeding programs, help identify genome variants linked to economically important traits and preserve population health. Next-generation sequencing (NGS) has identified millions of single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELs), enabling detection of genome-wide regions associated with selection traits. Recent studies have pinpointed such regions using broiler lines, laying hen lines, or pooled genomic data. This review discusses advances in chicken genomic and transcriptomic research focused on traits enhancing meat breed performance and reproductive abilities. Special attention is given to transcriptome studies revealing regulatory mechanisms and key signaling pathways involved in artificial molting, as well as metagenome studies investigating resistance to infectious diseases and climate adaptation. Finally, a dedicated section highlights CRISPR/Cas genomic editing techniques for targeted genome modification in chicken genomics.

SARS-CoV-2 entry into host cells involves multiple steps and is a highly orchestrated process. Both the host protease TMPRSS2 and the HR1/HR2 segment within the spike (S) protein play a crucial role in promoting viral invasion. Herein, we report a series of bifunctional SARS-CoV-2 entry inhibitors formed by covalently linking a TMPRSS2 inhibitor, Camostat (Cm), and an HR1-targeting peptide fusion inhibitor IPB19 via a poly (ethylene glycol) (PEG) linker. Among them, IP4X and IP4Z display potent inhibitory activities against SARS-CoV-2 with similar IC₅₀ values of 0.16 μM and 0.17 μM, respectively. The efficacy surpassed that of their parent inhibitors by approximately 28-fold relative to Camostat and 15-fold relative to IPB19. We confirm that IP4X and IP4Z exhibit a dual-targeting mechanism by binding to both TMPRSS2 and HR1 region of S protein. These findings highlight the potential of the bifunctional inhibitors for further development as a novel multitarget therapy against SARS-CoV-2 infection and enrich the understanding of S-mediated entry of SARS-CoV-2 into host cells.

<b>Background/Objectives</b>: Gut microbiota has been implicated in the pathogenesis of metabolic dysfunction-associated steatotic liver disease (MASLD) and cardiovascular disease (CVD). This study aimed to identify associations between gut dysbiosis and MASLD, regarding body mass index (BMI) and subclinical coronary atherosclerosis (SCA). <b>Methods</b>: We conducted a cross-sectional study of 202 patients with MASLD who had no previous history of CVD. The severity of MASLD was evaluated using a magnetic resonance imaging-based method, and SCA was measured by assessing coronary artery calcification (CAC). Gut microbiota was assessed in fecal specimens using sequencing targeting the V4 region of the <i>16S rRNA</i> gene. <b>Results</b>: Our results demonstrated that gut microbial profiles between low- and high-BMI groups (<30 vs. ≥30 kg/m²) differed significantly in beta-diversity, but not in alpha-diversity indices. At the genus level, we identified <i>Megamonas</i>, <i>Sutterella</i>, <i>Catenibacterium</i>, and <i>Odoribacter</i>, enriched in the high BMI group. Compared to the low CAC group (<100 AU), MASLD patients with high CAC scores (≥100 AU) exhibited enrichment in <i>Ruminococcus gnavus</i>, <i>Bacteroides</i>, and <i>Lachnoclostridium</i>, along with depletion of several short-chain fatty acid (SCFA)-producing bacteria, such as <i>Faecalibacterium</i>. Multivariate analysis demonstrated that older age, the presence of diabetes, high BMI, fibrosis stage F3-F4, and high plasma trimethylamine oxide (TMAO) levels were independently associated with a high CAC score in patients with MASLD. <b>Conclusions</b>: These data indicated that gut dysbiosis and related metabolites, in association with advanced liver disease, were potential contributors to the progression of SCA in obese patients with MASLD.

<h4>Background</h4>Lysine acetylation plays a critical role in prostate cancer (PCa) by modulating androgen receptor (AR) signaling. However, the exact mechanisms by which lysine acetylation impacts PCa prognosis remain unclear. The aim of this study was to investigate the mechanism by which lysine acetylation affects PCa prognosis by modulating the AR signaling pathway.<h4>Methods</h4>Data from The Cancer Genome Atlas-Prostate Adenocarcinoma (TCGA-PRAD), GSE54460, and lysine acetylation-related genes (LARGs) were obtained from public databases and literature. Differentially expressed genes (DEGs) were identified in TCGA-PRAD, and key module genes associated with LARGs were selected using weighted gene co-expression network analysis (WGCNA). Candidate genes were identified by overlapping DEGs and key module genes. A biochemical recurrence-free (BCR-free) prognostic model was constructed and validated using BCR-free survival data from patients with PCa. Prognostic genes were further confirmed through machine learning. PCa samples were stratified into high- and low-risk subgroups based on the median risk score. A nomogram model was developed integrating clinicopathological features and risk scores to identify independent prognostic factors. Enrichment analysis, tumor microenvironment profiling, and drug sensitivity assessments were performed for the two risk subgroups.<h4>Results</h4>A total of 2,658 DEGs and 723 key module genes were analyzed, yielding 105 overlapping candidate genes. Five genes-<i>UBXN10</i>, <i>ACOX2</i>, <i>PLCL1</i>, <i>PLS3</i>, and <i>SLIT3</i>-were identified as BCR-free-related prognostic markers in TCGA-PRAD. The prognostic risk model revealed significantly lower BCR-free survival rates in the high-risk subgroup compared to the low-risk subgroup. A nomogram incorporating Gleason score, tumor stage (T stage), and risk score effectively predicted BCR-free survival in patients with PCa. Notably, natural killer (NK) cells, myeloid dendritic cells, endothelial cells, and fibroblasts were significantly correlated with PLS3 (|Cor| >0.3, P<0.05). Drugs such as cisplatin, MK-1775, and ulixertinib were identified as potential therapeutic agents for PCa.<h4>Conclusions</h4>Five BCR-free-related prognostic genes were identified as potential therapeutic targets. Additionally, a BCR-free-related prognostic risk model was developed, offering a robust tool for predicting BCR-free survival in patients with PCa.

Traumatic experiences elicit a wide range of cognitive responses in both humans and animals, leading to diverse outcomes such as enhanced performance, cognitive impairment, or the development of mood and anxiety disorders like posttraumatic stress disorder (PTSD). A key challenge in understanding these varied responses is to decipher the underlying biological mechanisms that contribute to individual variability in trauma resilience or susceptibility. The purpose of this study was to elucidate the molecular bases for these differences, focusing on the amygdala and hippocampus-brain regions integral to stress responses. We exposed adult, male rats to an acute, severe stressor and profiled persistent anxiety-like behavior outcomes 7 days later. We investigated the transcriptional signatures in the basolateral amygdala and hippocampal dentate gyrus via bulk RNA sequencing from animals with behavioral outcomes indicative of stress resilience or vulnerability. Our results suggest that the basolateral amygdala and dentate gyrus display distinct transcriptomic changes following acute, severe stress. Furthermore, we identified specific region-dependent genes related to insulin signaling, neural plasticity, and stress responses that correlate with resilient and vulnerable phenotypes. Notably, a larger number of genes separated stress-resilient animals from both control and stress-susceptible animals, underscoring that an active molecular response, particularly in the hippocampus, facilitates protection from the long-term consequences of severe stress. These findings provide novel insight into the mechanisms that engender individual variability in the behavioral responses to stress and offer new targets for the advancement of therapies for stress-induced neuropsychiatric disorders.

Liquid-liquid phase separation (LLPS) is now recognized as one of the key mechanisms underlying the formation of membraneless organelles. Typically, condensates formed through LLPS are dynamic and play a crucial role in the spatiotemporal regulation of essential cellular processes. In some cases, however, condensates can undergo an aberrant liquid-to-solid transition, which is now recognized as being related to the onset of cancers and neurodegeneration. The microtubule-associated protein Tau, the aberrant aggregation of which is implicated in neurodegenerative disorders like Alzheimer's and Parkinson's, has been found to undergo LLPS. The Tau condensates formed through LLPS are considered to be intermediate on-pathway precursors of amyloid aggregates. Unlike other known phase-separating proteins (e.g., FUS or TDP-43) that have low-complexity domains (LCDs), Tau is intrinsically disordered. Thus, Tau exhibits a highly flexible structure that can be modulated by changes in environmental changes. The intricate relationship between different conformations of full-length Tau and its phase behavior remains poorly understood. To bridge this gap, here, by employing a combination of single-molecule FRET and molecular dynamics simulations, we demonstrate that Tau undergoes conformational transitions from compact to extended states during LLPS, irrespective of diverse driving forces. Moreover, we show that intramolecular interactions responsible for stabilizing the compact conformations of monomeric Tau correlate with the intermolecular interactions driving the LLPS of Tau, thereby facilitating the formation of dynamic networks. These findings provide crucial mechanistic insights into how the conformational state of Tau governs its propensity for phase separation, shedding light on sequence-encoded structural processes that ultimately drive biological phase separation.

Lyngbyabellins O and P are complex natural products derived from non-ribosomal peptide synthetase/polyketide synthase (NRPS/PKS) biosynthetic pathways and have been isolated from marine cyanobacterial sources. Both metabolites are characterized by the presence of two thiazole rings and a distinctive dichlorinated <i>β</i>-hydroxy acid side chain. Notably, lyngbyabellin P is further distinguished by the incorporation of a (3<i>R</i>,4<i>S</i>)-statine moiety. Herein, we report the first total syntheses of lyngbyabellins O and P, which are achieved through the convergent coupling of three key synthetic fragments, namely, two enantiomerically enriched thiazole subunits and a hydroxycarboxylic acid derivative, the latter constructed via a stereoselective aldol reaction. The total syntheses were completed in 12 and 13 longest linear steps (LLSs) for lyngbyabellins O and P, respectively, furnishing the natural products in overall yields of 5.6% and 2.5%.

Tuberculosis (TB) remains a formidable global public health challenge. The rising prevalence of drug-resistant TB and increased human immunodeficiency virus(HIV) co-infection further exacerbate TB control efforts. Mycobacterium tuberculosis (Mtb) achieves highly heterogeneous infection outcomes (active disease, latency, or clearance) through immune evasion and host metabolic reprogramming. While conventional diagnostic techniques offer cost-effectiveness and accessibility without complex infrastructure, they are constrained by low sensitivity, prolonged turnaround times, and an inability to distinguish latent TB infection (LTBI) from active TB disease (ATB). Recent research into host-derived biomarkers provides a promising strategy to overcome diagnostic bottlenecks by deciphering characteristic molecular changes in host-pathogen interactions. This review systematically reviews advances in host-derived biomarkers for TB diagnosis, critically discussing the clinical potential, translational challenges, and future research directions of integrated multi-omics biomarker panels to enhance diagnostic sensitivity and specificity, differentiate ATB from LTBI, and guide precision therapy.

In recent times, neurodegenerative diseases have become a global health concern, particularly among the elderly. This may be attributed to the increased risk of neuronal death due to age. Moreover, the underlying mechanisms of neurodegeneration are largely driven by age-related processes that include oxidative stress, mitochondrial dysfunction, and inflammation. Despite extensive research efforts, however, neurodegenerative disorders still remain incurable as current therapeutic strategies provide limited efficacy as well as severe side effects. For these reasons, dietary phytochemicals are being considered as preventive strategies because they have potential neuroprotective functions against age-related neurodegeneration. This review summarizes the mechanisms underlying age-related neurodegeneration and highlights the current challenges in their treatment and management. It also discusses the potential of dietary phytochemicals as complementary interventions, focusing on their neuroprotective functions and mechanisms of action. Finally, challenges surrounding the use of dietary phytochemical interventions in controlling age-related neurodegenerative disorders are addressed and solutions to these challenges based on available research are discussed.

Lung adenocarcinoma (LUAD) remains a leading cause of cancer-related mortality. We performed multi-omics approaches on LUAD samples spanning different stages, establishing a detailed epigenetic landscape. We identified 1,416 regions characterized by hypo-DNA methylation and hyperchromatin accessibility associated with high mortality and recurrence rates, serving as a biomarker panel for LUAD staging and diagnosis (AUC of 0.89 and 0.87 for two-class and multi-class models). Epigenetics-related transcription factors, especially the AP-1 family, were observed at these loci, and transcriptomic profiling indicated persistent activation of the mitogen-activated protein kinase (MAPK) signaling pathway. Hypomethylation and hyperaccessibility of AP-1 binding sites enhance EGFR and FGFBP1 expression, activating the MAPK pathway. This reveals an epigenetic-mediated positive feedback loop, MAPK→AP-1/Epigenetic Modifications→EGFR/FGFBP1→MAPK, in LUAD, highlighting the complex interplay between epigenetic modifications and LUAD progression, offering potential detection and treatment targets. These findings define a high-resolution, multi-stage epigenetic landscape of LUAD, providing a resource for biomarker discovery and mechanistic insight.

Autophagy is a process of engulfing cytoplasmic proteins or organelles, thereby fulfilling cells' metabolic needs and the renewal of specific organelles. Given its key roles in tumor progression, autophagy has attracted tremendous attention in cancer therapies. Notably, there is a megatrend to integrating autophagy regulation into mainstream treatments. This review focuses on autophagy-targeting nanomedicine (ApT-NM) to modulate autophagy in tumor therapy, including the unmodified and functionalized nanoparticles that target tumors by carrying autophagy modulators. On the one hand, it can reverse treatment resistance by inhibiting protective autophagy, and on the other hand, it can promote the death of cancer cells through type II apoptosis by inducing autophagy. Moreover, advanced nanoplatforms combining various treatments (such as chemotherapy, radiotherapy, photothermal therapy, and photodynamic therapy, etc.) have also been summarized. Last, the future perspectives and directions for ApT-NM research are provided, hoping to emphasize this rising filed and promote the development of ApT-NM.

No abstract available.

No abstract available.

No abstract available.

No abstract available.

<h4>Background</h4> The iron overload disease haemochromatosis is primarily caused by HFE p.C282Y homozygosity, yet penetrance of clinical outcomes (including liver disease/cancer) varies. We estimated the effect of genetic and lifestyle factors on disease penetrance and expressivity in C282Y homozygotes. <h4>Methods</h4> We analysed 2,893 C282Y homozygous UK Biobank participants (n=1,295 male). We ascertained haemochromatosis from medical records, liver disease/cancer, osteoarthritis, joint replacement surgeries, and dementia diagnoses. We derived polygenic scores (PGS) for iron biomarkers including hepcidin and transferrin saturation (TSAT). Sex-stratified logistic regression assessed associations with clinical outcomes. We used time-to-event regression estimating effects of age, lifestyle, and PGS. We estimated effects of rare HFE variants using whole genome sequencing data. <h4>Results</h4> In male HFE C282Y homozygotes, higher TSAT PGS increased likelihood of diagnosis of haemochromatosis, and separately any clinical consequence (OR top-vs-bottom-PGS-quintile =1.83, 95%CI: 1.26-2.66, p=0.001). Cumulative incidence of assessed haemochromatosis clinical outcomes in men by age 80 was 64.5% (highest quintile) versus 51.6% (lowest) (p-value for difference=0.025). In females, TSAT PGS increased haemochromatosis likelihood (cumulative incidence: 45.3% vs. 23.3% [highest/lower quintile], p=0.00001) but not liver disease. PGS for other iron biomarkers were not significantly associated with clinical outcomes. Rare heterozygous predicted loss-of-function variants in HFE increased haemochromatosis likelihood in non-C282Y homozygotes (aggregate-OR=14.8, 95%CI 4.7-41.1, p=0.003), highlighting the importance of sequencing undiagnosed individuals to find rare causes of haemochromatosis. <h4>Conclusion</h4> Higher genetically predicted transferrin saturation significantly increased risk of clinical outcomes in HFE C282Y homozygotes. Combined with modifiable lifestyle factors, genetic information could refine risk stratification and personalise iron monitoring, following validation.

<h4>Purpose</h4>Implant stability, related to mechanical (primary) and biological (secondary) bone-to-implant interactions, is essential for osseointegration. Implant surface bioactivation is a process designed to accelerate and enhance surface-cell interaction.The purpose of this systematic review was to determine whether a beneficial effect of bioactive (BS) over traditional surfaces (TS) can be identified.<h4>Materials and methods</h4>An electronic search of Pubmed, Scopus, and CENTRAL databases was performed to identify randomized (RCT) and non-randomized controlled trials comparing BS and TS implants. Risk of bias was assessed using the Cochrane Collaboration tool for RCTs and the Joanna Briggs Institute tool for non-RCTs. Outcome variables were implant stability quotient (ISQ) measured through resonance frequency analysis from placement to prosthetic loading, one-year implant survival rate, and marginal bone loss (MBL). Meta-analysis was performed where possible.<h4>Results</h4>Of the 6920 records identified, 13 RCTs and two non-RCTs were included, reporting on 1256 implants (49.8% TS and 50.2% BS) in 596 patients. Four of the studies had a low risk of bias, three had a moderate risk and eight had a high risk. The meta-analysis showed no evidence of an effect of implant surface on survival rate (p = 0.99, 10 studies) and MBL (p = 0.86, 5 studies). At baseline (10 studies) and at one month (9 studies) the ISQ did not differ statistically significantly different between groups. A statistically significantly greater increase in ISQ was found for the BS implants compared to the TS implants (p = 0.04) at three months after placement (9 studies).<h4>Conclusion</h4>An advantage of BS over TS during the early osseointegration phase could not be demonstrated, but a positive effect on implant stability seems to occur after three months of placement. The statement that bioactive surfaces may safely allow early and immediate implant loading is insufficiently supported by the current evidence.

<h4>Background</h4>Esophageal carcinoma (EC) is the eighth most common cancer, which is a major health problem on a global scale. Comprehensive treatments such as surgery, radiotherapy and chemotherapy have demonstrated efficacy in certain patients with early-stage esophageal carcinoma. However, it is crucial to explore improved treatment modalities for individuals with advanced, recurrent and metastatic esophageal carcinoma.<h4>Methods</h4>The eQTL datasets were obtained from Drug-Gene Interaction Database (DGIdb V.4.2.0) and an interesting review about druggable genes. The two-sample MR was used to identify potential drug targets. Co-localization analysis was applied to evaluate if the genes expression single nucleotide polymorphisms (SNPs) drove and was associated with EC risk. And drug prediction and molecular docking were used to verify the pharmaceutical value of potential clinical drugs.<h4>Results</h4>The present study found thirty potential drug targets that were remarkably correlated with the risk of developing esophageal carcinoma at least in two eQTL datasets by two-sample MR analysis. Five genes colocalized with esophageal carcinoma were identified through colocalization analysis and the medicinal value of four potential clinical drugs was testified by molecular docking.<h4>Conclusions</h4>We identified five drug targets by MR analysis and co-localization analysis and four drugs by drug prediction and molecular docking analysis respectively, which provide new insights for treatment of esophageal carcinoma.

With the development of medical and health care, esophageal cancer (EC) has become a disease of concern to the scientific research community. At present, among all treatment regimens for EC, surgical resection is conducive to the prognosis of early patients neoadjuvant therapies are recommended for advanced patients. However, treatments now are not satisfactory in suppressing the progression of EC. Ferroptosis is one distinctive cell death mode, noted for the accumulation of iron as well as lipotoxicity, which induce cell membrane to breakdown. As a star protein of ferroptosis related pathway, GPX4 is related to the homeostatic imbalance of tumor immune microenvironment (TIME) of EC, thereby regulating the onset as well as progression of the cancer. In our manuscript, we present the mechanisms involved in ferroptosis, the functions of ferroptosis in the TIME. We also focused on the progression about ferroptosis in EC, as well as targeting ferroptosis-related pathways to delay the development of EC. We expect that these contents can expand fresh insights and aim for EC therapeutic strategy in clinical practice.

Chimeric antigen receptor (CAR) T cell therapy targeting CD19 elicits remarkable clinical efficacy in B cell malignancies, but many patients relapse owing to failed expansion and/or progressive loss of CAR-T cells. We recently reported a strategy to potently restimulate CAR-T cells in vivo, enhancing their functionality by administration of a vaccine-like stimulus comprised of surrogate peptide ligands for a CAR linked to a lymph node-targeting amphiphilic PEG-lipid (amph-vax). Here we demonstrate a general strategy to discover and optimize peptide mimotopes enabling amph-vax generation for any CAR. We use yeast surface display to identify peptide binders to FMC63 (the scFv used in clinical CD19 CARs), which are then subsequently affinity matured by directed evolution. CAR-T vaccines using these optimized mimotopes triggered marked expansion and memory development of CD19 CAR-T cells in both syngeneic and humanized mouse models of B-acute lymphoblastic leukaemia/lymphoma, and enhanced control of disease progression compared with CD19 CAR-T-only-treated mice. This approach enables amph-vax boosting to be applied to any clinically relevant CAR-T cell product.

Lichen simplex chronicus (LSC), also known as neurodermatitis, is a common chronic pruritic dermatosis defined by lichenified plaques resulting from persistent scratching. Though often secondary to underlying dermatologic, systemic, or psychological triggers, LSC represents a distinct clinical entity with significant morbidity. The hallmark itch-scratch cycle contributes not only to visible skin changes but also to substantial sleep disruption, emotional distress, and functional impairment. Psychological stress, anxiety, and depression are frequent comorbidities and can further perpetuate disease chronicity. This review provides a comprehensive summary of the evolving understanding of LSC, from its neuroimmune-driven pathogenesis to the wide spectrum of therapeutic strategies currently available. In addition to topical corticosteroids, novel approaches including immunomodulators, neuromodulators, Janus kinase (JAK) inhibitors, and biologics are being increasingly explored. Procedural therapies such as cryotherapy, fractional laser resurfacing, and botulinum toxin injections, have also emerged as valuable tools, particularly in treatment-refractory cases. Recent insights into type 2 inflammation and dysregulated sensory pathways have informed the rationale for these targeted strategies. In anatomically sensitive areas such as the genital region, where topical agents may be poorly tolerated, systemic treatments may be required. Given this complexity, individualized, multimodal treatment plans are critical to optimizing management and improving quality of life (QoL) in patients with LSC. By synthesizing current data on pathophysiology, diagnosis, and both established and emerging therapies, this review aims to guide clinicians in optimizing care for patients with LSC and addressing its far-reaching psychosocial burden.

<h4>Background</h4>Dysregulation of the endocannabinoid system (eCBS) and the loss of CB1 receptors (CB1R) in the basal ganglia are well-established hallmarks of Huntington's disease (HD). As a result, significant research efforts have focused on targeting the eCBS to alleviate motor disturbances associated with the disease. Beyond its role in motor control, the eCBS is a complex signaling network critically involved in regulating learning and memory. Despite this, the potential involvement of eCBS dysfunction in the cognitive decline characteristic of HD, often manifested well before motor dysfunction, has remained largely unexplored.<h4>Methods</h4>CB1R expression in the hippocampus was evaluated in both human HD samples and HD mouse models (R6/1 and Hdh^Q7/Q111 models, including both sexes) using Western blotting, immunohistochemistry, and radioligand binding assays. To restore CB1R function, CB1R agonist WIN-55212-2 was systemically administered, or viral vectors encoding CB1R were locally infused into the hippocampus of HD mice. A multidisciplinary approach combining behavioral, biochemical, electrophysiological, and morphological analyses, was employed to investigate the molecular mechanisms underlying the effects of CB1R activation in the context of HD-related cognitive dysfunction.<h4>Results</h4>In both human HD samples and HD mouse models, CB1R protein levels were reduced in the hippocampus, accompanied by structural synaptic alterations and impairment in spatial, recognition and working memory. Moreover, hippocampal depolarization-induced suppression of inhibition was significantly disrupted in R6/1 mice. Administration of WIN-55212-2 successfully restored these synaptic and cognitive deficits. Immunohistochemical analysis revealed that the CB1R decrease was specifically localized to GABAergic interneurons within the hippocampus. Notably, targeted restoration of CB1R expression in these interneurons via viral vector delivery was sufficient to rescue hippocampal-dependent memory deficits in HD mice.<h4>Conclusion</h4>This study suggests that impaired CB1R function in hippocampal GABAergic interneurons contributes to memory dysfunction in HD.

<h4>Background</h4>Glucocorticoids (GCs) are key regulators of hematopoiesis, but the effects of chronically elevated endogenous GC levels on hematopoietic stem cell (HSC) function and immune cell development remain poorly understood.<h4>Methods</h4>We used a mouse model with adrenocortical cell-specific deletion of hypoxia-inducible factor-1 alpha (HIF1α; P2H1^Ad.Cortex), which results in sustained and systemic elevation of GC. Hematopoietic stem and progenitor cell (HSPC) populations were analyzed phenotypically and functionally. Transplantation assays assessed the regenerative capacity of HSCs. To determine the role of glucocorticoid receptor (GR) signaling, bone marrow from GR-deficient or wild-type donors was transplanted into P2H1^Ad.Cortex or wild-type (WT) recipients.<h4>Results</h4>Chronic GC exposure in P2H1^Ad.Cortex mice resulted in HSPC expansion and promoted HSC quiescence and metabolic restraint. Functionally, these HSCs showed enhanced regenerative capacity with superior donor chimerism upon transplantation. There was a marked increase in myeloid progenitors and their progeny, including monocytes and granulocytes. In contrast, B-cell development was significantly impaired, with a developmental block at the pre-pro-B-cell stage. Transplantation studies confirmed that these effects were dependent on GR signaling.<h4>Conclusions</h4>Our study reveals a critical role for chronic GC-GR signaling in modulating HSC function, promoting myeloid output, and impairing B-cell development. The P2H1^Ad.Cortex mouse model provides a valuable system to study the hematopoietic consequences of prolonged endogenous glucocorticoid exposure and may aid in understanding the hematologic complications of chronic GC therapy.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder that leads to dementia. Many cases are diagnosed annually and there is no currently available cure. Understanding the underlying disease biology of AD through the study of molecular networks, particularly by mapping clinical variants to tissue-specific interactomes and regulatory macromolecular assemblies, offers a promising avenue to elucidate altered disease pathways. In this study, we applied differential interactome analysis using a manually curated AD dataset to identify how disease-associated mutations alter both transient and stable protein interactions. By focussing on variant-specific associations detected in brain-relevant tissues, we mapped disruptions in stable macromolecular assemblies and performed Reactome enrichment analysis to uncover perturbed pathways unique to each variant. Additionally, we explored therapeutic insights through the analysis of amyloid precursor protein (APP) physical interactors, identifying potential intervention points that influence amyloidogenic processing. Complementing protein-level data, we integrated microRNA (miRNA)-mediated regulatory interactions, revealing an additional layer of posttranscriptional control over key AD genes. Together, this multilayered strategy provides a framework for precision therapeutics in AD.

Cellular maturation is a crucial step for tissue formation and function, distinct from the initial steps of differentiation and cell fate specification. In the central nervous system, failure of oligodendrocyte maturation is linked to diseases such as multiple sclerosis. Here, we report a transcriptional mechanism that governs the timing of oligodendrocyte maturation. After progenitor cells differentiate into immature oligodendrocytes, the transcription factor SOX6 redistributes from super-enhancers to cluster across specific gene bodies. These sites exhibit extensive chromatin decondensation and transcription, which abruptly turn off upon maturation. Suppression of SOX6 deactivates these immaturity loci, accelerating the transition to mature, myelinating oligodendrocytes. Notably, cells harboring this immature SOX6 gene signature are enriched in multiple sclerosis patient brains and antisense oligonucleotide-mediated Sox6 knockdown drives oligodendrocyte maturation in mice. Our findings establish SOX6 as a key regulator of oligodendrocyte maturation and highlight its potential as a therapeutic target to promote myelination in disease.

<h4>Introduction</h4>Cryptosporidium spp. is a protozoan parasite known to cause gastrointestinal infections, particularly in immunocompromised individuals such as cancer patients. It typically infects the small intestine but has been reported in extraintestinal sites, including the biliary tract, and lungs. While Cryptosporidium has been implicated in the pathogenesis of several gastrointestinal cancers, there is very limited data its association with pancreatic ductal adenocarcinoma (PDAC). This study aimed to investigate the presence of Cryptosporidium in pancreatic tissue and saliva samples from PDAC patients using digital PCR (dPCR), a highly sensitive diagnostic tool, and to compare findings with those in distal cholangiocarcinoma (DCC) patients and healthy controls.<h4>Methods</h4>A total of 50 participants were enrolled, including 20 PDAC patients, 10 DCC patients, and 20 healthy controls. Pancreatic tissue and saliva samples were collected from PDAC and DCC patients, while only saliva was collected from healthy controls. DNA was extracted from all samples, and the presence of Cryptosporidium was investigated using both quantitative PCR (qPCR) and dPCR. Cancer patients were also analyzed for clinical findings and patients positive for Cryptosporidium spp. in saliva and tissue samples were evaluated clinically.<h4>Results</h4>Cryptosporidium DNA was detected by qPCR in the saliva of one DCC patient. In contrast, dPCR revealed Cryptosporidium spp. in the saliva of one PDAC and two DCC patients (one overlapping with qPCR), and in the pancreatic tissue of two PDAC patients (whose saliva was negative). No Cryptosporidium spp. was detected in the healthy control group. Although the differences between groups were not statistically significant, the presence of Cryptosporidium in pancreatic tissue was demonstrated for the first time using dPCR. Some positive patients showed respiratory symptoms or were asymptomatic, raising the possibility of subclinical or extraintestinal infection.<h4>Conclusion</h4>This study provides the first dPCR-based evidence of Cryptosporidium in pancreatic tissue, suggesting its potential for extraintestinal dissemination in cancer patients. The findings highlight the superiority of dPCR over qPCR for detecting low-abundance pathogens in clinical samples. Although no direct causal link with PDAC was established, the detection of Cryptosporidium in pancreatic tissues warrants further investigation into its potential role in pancreatic carcinogenesis.

Protein mislocalization and aggregation are hallmark features in neurodegeneration. As proteins mislocalize, proteostasis deficiency and protein aggregation typically follow. Autophagy is a crucial pathway for the removal of protein aggregates to maintain neuronal health, but is impaired in various neurodegenerative diseases, including Huntington disease (HD). We identified S-acylation, a reversible lipid modification of proteins, as an important regulator in protein trafficking and autophagy. SQSTM1 (sequestosome 1/p62) is an essential selective autophagy receptor for the sequestration of ubiquitinated cargoes within autophagosomes and subsequent delivery into lysosomes for degradation. Recently, we reported that S-acylation of SQSTM1 at the di-cysteine motif C289,290 directs SQSTM1 to lysosomes. We further showed that SQSTM1 S-acylation is significantly reduced in brains from both HD patients and mouse HD model, which may result in the cargo sequestration defect within autophagosomes in HD. Treatment with palmostatin B, a deacylation inhibitor, significantly increases SQSTM1 localization to lysosomes. Our work highlights SQSTM1 S-acylation as a novel potential therapeutic strategy in HD. As a crucial autophagy component, our work suggests S-acylation of SQSTM1 may have a broader role in neurodegeneration.

Breakthrough discoveries in RNA biology have led to a paradigm shift in our understanding of RNA-from passive intermediates to active regulators of gene expression. Technological innovations and deeper insights into RNA regulation have transformed the field, positioning RNA as a powerful tool for therapeutic development. Recent advances in RNA technologies have revolutionized medicine by enabling the precise targeting of specific mRNAs to modulate aberrant transcripts, correct genetic defects, and reprogram cellular behavior. This review provides an overview of the coordinated regulation of mRNA processing and its application to RNA-based therapeutics, including antisense oligonucleotides (ASOs), splice-switching oligonucleotides (SSOs), small interfering RNAs (siRNAs), and Aptamers. We focus on clinically approved RNA therapeutics, emphasizing their biological mechanisms such as RNA stability and splicing regulation. The expanding repertoire of RNA technologies underscores the translational potential of RNA biology and its growing clinical impact. Future developments are expected to yield highly specific, modular, and programmable RNA medicines capable of treating a wide range of previously intractable diseases.

<h4>Background</h4>Whether very preterm infants benefit from provision of respiratory support prior to cord clamping has been an important knowledge gap. Four randomised controlled trials have recently been published on this topic and have provided new insights. These 4 studies and 2 previous ones included preterm infants less than 32 weeks of gestational age receiving deferred cord clamping (DCC). The intervention consisted of providing respiratory support in the form of continuous positive airways pressure or positive pressure ventilation via T piece. The intervention group had cord clamping performed when preset stability criteria were met in one study or after an elapsed time of up to 120 s was achieved in the other 5 studies (50-120 s range). The control group had time-based cord clamping after 30-60 s without respiratory support (5 studies) or cord milking (1 study).<h4>Summary</h4>Outcome measures based on important neonatal outcomes such as death, severe intraventricular haemorrhage (sIVH), and bronchopulmonary dysplasia (BPD) were reported as were other outcomes such as admission temperature and transfusions. Overall outcomes (death, sIVH, and BPD) were similar in intervention and control groups in these studies with moderate certainty of evidence to conclude there was no benefit for these outcomes. Maintaining normothermia was difficult, and the mean difference in admission temperature was significantly lower in the intervention group, although hypothermia was generally mild.<h4>Key message</h4>Overall, the provision of respiratory support during DCC did not improve important neonatal outcomes (moderate certainty of evidence). Although we do not currently recommend the procedure as routine practice, we acknowledge there is room for further studies.

<h4>Objective</h4>Wool serves as an important textile raw material, with fiber diameter being a major determinant influencing the economic value and quality of wool products. Investigating the regulatory mechanisms that influence wool fiber diameter is necessary for formulating aimed at improving wool fineness. We used methylationomics to examine the skin tissue of individuals with different fiber diameters, and investigated the regulation mechanisms influencing wool fiber diameter.<h4>Methods</h4>In this study, we analyzed the transcriptome and m6A methylome of skin tissues from individual Alpine Merino sheep, classified into three groups based on wool fiber diameters, to identify key methylated RNAs and explore the role of m6A methylation in regulating this trait.<h4>Results</h4>A total of 54,057 methylated peaks, 4,273 differentially methylated genes, 139 differentially methylated lncRNAs, and 2,992 differentially methylated circRNAs were found in the three comparisons. These gene loci showed enrichment in the Wnt, Notch, and TGF-β signaling pathways, as determined through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses. RNA correlation analyses revealed key RNAs, such as CACNA1E, FOS, CAMK2B, RNF43, circ-0317, circ-4794, TCONS-00020832, and TCONS-00020845, indicating that hypermethylation may play an important role in affecting wool fiber diameter.<h4>Conclusion</h4>The findings elucidate the molecular regulatory mechanisms underlying wool fiber diameter and provide a theoretical foundation for advancements in the wool industry.

<h4>Objective</h4>This study aims to investigate the effects of time and light variation on the growth and reproductive performances of breeder geese.<h4>Methods</h4>A total of 32 ganders and 96 dames in their first laying season, with an average age of 10 months, were assigned to eight rooms, each containing four ganders and twelve dams. A split-plot design was used, incorporating two prelay photoperiod treatments: a 7-hour light period (P7H), and a gradually decreasing light (GDL) group. Two variable photoperiod schedules were applied as sub-treatments and randomized across the rooms. In one treatment, the egg production rate in breeder geese decreased to an average below 30%, after which the photoperiod increased by 15 minutes each week (change time). A fixed photoperiod of nine hours was maintained (fixed time) until the end of the egg-laying period.<h4>Results</h4>Geese in the GDL light group had a longer laying duration than P7H light group (243.75 vs 191.75 days; p<0.01). Number of eggs per goose in the GDL light group showed a trend toward higher values than P7H light group (81.82 vs 55.45 egg/bird; p = 0.0779). However, the fertility and hatchability in GDL light group were significantly lower than the P7H light group at all periods, respectively (48.35 vs. 62.57% and 42.80 vs. 53.17%; p<0.05). Income over feed cost for the GDL and P7H light groups was 3,069.6 and 2,535.5 NT$/bird, respectively.<h4>Conclusion</h4>Geese exposed to a 12-hour light (12L:12D) regimen during the pre-laying period exhibited a longer laying duration and higher egg production per bird. However, maintaining a fixed lighting schedule of 9 hours of light and 15 hours of darkness (9L:15D) after the peak laying period is recommended to optimize production profitability by supporting better fertility and hatchability.

<h4>Background</h4>Colorectal cancer (CRC) is a major cause of cancer deaths globally, mainly due to treatment resistance. Neddylation, a key post-translational modification, is linked to tumor growth and immune response, offering potential therapeutic targets, though its role in CRC is not well-explored.<h4>Methods</h4>We examined neddylation-related genes (NRGs) across cell subtypes using CRC scRNA-seq data from the TISCH database. Unsupervised clustering of TCGA and GEO bulk RNA-seq data identified various neddylation patterns. A neddylation-related gene signature (NRGS) was developed using ten machine-learning algorithms and validated externally. The study compared biofunctions, including functional analysis, immune cell infiltration, genomic mutations, enrichment analysis, and responses to immunotherapy and chemotherapy, between high- and low-risk groups defined by the NRGS model.<h4>Results</h4>scRNA-seq analysis showed that the high neddylation score group had more malignant and diverse immune and stromal cells, with activated pathways aiding tumor growth and spread. We identified two neddylation patterns: Cluster A and Cluster B. Cluster B, associated with worse survival, had more immunosuppressive cells and increased tumor progression. We developed a neddylation-related gene signature (NRGS) using ten machine-learning algorithms, which accurately predicted outcomes. Higher risk scores correlated with poorer survival, with AUCs of 0.979, 0.989, and 0.996 for 1-year, 2-year, and 3-year OS in the training cohort. The NRGS was linked to higher recurrence or metastasis, advanced disease stage, and independently predicted OS risk. Patients with high NRGS may resist immunotherapy and standard chemotherapy.<h4>Conclusion</h4>The NRGS could predict outcomes and responses to immunotherapy and chemotherapy in CRC patients, aiding personalized treatment, though further validation is needed.

<h4>Background</h4>MicroRNA-491-5p (miR-491-5p) is a key regulator of cell proliferation and inflammation, but its role in asthma pathogenesis remains unclear. This study aimed to investigate the mechanistic involvement of miR-491-5p in airway remodeling and inflammation, focusing on its downstream target, B4GalT5, and oxidative stress pathways.<h4>Methods</h4>MicroRNA sequencing of airway smooth muscle (ASM) tissues from asthma patients revealed significant downregulation of miR-491-5p, and bioinformatic prediction combined with dual-luciferase reporter assays identified B4GalT5 as a direct downstream target. Clinical correlation analyses assessed the relationship between B4GalT5 expression, Ki-67 (a marker of cell proliferation), and percentage of airway wall area to total tracheal area (WA%). To explore the functional role of miR-491-5p, an ovalbumin (OVA)-induced asthma model was established in C57BL/6 mice, followed by intratracheal instillation of AAV-miR-491-5p to restore its expression in vivo. Airway inflammation and remodeling were evaluated using ELISA, and histological staining (HE, Masson and PAS staining). Oxidative stress markers, including reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and adenosine triphosphate (ATP), were also quantified. In vitro, airway smooth muscle cells (ASMCs) were stimulated with TNF-α to mimic the inflammatory microenvironment and then transfected with a miR-491-5p mimic to assess effects on cell proliferation, cytokine secretion, mitochondrial morphology, ROS production, and Ca²⁺ homeostasis.<h4>Results</h4>MiR-491-5p was significantly downregulated in ASM tissues from asthma patients. B4GalT5 was identified and validated as a direct target of miR-491-5p. Clinically, B4GalT5 expression was positively correlated with Ki-67 and WA%, suggesting a role in airway remodeling. In the OVA-induced asthma model, miR-491-5p overexpression markedly alleviated airway inflammation, mucus hypersecretion, collagen deposition, and structure remodeling, accompanied by reduced oxidative stress (decreased ROS and MDA, increased SOD activity and ATP levels). In TNF-α-stimulated ASMCs, miR-491-5p mimic suppressed excessive proliferation, cytokine release, mitochondrial dysfunction, ROS elevation and intracellular Ca²⁺ homeostasis. Mechanistic studies confirmed that miR-491-5p directly binds to the 3'-UTR of B4GalT5, inhibiting its expression and downstream oxidative stress pathways.<h4>Conclusion</h4>These findings demonstrate that miR-491-5p attenuates asthma-related airway remodeling and inflammation by targeting B4GalT5 and modulating oxidative stress, providing a novel target for therapeutic intervention.

Mast cell leukemia (MCL) is an exceedingly rare and aggressive variant of systemic mastocytosis (SM). MCL is classified as primary, occurring <i>de novo</i> without prior mast cell (MC) disorders or secondary, from a pre-existing SM, and acute aggressive form with C-findings that indicate organ damage or chronic indolent form without organ damage. Of the cases, 60-65% are aleukemic with < 10% circulating MCs in the peripheral blood, and the rest of the cases are leukemic with > 10% MCs. Diagnosis is typically confirmed by bone marrow biopsy revealing greater than 20% atypical or immature MCs in the smear. Specific MCL-targeted treatments are limited, and multiple treatment modalities used for SM and acute myeloid leukemia (AML) have been tried in MCL with limited success and variable survival benefit. The management has significantly advanced since the implication of the receptor tyrosine kinase type III (KIT) D816V mutation in the pathogenesis of SM and MCL. The two targeted therapies approved for MCL are midostaurin, a multikinase inhibitor, and avapritinib, a selective KIT D816V mutation-targeted tyrosine kinase inhibitor. Multiple drugs are being evaluated in clinical trials for managing MCL. MCL has a poor prognosis with a median overall survival (OS) of around 1.5 years. Further advancements and research are essential to develop treatments that may enhance median OS. In this article, we conducted a comprehensive yet simplified review of MCL, focusing primarily on its clinical manifestations and recent updates on management. We also identified the areas that require further research and emphasized the aggressive nature and poor prognosis associated with this disease.

In the cardiovascular system, elastic fibres exert a fundamental role providing the long-range elasticity required for physiological functions. Elastic fibres are complex in composition and structure containing, in addition to elastin, a wide range of matrix components, such as microfibrillar proteins, calcium-binding proteins and glycosaminoglycans. Changes in composition and/or structure can affect the biomechanics of the tissue as well as the intrinsic affinity of elastin for Ca²⁺ ions. Mineralization of elastic fibres can occur in genetic as well as in age-related chronic diseases. In cardiovascular diseases, for instance, calcification represents an integral part of the pathogenetic process, although the regulatory mechanisms are not completely understood. Therefore, a focus is given on elastin synthesis and assembly, on elastic fibre components and on elastin degradation. Moreover, the role and the impact of altered composition and supramolecular organization of elastic fibres are described in the context of the calcified cardiovascular system. Finally, some <i>in vitro</i> and <i>in vivo</i> models of elastic fibres calcification are presented and discussed.

Alginates have been modified to become a suitable platform for targeted drug release, triggering advances in targeted and controlled-release systems. Alginate chemistry and various structural modifications, including ionic crosslinking, cyclodextrin-linking, and chemical carboxyl and hydroxyl group modification, such as esterification, amidation, and sulfation, are covered in this review. These modifications are aimed at enhancing drug release mechanisms, like dissolution, diffusion, water penetration, and ion exchange-based systems. Application of modified alginates in controlled-release systems is extensive, from hydrogels, bioprinting approaches, nanofiber-based systems, in-situ gelling systems, to nanoparticle systems. Targeted drug delivery utilizes passive and active targeting strategies for numerous applications, including the treatment of cancer, inflammatory disease therapy, infectious disease, and wound healing. Emerging platforms such as hydrogels and nanoparticles provide evidence of the versatility of modified alginates. However, issues regarding scalability and biocompatibility remain as obstacles. In this review, the capabilities of alginate-based delivery systems and their role in making various release mechanisms for targeted delivery are discussed in relation to the future directions for precision medicine development.

Cirrhotic cardiomyopathy (CCM) is a cardiac dysfunction in patients with cirrhosis, occurring in the absence of structural heart disease. It increases perioperative risk, especially in liver transplantation, and may contribute to hepatorenal syndrome. Despite its clinical significance, CCM remains poorly understood and lacks effective treatments. This review aims to summarize recent findings on the pathogenesis of CCM and highlight potential therapeutic targets. A focused literature review was conducted using PubMed, Scopus, and Clarivate databases, selecting studies from the last five years. Included studies investigated molecular, cellular, and receptor-mediated mechanisms involved in CCM. Results: CCM results from neurohumoral, inflammatory, and electrophysiological disturbances. Key mechanisms involve dysfunction of β-adrenergic and muscarinic receptors, altered ion channels (potassium, L-type calcium), impaired sodium-calcium exchange, and suppression of the P2X7 receptor (P2X7R). Dysregulation of the CD73 (5'-nucleotidase, ecto-5'-nucleotidase)-A2 adenosine axis, along with effects from endocannabinoids, nitric oxide (NO) inhibition by tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6), carbon monoxide (CO), and elevated galectin-3 (Gal-3), further contribute to myocardial dysfunction. Conclusions: CCM is a multifactorial condition linked to systemic and myocardial effects of cirrhosis. A deeper understanding of its mechanisms is essential for developing targeted therapies. Further research is needed to improve patient outcomes.

Segmental bone defects often present as new irregularly growing bones that fail to bridge gaps. Optimizing the scaffold design for direct bone growth can enhance bone defect reconstruction. We evaluated three carbonate apatite honeycomb-structured scaffolds with uniaxial macropores of 280, 440, and 640 μm (labeled P280, P440, and P640, respectively) implanted into critical-sized ulnar defects in rabbits. The scaffold performance was assessed at 4 and 12 weeks postimplantation. P280 formed small volumes of oriented bone, which decreased the bone strength. P440 formed large volumes of oriented bone and restored bone strength. Meanwhile, P640 formed abundant volumes of disorganized bone and failed to enhance bone strength. The 440 μm macropores effectively guided new bone and enhanced the bone strength. These findings suggest that the macropore size is crucial for designing scaffolds for effective segmental bone defect reconstruction.

<h4>Background</h4>Limited evidence and contradictory results exist regarding lymph node metastasis (LNM) and prognosis in early gastric cardia cancer (EGCC) and early gastric non-cardia cancer (EGNCC). This study aims to compare the clinicopathological features, LNM patterns, and survival outcomes between EGCC and EGNCC using a large population-based dataset.<h4>Methods</h4>This study utilized data from the Surveillance, Epidemiology, and End Results (SEER) population and employed multivariate analysis, Kaplan-Meier method, propensity score matching (PSM), and nomogram analysis to achieve comprehensive insights.<h4>Results</h4>EGCC tended to be younger in age, intestinal type, smaller tumor size, and well-differentiated type (P<0.05). No positive association was found between LNM and tumor location after adjusting for other risk factors [odds ratio (OR): 0.87; 95% confidence interval (CI): 0.60-1.25; P=0.44]. Moreover, patients with EGNCC showed a better prognosis compared with EGCC patients [5-year disease-specific survival (DSS): 87.3% <i>vs.</i> 80.3%, P<0.001 for log-rank test]. Patients with early gastric cancer (EGC) were further divided by invasion depth. When EGC patients were limited to the mucosa, EGCC patients had a similar overall survival (OS) to EGNCC patients (P=0.26). As the depth of infiltration reached the submucosa, EGCC had a significantly worse DSS compared to EGNCC (5-year DSS: 73.9% <i>vs.</i> 85.7%, P<0.001 for log-rank test). PSM further proved that our analysis was credible and reliable.<h4>Conclusions</h4>The risk of LNM in EGCC is comparable to that in EGNCC. However, EGCC exhibits poorer survival outcomes compared to EGNCC. This discovery underscores the importance of enhanced monitoring and individualized treatment approaches for patients with EGCC to improve their prognosis and survival.

<h4>Purpose</h4>This review mainly focuses on the aetiology of attention-deficit/hyperactivity disorder (ADHD) including genetic, neuro-biological, biochemical and environmental factors. It also emphasizes the possible management of the disorder. This knowledge will be essential for developing therapeutic interventions and prevention measures.<h4>Views</h4>ADHD is a neurodevelopmental psychiatric disorder, characterized by pervasive patterns of inattention, hyperactivity, or impulsivity. Globally, it affects up to 7-10% of elementary school children, and symptoms frequently persist into adulthood. Among adults, 2-5% experience ADHD symptoms worldwide. The aetiology and developmental trajectory of the disorder remain unknown. ADHD is caused by a combination of genetic, psychological, physiological, biochemical, and environmental factors. All of these factors are thought to permanently change the development of the dopaminergic and noradrenergic system, which is a fundamental neurobiological mechanism behind the disorder.<h4>Conclusions</h4>Untreated ADHD can have long-term effects on individuals throughout their lifetime and cause adverse life trajectories such as poor educational and occupational outcomes, substance addiction, accidents, suicide, criminal activity, and co-occurring mental and physical health disorders. Principal management options include psychopharmacological treatments such as group cognitive behavioural therapy and social skills training, parent education programmes, and school-based initiatives, as well as medication employing stimulants and non-stimulants.

Iron represents an essential element required for normal physiologic processes throughout organ systems. A vast network of transporters is involved not only in uptake of this element but in processing, oxidation, and recycling to maintain it in a tight balance to avoid excess storage. This complex network of transporters, including heme and ferroportin, among many others, are responsible for facilitating inter-organ tissue iron exchange and availability, contributing to overall heme homeostasis. However, exposure to high levels of iron can overwhelm compensatory mechanisms that result in its accumulation and toxicity. This is the case of patients with genetic diseases such as hemoglobinopathies who suffer from chronic anemia and require, in most instances, a lifetime of red blood cell transfusions to overcome disease crises. Thus, in light of the extensive role of iron in the body, the aim of this review is to present important metabolic pathways involved in iron homeostasis across the cardiovascular, reproductive, hematopoietic, urinary, respiratory, endocrine, and central nervous systems while contrasting these against negative effects caused by iron excess.

Type 2 diabetes mellitus (T2DM) is frequently complicated by atherosclerosis (AS), with substantial overlap in their underlying pathophysiological mechanisms, posing serious threats to patient health. Tirzepatide, a novel dual agonist of glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptors, has demonstrated remarkable efficacy in glycemic control, weight reduction, and cardiometabolic improvement, making it a promising candidate for managing T2DM comorbid with AS. However, substantial interindividual variability in treatment response suggests a role for genetic determinants. This review systematically summarises current evidence on pharmacogenomic variants influencing the efficacy and toxicity of tirzepatide, explores the interplay between drug response genes and genetic susceptibilities to T2DM and AS, and highlights the potential of pharmacogenomics in guiding precision subtyping and individualised therapy. Finally, we highlight key challenges and future directions in the clinical translation of tirzepatide pharmacogenomics, aiming to inform personalized, genomics-guided therapy for cardiometabolic disease.

Equine semen preservation is fundamental to modern equine reproduction, supporting breeding programs, genetic conservation, and industry sustainability. However, significant challenges persist, including temperature sensitivity, oxidative stress, bacterial contamination, individual variability, and lack of standardized preservation protocols. These factors contribute to reduced sperm viability and fertility following cryopreservation. This review examines critical obstacles in equine semen preservation, focusing on cryopreservation sensitivity, molecular damage mechanisms, economic constraints, and seasonal quality variations. We analyze the molecular and structural alterations (e.g., oxidative stress, membrane damage, and DNA fragmentation) and their impact on cryopreservation success. The review evaluates evidence-based enhancement strategies, including nutritional supplementation and genetic approaches, for improving semen quality. Nutritional interventions that utilize antioxidants, polyunsaturated fatty acids (PUFAs), and nutraceuticals have demonstrated promising results in enhancing sperm motility, preserving membrane integrity, and improving overall semen quality. Additionally, we discuss key candidate genes associated with equine semen-quality traits, including sperm motility, viability, and cryotolerance. The integration of nutritional supplementation and genetic selection strategies presents viable pathways for optimizing equine semen preservation techniques. These combined approaches offer potential solutions for overcoming current limitations, ultimately supporting sustainable breeding programs and advancing genetic conservation efforts in the equine industry.

A significant challenge of mRNA-based protein replacement therapies is the diminishing efficacy and escalating toxicity associated with repeated dosing of lipid nanoparticles (LNPs). Many existing lipid formulations were originally designed for vaccine and are not optimized for therapeutic applications. We developed two libraries of ionizable lipids-one based on piperazine and the other on a newly introduced cyclohexane structure-with variations in linker and tail groups to enhance molecular diversity. GC Biopharma's cyclohexane- and piperazine-based LNPs (GCP LNPs) supported stable and high-level expression without inducing liver toxicity under repeated dosing regimens. These LNPs effectively corrected disease markers in mouse models of phenylketonuria (PKU) and succinic semialdehyde dehydrogenase (SSADH) deficiency. Specially, we observed that the rigid structure and chemical stability of cyclohexane-based lipids contributed to sustained delivery performance. These findings offer a promising direction for the development of LNPs suitable for chronic mRNA-based therapies.

<h4>ABSTRACT</h4> Various synthetic microcompartment systems have been developed to mimic key features of living cells. Here, we focus on artificial cells that capture their capacity to serve as vessels for Darwinian evolution. We assemble micro-compartmentalized In Vitro Transcription-Translation-Replication systems containing a minimal genome, a basic metabolic pathway, a reconstituted protein expression machinery, and a simple DNA replication module, wired in a positive feedback loop. The minimal genome encodes the enzyme deoxyribonucleoside kinase (DNK) whose expression, and then metabolic activity, is required for the genome’s replication. We show that these compartments act as minimal Darwinian elements by filtering out non-functional genotypes. We track individual replicators from a library of 42 genetic variants to reveal the system’s dynamics at both the population and the single replicator levels. At the population level, we extract the fitness function, which links a genome’s metabolic efficiency to its selective success, considering co-encapsulation and hitch-hiking effects. At the individual replicator level, we observe a bimodal distribution of replication yields and propose a mixed model with an inter-droplet heterogeneity with presence or absence of a metabolic feedback loop on the replicator. In addition, we leverage this autonomous self-selection loop to generate a high-resolution mutational map of the DNK enzyme.

<h4>Introduction</h4>Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss among people over 55 years of age globally, being neovascular AMD (nAMD) its most aggressive form. Its treatment consists of the use of drugs that block vascular endothelial growth factor (anti-VEGF). Proteomics may allow the identification of differentially expressed proteins between responders and non-responders to each anti-VEGF drug. Thus, the objective of Pharmacoproteomics in the development of personalised medicine in Age-related Macular Degeneration (PHARPRO-AMD) is to find new proteomic biomarkers, predictive of response to antiangiogenic treatment in patients with nAMD.<h4>Methods and analysis</h4>PHARPRO-AMD is a nationwide, multicentre, prospective, observational study. Treatment-naïve patients with nAMD starting anti-VEGF therapy will be enrolled and followed up for 2 years. During this period, clinical variables will be gathered to classify treatment response. In addition, blood, tear and vitreous and aqueous humour samples will be collected and will undergo a ZenoSWATH proteomic analysis. Relevant biomarkers identified and response classification will be used to perform a multivariate logistic regression and construct receiver operating characteristic curves.<h4>Results</h4>The study is expected to identify a panel of proteomic biomarkers predictive of anti-VEGF treatment response. Integrating data from invasive and non-invasive biological samples may enhance clinical applicability. Once validated, these biomarkers could support the design of future clinical trials on biomarker-guided therapies, helping to optimise treatment regimens and improve visual outcomes.<h4>Conclusions</h4>The PHARPRO-AMD study aims to provide proof-of-concept for biomarker-guided anti-VEGF therapy in nAMD, potentially improving vision outcomes. A notable limitation is the exclusion of patients with visual acuity above 73 Early Treatment of Diabetic Retinopathy Study letters, a criterion chosen to reduce potential ceiling effects and improve response assessment accuracy.<h4>Ethics and dissemination</h4>Approved by the Galician Network of Ethics Committees, with nationwide validity. Anonymised data will be deposited in open-access repositories and published in peer-reviewed journals.<h4>Trial registration number</h4>Spanish Clinical Studies Registry (REec) (0033-2024-OBS).

Nt17, the N-terminal domain of the huntingtin protein (htt), has garnered significant attention for its role in htt's membrane binding and aggregation processes. Previous studies have identified a nuclear export sequence within the Nt17 domain and demonstrated its localization at various cellular organelles. Recent evidence suggests that, like other amphipathic helices, Nt17 can sense and preferentially bind to curved membranes. Gaining deeper insight into this behavior is essential to fully understand the function of this domain. In this study, we combine coarse-grained molecular dynamics simulations with circular dichroism (CD) spectroscopy to investigate the mechanism behind Nt17's curvature sensing. We generated a unique hemispherical-planar membrane model, where 36% of the upper leaflet surface is curved, allowing us to evaluate Nt17's binding preferences. Our findings show that Nt17 exhibits a strong preference for curved regions, with approximately 78 ± 7% of peptides binding to these areas. This interaction is primarily mediated by the terminal Phe residues, indicating that Nt17's curvature sensing is driven by its ability to detect lipid packing defects. Furthermore, Nt17 not only senses these defects but also amplifies them by coalescing into smaller pockets. Mutating the Phe residues to methionine, a smaller hydrophobic residue, significantly reduces Nt17's curvature sensitivity, resulting in equal binding to both curved and planar regions. CD spectroscopy corroborates these results, showing that Nt17 binds more strongly to highly curved small unilamellar vesicles (SUVs) compared to larger, less curved large unilamellar vesicles (LUVs).

RNA-targeting small molecules represent a transformative frontier in drug discovery, offering novel therapeutic avenues for diseases traditionally deemed undruggable. This review explores the latest advancements in the development of RNA-binding small molecules, focusing on the current obstacles and promising avenues for future research. We highlight innovations in RNA structure determination, including X-ray crystallography, nuclear magnetic resonance spectroscopy, and cryo-electron microscopy, which provide the foundation for rational drug design. The role of computational approaches, such as deep learning and molecular docking, is emphasized for enhancing RNA structure prediction and ligand screening efficiency. Additionally, we discuss the utility of focused libraries, DNA-encoded libraries, and small-molecule microarrays in identifying bioactive ligands, alongside the potential of fragment-based drug discovery for exploring chemical space. Emerging strategies, such as RNA degraders and modulators of RNA-protein interactions, are reviewed for their therapeutic promise. Specifically, we underscore the pivotal role of artificial intelligence and machine learning in accelerating discovery and optimizing RNA-targeted therapeutics. By synthesizing these advancements, this review aims to inspire further research and collaboration, unlocking the full potential of RNA-targeting small molecules to revolutionize treatment paradigms for a wide range of diseases.

<h4>SUMMARY</h4> Inherited mutations in VPS35 and the kinase LRRK2 lead to hyperphosphorylation of Rab GTPases and promote the formation of phospho-Rab signalling complexes. A subset of RH2 domain-containing proteins from the RILP-homology family, including RILP, RILPL1, RILPL2, JIP3, and JIP4 are Rab effectors that recognize the LRRK2-phosphorylated switch 2 threonine of phospho-Rab8A and phospho-Rab10. More recently, phospho-Rabs have been found on lysosomal membranes within multi-protein assemblies involving TMEM55B and RILPL1. TMEM55B is a 284-residue lysosomal membrane protein with no homology to known proteins. It comprises a 218-residue cytosolic N-terminal region and two predicted transmembrane α-helices. Residues 80– 160, which face the cytosol, mediate binding to a C-terminal motif of RILPL1, formed after RILPL1 associates with phospho-Rab8A. Here, we report the crystal structures of TMEM55B alone and in complex with a C-terminal RILPL1 peptide, encompassing the TMEM55B interaction region, which we define as the TMEM55B Binding Motif (TBM). The cytosolic domain of TMEM55B adopts a rigid architecture of two tandem RING-like domains, each forming a Zn²⁺-stabilized 40-residue β-sandwich. TBM binding is mediated primarily by backbone hydrogen bonding and anchored by two glutamate residues from RILPL1. These findings support a model in which RILPL1 is recruited to phospho-Rab8A–positive lysosomes prior to TMEM55B engagement. Further co-immunoprecipitation and mutational analyses indicate that TMEM55B forms complexes independently of phospho-Rabs with proteins containing a conserved TBM, like that of RILPL1, including JIP3, JIP4, OCRL, WDR81, and TBC1D9B. Together, these findings uncover previously unrecognized regulatory networks associated with TMEM55B and lysosomal function and suggest that TMEM55B serves as a central hub for adaptor recruitment at the lysosomal membrane.

Colorectal cancer (CRC) arises in the colorectal tissue driven by genetic disorder or the accumulation of somatic mutations, leading to abnormal epithelial cell growth. In this study, we employed single-nucleus multi-omics analysis, including single-nucleus RNA-seq and single-nucleus ATAC-seq, on over 100,000 high-quality nuclei to investigate the molecular landscape of both primary tissue and patient-derived organoids (PDOs). Our analysis showed that normal PDOs (N-PDOs) derived from tissue adjacent to tumors replicate the cellular composition and differentiation trajectory of colorectal crypts. In contrast, tumor PDOs (T-PDOs) showed patient-specific transcriptomic and epigenomic heterogeneity yet consistently maintained a stem cell-like state. T-PDOs retained the somatic mutation profile of the primary tumor while also exhibiting de novo mutations not detected in either the primary tumor or N-PDOs. Notably, inferred cell-cell interaction analysis highlighted the activin signaling pathway as a potential unique feature of fibroblast-epithelial interactions within the tumor microenvironment. This study provides a comprehensive view of the transition from normal to malignant colorectal epithelium and underscores the utility of PDOs as a faithful model for capturing both conserved and patient-specific features of colorectal cancer.

Sex-biased longevity is observed across a wide range of animal taxa, including bats, for reasons not well understood. Patterns of cytosine methylation vary predictably with age in many organisms, offering a valuable means to investigate differences in patterns of aging at the molecular level. We tested sex differences in cytosine methylation across 14 bat species and compared patterns of age-associated variation. Sex differences were overrepresented on the X chromosome, showing a strong pattern of female hypermethylation within promoter regions. Sex- and age-associated differences in methylation were nonrandomly distributed with respect to proximity to putative sex hormone receptor binding sites, with sites hypermethylated in males and females tending to be underrepresented near androgen and estrogen receptor binding sites, respectively. Across species, we observed that the relative steepness of male versus female slopes of age-associated variation appeared to be associated with the strength of precopulatory sexual selection, with especially strong trends toward male-biased age-associated slopes in two harem-polygynous species that exhibit female-biased longevity. Our results offer insights into how patterns of methylation differ across sexes and ages, and raise intriguing questions for future research, such as whether sex differences in molecular aging reflect sex-biased longevity, for which records in bats are sparse.

This study investigated the effect of smoking and regular physical activity (PA) on the composite risk of cardiovascular disease (CVD) in patients with type 2 diabetes mellitus (T2DM) across different steatotic liver disease (SLD) categories. We analyzed data for 1,921,310 patients with T2DM aged 20 years and older. The participants were categorized by SLD status, with hepatic steatosis (fatty liver index ≥ 30). Among current smokers, the adjusted hazard ratios (HR) for composite CVD events significantly increased from the no steatosis to metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction and alcohol-related steatotic liver disease (MetALD), and alcohol-related liver disease (ALD) groups, with the highest HR observed in the ALD group (aHR, 2.14; 95% confidence interval [CI], 2.04-2.24). The ALD group without regular PA had the highest risk of composite CVD events (aHR, 1.33; 95% CI, 1.28-1.38). The highest risk of composite CVD events was found among current smokers without regular PA, with the aHR increasing in a stepwise manner from no steatosis to MASLD, MetALD, and ALD groups, with the latter showing the highest risk. Smoking and physical inactivity significantly increased the CVD risk in patients with T2DM, with the highest risk in the ALD group.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the hepatic manifestation of metabolic syndrome. Hepatic lipotoxicity and inflammation are two key factors driving progression of steatosis to metabolic dysfunction-associated steatohepatitis (MASH). The presence of MASH increases the risk of cardiovascular events, cirrhosis, hepatocellular carcinoma (HCC) and non-liver malignancies. Although MASLD and lipid species have been extensively examined in persons with type 2 diabetes, much less is known in type 1 diabetes. We examined the association of key lipid species with MASLD in individuals with type 1 diabetes. We designed a cross-sectional study of 30 participants with type 1 diabetes recr1uited from our institutional diabetes clinics. All participants had fasting blood drawn for targeted lipidomics and underwent a FibroScan. Those with steatosis score of ≥ 248 as determined by controlled attenuation parameter (CAP) were categorized as cases (n = 17); those with steatosis score < 248 were categorized as controls (n = 13). BMI was significantly higher in cases than controls (P = 0.0007) and used significantly higher 24-h insulin doses than controls (P = 0.004). Cases displayed significantly higher circulating levels of total ceramides (P = 0.02), diacylglycerols (P = 0.0009) and triacylglycerols (P = 0.0004). The two groups displayed similar levels of hexosylceramides, dihydrosphingomyelins, sphingomyelins, and phosphatidylcholines. Similar to previous findings, numerous sphingolipids species, diacylglycerols, and triacylglycerols were found to correlate positively with higher BMI and 24-h insulin dose. Total circulating dihydroceramides, ceramides, diacylglycerols, and triacylglycerols levels significantly correlated with steatosis score (P < 0.05). None of the lipid species correlated with fibrosis score. These results suggest that persons with type 1 diabetes and MASLD have a higher BMI, are likely to be insulin resistant, and display elevated circulating levels of dihydroceramides, ceramides, diacylglycerols, and triacylglycerols, which are strongly associated with the pathogenesis of steatotic liver disease.

Biomolecular phase separation has emerged as a fundamental mechanism governing intracellular spatial organization and functional compartmentalization, and is increasingly recognized as a critical factor in tumor initiation and progression. Through multivalent molecular interactions, biomolecular phase separation contributes to the formation of condensates that mediate the assembly of membraneless organelles, coordination of signaling pathways, and transcriptional programs. Under physiological conditions, condensation contributes to the maintenance of gene expression homeostasis, stress adaptation, and metabolic balance. In cancer cells, however, biomolecular condensates (BMCs) often exhibit aberrant behavior, accompanied by alterations in their structure, components, and regulatory mechanisms. Such perturbations may disrupt cellular homeostasis and influence key biological processes including gene regulation, signal transduction, metabolic reprogramming, and immune responses, thereby modulating various cancer hallmarks. Although the mechanistic understanding of BMCs remains incomplete, their intrinsic plasticity and environmental sensitivity make them attractive therapeutic targets for cancer treatment. This review provides a comprehensive overview of the regulatory factors and functional mechanisms of BMCs in cancer biology, with a particular focus on their involvement in diverse cancer hallmarks. This review further summarizes emerging therapeutic strategies targeting condensation, aiming to inspire novel treatment opportunities.

Lanthanide/transition metal-doped luminescent materials are advanced materials with broad application potential. This type of material achieves control and optimization of luminescence performance by introducing lanthanide/transition metal ions into the host material and utilizing its unique electronic structure and optical properties. Luminescent materials are suitable for optical communication devices, biological imaging, and photodetectors. The combination of lanthanide/transition metals with various matrix materials provides a new platform for creating new chemical and physical properties in materials science and device applications. In this paper, we summarize the latest progress in the research of lanthanide/transition metal-doped luminescent materials and explain their roles in biological imaging, sensing, and optoelectronic applications. It starts with various synthesis techniques and explores how to cleverly incorporate rare earth/transition metals into various matrices, thereby endowing them with unique properties. Then, the advantages and disadvantages of each synthesis technique are discussed. Subsequently, the focus will be on functional strategies and their applications. Finally, strategies for lanthanide/transition metal ion-doped luminescent materials to address challenges are proposed, and insights from each section are summarized.

Chicken meat represents the most widely consumed source of animal protein globally. The identification of non-coding RNAs (ncRNAs) that affect muscle development provides new selection targets for poultry breeding. In this study, muscle samples from high- and low-breast-weight chickens were collected and sequenced for long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and mRNAs. Using weighted gene co-expression network analysis, we found 95 lncRNAs and 46 circRNAs that were significantly associated with breast muscle traits. Subsequently, 51 candidate lncRNAs and 22 candidate circRNAs were screened through differential expression analysis. Finally, by constructing an ncRNA-mRNA regulatory network and performing pathway enrichment analysis, we identified four lncRNAs (e.g., MSTRG.9172.1) and seven circRNAs (e.g., novel_circ_009419) as key regulatory molecules. Functional analysis revealed that these molecules modulate genes such as <i>CD28</i>, <i>CCND2</i>, <i>TIAM1</i>, and <i>RRM2</i> through pathways including the actin cytoskeleton, p53 signaling pathway, and other pathways. In conclusion, this study provides clearer insight into the epigenetic regulatory network involved in chicken breast muscle development and offers important molecular markers for chicken genetic selection.

This review aims to provide a comprehensive overview of how oxidative stress drives inflammation, structural remodeling, and clinical expression of childhood asthma, while critically appraising emerging redox-sensitive biomarkers and antioxidant-focused preventive and therapeutic strategies. Oxidative stress arises when reactive oxygen species (ROS) and reactive nitrogen species (RNS) outpace airway defenses. This surplus provokes airway inflammation: ROS/RNS activate nuclear factor kappa-B (NF-κB) and activator protein-1 (AP-1), recruit eosinophils and neutrophils, and amplify type-2 cytokines. Normally, an antioxidant network-glutathione (GSH), enzymes such as catalase (CAT) and superoxide dismutase (SOD), and nuclear factor erythroid 2-related factor 2 (Nrf2)-maintains redox balance. Prenatal and early exposure to fine particulate matter <2.5 micrometers (µm) (PM_2.5), aeroallergens, and tobacco smoke, together with polymorphisms in glutathione S-transferase P1 (GSTP1) and CAT, overwhelm these defenses, driving epithelial damage, airway remodeling, and corticosteroid resistance-the core of childhood asthma pathogenesis. Clinically, biomarkers such as exhaled 8-isoprostane, hydrogen peroxide (H₂O₂), and fractional exhaled nitric oxide (FeNO) surge during exacerbations and predict relapses. Therapeutic avenues include Mediterranean-style diet, regular aerobic exercise, pharmacological Nrf2 activators, GSH precursors, and mitochondria-targeted antioxidants; early trials report improved lung function and fewer attacks. Ongoing translational research remains imperative to substantiate these approaches and to enable the personalization of therapy through individual redox status and genetic susceptibility, ultimately transforming the care and prognosis of pediatric asthma.

Saline-alkaline water resources are globally widespread, and their rational development offers significant potential to alleviate freshwater scarcity. Saline-alkaline water aquaculture farming not only affects fish growth and survival but also impairs reproductive and developmental functions. Largemouth bass (<i>Micropterus salmoides</i>), an economically important fish, has demonstrated excellent high tolerance to such environments, in order to investigate the effects of alkaline water aquaculture environments on its growth performance, sex hormone levels, gonadal development, and molecular adaptation mechanisms. In this study, largemouth bass were chronically exposed to freshwater (0.55 mmol/L), low alkalinity (10 mmol/L), or high alkalinity (25 mmol/L) and cultured for 80 days. Alkalinity exposure more severely impacted the growth rate of females. High alkalinity significantly increased the hepatosomatic index and decreased the gonadosomatic index in both sexes; moreover, it induced oxidative stress in both sexes, evidenced by reduced superoxide dismutase (SOD), catalase (CAT), and total antioxidant capacity (TAOC) levels and elevated malondialdehyde (MDA) content. Furthermore, the levels of sex hormones Serum estradiol (E2), 11-ketotestosterone (11-KT), and testosterone were significantly reduced, accompanied by either an elevated ratio of primary oocytes and follicular atresia, or by reduced spermatogenesis. Apoptotic signals appeared in gonadal interstitial cells, with upregulated expression of genes <i>P53</i>, <i>Bax</i>, <i>Casp3</i>, and <i>Casp8</i>. Ultrastructural damage included fewer mitochondria and cristae blurring, further indicating tissue damage causing dysfunction. Transcriptome results showed that oxidative stress damage and energy metabolism imbalance caused by carbonate alkalinity were key to the delayed gonadal development, which was mainly manifested in enrichment of the ECM-receptor interaction and PI3K-Akt signaling pathways in females exposed to low alkalinity, and the GnRH secretion and chemokine signaling pathways in males. Glycosphingolipid biosynthesis and Ferroptosis pathway were enriched in females exposed to high alkalinity, and the Cortisol synthesis and secretion pathway were enriched in males. Overall, high-alkalinity exposure significantly delayed gonadal development in both sexes of largemouth bass, leading to reproductive impairment.

<b>Background:</b> Trimetazidine is a clinically established cardioprotective agent with anti-ischemic and antioxidant properties, widely used in the management of coronary artery disease. Combining its metabolic and cytoprotective effects with the potent anti-inflammatory activity of profens presents a promising therapeutic strategy. <b>Methods:</b> Five novel trimetazidine-profen hybrid compounds were synthesized using <i>N</i>,<i>N</i>'-dicyclohexylcarbodiimide-mediated coupling and structurally characterized by NMR and high-resolution mass spectrometry. Their antioxidant activity was evaluated by hydroxyl radical scavenging assays (HRSA), and the anti-inflammatory potential was assessed via the inhibition of albumin denaturation (IAD). Lipophilicity was determined chromatographically. Molecular docking and 100 ns molecular dynamics simulations were performed to investigate the binding modes and stability in human serum albumin (HSA) binding sites. The acute toxicity of the hybrid molecules was predicted in silico using GUSAR software. <b>Results:</b> All synthesized hybrids demonstrated varying degrees of biological activity, with compound <b>3c</b> exhibiting the most potent antioxidant (HRSA IC₅₀ = 71.13 µg/mL) and anti-inflammatory (IAD IC₅₀ = 108.58 µg/mL) effects. Lipophilicity assays indicated moderate membrane permeability, with compounds <b>3c</b> and <b>3d</b> showing favorable profiles. Docking studies revealed stronger binding affinities of <i>S</i>-enantiomers, particularly <b>3c</b> and <b>3d</b>, to Sudlow sites II and III in HSA. Molecular dynamics simulations confirmed stable ligand-protein complexes, highlighting compound <b>3c</b> as maintaining consistent and robust interactions. The toxicity results indicate that most hybrids, particularly compounds <b>3b</b>-<b>3d</b>, exhibit a favorable safety profile compared to the parent trimetazidine. <b>Conclusion:</b> The hybrid trimetazidine-profen compounds synthesized herein, especially compound <b>3c</b>, demonstrate promising dual antioxidant and anti-inflammatory therapeutic potential. Their stable interaction with serum albumin and balanced physicochemical properties support further development as novel agents for managing ischemic heart disease and associated inflammatory conditions.

<h4>Objective</h4>HTT, encoding a protein involved in axonal trafficking, contains a key region of CAG repeats. When expanded beyond 39 repeats, this region leads to Huntington's disease (HD). However, several studies have suggested that increasing the number of CAG repeats below the pathological threshold may confer functional advantages by enhancing HTT activity. In the present study, we aim to investigate the association between CAG repeat length below the pathological threshold and neurodegeneration biomarkers in prodromal Alzheimer's disease (AD).<h4>Methods</h4>Ninety-five patients (36 with SCD and 59 with MCI) underwent blood collection for NfL measurement and HTT genetic analysis. Cerebrospinal fluid was collected for the measurement of Aβ₄₂, Aβ₄₀, total tau, and phosphorylated tau, and/or amyloid PET imaging was performed. Thirty-nine patients who were positive for both Aβ and phosphorylated tau biomarkers were classified as "A+/T+", while 56 patients who were either negative for both markers or positive for only one were classified as "isolated Aβ/non-AD."<h4>Results</h4>In the A+/T+ group, quadratic models described the association between CAG repeat length with NfL concentrations and 18F-FDG uptake. In particular, a concave curve was observed in the medial and middle frontal gyri, while a convex curve was found in the parahippocampal and fusiform gyri.<h4>Interpretation</h4>Among individuals with SCD and MCI who show evidence of AD pathology, CAG repeat length in the HTT gene below the HD pathological threshold is associated with biomarkers of neurodegeneration in a region-specific and U-shaped manner. These findings suggest a modulatory role of HTT in prodromal AD.

Water droplets, acting as natural bioreactors and optical whispering-gallery-mode (WGM) resonators, hold the potential for laser-assisted analysis. However, water/aqueous droplet lasers can only survive in air with a limited lifespan (<100 s) due to rapid evaporation, restricting their applications in bioreactions. To address this challenge, we introduce laser-emitting aqueous bioreactors (LEABs) in fluorocarbon oils. These LEABs enable stable laser emission and extend a droplet lifespan over 1,000-fold. LEABs enable the encapsulation of bioactive materials for long-term analysis with unique lasing characteristic fingerprints. The reactions within LEAB can interact with the most resonating light, enhancing detection sensitivity by over 100-fold compared to conventional WGM sensors. By integrating LEABs with microfluidic droplet technology, we demonstrated their application in monitoring enzyme activity and cellular metabolism at single-cell and multicellular levels. Furthermore, we showed the laser threshold-gated screening of single yeast. This platform can bridge the gap between laser technology and biochemical applications, broadening the scope of laser-based analysis.

Eukaryotic cells direct toxic misfolded proteins to various quality control pathways based on their chemical properties and aggregation status. Aggregated proteins are targeted to selective autophagy or specifically sequestered into the "aggresome", a perinuclear inclusion at the microtubule-organizing center (MTOC). However, the mechanism for selective aggresome recruitment remains unclear. To investigate this process, here we reconstitute MTOC-directed aggregate transport in Xenopus laevis egg extract using AgDD, a chemically inducible aggregation system. High-resolution single-particle tracking reveals that dynein-mediated aggregate transport is highly episodic, with average velocity positively correlating with aggregate size. Mechanistic modeling suggests that recurrent formation of the dynein transport complex biases larger aggregates towards active transport, compensating for the slowdown due to viscosity. Both episodic transport and positive size selectivity are conferred by aggresome-specific dynein adapters. Coupling an aggresome adapter to polystyrene beads recapitulates positive size selectivity in transport, while recruiting conventional dynein adapters to protein aggregates perturbs aggresome formation and reverses the size selectivity.

Dysregulated expression of circular RNAs (circRNAs) has been implicated in the initiation and progression of various diseases, including cancer. In this study, we investigated the role of hsa_circ_0005571 in modulating the biological characteristics of colorectal cancer (CRC) cells. Initially, the circRNA expression profiles of CRC tissues and corresponding normal tissues were analyzed using bioinformatics to identify differentially expressed circRNAs. Subsequently, quantitative real-time PCR (qRT‒PCR) was used to validate the expression levels of hsa_circ_0005571 in both CRC tissues and cell lines. In vitro assays, including the Cell Counting Kit-8 (CCK-8), colony formation, 5-ethynyl-2'-deoxyuridine (EdU) incorporation, wound healing, and Transwell invasion assays, were employed to assess the effects of hsa_circ_0005571 on CRC cell proliferation and invasion. Additionally, tumorigenic potential was examined through in vivo tumorigenesis experiments in nude mice. Bioinformatic predictions and experimental evidence indicated that hsa_circ_0005571 may function as a sponge for miR-520f-3p, which was found to be downregulated in CRC cells. Moreover, LRP8 was predicted and confirmed as a direct downstream target of miR-520f-3p, with both the mRNA and protein levels of LRP8 significantly elevated in CRC cells. Overall, our results suggest that hsa_circ_0005571 enhances CRC proliferation and invasion by modulating the miR-520f-3p/LRP8 axis. Furthermore, elevated hsa_circ_0005571 expression in CRC tissue samples and cell lines relative to that in normal controls was positively correlated with metastasis (p = 0.029) and advanced clinical stage (p = 0.006). Survival analyses revealed that CRC patients with high levels of hsa_circ_0005571 expression had significantly lower overall survival rates than did those with low expression (p < 0.05). Functional assays confirmed that hsa_circ_0005571 facilitates CRC cell proliferation and migration, whereas its knockdown results in reduced migration, proliferation, and tumorigenic potential in vivo. Finally, Western blot analyses demonstrated that LRP8 expression varied in accordance with the levels of hsa_circ_0005571 and miR-520f-3p, further confirming the involvement of the hsa_circ_0005571/miR-520f-3p/LRP8 regulatory axis in CRC. Our study suggested that hsa_circRNA_0005571 promotes proliferation and migration through the miR-520f-3p/LRP8 axis in CRC. Consequently, hsa_circ_0005571 may represent a promising novel gene target for the diagnosis and treatment of CRC.

<h4>Background</h4>Plasma is the most used clinical specimen, yet diurnal variation in plasma proteins remains largely unexplored. We aimed to identify diurnally-regulated proteins in healthy individuals and assess their potential diagnostic implications, and highlight how diurnal awareness can advance future biomarker research.<h4>Methods</h4>Twenty-four healthy young individuals were studied under highly controlled conditions. Venous blood was drawn every three hours over a 24-h period, yielding 216 samples, of which 208 high-quality plasma samples were analyzed via high-throughput mass spectrometry. The missing data were filtered and imputed, and rhythmicity was assessed using Cosinor-based modeling with Benjamini-Hochberg correction. Tissue and pathway enrichment analyses were performed using the DAVID functional annotation tool.<h4>Findings</h4>Of 523 proteins that passed quality thresholds, 138 (~ 26%) exhibited significant diurnal oscillations. Tissue enrichment analysis revealed that most rhythmic proteins originated from the liver and platelets, with additional enrichment in a variety of tissue types. Pathway enrichment showed diurnal regulation of hemostasis, immune signaling, integrin-mediated processes, glucose metabolism, and protein synthesis. Notably, 36 clinically utilized biomarkers, including albumin, amylase, and cystatin C exhibited diurnal variation, suggesting that failing to account for temporal fluctuations may reduce diagnostic precision.<h4>Interpretation</h4>These findings demonstrate that over one-quarter of the human plasma proteome is under diurnal control. Such oscillations might have direct clinical implications, as the time-of-day may alter biomarker accuracy. Incorporating diurnal timing into diagnostic and research protocols, through standardized sampling or time-sensitive reference intervals, could improve patient care and inform future biomarker discoveries. Further research in larger, more diverse populations is needed to generalize these results and streamline practices in a way that takes diurnal variation into account.

Iron is indispensable to most lifeforms, underpinning a myriad of physiological processes. Dysregulation of iron homeostasis underlies a broad spectrum of biological phenomena and pathological conditions. Notably, excessive iron overload acts as a key driver of ferroptosis, a unique form of regulated cell death. Consequently, through the lens of ferrology, targeted modulation of iron balance and ferroptosis has emerged as a compelling avenue for the prevention and treatment of major diseases. Herein, we review the molecular mechanisms governing iron homeostasis, the roles of iron metabolism disorders and ferroptosis in disease pathogenesis, and the latest breakthroughs in iron-regulated therapeutic agents.

Atherosclerosis is characterized by the accumulation of lipids and immune cells in the arterial wall, leading to the narrowing and stiffening of blood vessels. Innate and adaptive immunity are involved in the pathogenesis of human atherosclerosis. However, spatial organization and roles of immune cells during disease progression remain poorly understood. A better understanding of the immune response's contribution to atherosclerosis progression could unveil novel therapeutic targets to mitigate plaque development and rupture, ultimately reducing cardiovascular events burden. Here, we utilised GeoMx® and CosMx™ technologies to analyse serial sections of human coronary arteries from patients with varying degrees of atherosclerotic lesion severity. Our work comprises a series of investigations and integrates findings from both datasets, including pathway analyses, cell typing, and neighbourhood analysis. This workflow highlights the power of combining these spatial transcriptomics platforms to elucidate biological processes at the single-cell level. Our approach unbiasedly identifies molecules and pathways of relevance to support the understanding of atherosclerosis pathogenesis and assess the potential for novel therapies.

Patients with the rare genetic disorder Hyaline Fibromatosis Syndrome (HFS) often succumb before 18 months of age due to severe diarrhea. As HFS is caused by loss-of-function mutations in the gene encoding capillary morphogenesis gene 2 (CMG2), these symptoms highlight a critical yet unexplored role for CMG2 in the gut. Here, we demonstrate that CMG2 knockout mice exhibit normal colon morphology and no signs of inflammation until the chemical induction of colitis. In these conditions, the colons of knockout mice do not regenerate despite previously experiencing similarly severe colitis, due to an inability to replenish their intestinal stem cell pool. Specifically, CMG2 knockout impairs the transition from fetal-like to Lgr5+ adult stem cells, which is associated with a defect in ß-catenin nuclear translocation. Based on our findings, we propose that CMG2 functions as a context-specific modulator of Wnt signaling, essential for replenishing the pool of intestinal stem cells following injury. This study provides new insights into the molecular mechanisms underlying lethal diarrhea in HFS and offers a broader understanding of fetal-like regenerative responses.

Oxidative stress is a pathological condition of redox signaling dysregulation and macromolecular oxidative damage arising from elevated ROS levels. Oxidative stress interacts with tumor cell growth regulation and tumor microenvironment remodeling, and has been a critical hallmark of cancer. Targeting oxidative stress has garnered great attention in cancer therapy development. However, it is still challenging due to the complexity and heterogeneity of oxidative stress regulation across different cancers, and this encourages a comprehensive understanding of the oxidative stress network in cancers to overcome this obstacle. Therefore, we introduced the oxidative stress generation and regulatory network within tumor cells and discussed their roles in both tumor cells and the tumor microenvironment. Subsequently, we summarized the current therapeutic strategies and highlighted emerging clinical applications, providing an up-to-date overview of oxidative stress-based approaches. Particularly, their cross-application with immunotherapy and nanomedicine has provided an excellent opportunity to integrate multiple effects, exhibiting surpassing advantages. This review elaborates on oxidative stress in cancer biology and its therapeutic implications. By integrating current knowledge and the emerging coordination with immunotherapy and nanomedicine, we underscore the potential of oxidative stress-targeting approaches. Future research on overcoming therapeutic resistance and developing compatible platforms to combine multiple approaches will pave the way to cancer elimination.

Cardiac fibrosis, a condition characterized by the deposition of excess collagen in the cardiac tissue, is a major complication of various cardiovascular diseases, including myocardial infarction, hypertension, and different types of cardiomyopathies. CAR T-cell therapy, a form of immunotherapy that involves the genetic modification of T cells to recognize and target specific antigens, has shown promise in the treatment of various cancers and autoimmune diseases. The rationale behind using CAR T-cell therapy to treat cardiac fibrosis lies in the fact that fibrosis is often driven by the activation of pro-fibrotic immune cells, such as myofibroblasts. By targeting these pro-fibrotic cells with CAR T-cells, it may be possible to reduce the severity of cardiac fibrosis. Enhancing CAR T-cell therapy through innovative nanoparticle delivery systems provides a comprehensive approach to treating cardiac fibrosis, with experimental evidence indicating potential in reducing fibrosis and improving cardiac function. Despite these benefits, significant challenges such as cardiotoxicity and cardiovascular complications remain. Therefore, this review explores the molecular mechanisms underlying cardiac fibrosis and the effects of CAR T-cell therapy on the heart, elucidating both its antifibrotic properties and associated cardiotoxic effects based on findings from recent studies.

Metastasis is the main cause of death from solid tumors. Therefore, identifying the mechanisms that govern metastatic growth poses a major biomedical challenge. Tumor microenvironment signals regulate the fate and survival of disseminated tumor cells (DTCs) in secondary organs. However, very little is known about the role of nervous system mediators in this process. We have previously reported that neuropilin-2 (NRP2) expression in breast cancer correlates with poor prognosis. Here, we show that NRP2 positively regulates the proliferation, invasion, and survival of breast and head and neck cancer cells in vitro. NRP2 deletion in tumor cells inhibits tumor growth in vivo and decreases the number and size of lung metastases by promoting lung DTCs quiescence. NRP2 deletion upregulates dormancy and cell cycle regulators expression and promotes DTCs reprograming into quiescence. Moreover, lung fibroblasts and macrophages induce NRP2 upregulation in DTCs through the secretion of TGFβ1. NRP2 facilitates lung DTC interaction with the extracellular matrix and promotes lung DTCs activation and metastasis. Therefore, we conclude that the TGFβ1-NRP2 axis is a new key dormancy-awakening inducer that promotes DTCs proliferation and lung metastasis development.

Astrocytes, the most prevalent glial cells in the central nervous system (CNS), play crucial roles in maintaining CNS homeostasis and responding to various pathological stimuli. They play key roles in neural development, neurotransmission, neuroinflammation, metabolic support, and tissue repair. Recent advancements in single-cell sequencing have revealed the remarkable heterogeneity of astrocytes, with distinct subpopulations differentially contributing to disease progression in neurological disorders, including Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, ischemic stroke, intracerebral hemorrhage, and multiple sclerosis. In addition, they play an important role in various behavioral neuropsychiatric disorders. This review highlights the dual roles of astrocytes in disease progression, driven by their diverse molecular profiles and functions. It outlines the key molecular mechanisms underlying astrocyte heterogeneity and their impact on neuroinflammation, neuronal support, and ionic balance regulation. Additionally, the review discusses potential therapeutic strategies targeting astrocytes to modulate these processes, aiming to improve treatment outcomes in neurological diseases. By elucidating the specific roles of astrocyte subsets in disease, this review seeks to advance the development of precision medicine for astrocyte-related neurological disorders.

<h4>Background</h4>Premature ovarian insufficiency (POI) is a persistent condition in young women characterized by early follicular development disorders and reduced fertility. Research has found that exosomes derived from human umbilical mesenchymal stem cells (hUCMSC-Exo) have significant tissue repair effects. This study aims to investigate the therapeutic effect and potential molecular mechanism of hUCMSC-Exo on POI.<h4>Methods</h4>In vivo experiments were conducted by intraperitoneally injecting the chemotherapy drug cyclophosphamide (CTX) to establish a 14-day POI rat model. Serum hormone levels were measured using an enzyme-linked immunosorbent assay, and changes in ovarian tissue structure were analyzed using hematoxylin-eosin (HE) staining. Perls staining and transmission electron microscopy were used to assess changes in ovarian ferroptosis. In vitro experiments involved exposing theca interna cells (TICs) treated with CTX to normal and miR-26a-5p inhibitor-treated hUCMSC-Exo. The expression changes of PTEN, Nrf2, and GPX4, which are associated with ferroptosis, were analyzed using immunofluorescence, Western blot, and quantitative reverse-transcription polymerase chain reaction.<h4>Results</h4>hUCMSC-Exo intervention can significantly repair the ovarian tissue structure and functional abnormalities in the model rats, especially ferroptosis. Further bioinformatics analysis revealed that the inhibition of the PTEN/GPX4 pathway-mediated ferroptosis in TICs might be the main mechanism through which exosomes exert their regulatory/therapeutic effects. In vitro experiments, where exosome miR-26a-5p was inhibited, further confirmed that the delivery of miR-26a-5p is crucial for the regulatory effect of exosomes.<h4>Conclusion</h4>In conclusion, our results suggest that hUCMSC-Exos alleviates POI-related dysfunction of ovarian structure and function. The mechanism could be related to the transfers of miR-26a-5p and suppression of PTEN/GPX4 axis signaling-mediated autophagy of TICs. It provides a new perspective for developing treatment methods for patients with metabolic abnormalities related to POI.

Hypergonadotropic hypogonadism is usually caused by the impairment of the structure and function of the gonads, but there are cases caused by reduced stimulation by the follicle-stimulating hormone (FSH) as a result of abnormal variants of genes encoding the follicle-stimulating hormone or its receptor (FSHR). We present the case of an elite athlete with the c.2039A>G variant in one allele of the FSHR gene resulting in hypergonadotropic hypogonadism, low testicular volume, and reduced semen parameters, placing particular emphasis on the diagnostic process and the importance of correct diagnosis in the context of possible treatment with gonadotropins, which can significantly improve fertility, increase testosterone levels, and, in the case of athletes, obtain approval from the anti-doping organization for treatment that increases testosterone levels.

<h4>Objective</h4>The diagnosis of precancerous lesions of colorectal cancer (CRC) presents significant challenges in clinical practice. In this study, we conducted a clinical investigation using the UCAD technique after analyzing chromosomal copy number variations (CNVs) in formalin-fixed, paraffin-embedded (FFPE) samples from various pathological stages, aiming to evaluate the value of detecting chromosomal instability (CIN) in CRC diagnosis.<h4>Methods</h4>Based on colonoscopic pathological findings, we selected 39 FFPE specimens of tubular adenomas, 8 FFPE specimens of villous adenomas, 16 cases diagnosed as tubular-villous adenomas, and 14 cases without defined pathological subtype classification. The UCAD technique was employed to analyze these specimens, with the objective of delineating differences in chromosomal instability among the various pathological subtypes.<h4>Results</h4>UCAD analysis confirmed that among 39 patients diagnosed with tubular adenomas, 12 (30.76%) exhibited CIN positivity, primarily characterized by amplifications of chromosomal segments on 13q, 7, and 8, and losses on 18q and 14q. In the 8 patients diagnosed with villous adenomas, 6 (75%) were CIN-positive, displaying amplifications at 13q, 7, 8q, and 20, along with losses at 18q and 14q. Among 16 patients diagnosed with tubular-villous adenomas, 8 (50%) demonstrated CIN positivity. Additionally, 8 out of 14 cases lacking a defined pathological subtype were CIN-positive.<h4>Conclusion</h4>The assessment of CIN correlates with both pathological subtypes and disease progression. UCAD-based detection of CIN contributes to the diagnosis of colorectal adenomas (CRA), with aberrations in chromosomes 7 and 8 potentially being closely associated with PLCRA.

Ferroptosis is a regulated, non-apoptotic form of cell death marked by the accumulation of iron-dependent lipid peroxides. This process causes rapid rupture of the plasma membrane and the release of intracellular contents. Ferroptosis acts as an intrinsic tumor-suppressive mechanism. It plays a crucial role in tumor progression, metastasis, and resistance to standard therapies, including chemotherapy and radiotherapy. Its unique molecular mechanisms confer significant therapeutic potential. In recent years, multiple experimental therapies aiming to induce ferroptosis have been developed for cancer treatment. Although these therapies show promise in controlling tumor growth, their effects on the tumor microenvironment (TME) require further investigation. Recent studies indicate that distinct cell populations within the TME have different sensitivities to ferroptosis. This variability may lead to unintended effects, such as damage to normal cells or increased inflammation, resulting in toxicity. Cells in the TME can either undergo ferroptosis or modulate its regulation through intercellular signaling and interactions. Notably, ferroptosis-related interactions between tumor cells and other components of the TME, such as immune cells, stromal cells, and endothelial cells, are central to TME remodeling. This mini-review summarizes recent advances in ferroptosis mechanisms and highlights the dynamic interplay between ferroptosis and the TME. It also discusses the prospects and challenges of ferroptosis-based cancer therapies.

<h4>Introduction</h4>Endothelial-to-mesenchymal transition (EndoMT), cell death, and fibrosis are increasingly recognized as contributing factors to Alzheimer's disease (AD) pathology, but the underlying transcriptomic mechanisms remain poorly defined. This study aims to elucidate transcriptomic changes associated with EndoMT, diverse cell death pathways, and fibrosis in AD using the 3xTg-AD mouse model.<h4>Methods</h4>Using RNA-seq data and knowledge-based transcriptomic analysis on brain tissues from the 3xTg-AD mouse model of AD. This included pathway-level analysis of gene expression changes across multiple brain cell types. Mechanistic insights were further validated using single-cell RNA sequencing (scRNA-Seq) dataset from human AD brain.<h4>Results</h4>Our analysis showed that in the 3xTg-AD model: (i) multiple brain cell type genes are altered, promoting EndoMT through upregulation of RGCC and VCAN; (ii) genes related to various types of cell death, including apoptosis, ferroptosis, necrosis, anoikis, mitochondrial outer membrane permeability programmed cell death, mitochondrial permeability transition-driven necrosis, NETotic, and mitotic cell death, are upregulated in the several brain cell types; (iii) fibrosis-related genes are upregulated across multiple brain cell types. Further mechanistic analysis revealed: (1) mitochondrial stress through upregulation of mitochondrial genes in the brain cells; (2) upregulation of cellular, oxidative, and endoplasmic reticulum (ER) stress genes; (3) nuclear stress via upregulation of nuclear genes, transcription factors (TFs), and differentiation TFs FOSB and MEOX1; (4) metabolic reprogramming/stress through the upregulation of genes related to lipid and lipoprotein metabolism, fatty acid oxidation (FAO), glucose metabolism, and oxidative phosphorylation (OXPHOS); (5) catabolic stress via upregulation of catabolic genes. Single-cell RNA-Seq data indicated that many of these were also increased in AD patients' brain cells. These changes were reversed by knockdown of the ER stress kinase PERK (EIF2AK3) and deficiencies in FOSB and MEOX1.<h4>Discussion</h4>This study uncovers previously unrecognized molecular signatures of organelle stress and bioenergetic reprogramming that drive EndoMT, cell death, and fibrosis in AD. The reversal of these changes via PERK, FOSB, and MEOX1 inhibition highlights potential therapeutic targets for mitigating neurodegenerative processes in AD.

S-glutathionylation (SSG), a redox-sensitive post-translational modification mediated by glutathione, regulates protein structure and function through reversible disulfide bond formation at cysteine residues. Glutaredoxins (GRXs), pivotal antioxidant enzymes, catalyze SSG dynamics to maintain thiol homeostasis. Recent advances in redox proteomics have revealed that SSG dysregulation is intricately linked to neurodegenerative, cardiovascular, pulmonary and malignant diseases. Notably, GRX isoforms (GRX1 and GRX2) play compartment-specific roles in disease pathogenesis: GRX1 modulates hepatic lipid metabolism and pulmonary fibrosis, while GRX2 sustains mitochondrial redox balance and Fe-S cluster assembly. Notably, SSG functions as a 'double-edged sword' in programmed cell death (PCD). While moderate SSG protects against irreversible cysteine oxidation, persistent SSG accumulation due to GRX dysfunction triggers apoptosis, necroptosis and ferroptosis by disrupting redox-sensitive targets, such as caspases, BAX and glutathione peroxidase 4. The present review summarizes, for the first time, at least to the best of our knowledge, the association of SSG with distinct PCD subtypes, and highlights therapeutic strategies targeting GRX activity or site-specific SSG modulation (e.g., pyruvate kinase M2 Cys423/424). Emerging approaches, including GRX mimetics and thiol-targeted drugs, hold promise for precision medicine in redox-related pathologies.

<h4>Background</h4>Diabetic retinopathy (DR), a leading cause of blindness among working-age adults, lacks targeted therapies besides glucose management. Early retinal lesions are linked to serum metabolites, but the underlying peripheral regulatory networks is unclear.<h4>Methods</h4>We first established a streptozotocin (STZ)-induced mouse model of early DR exhibiting retinal inflammation characteristics. This study employed an integrative approach, combining retinal and serum transcriptomic and metabolomic profiles with genome-wide association study (GWAS) data, to identify peripheral metabolites potentially linking early retinal lesions.<h4>Results</h4>STZ-induced mice exhibited retinal inflammation and metabolic dysregulation. Metabolites including glucose, sorbitol, and mannitol were altered in both serum and retina, implicating their potential involvement in retinal inflammation. Utilizing GWAS data of diabetic patients, we further explore the potential the upstream regulation of shared metabolites and their peripheral pathways potentially instigating early retinal inflammation through metabolite-related genes correlated with single nucleotide polymorphisms. Key enzyme genes including HK1, HKDC1, AKR1B1 in hyperglycemic pathway, CEL and HMGCR in cholesterol pathway, and ACSL1, PPT2 in palmitic acid pathway, may connect the metabolic network of hyperglycemia, hyperfructosemia and disrupted lipid metabolism to retinopathy.<h4>Conclusion</h4>This study elucidates the upstream regulatory network of peripheral serum metabolites associated with early retinal lesions. Specifically, the SNPs in key peripheral enzyme genes may exert remote effects on retinal inflammation in DR. This finding provides insights into the systemic metabolic management and offering peripheral precise early detection and treatment.

Parasitic infections, such as those caused by the myxozoans <i>Myxobolus cerebralis</i> and <i>Tetracapsuloides bryosalmonae</i>, pose major threats to wild and farmed salmonids due to severe tissue damage and impairment of the host immune system. While individual infections have been studied, limited information is available on the host response during co-infection. This study investigated the transcriptomic immune response of rainbow trout (<i>Oncorhynchus mykiss</i>) during single and sequential co-infections with <i>M. cerebralis</i> and <i>T. bryosalmonae</i> using RNA-seq. Trout were exposed to single infections (Mc or Tb) followed by co-infections (Mc⁺ or Tb⁺). Fish were sampled at 31 days post-single infection (1 day post-co-infection). RNA from gill and caudal fin (portal of parasite entry) was sequenced, followed by differentially expressed genes (DEGs) identification and GO and KEGG enrichment. In the caudal fin, Mc⁺ (1 day after co-infection with <i>T. bryosalomne</i>) fish showed mild immune activation with <i>C4B</i> upregulation, while Tb⁺ fish exhibited a stronger response involving <i>IFI44, ISG15, RSAD2, and TLR7</i> signaling. In gills, Mc⁺ fish showed moderate cytokine-related gene upregulation, while Tb⁺ (1 day after co-infection with <i>M. cerebralis</i>) fish displayed increased expression of humoral response genes (<i>C3</i>, immunoglobulin pathways) but suppression of genes involved in B cell development. These results indicate that the order of infection shapes the outcome of the host immune response, offering candidate targets at the host-pathogen interface.

Dental enamel discoloration, extrinsic staining, and periodontal biofilms remain persistent challenges in oral health. This study explores the in vitro, dual-functional potential of <i>Magnolia figo</i> flower extract (FMO), a sesquiterpene-rich botanical active phytochemical ingredient (API), for aesthetic and antimicrobial oral applications. FTIR identified characteristic terpenoid and long-chain fatty acid functional groups, including β-elemene, γ-elemene, and caryophyllene oxide. Whitening efficacy on coffee-stained bovine enamel was quantified using CIELAB colorimetry. The 0.5% FMO treatment achieved ΔE* = 8.49, which was within the clinical perceptibility threshold and the optimal biocompatibility balance. SEM confirmed no demineralization on the enamel surface after immersion in 3.0% FMO for 12 h. Antimicrobial assays demonstrated inhibition of <i>Porphyromonas gingivalis</i>, with MIC and MBC values of 0.25% and 0.5%, respectively. Biofilm formation was reduced by over 50% at a 0.148% concentration. Cytocompatibility assays using HGF-1 cells with various concentrations of FMO showed reduced cell viability at higher concentrations. When exposed for 5 min (simulating daily oral care) or 2 h, 0.5% FMO exhibited greater biocompatibility with L929 cells compared to toothpaste and peroxide-based agents. These findings suggest that FMO may serve as a natural candidate for dual-function oral care; however, further in vivo and clinical investigations are needed to validate its potential use within oral care treatments.

Huntington's Disease (HD) originates from the expansion of a polyglutamine (PolyQ) tract in the huntingtin protein (Htt), which can assume a coiled-coil fold (Cc). We previously found that Cc structures mediate the aggregation and toxicity of polyQ Htt. Since polyQ Htt aggregates were previously found to be internalized by cells, here we hypothesize that Cc structures might be implicated in the intercellular propagation of Htt aggregates. To test this hypothesis, we performed experiments using human cell lines expressing Htt proteins with different probabilities to acquire a Cc fold. We found that Htt with reduced Cc structures were released significantly less compared to Htt with intact Cc structures. We also found that Cc structures mediate the internalization of Htt proteins in recipient cells. Together, these results underline the importance of the Cc structure in the process of intercellular propagation of Htt polyQ aggregates and suggest that interfering with Cc formation might be a therapeutic strategy for HD.

The progression of metabolic dysfunction-associated steatotic liver disease (MASLD) to severe forms, including metabolic dysfunction-associated steatohepatitis (MASH) and liver fibrosis, involves metabolic dysfunction, genetics, and gut dysbiosis. People with HIV (PWH) represent a high-risk group for MASLD, but the role of gut microbiota alterations in disease severity within this population remains poorly understood. We prospectively recruited PWH with MASLD, defined as the controlled attenuation parameter (CAP) ≥ 238 dB/m, and excluded those with viral hepatitis coinfection or alcohol abuse. Severe MASLD was defined as the presence of MASH (cytokeratin-18 ≥ 130.5 U/L) and/or significant liver fibrosis (liver stiffness ≥ 7.1 kPa). Stool samples were collected for 16S rRNA gene sequencing to characterize gut microbiota composition. Functional predictions were generated using PICRUSt. The differential abundance of bacterial taxa and predicted functions were analyzed using a generalized linear model with a negative binomial distribution. Among 34 PWH with MASLD, 18 (53%) met the criteria for severe MASLD. Microbiota profiling revealed significant differences in bacterial genera between the PWH with and without severe MASLD. Enrichment was observed in the <i>Ruminococcus gnavus</i> group, <i>Negativibacillus</i>, <i>Holdemanella</i>, <i>Subdoligranulum</i>, the <i>Eubacterium hallii</i> group, and <i>Butyricicoccus</i>, while depletion was seen in <i>Prevotella</i>, <i>Alloprevotella</i>, <i>Dialister</i>, <i>Catenibacterium</i>, the <i>Christensenellaceae R 7</i> group, <i>Clostridium sensu stricto</i>, <i>Olsenella</i>, <i>Oscillospiraceae UCG-005</i>, <i>Libanicoccus</i>, and the <i>Eubacterium siraeum</i> group. Predicted functional pathways related to fatty acid degradation, folate biosynthesis, and amino acids metabolism did not differ between groups. MASLD severity in PWH is associated with a distinct gut microbiota signature, though not with functional pathway alterations. Microbial profiling may complement existing non-invasive biomarkers for risk stratification in this high-risk population.

Mitochondria are dynamic organelles integral to cellular energy metabolism and homeostasis. Beyond their traditional roles, a growing body of evidence underscores the importance of mitochondria as pivotal regulators of innate immune signaling pathways. Recently, mitochondrial RNA (mtRNA) has been identified as a novel modulator of inflammatory responses. mtRNA is detected by intracellular pattern recognition receptors (PRRs), which subsequently activate the mitochondrial antiviral-signaling protein (MAVS) and the interferon regulatory factor 3 (IRF3)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling axis, as well as inflammasome pathways. This activation leads to the production of type I interferons and pro-inflammatory cytokines. Furthermore, mtRNA facilitates the propagation of inflammatory signals through exosome-mediated intercellular transfer. Among the various forms of mtRNA, mitochondrial double-stranded RNA (mt-dsRNA) is particularly prone to activating inflammatory responses due to its distinctive double-helical structure. The aberrant accumulation of mt-dsRNA is strongly linked autoimmune diseases, degenerative disease, Liver Disease, kidney disease, cancers, cardiovascular diseases, and respiratory ailments. This review proposes innovative therapeutic strategies aimed at degrading pathological mtRNA or interrupting inflammatory pathways by targeting critical regulatory nodes in mtRNA metabolism and its downstream inflammatory processes.

Lipid-derived prodrugs can enhance the oral bioavailability of therapeutic agents by promoting uptake into the lymphatic system. Chylomicrons (CMs) are lipoproteins that transport dietary lipids into the lymphatics. However, their role in the systemic distribution of orally administered lipid prodrugs remains understudied. We developed an <i>in vitro</i> assay demonstrating that human intestinal enterocyte-like cell-derived CMs incorporate lipid prodrugs, and that the efficiency of this incorporation varies as a function of the lipid promoiety. We synthesized a series of lipid prodrugs of tenofovir (TFV), an important but poorly bioavailable anti-HIV agent. These lipid prodrugs, featuring a benzyloxyglycerol (BOG) motif and/or an ω-CF₃ group, demonstrated improved metabolic stability and favorable antiviral activity <i>in vitro</i>, relative to unfunctionalized lipid conjugate TFV exalidex (TXL). Additionally, the ω-CF₃ and BOG modifications significantly increased prodrug uptake into CMs <i>in vitro</i>. Subsequent mouse pharmacokinetic (PK) studies revealed higher systemic drug levels of orally dosed ω-CF₃ BOG prodrugs relative to TXL, as well as substantially enhanced lung distribution. This study is the first to quantify drug incorporation into human intestinal enterocyte-like cell-derived CMs using LC-MS/MS. In conclusion, highly lipophilic TFV prodrugs efficiently incorporate into CMs <i>in vitro</i>, and mouse PK data is consistent with lymphatic absorption <i>in vivo</i>, providing a framework for the rational design and screening of lipid-based prodrugs for optimization of drug distribution.

Reverse Shapiro syndrome (RSS) is an exceptionally rare neurological disorder characterized by recurrent episodes of hyperthermia in the context of agenesis of the corpus callosum (ACC), in contrast to the hypothermic episodes seen in classic Shapiro syndrome (SS). The exact pathophysiology remains unclear; however, hypothalamic dysregulation, neurotransmitter imbalances, and melatonergic involvement are believed to play key roles. Herein, we present a case of RSS in a 33-year-old male patient with a medical history of partial agenesis of the corpus callosum and spastic tetraparesis with persistent episodes of unexplained high fever leading to multiple hospitalizations. Endocrine and infectious causes were excluded. Both antibiotics and supportive management failed to resolve the symptoms, whereas levodopa administration led to complete remission of symptoms. Fewer than 10 cases have been reported in the literature to date. Management in reported cases was largely supportive, with mixed responses to pharmacological agents, such as dopamine agonists and serotonin antagonists. Given the potential for diagnostic delay and its impact on patients' quality of life, awareness of RSS is essential in cases of fever of unknown origin, particularly in patients with known or suspected congenital brain anomalies.

The BCR-ABL1 fusion protein is a critical therapeutic target in Chronic Myeloid Leukemia (CML). Current monotherapy approaches involve types of inhibitors that can be categorized into ATP competitive inhibitors and allosteric inhibitors. However, resistance mutations in the tyrosine kinase domain of BCR-ABL1 have limited the effectiveness of these drugs. Research indicates that dual inhibition of BCR-ABL1 by combining these two types of inhibitors effectively addresses the issue of drug resistance as there are no overlapping resistance mechanisms. However, the underlying reasons for the observed synergistic effects have not yet been thoroughly elucidated. In this study, we employed molecular dynamics simulation to observe the synergistic interactions of BCR-ABL1 by the allosteric inhibitor asciminib and ATP competitive inhibitors nilotinib and ponatinib. Our study reveals that when asciminib binds to BCR-ABL1, nilotinib and ponatinib exhibit more substantial binding stability compared to monotherapy. At the atomic level, we have elucidated the reasons for the enhanced binding affinity of nilotinib and ponatinib when using a co-inhibition therapy. Our study reveals the allosteric communication pathway between asciminib and ponatinib, providing more detailed insights into the effectiveness of combination therapy. These findings provide valuable insights into combination therapies, aiding in the rational use of medications and guiding the design of novel inhibitors.

Graphene, owing to its exceptionally high specific surface area, abundant surface functional groups, and outstanding biocompatibility, exhibits tremendous potential in the development of nanodrug delivery systems. This review systematically outlines the latest research advancements regarding graphene and its derivatives in drug loading, targeted delivery, and smart release. It covers delivery strategies and mechanisms for various types of drugs, including small molecules and macromolecules, with a particular emphasis on their applications in major diseases such as cancer, neurological disorders, and infection control. The article also discusses stimulus-responsive release mechanisms, such as pH-responsiveness and photothermal responsiveness, and highlights the critical role of surface functionalization of graphene and its derivatives in enhancing therapeutic efficacy while reducing systemic toxicity. Furthermore, the review evaluates key challenges to the clinical translation of graphene-based materials, including safety, toxicity, and metabolic uncertainties. It points out that future research should focus on integrating structural modulation of materials with biological behavior to construct intelligent nanoplatforms featuring biodegradability, low immunogenicity, and precise therapeutic targeting. The aim of this paper is to provide theoretical insights and technical guidance for the customized design and precision medicine applications of graphene and its derivative-based drug delivery systems.

<h4>Background & aims</h4>The impact of metabolic dysfunction-associated steatohepatitis (MASH) on patient-reported outcomes is poorly understood. We assessed work productivity burden in a real-world population with suspected/confirmed MASH, comparing work productivity and symptoms across subgroups (risk status, age, BMI, and comorbidities) in the USA using secondary data from Adelphi Real World MASH Disease Specific Programmes™, cross-sectional surveys of physicians and consulting patients in 2019 and 2022.<h4>Methods</h4>Physicians (hepatologists, gastroenterologists, and endocrinologists) reported sociodemographic data and signs/symptoms for eight or fewer consecutive participants with MASH. Participants voluntarily completing questionnaires that assessed work productivity, health status, and quality of life/symptoms were categorized with low-, indeterminate-, or high-risk MASH using physician-stated fibrosis stage and derived risk categories. Principal components factor analysis identified factors from MASH signs and symptoms. Elastic net regression determined features associated with work productivity impairment.<h4>Results</h4>In total, 87 physicians and 429 individuals with MASH provided data. The impact of MASH on activities was greater in high-versus low-risk MASH (activity impairment score: high risk, 30.8%; low risk, 17%; <i>p</i> <0.001 across all risk categories). Overall work impairment scores were 18.8% in low-risk and 19.9% in high-risk MASH. Of the 14 physician-reported signs/symptoms, 11 were significantly associated with physician-stated fibrosis stage and/or derived risk category. Two clusters of signs and symptoms ('memory loss/swelling legs/abdomen' and 'fatigue/sleep disturbance/insomnia/general weakness') and female sex were the strongest work impact predictors.<h4>Conclusions</h4>Activity impairment was greater in participants with high-risk MASH, whereas overall work impairment was comparable, almost 20%, in low- and high-risk MASH. Non-specific symptoms of fatigue, sleep disturbance, and general weakness associated with MASH and other conditions predicted work impact in low-risk MASH. Thus, early detection and management of MASH could ameliorate work impairment.<h4>Impact and implications</h4>Even in its early stages, when otherwise asymptomatic, metabolic dysfunction-associated steatohepatitis (MASH) may impact the ability to work. Using the Work Productivity and Activity Impairment: Specific Health Problem (WPAI:SHP) questionnaire, we identified activity and overall work impairment in people with high- and low-risk MASH. We found a link between difficulty working and other signs, such as tiredness, sleep disturbance, and general weakness, in low-risk MASH that are not specific to MASH but can be associated with liver health. These general, non-specific signs could represent possible early warning signs preceding reduced productivity in patients with MASH.

Granular hydrogels represent an important advancement in hydrogel biomaterials for tissue engineering. These granular materials hold advantages over the traditionally formulated hydrogel because the constituent microgels add modularity and a high degree of porosity upon assembly. While granular hydrogels have shown great promise in tissue engineering, their increased porosity is still randomly distributed, unlike the structure of tissues like nerve which possess an anisotropic or hierarchical degree of porosity. We have developed and utilized a technique termed magnetic templating which allows the micropatterning of aligned sacrificial magnetic porogens that can be then removed upon hydrogel crosslinking, leaving an aligned pore architecture. Here we demonstrate the feasibility of magnetic templating of scaffolds consisting of synthetic polymer within a granular hydrogel system. To do so, we evaluated the extent to which microgel concentration impacts the rheology of jammed granular gels and utilized nano computed tomography to evaluate its concomitant influence on the degree of porogen chain alignment. Lastly, we show that the porogens are effectively cleared from templated granular hydrogels. This work establishes proof-of-concept for using magnetic templating to impart highly anisotropic structure within granular PEG hydrogel composites, with potential applications in regenerative medicine and tissue engineering.

Diet, microbiota, and other exposures make the intestinal epithelium a nexus for evolutionary change; however, little is known about genomic changes associated with adaptation to a distinctly human environment. In this work, we interrogate the evolution of cell types in the developing human intestine by comparing tissue and organoids from humans, chimpanzees, and mice. We find that recent changes in primates are associated with immune barrier function and lipid and xenobiotic metabolism and that human-specific genetic features affect these functions. Enhancer assays, genetic deletion, and in silico mutagenesis resolve evolutionarily important enhancers of lactase (<i>LCT</i>) and insulin-like growth factor binding protein 2 (<i>IGFBP2</i>). Altogether, we identify the developing human intestinal epithelium as a rapidly evolving system and show that great ape organoids provide insight into human biology.