Gene Literature Dashboard

BACKGROUND: Liver steatosis, fibroinflammation, and iron overload, are growing global health concerns, yet the genetic architecture and causal pathways linking liver pathology to systemic disease remain incompletely understood. METHODS: We analysed MRI-derived liver traits—corrected T1 (cT1), proton density fat fraction (PDFF), and liver iron—in 37,626 UK Biobank participants. Genome-wide (GWAS), transcriptome-wide (TWAS), and cis-protein Mendelian randomisation (cis-MR) analyses were used to identify genes and proteins influencing these traits. We applied two-sample MR to assess bidirectional causal relationships with metabolic and vascular traits and used Multi-Trait Analysis of GWAS (MTAG) to enhance discovery by leveraging genetic correlations. RESULTS: GWAS identified 18 loci for cT1, 15 for PDFF, and 5 for liver iron, including six not previously reported. TWAS, cis-MR, and proteome-wide analyses prioritised genes (e.g., FADS1, GPAM, MBOAT7, RAD51C) and proteins (e.g., RAB2B, GPN1, GSTM4) with putative mechanistic roles. Fine-mapping refined several signals (GSTM1, TMPRSS6) to single-variant credible sets. Cell-type enrichment revealed distinct tissue contributions: hepatocytes and intestinal mucosa for cT1, adipose tissue for PDFF, and gastrointestinal tissues for liver iron. MR suggested causal effects of higher liver PDFF and cT1 on obesity-related traits, and inverse genetic associations between liver iron and coronary artery disease. MTAG identified seven additional loci (three for cT1, four for PDFF) not previously reported. CONCLUSIONS: This integrative imaging-genetics study reveals 13 potentially novel genes and several protein candidates implicated in hepatic steatosis, inflammation, and iron homeostasis. These findings enhance understanding of liver disease biology and may help identify new targets for early detection or treatment. IMPACT AND IMPLICATION: This large imaging-genetics study in over 37,000 people identifies genetic and protein factors linked to liver fat, fibroinflammation, and iron levels. It shows that higher liver fat and inflammation are associated with increased cardiometabolic risk, while higher liver iron appears inversely linked to risk of heart disease. These findings highlight molecular targets such as FADS1, FUT2, TMC4, RAB2B, and GPN1, which could inform future efforts to improve early detection or treatment of liver disease and its complications in people with obesity or metabolic syndrome.

Adrenergic-driven thermogenic activation of brown adipose tissue requires high amounts of nutrients including iron to support mitochondrial biogenesis. This is governed by rapid gene expression changes in ex vivo differentiated human cervical-derived brown adipocytes. Transferrin receptor 1 (TFRC) is upregulated in response to dibutyryl-cAMP. We aim to investigate the mechanism of facilitated iron uptake when thermogenesis is activated. Pharmacological inhibition and siRNA-mediated knock-down of TFRC during stimulation decrease intracellular iron content and prevent elevation of oxygen consumption and induction of thermogenic markers. Deferoxamine-mediated iron chelation also shows comparable effects. Contrarily, the expression of ferroportin exporter is suppressed during activation; however, its inhibition does not increase thermogenesis. Brown adipocytes constitutively express and secrete high amounts of transferrin, while melanotransferrin expression and release are upregulated only in activated adipocytes. In silico analysis suggests that melanotransferrin interacts with the helical domain of TFRC. Our findings support that iron is critical in stimulating adipocyte thermogenesis.

BACKGROUND: Receptor interacting protein kinase 1 (RIPK1) is crucial in the regulation of apoptosis; however, its significance in clear cell renal cell carcinoma (ccRCC) is not well understood. METHODS: Utilizing data collected from public databases, RIPK1 expression level was compared between ccRCC and control groups. The diagnostic capability of RIPK1 was confirmed by drawing receiver operating characteristic (ROC) curve. A nomogram was created based on the RIPK1 and primary clinical features of ccRCC patients. Finally, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed to confirm the expression of RIPK1, and cell experiments including colony formation, cell counting kit8 (CCK-8), cell scratch, and Transwell assays were conducted to explore the role of RIPK1. RESULTS: The expression of RIPK1 was higher in ccRCC samples compared to normal samples (Fold-change = 0.24), which was confirmed in in vitro verification experiments. And the area under the ROC curve (AUC) values in the TCGA-ccRCC, GSE53757, and GSE66272 datasets were all greater than 0.7, indicating that RIPK1 had acceptable diagnostic ability for ccRCC patients. Then, clinical parameters such as stage, grade, and age may serve as independent prognostic indicators for ccRCC, and a nomogram was developed based on these clinical attributes and RIPK1. The AUC values of the nomogram at 1, 3, and 5 years were 0.904, 0.841, and 0.779, respectively, proving that it can effectively distinguish patients with different prognostic outcomes. According to the results of cell experiments, the RIPK1 overexpression (OE) group was associated with increased cell viability, proliferation, invasion, and migration capabilities compared to the control group. In contrast, these capabilities were markedly decreased in cells subsequent to RIPK1 knockdown (KD). CONCLUSION: The elevated expression of RIPK1 may be involved in enhancing the viability of ccRCC cells and promoting their proliferation and migration capabilities, therefore establishing a basis for clarifying the function of RIPK1 in the advancement of ccRCC.

BACKGROUND: Despite evidence for a genetic component, few genetic associations with lung function decline have been identified. We aimed to evaluate genome-wide associations and putative downstream functionality of genetic variants for lung function decline. METHODS: We conducted genome-wide association study (GWAS) analyses of decline in FEV1, FVC, and FEV1/FVC in 52,056 White (N = 44,988), Black (N = 5,788), Hispanic (N = 550), and Chinese American (N = 730) participants across seven general population cohorts. GWAS analyses were stratified by cohort, ancestry, and sex. Results were combined in cross-ancestry and ancestry-specific meta-analyses. Significant variants available in two independent COPD-enriched cohorts were tested for replication. RESULTS: We identified 361 distinct genome-wide significant (p < 5E-08) variants for one or more of the FEV1, FVC, and FEV1/FVC decline phenotypes, which overlapped with previously reported genetic signals for pulmonary traits. Four variants, or 10.3% of variants available for replication testing, were nominally associated (p < 0.05) with at least one decline phenotype in COPD-enriched cohorts. Gene-level analysis of GWAS results implicated 38 genes, many with consistent associations across ancestries or decline phenotypes. Annotation class analysis revealed enrichment of regulatory processes for corticosteroid biosynthesis and metabolism. Drug repurposing analysis identified 43 approved compounds targeting eight implicated genes. CONCLUSIONS: Our GWAS meta-analyses identified numerous genetic loci associated with lung function decline. These findings contribute knowledge to the genetic architecture of lung function decline, provide evidence for a role of corticosteroids in the etiology of lung function decline, and identify drug targets meriting further study for potential repurposing to slow lung function decline and mitigate lung disease.

BACKGROUND: Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide. Serine protease inhibitors (SERPINs) have emerged as potential regulators in tumor progression, yet the specific function and regulatory mechanisms of SERPINA4 in CRC remain poorly characterized. METHODS: We integrated transcriptomic data from multiple CRC cohorts and conducted survival, pathway, and immune correlation analyses. Functional validation was performed using in vitro proliferation, migration, and invasion assays, as well as in vivo xenograft models. Post-transcriptional regulation of SERPINA4 via m6A modification was examined through MeRIP-qPCR, RNA immunoprecipitation, and decay assays. RESULTS: High expression of SERPINA4 was significantly associated with poor prognosis across multiple survival endpoints. SERPINA4 overexpression promoted CRC cell growth and metastasis in vitro and enhanced tumor formation in vivo. Mechanistically, ALKBH5-mediated m⁶A demethylation reduced SERPINA4 transcript stability by weakening IGF2BP3 association. Rescue experiments with methylation-deficient mutants confirmed the necessity of m6A for SERPINA4-driven oncogenicity. Moreover, SERPINA4 expression correlated with immunosuppressive signatures and reduced CD8 + T cell infiltration. CONCLUSIONS: Our findings reveal that SERPINA4 facilitates CRC progression through both tumor-intrinsic mechanisms and modulation of the immune microenvironment. These findings support an epitranscriptomic regulatory mechanism involving an ALKBH5–m6A–IGF2BP3 axis in CRC. SERPINA4 may serve as a prognostic biomarker and therapeutic target in precision oncology.

The global annual production of animal by-product (ABP)-derived bone, estimated at 95‒126 million tonnes, presents both an environmental challenge and an opportunity for sustainable resource utilization. We estimate that bone char (BC) could theoretically replace 13‒32% of the global phosphorus (P) fertilizer market. BC, produced through the pyrolysis of animal bones, has emerged as a promising material for use in a range of agricultural applications related to soil fertility and water quality. The conversion of ABP-derived bone into BC through pyrolysis not only eliminates potential human and animal pathogens (e.g., prions, viruses, bacteria), but also creates a valuable resource rich in P, calcium, and magnesium. This review synthesizes current research on the potential applications of BC in agriculture, focusing on its multifunctional role as a slow-release P fertilizer, a carbon (C) storage material, and an effective adsorbent for remediating contaminated soils. Field and laboratory studies demonstrate that BC's performance is strongly influenced by pyrolysis conditions, with optimal temperatures between 300 and 500 °C for nutrient release applications and above 600-800 °C for enhanced surface area and contaminant remediation. Its hydroxyapatite structure enables gradual P release and potential toxic element (PTE) immobilization, while its porous nature can provide new habitat niches for soil microorganisms and improve soil water retention. In comparison to most conventional inorganic fertilisers, BC can enhance soil fertility by releasing P slowly, thereby improving plant growth and productivity, particularly in acidic soils. The low cost, renewable nature, and ease of regeneration of BC further enhance its appeal as a viable solution for mitigating environmental pollution and promoting sustainable resource management practices. Beyond its established applications, this review identifies critical knowledge gaps, including the need to investigate BC's long-term impacts on soil health, microbial communities, and greenhouse gas emissions. We also discuss opportunities for optimizing production methods and expanding applications beyond agriculture. Given BC's potential to address multiple agricultural and environmental challenges, we emphasize the importance of interdisciplinary research to evaluate implementation barriers, including economic viability, social acceptance, and regulatory frameworks.<h4>Graphical abstract</h4>

Poly (ADP-ribose) polymerase (PARP) inhibitors have emerged as a paradigm-shifting therapeutic strategy in oncology, demonstrating significant synthetic lethality in tumors characterized by homologous recombination deficiency. Due to their substantial efficacy, PARP inhibitors (PARPi) have received approval for the treatment of four malignancies: ovarian cancer, breast cancer, prostate cancer, and pancreatic cancer. However, the specific clinical indications and optimal utilization settings vary among these types of cancers. Identifying novel biomarkers is thus essential for accurately predicting patients’ responses to PARPi, therefore enhancing effective patient stratification. Notably, the emergence of acquired resistance to PARPi presents a considerable challenge to their practical implementation in clinical practice. Both preclinical and clinical studies have elucidated numerous molecular alterations contributing to this resistance, offering valuable insights into potential strategies for overcoming it. This review comprehensively summarizes landmark clinical trials involving both PARPi monotherapy and various combination strategies, and outlines future research directions. We compare existing predictive tools for resistance to PARPi, aiming to refine future clinical applications and identify critical gaps requiring further investigation. This review also presents new insights into primary and acquired resistance by updating mechanisms of PARPi action and summarizing the biological processes involved in PARPi resistance. Additionally, we discuss potential strategies designed to overcome these mechanisms.

<h4>Background and aims</h4>Despite growing interest in the role of vitamin D in various infectious diseases, limited evidence exists regarding its association with platelet (PLT) parameters in patients with COVID-19 and Hepatitis C Virus (HCV) infection. This study aims to assess the relationship between serum vitamin D levels and PLT indices in patients with COVID-19 and chronic HCV.<h4>Methods</h4>The study included 113 patients with confirmed COVID-19 and 97 patients with chronic HCV infection. All participants were classified into two subgroups based on their vitamin D status (deficient vs. non-deficient). The chi-square test (χ2) or Fisher exact test was used to compare the results between the two groups.<h4>Results</h4>Patients with COVID-19 had significantly lower levels of vitamin D compared with HCV patients (<i>p</i> < 0.001). PLT counts were lower in the COVID-19 group, and MPV, PDW, and inflammatory markers (PLR, MPR, and MLR) were significantly higher. Also, a significant negative correlation was observed between vitamin D levels and MPV in both groups (<i>p</i> < 0.001). In patients with vitamin D deficiency, PLT and inflammatory changes were more pronounced in COVID-19 than in HCV.<h4>Conclusion</h4>The results of this study showed that COVID-19 patients had lower vitamin D levels, lower PLT counts, and higher indices of platelet activation compared with HCV patients. These changes are associated with the severity of the inflammatory response in COVID-19 and could be part of the mechanisms that cause thrombotic complications in this disease.

No abstract available.

The autonomic nervous system (ANS) plays a key role in cardiovascular regulation by maintaining hemodynamic and metabolic homeostasis through balanced sympathetic and parasympathetic activity. While autonomic dysfunction is classically associated with diabetes, neurodegenerative diseases, autoimmune neuropathies, and chronic cardiovascular conditions, growing evidence suggests that disturbances in iron metabolism represent an underrecognized contributor to cardiac autonomic dysregulation. This narrative review summarizes data from 107 studies on ANS disorders, including 49 investigating cardiovascular involvement. Reported abnormalities included reduced heart rate variability and baroreflex sensitivity, prolonged P-wave duration and QT dispersion, and deviations in non-invasive autonomic testing parameters. In iron overload states, these changes appear to be driven primarily by oxidative stress, whereas in iron deficiency they are likely mediated by tissue hypoxia. Importantly, several studies indicate that normalization of iron homeostasis may partially reverse autonomic dysfunction. This potentially reversible component underscores the clinical relevance of screening for and correcting iron imbalance not only to improve hematological status but also to reduce cardiovascular risk. Large-scale, multicenter studies using standardized autonomic assessment protocols are required to clarify prognostic implications and inform evidence-based clinical guidelines.

The heterogeneity of colorectal cancer (CRC) represents a great challenge in therapy. We integrated multiomics and machine learning, interpreted by SHAP models to provide a clinical rationale, to identify Calcineurin B Homologous Protein 2 (CHP2) as a core candidate, which was further validated via in vitro and zebrafish models. The expression of CHP2 are decreased in CRC, which is associated with a poor prognosis and an immune suppressed "cold" TIME. Functionally, CHP2 overexpression inhibits cell growth and invasion by inducing PANoptosis. Clinically, specific CHP2 expression profiles discriminate patients at high risk that are resistant to standard chemotherapy (e.g., 5-FU) but sensitive to targeted inhibitors. CHP2 is a powerful dual-function biomarker-prognostic for survival and predictive for the response to therapy-that could lead to a personalized approach in treating drug-resistant CRC.

Age-related macular degeneration (AMD) remains a leading cause for visual impairment in the elderly. We recently showed that activated mammalian target of rapamycin complex 1 (mTORC1) in photoreceptor cells causes AMD-like pathologies in mouse. Employing mouse genetics, we dissect the mTORC1 pathway and identify ribosomal protein S6 kinase beta-1 (<i>S6k1</i>) as a key component required for disease onset in our mouse model. Using a previously identified fully chemically modified tetravalent small interefing RNA (siRNA) that enriches in photoreceptors, we target <i>S6k1</i> in mouse, pigs, and non-human primates (NHP) by intravitreal injection. We find that <i>S6k1</i> silencing in diseased mice reverses phospholipid changes induced by activated mTORC1, restores lysosomal activity of retinal-pigmented epithelium cells, and reduces lipoprotein buildup at Bruch's membrane (BM). In pigs, which do not develop disease, we find a similar shift in phospholipids as in mouse, indicating a conserved role for <i>S6k1</i> in photoreceptor phospholipid metabolism. In aged NHPs with macular drusen, the lipoprotein-rich BM deposits that are a hallmark of human AMD, <i>S6k1</i> silencing slows drusen growth over a 6-month period. These findings establish <i>S6k1</i> as modifier of lipoprotein buildup at the BM and support our siRNA platform as a potential treatment modality for AMD and other retinal diseases.

<b>Background</b>: Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is an indolent B-cell lymphoma with long-term survival and a tendency for late relapse. Hepatic manifestations of varying etiologies have been described in lymphoproliferative disorders. However, paraneoplastic hepatitis is rare, and reports typically describe acute presentations. We describe an unusual case of paraneoplastic hepatitis with an indolent and progressive clinical course occurring in the setting of relapsed NLPHL. <b>Case Presentation</b>: A 32-year-old man with a history of NLPHL was found to have marked transaminase elevation with preserved liver function during routine follow-up. Extensive evaluation excluded viral, autoimmune, and metabolic causes of liver disease. Liver biopsy demonstrated confluent and bridging necrosis with lymphoplasmacytic infiltrates, without evidence of direct lymphoma involvement. Excisional biopsy of a cervical lymph node revealed relapse of NLPHL without histologic transformation. Treatment with corticosteroids resulted in partial biochemical improvement, and subsequent rituximab monotherapy achieved lymphoma remission. Despite this, low-grade transaminase elevation persisted, and follow-up imaging and liver biopsy demonstrated progression to fibrosis, suggesting a tendency towards chronicity. <b>Conclusions</b>: Paraneoplastic hepatitis should be considered in patients with NLPHL who present with unexplained liver abnormalities. This report illustrates a fibrosing form of paraneoplastic hepatitis associated with NLPHL and broadens the clinical spectrum of paraneoplastic hepatic injury. Early recognition, histological confirmation, and tailored immunosuppressive management are critical to optimizing hepatic and lymphoma-related outcomes.

Colorectal cancer (CRC) is a complex ecosystem shaped by bidirectional interactions between epithelium and the tumor microenvironment, prominently mediated by TGFβ signaling. Cancer-associated fibroblasts (CAFs) are regulators of epithelial plasticity and immune cell recruitment; yet, their diversity has impacted translationally applicable spatial analysis. Here, we distil the fibroblast continuum into two overarching CAF populations that are largely transcriptomically distinct and are marked by PDGFRA⁺ and ACTA2⁺ expression, enabling robust spatial identification using single immunohistochemical markers. We show that TGFβ signaling drives dynamic transitions between these states. In a preclinical model, selective ALK5 inhibition remodels CAF composition <i>in vivo</i>, reconfiguring local immune neighborhoods and indirectly altering epithelial stem cell states. Finally, we demonstrate that multiscale spatial analysis provides a quantitative readout of stromal-immune-epithelial remodeling following therapy. These findings establish a simplified, translationally relevant CAF framework and highlight spatially resolved stromal dynamics as measurable indicators of therapeutic response in CRC.

<h4>Background</h4>Canine parvovirus (CPV) causes acute hemorrhagic gastroenteritis in dogs, whereas canine circovirus (CanineCV), an emerging virus, is associated with immunosuppression and gastrointestinal disease. However, data on CPV-CanineCV coinfections are limited in Vietnam.<h4>Aim</h4>The aim of this study was to investigate the prevalence and clinical, hematological, biochemical, and pathological features of CPV-CanineCV coinfections in dogs with enteric disease in Hanoi, Vietnam.<h4>Methods</h4>The fecal and blood samples from suspected CPV cases were tested using molecular diagnostics. Clinical and laboratory findings were compared between the CPV mono- and coinfected groups. Necropsies were performed on the deceased animals.<h4>Results</h4>CPV infection was confirmed in 94.68% (89/94) of the cases, with 21.28% (19/89) showing CanineCV coinfection. Diarrhea, vomiting, and anorexia were observed in both groups, but coinfected dogs exhibited more severe symptoms, higher mortality, longer treatment, and worse recovery. The hematological findings included marked leukopenia, lymphopenia, and thrombocytopenia. Biochemical changes, such as hypoglycemia, hypoproteinemia, hypoalbuminemia, and mild liver enzyme elevations, along with notable hypokalemia and mild metabolic acidosis, were observed. Pathological findings showed a more extensive intestinal and systemic damage in patients with co-infections.<h4>Conclusion</h4>CPV-CanineCV coinfection results in more severe disease than CPV alone. Routine Canine CV screening in CPV-positive dogs is recommended for better clinical management.

<h4>Background: </h4> The diagnosis of autonomic dysfunction underlying Hereditary Coproporphyria (HCP) represents a major clinical challenge. Given the nonspecific and episodic nature of its crises, neurological and gastrointestinal symptoms are frequently misinterpreted and misdiagnosed as anxiety disorders or psychosomatic syndromes, severely delaying proper therapeutic management. Although HCP is characterized by the accumulation of porphyrin precursors, the high variability in tissue damage—ranging from refractory Small Intestinal Bacterial Overgrowth (SIBO) to inflammatory oncological processes (such as cholangiocarcinoma and cutaneous lymphomas)—suggests the necessity of an external catalyst. I postulate that abnormal iron metabolism is the primary driver of this pathological progression. Hypothesis: Iron overload mediated by the HFE mutation, combined with low ferroxidase activity (ceruloplasmin), generates an excess of non-transferrin-bound iron (NTBI) in the ferrous state (Fe2+). Furthermore, the high demand for heme synthesis driven by elevated hemoglobin forces the defective CPOX enzymatic pathway, massively increasing the accumulation of isomer III precursors. Free Fe2+ then acts as an oxidizing agent, transforming the inert coproporphyrinogen III into highly reactive and toxic coproporphyrin III. Mechanism: The oxidation of these isomers triggers systemic oxidative stress. Locally, oxidized porphyrins induce autonomic neuropathy in the myenteric plexus, paralyzing gastrointestinal motility and promoting intractable SIBO. Systemically, the massive biliary excretion of these reactive compounds causes chronic chemical inflammation in the bile ducts, increasing the risk of cholangiocarcinoma. Concurrently, cutaneous deposition of oxidized porphyrins generates chronic antigenic stimulation in the dermis, potentially acting as a trigger for lymphoproliferative neoplasms, such as marginal zone B-cell lymphoma. Clinical Relevance: This model suggests that patients with overlapping HFE and CPOX mutations face both autonomic dysfunction (SIBO) and elevated oncological risk due to chronic inflammation. Early management through therapeutic phlebotomies to reduce hemoglobin-driven heme demand and halt Fe2+-mediated oxidation, alongside intestinal binders, could prevent both neurogastrointestinal damage and long-term malignant transformation.

One of the great mysteries of Huntington disease (HD) is its adult onset. HD is caused by a CAG repeat expansion in the <i>HTT</i> gene, which encodes a protein (HTT) that is crucial for intracellular transport and neuronal growth, so one would expect development to be altered in some way. Human fetal samples, cell and animal model studies have shown that neurodevelopment is indeed altered in HD. Structural and functional abnormalities can be detected during the presymptomatic phase, and mutation carriers may display subtle cognitive deficits, psychiatric disturbances, or motor impairments years before clinical diagnosis. However, despite these early developmental alterations and the lifelong presence of mutant huntingtin (mHTT), HD carriers typically experience a long asymptomatic period. In this review, we provide an overview of the functions of HTT and the dysfunctions caused by mHTT or HTT loss during development, from gastrulation to neurulation, and examine the specific contribution of the cerebral cortex in HD. Drawing on nearly 25 years of research, we integrate studies ranging from in vitro to in vivo studies, highlighting the molecular mechanisms that contribute to HD neuropathology. Understanding these early developmental processes may define a window for therapeutic intervention, a period of plasticity during which the brain may still correct or compensate for developmental errors. This could strengthen the foundations to extend the presymptomatic phase and delay neurodegeneration.

Chagas disease is classified as a neglected tropical disease (NTD) and is predominantly endemic in South America. Despite advances in modern medicine, more than 10 million people worldwide are currently infected, and approximately 12 000 deaths are reported annually. The World Health Organization (WHO) aims to eliminate Chagas disease by 2030. Although more than 100 years have passed since the discovery of Chagas disease, no effective vaccine has yet been developed. The only available chemotherapeutic agents are nifurtimox and benznidazole, both of which were introduced more than half a century ago and are included in the WHO's list of essential medicines. However, because these drugs are associated with severe adverse effects and are effective only during the early phase of infection, they have not achieved eradication of Chagas disease. In this study, a scalable synthetic method was developed for noncanonical cyclic peptides bearing a rare N-acylindole linkage, a structural motif not typically observed in conventional cyclic peptides. This strategy enabled the synthesis of bulbiferamide A, a potent antitrypanosomal agent reported to be effective against Chagas disease, as well as a variety of related analogues that had previously been difficult to access synthetically.

No abstract available.

<h4>Introduction</h4>Differentiating frontotemporal lobar degeneration (FTLD) from Alzheimer's disease (AD), particularly when presenting with overlapping behavioural symptoms, remains clinically challenging. This study assessed the utility of Addenbrooke's Cognitive Examination-III (ACE-III)-derived ratios in distinguishing these conditions.<h4>Methods</h4>A retrospective cohort of 115 patients (n = 52 biomarker-confirmed AD, n = 63 FTLD) with overlapping behavioural symptoms was analysed. ACE-III scores and behavioural profiles were examined, and novel ratios were tested.<h4>Results</h4>The novel Phonemic Fluency/Orientation-Memory (PFOM) ratio outperformed existing ACE-III metrics, achieving an area under the curve of 0.85 (sensitivity 84.1%, specificity 73.1%) for differentiating FTLD from AD. Notably, longer symptom duration in AD, but not in FTLD, was associated with worsening frontal and cognitive symptoms. Patients with AD were significantly older than those with FTLD.<h4>Conclusion</h4>These novel ratios showed robust diagnostic performance, regardless of disease duration, and better reflect FTLDs cognitive profile. They may offer improved clinical utility over traditional ACE-III measures.

Diversity Outbred (DO) mice are a powerful model system for mapping complex traits due to their high genetic diversity and mapping resolution. However, while there are extensive tools available for standard genetic analysis in DO mice, fewer techniques have been implemented to facilitate integrated, cross-study analysis. Here, we implement Haseman-Elston regression to estimate genetic correlations among 7,233 phenotypes measured across eleven independent DO mouse studies. We used this network of genetic correlations to cluster phenotypes according to shared genetics, which enhanced the power to detect quantitative trait loci (QTL). This approach empowered the detection of 884 QTL for 383 meta-phenotypes, explaining an average of 40.36% of the total genetic variance per mega-analysis. We leveraged this network for insights into specific areas of biology, including lifespan, frailty, immune composition, histological and functional lung phenotypes, and histological phenotypes of the aorta. We found the genetics of lifespan to share limited correlation with the genetics of frailty but stronger correlation with the genetics of immune cell composition. Additionally, mega-analyses driven by genetic correlations identified candidate genes (e.g., Cdkn2b) associated with degraded extracellular matrix in the aorta. Finally, an ensemble of genetic analyses implicated pulmonary neuroendocrine cell signaling and/or differentiation as a key driver of multiple lung pathophenotypes.

A plausible hypothesis for the origin of biological homochirality invokes chiral symmetry breaking, transfer, and amplification driven by autocatalysis. Here we experimentally demonstrate that crystallization-driven template autocatalysis (CDTA) induces mirror symmetry breaking and amplification in helices. CDTA enables the reductive cyclotetramerization of hydrogen-bonded naphthalonitrile precursors into crystalline fibers of naphthalocyanine derivatives. In achiral or racemic systems, a kinetically controlled right-handed helical bias emerges during secondary nucleation and develops into P-helical dominance as the fibers elongate, achieving mirror symmetry breaking. CDTA also transfers the single handedness of chiral seeds formed from enantiopure analogs to achiral naphthalocyanines through template-assisted replication, resulting in chiral amplification. A key mechanistic step involves the preorganization of naphthalonitrile molecules in a counterclockwise direction at the termini of P-helical fibers via J-type π-π stacking and hydrogen-bonding interactions for autocatalytic transformation. Thus, once mirror symmetry is broken, the resulting chiral imbalance is amplified in a self-replicating manner.

Since 2018, truncating and missense mutations in the IRF2BPL gene have been associated with the neurodevelopmental NEDAMSS syndrome and other IRF2BPL-related disorders. These mutations mainly affect the gene region encoding the central portion of the IRF2BPL protein, whose physiopathological roles are poorly understood. Here we show that this region contains three low-complexity regions (LCR_1-3) and a higher-complexity domain (HCD) similar to DNA-binding domains. The polyA/polyQ-rich LCR₁, together with an upstream zinc finger domain, drives IRF2BPL liquid-liquid phase separation (LLPS) in both neuronal and non-neuronal cells. Disease-related truncating and missense mutations disrupt the physiological IRF2BPL LLPS in cells, leading to the formation of aberrant cytoplasmic condensates sequestering wild-type IRF2BPL from the nuclear compartment, activating molecular disease mechanisms, i.e., WNT1 upregulation, and altering neuronal cell electrophysiological properties. These findings identify aberrant LLPS as a key biophysical mechanism underlying the nuclear depletion of wild-type IRF2BPL, a fundamental pathogenic alteration underlying NEDAMSS, triggering downstream cellular and molecular disease mechanisms.

Cyclooxygenase (COX) inhibitors (COXIBs) have shown preventive and therapeutic potential for colorectal cancer (CRC). In addition to inhibiting COX, approved COXIBs also target COX-independent pathways, including NF-κB and AKT. We evaluated how inhibition of various COXIB targets affects cell proliferation and apoptosis using a Boolean model incorporating the effect of several genetic mutations linked to CRC. The mutations activate two positive feedback loops and cause increased cell proliferation and survival. When simulating loss of function of APC, proliferation and apoptosis can be restored to healthy rates by inhibiting COX2. In simulations reflecting other genetic mutations, none of the investigated targets restore apoptosis and proliferation is reduced by inhibiting AKT or NF-κB, but not by any of the other COX targets tested. These results show that the off-target effects of COXIBs are crucial for their efficacy in the treatment of sporadic CRC.

Responsive optical materials offer real-time visual feedback to external stimuli by converting them into visible light signals, holding great promise in display, sensing, and information encryption. Typically, heterogeneous Janus droplets can exhibit vivid coloration induced by total internal reflection and interference (TIRI) at concave liquid-liquid interfaces. Yet, achieving stimuli-responsive TIRI-based structural colors remains a challenge due to the difficulty in controlling interface curvature and reflective behavior under external stimuli. Here, we report the fabrication of Janus droplets with tunable TIRI structural colors that respond to both surfactant composition and temperature. By tailoring surfactant formulation, the droplet morphology and interface curvature can be adjusted, enabling reversible modulation of structural color appearance, including both iridescent and non-iridescent modes depending on incident light conditions. Furthermore, the structural color can be reversibly or irreversibly switched on/off in response to temperature changes, due to phase transitions that alter light reflection at the concave interface. Owing to their temperature-dependent optical response and real-time visual signaling, these droplets can serve as single-use security labels for monitoring freeze history of freeze-sensitive pharmaceuticals, providing reliable indication of product integrity.

​Integrated stress response (ISR) genes are implicated in lung adenocarcinoma (LUAD) prognosis, but their clinical utility remains unexplored. This study aims to identify ISR-related prognostic genes and construct a risk model for LUAD survival prediction.​ LUAD transcriptomic and clinical data were retrieved from public databases. ISR-related genes (ISR-RGs) were screened via differential expression and regression analysis. A risk model and a nomogram integrating clinical indicators were built and validated. Functional enrichment, immune cell infiltration, and RT-qPCR in clinical samples were performed.​​​ Five prognostic genes (AGER, GPX3, CCNA2, KCNK3, and CHEK1) were identified. High-risk patients exhibited poorer survival. The nomogram was able to forecast the survival of LUAD well. Genes were functionally linked to cell cycle and DNA replication and correlated with immune cells (e.g., CCNA2 positively with CD4⁺ T cells [cor = 0.52]; CHEK1 negatively with memory B cells [cor = - 0.40]). RT-qPCR confirmed dysregulation: AGER, GPX3, and KCNK3 downregulated and CHEK1 and CCNA2 upregulated versus controls. The five prognostic genes pertinent to both LUAD and ISR were identified, and a risk model was constructed for the good prediction of LUAD survival, which offered a fresh outlook for alleviating the poor prognosis of LUAD.

BACKGROUND AND AIMS: The effects of caloric restriction (CR) during active colitis remain incompletely understood. We examined whether short-term CR, initiated during dextran sodium sulfate (DSS)-induced inflammation, modulates disease severity, host transcriptional responses, and gut microbiota composition. Cross-species transcriptomic analyses were used to contextualize these effects within established human ulcerative colitis (UC) signatures. METHODS: Male C57BL/6 mice were assigned to control, CR, DSS, or DSS with concurrent CR (DSS.CR) groups. Clinical disease activity, histopathology, hematology, anxiety-like behavior, colon and spleen transcriptomes (RNA-seq), and fecal microbiota (16S rRNA) were assessed. Public UC microarray datasets were used to compare DSS and DSS.CR transcriptional profiles with human active and inactive UC. RESULTS: Compared with DSS, DSS.CR mice showed modest but consistent improvements in stool consistency and bleeding, with limited effects on overall histopathological scores, and reduced splenic enlargement and disruption. Transcriptomic analysis revealed limited differential gene expression between DSS and DSS.CR, but gene set enrichment analysis indicated reduced activation of inflammatory pathways (e.g., NF-κB, IL-17, cytokine–cytokine receptor) and relative enhancement of epithelial renewal and proteostasis programs. CR also partially mitigated DSS-associated dysbiosis, modestly preserving microbial diversity and functional pathways linked to carbohydrate metabolism and detoxification. Human microarray meta-analyses confirmed that DSS recapitulates active UC signatures, while CR shifted these toward a remission-like profile. CONCLUSIONS: Short-term CR during acute DSS-induced colitis partially attenuated mucosal and systemic inflammation and modestly shifted transcriptional and microbial profiles toward epithelial repair and metabolic resilience. These findings highlight CR’s potential as a non-pharmacological adjunct for modulating intestinal inflammation and support further translational evaluation of feasible dietary restriction strategies in IBD.

SUMOylation is a critical post-translational modification of proteins. However, the roles of SUMOylation-related genes in regulating the tumor immune microenvironment and their potential as therapeutic targets in CRC remain unclear. We integrated transcriptomic data from TCGA and GEO to identify 200 SUMOylation-related genes associated with prognosis. Among these, 58 genes exhibiting significant survival differences were selected and classified into three distinct clusters. Cluster C patients have the worst prognosis, linked to increased tumor proliferation and weakened immune responses. The LASSO combined with Random Survival Forest (RSF) approach demonstrated optimal performance in ten machine learning algorithms, yielding the highest C-index (0.715). Subsequently, six key genes—SENP7, NUP85, UHRF2, BRCA1, TOPORS, and SENP5—were selected using the LASSO-RSF framework to develop a refined prognostic model. This model demonstrated high predictive accuracy, with 1-, 3-, and 5-year overall survival AUC values of 0.943, 0.961, and 0.981, respectively. Furthermore, the risk score derived from the model was significantly correlated not only with tumor metastasis and recurrence, advanced tumor stage, and patient age, but also with immune cell infiltration, tumor mutational burden, and sensitivity to anticancer drugs. Single-cell analysis revealed that the core gene UHRF2 is highly expressed in CD8 + exhausted T cells, but exhibits low expression in conventional CD8 + T cells and natural killer cells. Experimental data demonstrated that UHRF2 was upregulated in CRC and linked to poor prognosis. Silencing UHRF2 markedly inhibited both proliferation and migration of tumor cells in vitro, as well as tumor growth in vivo. In conclusion, the prognostic model derived from SUMOylation-related genes demonstrates robust predictive accuracy for patient outcomes. UHRF2 is associated with immunosuppression and promotes CRC progression, positioning it as a promising therapeutic target in CRC.

<h4>Background</h4>Genetic effects due to long term exposure to low doses of ionizing radiation (LDIR) in humans are not well understood. Human population living in high level natural radiation areas (HLNRAs) of Kerala coast in India are continuously exposed to chronic LDIR emanating from monazite containing beach sand for many generations. The background radiation level in this area varies from < 1.0 to 45mGy/year. The people residing in HLNRAs sometimes receives background radiation dose which is approximately 10-40 times higher than the people living in adjacent normal level natural radiation areas (NLNRAs). This population provides a unique opportunity to identify, if present, a mutational signature due to chronic low-dose radiation exposure in humans. We have employed whole exome sequencing approach to determine germline mutational changes in the lymphocytes of healthy individuals from HLNRAs (mean background dose: 31.8 ± 5.4 mGy/year, mean age: 43.0 ± 5.9 years) and compared them with healthy individuals from NLNRAs (mean background dose: 0.9 ± 0.2 mGy/year, mean age: 43.0 ± 11.3 years).<h4>Results</h4>Our results revealed that the overall number of single nucleotide variants (SNVs) and insertions/deletions (indels) were not significantly different in HLNRA (7744 SNVs, 880 indels) and NLNRA (7951 SNVs, 856 indels) groups. A similar number of protein affecting mutations (PAMs) were observed in HLNRA (1925) and NLNRA (2082) individuals. Interestingly, several unique SNVs were identified in both the groups. In HLNRA, unique SNVs were overrepresented in genes involved in important biological pathways such as DNA repair (EXO1, PARP2, DDB1, POLQ, LIG1), epigenetic modification (KDM5D, SETDB2, KMT2B, BRD8, SIRT1), cell cycle progression (CDK14, CCND1) etc. Furthermore, significant predominance of C > T transitions which were unique to HLNRA group was observed preferentially at CpG dinucleotide regions. Analysis with REVEL and AloFT tools did not show any increase in potentially pathogenic mutations including those involved in carcinogenesis in HLNRA individuals exposed to chronic radiation.<h4>Conclusion</h4>This study did not show any significant changes in genetic variants due to long term exposure to LDIR in human population living in HLNRAs of Kerala coast. However, presence of unique SNVs and C > T transitions in CpG islands of HLNRA individuals indicate the possible role of epigenetic mechanisms i.e. DNA methylation in response to chronic LDIR in this population. This study significantly enhances the current understanding of radiation induced genetic changes and associated cancer risk in human population.

Glioblastoma (GBM) remains a lethal tumor, largely due to robust mechanisms that prevent effective induction of cell death. Ferroptosis, a form of iron-dependent cell death, is a promising vulnerability in GBM. Here, we demonstrate that lipocalin-2 (LCN2) suppresses ferroptosis in GBM cells via the receptor tyrosine kinase AXL. LCN2 was elevated in GBM cells compared to lower-grade tumor and non-transformed cells, and Lcn2 knockdown impaired GBM cell fitness and growth in vitro and in vivo. Mechanistically, Lcn2 knockdown triggered ferroptosis, which was specifically rescued with ferroptosis inhibitors but not apoptosis or necroptosis inhibitors. Lcn2 knockdown reduced AXL phosphorylation, which was elevated in GBM patient tumors relative to non-tumor tissue. Notably, the combination of Lcn2 knockdown and pharmacological AXL inhibition extended survival compared to Lcn2 knockdown alone. Taken together, these data reveal a link between LCN2-mediated suppression of ferroptosis with AXL phosphorylation and support this axis as a potential therapeutic target for GBM.

<h4>Introduction</h4>We hypothesize that specific cognitive assessments and plasma biomarkers may exhibit heightened sensitivity during the stage of subjective cognitive decline (SCD). The integration of these plasma biomarkers and cognitive assessments could enhance the ability to predict beta-amyloid (Aβ) pathology in individuals with SCD.<h4>Methods</h4>A total of 231 participants, including 74 normal controls (NC) and 157 SCD, underwent Aβ and tau PET scans and blood testing for Aβ40, Aβ42, p-tau181, p-tau217, NfL, and GFAP. Cognitive assessments, plasma biomarkers, tau PET SUVr, and demographics were compared between Aβ+ and Aβ- groups within NC and SCD. The least absolute shrinkage and selection operator (LASSO) and logistic regression were employed to perform variable selection and develop predictive models.<h4>Results</h4>We observed significantly worse global cognition, visuospatial memory performance, executive function, and metamemory, as well as higher tau PET SUVr, elevated levels of p-tau217, p-tau181, and GFAP, and lower Aβ42/Aβ40 ratios in SCD Aβ+ compared to SCD Aβ-. The model incorporating BVMT-LD and p-tau217 achieved a slightly higher AUC than the model using p-tau217 and Aβ42/Aβ40 (0.94 vs. 0.93). Partial correlation analyses indicated that both auditory verbal memory (AVLT-LD) and visuospatial memory (BVMT-LD) were significantly negatively associated with p-tau217, whereas only AVLT-LD demonstrated a significant negative association with tau pathology severity.<h4>Conclusion</h4>Visuospatial memory deficit and plasma p-tau217 are powerful biomarkers for identifying Aβ+ in SCD. Auditory verbal memory links to tau pathology severity, while visuospatial memory is more sensitive to Aβ deposition, supporting early intervention to prevent AD progression.

A principal challenge in modern biology and biomedical research is the comprehensive elucidation of the molecular and cellular mechanisms that underpin human disease, with the ultimate aim of identifying novel and effective therapeutic targets. Embryogenesis represents a paradigm of tightly regulated biological complexity, orchestrated by elaborate networks of signaling pathways and transcriptional programs. This highly dynamic process, which initially unfolds within a seemingly disordered and heterogeneous microenvironment, is progressively structured through the establishment of intricate molecular equilibriums mediated by a diverse array of regulatory factors. Perturbations of these homeostatic balances at later stages frequently precipitate the emergence of various pathological conditions. Raf kinase inhibitor protein (RKIP) and Yin Yang 1 (YY1) are pleiotropic regulatory molecules implicated in a broad range of fundamental cellular processes, including proliferation, differentiation, migration, invasion, and epithelial-mesenchymal transition. Although these factors have conventionally been examined as discrete entities, emerging evidence indicates that RKIP and, YY1 may engage in significant molecular cross-talk that not only influences embryonic development but also contributes to the pathogenesis of diverse diseases, including cancer, autoimmune disorders, and immunological dysfunctions. Focusing on the pivotal developmental process of embryogenesis, this review explores the individual regulatory functions of RKIP and YY1, while emphasizing their potential points of convergence within signaling networks during both physiological development and pathological states.

Single-cell and single-nuclei RNA-seq (sc/snRNA-seq) have become a central approach in cancer research, and their widespread impact has been enabled by various computational tools developed specifically for sc/snRNA-seq analysis. Nevertheless, proper analysis and interpretation of sc/snRNA-seq data requires significant expertise, and the inadequate utility of certain computational methods may lead to dubious results. To mitigate these issues, it is important to recognize the limitations of sc/snRNA-seq data, the assumptions of common methods, and to perform robust analysis. Here, I describe common pitfalls in cancer sc/snRNA-seq analysis and discuss ways to overcome them. Among others, this includes a discussion of potential errors in statistical analysis, in inference of chromosomal aberrations, in trajectory analysis, and in signature-based analysis of bulk RNA-seq data. This review may help readers to avoid common pitfalls and to perform informative analysis and careful interpretation of sc/snRNA-seq datasets in cancer.

Neurological disorders such as Alzheimer's disease (AD) and Parkinson's disease (PD) pose a serious global public health threat, with complex etiologies involving genetic, environmental, and metabolic factors. Current data indicate that the prevalence of these disorders is rapidly increasing with the aging population, resulting in a growing economic and healthcare burden worldwide. In recent years, the imbalance of copper homeostasis has been increasingly implicated in the pathogenesis of neurological diseases. Copper overload can aggravate neuronal injury by inducing oxidative stress (OS), mitochondrial dysfunction, and protein misfolding, while copper deficiency disrupts the function of copper-dependent enzymes and leads to metabolic abnormalities. The mechanism of cuproptosis, proposed in 2022, describes a novel form of programmed cell death characterized by lipoylated protein aggregation and the loss of Fe-S cluster proteins, offering new insights into copper-related diseases. Multiple studies have demonstrated the crucial role of copper homeostasis and cuproptosis in the onset, progression, and treatment of neurological diseases. This narrative review summarizes the molecular mechanisms involved in copper homeostasis regulation and, on that basis, discusses the role of copper metabolism abnormalities in AD, PD, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Wilson's disease (WD), Menkes disease (MD), and stroke. Additionally, we highlight the mechanisms of existing copper-regulating drugs and their therapeutic potential in neurological disorders, while pointing out the limitations of current drug development.

Recent years have seen rapid progress in biological treatments for genetic diseases, as well as conditions like type 1 diabetes that lack an obvious genetic component. The authors sought to explain why this progress has emerged at this particular moment. The best way to illustrate this is by showcasing a wide range of therapies targeting diverse diseases. This progress has been driven by technological advances in genetically modified CAR-T and CAR-NK cells (e.g., using CRISPR or transgenes), which have led to significant improvements in cancer therapy. A key trend now is the emergence of "off-the-shelf" approaches aimed at generating cellular therapies compatible with a range of recipients by mitigating alloreactivity and immune rejection. Different diseases impose distinct biological and logistical limitations; thus, treatment of each patient requires an appropriate strategy. Emerging advances include the modification of therapeutic cells, either ex vivo or in vivo. Current options for transgene delivery mainly comprise lipid nanoparticles (LNPs), adeno-associated virus (AAV) vectors, and lentiviral vectors. Researchers also focus on selecting suitable promoters for specific expression in selected cell types. Altogether, these advances have led to remarkable progress in treating various diseases in recent years. This publication discusses the development of biological therapies, with particular emphasis on cell and gene therapies, illustrated by viable examples across various disorders. It covers implemented solutions for several types of cancer, as well as selected hereditary diseases and syndromes, including Huntington's disease, carbamoyl phosphate synthetase 1 (CPS1) deficiency, hemiplegia, epidermolysis bullosa, chronic granulomatous disease, and congenital deafness. Emerging applications in heart diseases and diabetes are also summarized, along with therapeutic strategies involving tRNA gene editing. Although numerous strategies exist, only the most representative, practical, and up-to-date examples are emphasized.

<b>Background/Objectives</b>: Primary ciliary dyskinesia (PCD) is a rare inherited disorder caused by dysfunction of motile cilia, leading to chronic respiratory disease. Diagnosis is challenging due to heterogeneous and non-specific clinical manifestations and the absence of a single definitive diagnostic test. Current diagnostic strategies rely on a combination of functional, ultrastructural, and genetic analyses. The objective of this study was to evaluate whether ciliary beat frequency (CBF), combined with ciliary beat pattern (CBP) assessment using digital high-speed video microscopy (DHSV), could serve as an effective first-line screening tool to identify patients requiring further diagnostic investigations. <b>Methods</b>: This single-center retrospective study included 65 patients (52 children and 13 adults) with clinical suspicion of PCD. Ciliary beat analysis was performed on nasal or bronchial samples using DHSV and Sisson-Ammons Video Analysis software. CBF and CBP were assessed and compared between patients with confirmed PCD and those in whom PCD was excluded based on transmission electron microscopy (TEM) and/or molecular genetic analysis. <b>Results</b>: Fifteen patients were diagnosed with PCD. Mean CBF was significantly lower in the PCD group compared with the non-PCD group (3.3 Hz vs. 8.1 Hz; <i>p</i> < 0.001). A CBF cut-off value of 5.25 Hz yielded a sensitivity of 78.6% and a specificity of 95.7%. Three patients with PCD had CBF values above this threshold; however, two of them exhibited abnormal CBP. Sample type, patient age, and the presence of airway pathogens did not significantly influence CBF measurements. <b>Conclusions</b>: CBF and CBP analysis using DHSV represents a useful first-line screening tool within a multifaceted diagnostic approach for PCD, allowing rapid identification of patients who should undergo further confirmatory testing.

Chronic consumption of high-fat diets (HFDs) induces obesity and metabolic dysfunction and is accompanied by progressive skeletal muscle wasting. Although aerobic exercise is generally considered less effective for maintaining muscle mass, accumulating evidence suggests that it can attenuate HFD-induced muscle wasting. However, the molecular mechanisms underlying this protective effect remain poorly defined. In this study, we investigated the effects of moderate-intensity continuous training (MICT) on HFD-induced muscle wasting and characterized the associated transcriptomic adaptations in the mouse gastrocnemius muscle. 21 weeks of HFD feeding increased body weight and serum glucose levels and induced marked muscle wasting, as evidenced by reduced gastrocnemius muscle index, impaired forelimb grip strength, decreased muscle fiber cross-sectional area, and excessive intramuscular lipid accumulation. These pathological alterations were significantly attenuated by an 8-week MICT intervention. RNA sequencing revealed that HFD predominantly induced a lipid-centered transcriptional program characterized by enhanced fatty acid uptake, trafficking, and β-oxidation. In contrast, MICT predominantly suppressed atrophy-associated genes (<i>Foxo1</i>, <i>Fbxo32</i>, and <i>Trim63</i>), while exerting minimal effects on myogenic genes (<i>Pax7</i>, <i>Myod1</i>, and <i>Myog</i>). Functional enrichment analyses further indicated that MICT modulated signaling pathways related to FoxO, PI3K-Akt, MAPK, and insulin signaling, together with biological processes associated with angiogenesis and calcium signaling. Collectively, these results suggest that MICT mitigates HFD-induced muscle wasting primarily by reprogramming the transcriptome from a lipotoxic, atrophic state toward a more insulin-sensitive, pro-angiogenic profile, with limited myogenic activation.

<h4>Introduction</h4>Soil salinization threatens global land use and food security, and halophytes combined with peat amendments are promising for saline-alkali soil remediation.<h4>Methods</h4>Here, we integrated transcriptomic and physiological analyses to investigate the adaptive responses of <i>Suaeda glauca</i> and <i>S. salsa</i> grown in saline-alkaline soils amended with peat at 0, 6, or 18 g/kg.<h4>Results and discussion</h4>Our results showed that a high peat concentration (18 g/kg) significantly improved salt tolerance and biomass accumulation in both species through distinct species-specific strategies. <i>S. glauca</i> upregulated growth-related pathways (e.g., nitrogen metabolism, and tricarboxylic acid cycle) mediated by bHLH and bZIP transcription factors (TFs), whereas S. salsa activated stress-mitigating secondary metabolism (e.g., flavonoids, phenylpropanoids, anthocyanins) regulated by MYB and NAC TFs. A conserved response across both species was the downregulation of genes involved in amino acid degradation, which helps conserve nitrogen for osmoprotection. RT-qPCR analysis confirmed the reliability of the RNA-seq data. This study identified 18 g/kg as the optimal peat concentration, uncovers species-specific adaptive mechanisms in halophytes, and lays a foundation for the precisely selection of halophyte-peat combinations in saline-alkaline soil remediation.

<h4>Background</h4>SET domain-containing 2 (SETD2), the sole histone H3 lysine 36 trimethyltransferase, has emerged as a critical tumor suppressor across multiple cancer types. Recent evidence suggests SETD2 orchestrates complex interactions between metabolic reprogramming and immune evasion in the tumor microenvironment.<h4>Methods</h4>Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)2020 guidelines, we systematically searched PubMed, EMBASE, Web of Science, and Cochrane databases from inception through April 2024. We included studies investigating SETD2's role in tumor metabolism and immunotherapy response. Meta-analysis was performed using random-effects models to assess the association between SETD2 status and clinical outcomes. Protocol was developed <i>a priori</i> but not registered due to the exploratory nature of this emerging field.<h4>Results</h4>Of 2,847 initially identified records, 78 studies met inclusion criteria, encompassing approximately 12,400 patients across 12 cancer types. SETD2 loss was associated with metabolic reprogramming (pooled OR: 2.34, 95% confidence interval (CI): 1.89-2.89, p < 0.001) and decreased immunotherapy response (hazard ratio (HR): 1.56, 95% CI: 1.32-1.84, p < 0.001). Substantial heterogeneity was observed (I-squared heterogeneity statistic (I²) = 52-68%) and explored through subgroup and sensitivity analyses. Mechanistically, SETD2 deficiency promoted glycolytic shift, lipid metabolism dysregulation, and immunosuppressive metabolite accumulation. Furthermore, SETD2 loss correlated with reduced CD8⁺ T cell infiltration and increased regulatory T cell presence.<h4>Conclusion</h4>This meta-analysis identifies SETD2 as an epigenetic regulator linking tumor metabolic reprogramming to antitumor immunity. SETD2 loss was associated with altered metabolic states and reduced clinical benefit from immune checkpoint inhibitors, with the strongest translational relevance observed in ccRCC and substantial evidence in NSCLC and CRC. These findings support further prospective validation and standardized assessment of SETD2, as well as exploration of rational metabolic-immunotherapy combination strategies in SETD2-deficient tumors.

<h4>Background</h4>Amino acid metabolism is integral to tumor proliferation, redox control, and immune regulation. Yet, studies in bladder cancer have largely centered on single amino acids, leaving the broader metabolic gene network insufficiently characterized.<h4>Methods</h4>Transcriptomic and clinical data from TCGA-BLCA, GSE13507, and GSE32894 were integrated with 32 MSigDB amino acid metabolism gene sets. Differential analysis, enrichment profiling, and consensus clustering defined metabolic subtypes. WGCNA and survival filtering identified candidates for a prognostic model, which was optimized using the MIME platform. Immune features and drug sensitivities were evaluated through multiple deconvolutions and pharmacogenomic resources. Single-cell data (GSE222315) were used to trace the cellular origin of model genes. <i>PSPH</i> expression and function were validated in tissues and bladder cancer cell lines.<h4>Results</h4>A total of 144 dysregulated amino acid metabolism-related genes were identified and used to define two distinct metabolic subtypes. One subtype was marked by coordinated upregulation of glutamine, branched-chain amino acid, tryptophan, and serine metabolic programs, accompanied by higher grade and stage, significantly worse survival, and dense but functionally impaired immune infiltration. From 24 candidate genes, a 16-gene metabolic signature was constructed and consistently validated across TCGA, GSE13507, and GSE32894, showing strong and stable prognostic performance superior to several published models. High-risk group displayed activation of cell-cycle, DNA-replication, <i>mTORC1</i>, and inflammatory-stress pathways, together with predicted sensitivity to <i>PI3K/mTOR</i> inhibitors, DNA-damaging agents, and selected epigenetic or cytoskeletal drugs. In the IMvigor210 cohort, the high-risk group showed a greater likelihood of responding to <i>PD-1/PD-L1</i> blockade. Single-cell profiling localized signature expression predominantly to malignant epithelial cells. <i>PSPH</i>, a core model gene, was overexpressed in tumor tissues and cell lines, and functional assays demonstrated its role in promoting proliferation, migration, invasion, and survival of bladder cancer cells.<h4>Conclusions</h4>This study highlights the central role of amino acid metabolic networks in shaping bladder cancer heterogeneity and provides a metabolically grounded framework for risk stratification and therapeutic development.

This article systematically reviews ferroptosis-an iron-dependent, lipid peroxidation-driven form of programmed cell death. It provides a detailed analysis of its core regulatory mechanisms, encompassing the drive from lipid peroxidation, the collapse of antioxidant defenses such as the glutathione peroxidase 4(<i>GPX4</i>)axis and alternative pathways like Ferroptosis Suppressor Protein 1 (<i>FSP1</i>), and the remodeling of iron and lipid metabolism. The interplay between ferroptosis and other cell death modalities, such as apoptosis and necroptosis, is also elucidated. The review focuses on the pivotal roles of key signaling pathways, including <i>NRF2, p53, and Hippo-YAP</i>, within the ferroptosis regulatory network. In the context of cancer therapy, the article emphasizes the potential of inducing ferroptosis for reversing drug resistance, inhibiting metastasis, and synergizing with immunotherapy. It systematically outlines direct induction strategies (e.g., small-molecule inducers, nanodelivery systems) and combination strategies with conventional therapies, targeted therapy, and immunotherapy. This review highlights that the bidirectional interplay between ferroptosis and the tumor immune microenvironment constitutes a novel therapeutic paradigm for combination therapy. Specifically, it elucidates how ferroptosis modulates immune cells such as CD8⁺ T cells and macrophages, reshaping the tumor immune microenvironment and offering new avenues for combination immunotherapy. We conclude by providing a roadmap for translating these insights into clinical practice, addressing current challenges, and outlining future directions for developing next-generation anticancer strategies.

The Regnase/MCPIP ribonucleases are involved in the regulation of immune homeostasis, by degrading RNA transcripts that encode inflammatory and regulatory proteins. This review highlights their molecular architecture, catalytic mechanisms, and intricate regulatory networks that orchestrate innate and adaptive immunity. This article presents a narrative review of the literature on distinct physiological roles of individual family members and how their dysfunction drives inflammatory, autoimmune, fibrotic, metabolic, and neoplastic disorders across multiple tissues. Although Regnase family members exhibit some functional redundancy, each also possesses distinct, non-overlapping roles. Regnase-1 restrains cytokine production and along with Regnase-2 modulates neuroinflammation. Both Regnase-3 and Regnase-4 possess homeostatic functions although they are also involved in orchestrating interferon and myeloid signaling and contribute to immune regulation and tumor suppression. We also examine emerging therapeutic strategies targeting Regnase activity, including antisense oligonucleotides to enhance Regnase-1 expression, gene- and RNA-based delivery approaches, and selective inhibition of Regnase-1 in T cells to boost cancer immunotherapy. Together, these findings underscore Regnase proteins as central post-transcriptional checkpoints in immunity and highlight their potential as targets for treating autoimmune disease, chronic inflammation, fibrosis, and cancer.

Chicken embryos and stem cells require precisely regulated levels of reactive oxygen species (ROS) to maintain self-renewal and pluripotency. However, excessive ROS induces oxidative stress, leading to DNA damage, chromosomal aberrations, loss of mitochondrial membrane potential, and ultimately, abnormal differentiation or cell death. This oxidative imbalance is a significant barrier to successful embryonic development and the in vitro culture of stem cells. Innate antioxidant defense systems exist within chicken and mammalian embryos to scavenge excess ROS, a finding that has prompted the strategic addition of antioxidants to culture media. The present review has two primary foci. Firstly, it seeks to expand the understanding of the antioxidant defense mechanisms of the chicken embryo. Secondly, it explores the role of exogenous antioxidant supplementation in the culture of various stem cell types, including embryonic stem cells (ESCs), primordial germ cells (PGCs), spermatogonial stem cells (SSCs), and induced pluripotent stem cells (iPSCs). Enhancing in vitro stem cell survival and directed differentiation through antioxidant supplementation holds significant promise for advancing fields such as tissue regeneration, organ transplantation, and the development of transgenic chicken models for improved production traits, vaccine development, and recombinant protein production.

Cell division is a highly regulated process that actively involves dynamic changes to the genetic material within the nucleus. DNA is faithfully replicated in the S-Phase of the cell cycle, being converted from loose, relaxed chromatin into tight, condensed chromosomes to be segregated in mitosis. In addition to scaffolding proteins that shape these mitotic chromosomes, post-translational modifications of histones within nucleosomes modulate chromosome dynamics throughout the cell cycle. In this review, we use a comparative approach to highlight some of the major epigenetic marks affected by the cell cycle during embryogenesis of <i>Caenorhabditis elegans</i>: H4K20me1, H3S10ph, H4S1ph, H2AS1ph, and H3T118ph. These five histone post-translational modifications will be specifically highlighted in the context of the mitotic cell cycle, as they are well documented in the <i>C. elegans</i> literature.

<b>Background/Objectives:</b> Pediatric sleep-disordered breathing (SDB) is influenced by craniofacial morphology and host susceptibility. Evidence integrating cephalometric airway features with targeted genetic variation in symptomatic skeletal Class II children remains limited. We explored whether children with skeletal Class II mandibular retrognathia and SDB symptoms harbor selected genetic variants and whether carriers show distinct cephalometric airway characteristics. <b>Methods:</b> This cross-sectional study included 48 children with skeletal Class II malocclusion, mandibular retrognathia, and snoring/mouth-breathing symptoms. Craniofacial and airway parameters were assessed on lateral cephalograms. SDB burden was evaluated by a baseline home sleep study (respiratory event index, REI). Targeted sequencing screened TNFRSF1A, PSTPIP1, SLC6A4 (5HTT), ACE, APOE, IRS1, and additionally PHOX2B and PMP22. Exploratory group comparisons used Student's <i>t</i>-test. <b>Results:</b> Variants were identified in 13/48 participants (27%) in TNFRSF1A, PSTPIP1, SLC6A4, ACE, APOE, and IRS1; none were detected in PHOX2B or PMP22. C3-H was higher in variant carriers (39.90 ± 6.40 vs. 36.48 ± 3.95 mm; <i>p</i> < 0.05). HH1 (perpendicular distance from the hyoid bone to the C3-RGN line) was higher but not significant (16.99 ± 7.58 vs. 14.61 ± 5.25 mm; <i>p</i> > 0.05). <b>Conclusions:</b> In this clinically screened pediatric skeletal Class II cohort with SDB symptoms, selected genetic variants co-occurred with specific hyoid-cervical cephalometric features. Given the cross-sectional design, absence of a control group, and small number of carriers, findings are exploratory and require replication in larger, controlled cohorts with standardized phenotyping.

<b>Background:</b> We sought to analyze the geographic distribution of <i>HFE</i> p.C282Y (homeostatic iron regulator c.845G>A; rs1800562) allele frequencies in Iberia. <b>Methods:</b> We analyzed published population/control cohorts of 50 or more subjects in mainland Spain and mainland Portugal and determined whether or not the p.C282Y genotypes in each cohort deviated from Hardy-Weinberg equilibrium (HWE) proportions. We defined combined p.C282Y allele frequencies from Spain and Portugal as representative of Iberia. We computed linear regressions (Pearson's correlations) of allele frequencies vs. latitudes and longitudes of cohort recruitment sites, defined significant regressions as allele frequency gradients, and mapped regional allele frequencies. <b>Results:</b> There were 34 Iberian cohorts: 25 Spanish (12,297 subjects; 11 autonomous communities) and 9 Portuguese (1024 subjects; five administrative regions). p.C282Y genotypes in one of 34 cohorts (2.9%) deviated significantly from HWE proportions. Aggregate allele frequency in Iberia was 0.0292 (778/26,642) [95% confidence interval: 0.0272, 0.0313]. The correlation of allele frequencies with latitude in Iberia was significant (r₃₄ = 0.4184; <i>p</i> = 0.0138). The correlation of allele frequencies with longitude was not significant (r₃₄ = 0.0014; <i>p</i> = 0.9936). The range of 16 regional allele frequencies in Iberia was 0.0068 (Murcia) to 0.5000 (Galicia). Frequencies were highest in regions adjacent to the north and northwest coasts (Cantabria, Galicia, Norte) and lowest in the south (Algarve, Murcia). <b>Conclusions:</b> There is a significant decreasing linear north-to-south gradient of <i>HFE</i> p.C282Y allele frequencies in Iberia. p.C282Y allele frequencies are highest in regions adjacent to the north and northwest coasts.

To gain new insights into the molecular underpinnings of coexisting rheumatoid arthritis (RA), cerebrovascular disease (cVD), and Alzheimer's disease (AD), we performed postmortem neuropathological examination and genetic screening of two individuals. The first individual (donor 1, D1) was a 74-year-old man who was diagnosed with both RA and AD and who also underwent hip replacement surgery bilaterally. The second individual (donor 2, D2) was a 90-year-old man with a reported diagnosis of RA, as well as two left hip replacements. A thorough histochemical (hematoxylin and eosin, H&E) and immunohistochemical (β-amyloid and tau protein) examination of D1 and D2 brains revealed the presence of AD-related pathology in both individuals, with AD stages being mild in D1 and intermediate in D2. The cVD-related pathology was also evident in both cases and was characterized by several microbleeds indicative of a compromised blood-brain barrier (BBB) integrity. Blood vessel wall thickening, a characteristic of arteriosclerosis, was significant in D1 but minor in D2. Therefore, the earlier RA diagnoses, along with the results of the neuropathological examination, indicated the coexistence in the donors of three major diseases: RA, cVD, and AD. The whole exome sequencing (WES) of DNA procured from D1 and D2 performed on the next-generation sequencing (NGS) Illumina platform (San Diego, CA) was followed by a very stringent bioinformatics analysis that yielded multiple genes with rare (minor allele frequency {MAF} ≤ 0.01) genetic pathological/deleterious variants associated with RA, cVD, and AD. Seven of those genes, <i>AQP7</i>, <i>ARSD</i>, <i>FAM160A1</i>, <i>HYDIN</i>, <i>IGSF3</i>, <i>OTOP1</i>, and <i>PRSS1</i>, were shared between D1 and D2, with all but <i>FAM160A1</i> having identical variants in both donors. Intriguingly, the subsequent analysis of the respective literature indicated that <i>FAM160A1</i>, <i>IGSF3</i>, and <i>PRSS1</i> were pleiotropic as they could be linked to all three coexisting diseases: RA, cVD, and AD. Altogether, the data presented herein are consistent with the notion that AD, cVD, and RA, when they coexist in humans, could be underpinned by a combination of polygenic and pleiotropic factors. Yet, a significant number of affected genes in the donors associated with bone and cartilage physiology point toward the possibility of joints being also damaged directly and independently of RA.

<h4>Background</h4>Glioblastoma (GBM) is the most common malignant brain tumor, and effective therapeutic strategies remain scarce. Therefore, the study aims to screen biomarkers to reveal the molecular mechanisms of cancer stem cells (CSCs) and disulfidptosis in GBM therapy.<h4>Methods</h4>The transcriptome data and clinical data for GBM were retrieved from The Cancer Genome Atlas (TCGA) database. The GSE74187 was obtained from the Gene Expression Omnibus (GEO) database. Firstly, the messenger RNA (mRNA)-based stemness index (mRNAsi) and disulfidptosis scores were computed, and weighted gene co-expression network analysis (WGCNA) was performed in TCGA-GBM using mRNAsi and disulfidptosis scores as traits to obtain the key module genes. Secondly, differentially expressed genes (DEGs) of the disease and normal groups in TCGA-GBM were screened, and DEGs were used to cross the key module gene to obtain intersection genes. Thirdly, univariate Cox, least absolute shrinkage and selection operator (LASSO), and multivariate Cox regression analyses were performed on intersection genes in TCGA-GBM to screen for biomarkers, and the biomarkers were used to construct survival risk score model. Meanwhile, clinical characteristics, immune infiltration, and drug sensitivity prediction of biomarkers were studied. To clinically validate the identified biomarkers, reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis was conducted on 20 samples.<h4>Results</h4>LOXL1, LOXL4, and SP6 were obtained by three regression analyses to build survival risk score model, and they were verified in GSE74187. Clinical pathological features analysis found that risk score and isocitrate dehydrogenase (IDH) status were independent prognostic factors for GBM. Immune-related analysis showed that the risk score had positive correlation with PDCD1, NRP1, TGFB1, and PVRL2. In silico drug sensitivity prediction suggested differential responses to 88 compounds between risk groups. Furthermore, preliminary molecular docking analysis indicated potential binding affinity of A_443654 and A_770041 with the biomarker proteins, highlighting these compounds as candidates for further experimental investigation. The RT-qPCR results revealed significant overexpression of LOXL1 and SP6 in the tumor group compared to the control group, while LOXL4 expression remained with no significant difference between the groups.<h4>Conclusions</h4>A risk prediction model based on LOXL1, LOXL4, and SP6 was constructed and validated. The model, rather than individual genes, provides valuable insights, providing valuable insights for the diagnosis of GBM in the field of CSC and disulfidptosis.

Pancreatic ductal adenocarcinoma shows early dissemination, stromal remodeling, and therapy resistance, but how tumor programs co-evolve with immune and stromal changes across stages remains unclear. We built a stage-resolved transcriptomic atlas using bulk RNA sequencing of FFPE samples from 443 untreated tumors spanning resectable, locally advanced, primary metastatic, and liver metastatic disease. Integrative modeling identified ten transcriptional programs with monotonic dynamics during progression. Early stages were enriched for epithelial differentiation and immune-stimulatory signals. Advanced stages showed increased cytoskeletal remodeling, vesicular trafficking, oxidative stress responses, and mitochondrial metabolism. Pathway analysis revealed enhanced PI3K-AKT-mTOR signaling and MYC target engagement in metastatic disease. Immune deconvolution showed loss of CD8⁺ T cells, dendritic cells, and M1 macrophages. Compact gene signatures stratified stage and survival and generalized to TCGA and ICGC cohorts. Functional assays confirmed greater invasiveness, altered redox balance, and mitochondrial dependence in advanced disease, suggesting stage-associated metabolic vulnerabilities.

Highly charged chains of poly(ADP-ribose) (PAR) are synthesized in the cell as part of their central role in DNA damage response. However, the effects of PAR on nucleosome structure and dynamics remain incompletely understood. Here we combine droplet-based microfluidic mixing with single-molecule Förster resonance energy transfer spectroscopy to resolve the kinetics of PAR-induced nucleosome decompaction in non-equilibrium measurements with millisecond time resolution. This approach avoids surface-adhesion and enables the tether-free observation of nucleosome remodeling. We find that PAR triggers nucleosome decompaction via a length-dependent kinetic threshold: Chains with less than ten ADP-ribose units act slowly and weakly, whereas longer PAR polymers induce efficient and rapid nucleosome opening. The extent and reversibility of decompaction further depend on PAR concentration and ionic strength, reflecting a mechanism dominated by electrostatic interactions. Enzymatic PAR digestion demonstrates that PAR can promote both reversible linker DNA opening and irreversible nucleosome disassembly. Coarse-grained molecular simulations suggest that these effects arise from a competition between PAR and DNA for histone tail binding. Altogether, our results establish PAR length as a key factor controlling chromatin accessibility during DNA repair and highlight droplet-based microfluidics as a powerful platform for studying such biomolecular interactions.

Recent genetic studies have shown somatic expansion of the CAG repeat is the key process driving Huntington’s disease (HD) pathogenesis. Recognition of insertion deletion loops (IDLs), lesions prone to form within the CAG repeat, by Mutsβ (MSH3/MSH2) is thought to be the primary event in the expansion process. This starts a cascade that leads to error prone repair and incorporation of additional CAG units into the repeat. In vitro data shows MSH3 binds IDLs through a DNA binding pocket formed by MSH3 residues Y245/K246. In this study, we investigated the significance of this DNA binding motif in CAG repeat expansion using cell lines harboring long, unstable HTT CAG repeats. Genetic disruption of the MSH3 Y245/K246 motif significantly reduced DNA interaction, exhibited MMR deficiency in a frameshift mutator assay and abrogated repeat expansion in a U2OS cell line expressing mutant HTT exon 1. Pharmacological blockade of this site using a small molecule targeting the DNA binding pocket similarly reduced DNA binding and repeat expansion in a U2OS cell line. Crucially, this molecule also slowed CAG repeat expansion in medium spiny neurons derived from HD patient-iPSCs. Targeting of the MSH3 IDL binding pocket may represent a possible therapeutic strategy.

Mitotic chromosome dimensions differ between species, and they differ between developmental stages within an organism. The physiological determinants of chromosome size remain poorly understood. Here, we investigate chromosome size determinants in the fission yeast Schizosaccharomyces pombe. Super-resolution microscopy and semi-automated measurements reveal that cell and nuclear volume in interphase, and the time spent in mitosis (both previously proposed chromosome size determinants), have little influence on resultant chromosome dimensions. Instead, levels of the chromosomal condensin complex affect chromosome size, with increasing condensin levels resulting in more compact (thinner and shorter) chromosomes. Our observations inform the understanding of how chromosome dimensions are controlled in an organism. They suggest that a chromosome-intrinsic mechanism sets chromosome size, more so than the environment in which chromosomes find themselves in.

<h4>Background</h4>Drought is one of the most severe and prevalent abiotic stresses affecting crop production. Foxtail millet (Setaria italica), an important cereal crop in the Poaceae family, is considered an ideal crop for future sustainable agriculture due to its remarkable drought tolerance, water-use efficiency (WUE), and strong adaptability to poor soils. The booting stage is widely acknowledged as a pivotal developmental phase in foxtail millet, and this consensus has been well established in existing research. However, the molecular mechanisms underlying the drought response of foxtail millet during this critical stage remain poorly elucidated. In the present study, we employed an integrated approach combining transcriptomic and metabolomic sequencing to identify the core regulatory mechanisms and key genes governing the drought stress response in foxtail millet at the booting stage.<h4>Results</h4>In this study, phenotypic variations, gene expression profiles, and metabolite accumulation patterns in foxtail millet were determined following drought treatments at different growth stages. Among these stages, drought stress imposed at the booting stage resulted in the most severe yield reduction of 34.94%, accompanied by the identification of 566 differentially expressed genes (DEGs) - the highest number across all tested stages. These findings indicated that the booting stage was the most vulnerable period to drought stress during foxtail millet growth. Furthermore, analyses of photosynthetic parameters and yield-related traits revealed that drought stress at the booting stage significantly inhibited photosynthetic capacity, which may represent a critical factor contributing to the yield decline observed in foxtail millet subjected to drought stress during the booting stage. Multi-omics integration analysis identified three core responsive pathways: phenylpropanoid biosynthesis, photosynthetic carbon fixation, and glyoxylate and dicarboxylate metabolism. Based on these findings, we constructed a drought resistance regulatory network model for the booting stage, revealing significant expression changes GAPA, ALDO, tktA, glpx, PRK, RPE, rbcS, maeB, GLUL, and gcvT. At the metabolic level, we observed significant accumulation of metabolites such as sinapyl alcohol, aconitic acid, citric acid, isocitric acid, and mesaconic acid, while the contents of methyl isoeugenol, eugenol, p-coumaric acid, chlorogenic acid, sedoheptulose, aspartic acid, and malic acid were markedly reduced. Most notably, this study revealed that downregulation of gcvT expression may restrict ammonia (NH₃) supply, thereby activating the methanesulfonate synthesis pathway as an alternative nitrogen (N) source. This discovery provided novel insights into N metabolic remodeling under drought stress in plants.<h4>Conclusions</h4>This study laid the foundation for preliminary exploration of the molecular mechanisms underlying drought stress response during the booting stage in foxtail millet. Furthermore, these findings had identified valuable key candidate genes that may contribute to the breeding of high-yield and drought-resistant foxtail millet varieties.

BACKGROUND: Lung cancer is the predominant contributor to cancer-induced mortality, with non-small cell lung cancer (NSCLC) constituting the majority. However, the intricacies of tumorigenesis and progression in this context remain incompletely understood. METHODS: Utilizing single-cell RNA sequencing data retrieved from the GEO database, we performed high-dimensional weighted gene co-expression network analysis to identify pivotal gene modules exhibiting the highest correlation with the clinical stages of NSCLC patients. Subsequently, we formulated a novel prognostic three-gene signature, subjecting it to thorough analysis using bulk RNA sequencing data obtained from the TCGA-LUAD dataset. RESULTS: A tumor-intrinsic prognostic signature, comprising SEC61G, NPTN, and ALDOA, emerged from our investigation. The risk score derived from this gene signature revealed significantly poorer overall survival among patients in the high-risk group. Patients in different risk groups exhibited distinct immune statuses, with those in the low-risk group likely to benefit more from immunotherapy. Furthermore, in vitro experiments demonstrated that SEC61G facilitates tumor proliferation and migration with the activation of the WNT/β-Catenin signaling pathway. CONCLUSION: Our study unveiled a novel tumor-intrinsic gene signature, shedding light on improved prognostication for NSCLC and facilitating risk-stratified management.

Huntington's disease (HD) is an inherited neurodegenerative disease characterised by progressive degeneration of GABAergic medium spiny neurons (MSNs) in the striatum. Neural precursor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) have been considered as a promising and scalable source for neuronal replacement and circuit restoration. In this study, we investigated the therapeutic effects of a clinical-grade, human leukocyte antigen (HLA)-homozygous iPSC line (YZWJ-s513) differentiated into NPCs (s513-NPCs) in a quinolinic acid (QA)-lesioned rat model of HD. Following intrastriatal transplantation, s513-NPCs not only survived for 12 weeks but also differentiated into neurons, astrocytes, and oligodendrocytes, while generating new DARPP32⁺ GABAergic MSNs. Specifically, graft-derived neurons projected to the host globus pallidus, indicating structural integration into the striato-pallidal pathways. Additionally, NPC-transplanted rats exhibited significant motor recovery across multiple tasks for up to 12 weeks, accompanied by reduced striatal atrophy and ventricular enlargement. Histological findings also revealed attenuated astrogliosis and microgliosis, along with a shift toward an anti-inflammatory milieu. Collectively, these results demonstrate that transplantation of clinical-grade, HLA-homozygous iPSC-derived NPCs can provide both neuronal replacement and modulation of the diseased microenvironment, supporting their potential as a regenerative therapy for HD. Key quality attributes and release criteria supporting the clinical-grade characterisation of the cell product used in vivo are summarised in Table S1.

Mitochondria are indispensable organelles that not only generate cellular energy through oxidative phosphorylation but also regulate calcium homeostasis, redox balance, and apoptotic signaling. Given the high metabolic demands of neurons, mitochondrial function and resilience mechanisms are essential for neuronal development, maturation, and survival; when these systems fail, pathological outcomes can arise. This review highlights the critical role of mitochondria in maintaining neuronal function, with discussion related to both the central (CNS) and enteric (ENS) nervous systems. We present how mitochondrial dysfunction, through impaired bioenergetics, oxidative stress, defective quality control, and altered dynamics, can drive neuronal cell loss. Furthermore, we highlight the link between mitochondrial defects and nervous system pathological outcomes in both primary mitochondrial disorders, such as mitochondrial neurogastrointestinal encephalomyopathy, and secondary mitochondrial disorders, such as Alzheimer, Parkinson, and Huntington disease, as well as amyotrophic lateral sclerosis. By integrating evidence from the CNS and ENS, this review highlights the central role of mitochondria in supporting and preserving neuronal health, as well as the potential of mitochondria as therapeutic targets in neurodegenerative disease.

No abstract available.

Adipocyte differentiation involves a metabolic transition from oxidative phosphorylation (OXPHOS) to aerobic glycolysis, allowing preadipocytes to meet the biosynthetic and energetic demands of maturation. Here, we show that nuclear myosin 1 (NM1), a chromatin-associated actomyosin motor, known to control transcription and chromatin accessibility, is required for metabolic homeostasis during adipocyte differentiation. Integrated ATAC-seq and RNA-seq profiling of NM1-deficient mouse embryonic fibroblasts (MEFs) revealed coordinated downregulation of key adipogenic and lipid-droplet machinery genes like Cebpa, Plin2, Abhd5, Agpat2, Pink1, and altered enhancer accessibility near adipogenesis-linked transcription factors (TFs) such as Klf6, Foxo3, Smad5, and Gata4. NM1 knockout (KO) mesenchymal stem cells (MSCs) exhibited impaired differentiation potential despite enlarged adipocyte morphology. In vivo, NM1-deficient mice developed progressive visceral obesity, accompanied by transcriptional reprogramming in epididymal white adipose tissue (eWAT), including repression of mitochondrial and adipogenic pathways and activation of inflammatory networks driven by IFNG, IL33, and TNF. Cross-species network analysis highlighted conserved regulatory architecture centered on MYO1C, implicating NM1/MYO1C as key chromatin-level regulators of adipose remodeling.

The self-renewal capacity of intestinal stem cells (ISCs) declines with aging, leading to a loss of homeostasis and an increased susceptibility to intestinal diseases. Despite the established significance of lipid metabolism and epigenetic regulation in ISC function, the molecular mechanisms that connect these processes to aging-related ISC dysfunction remain elusive. Here, we hypothesize that histone 3 lysine 36 trimethylation (H3K36me3) might act as a bridge between these processes. In this study, we demonstrate that H3K36me3 caused by SETD2 is critical for ISC stemness. H3K36me3 deficiency results in reduced ISC proliferation and differentiation, disrupts fatty acid oxidation (FAO), and promotes ISC senescence. Mechanistically, the loss of H3K36me3 triggers the activity of the SWI/SNF chromatin remodeling complex and leads to increased chromatin accessibility and enhancer activation, which alters FAO- and senescence-related gene expression. Importantly, our data demonstrate that metabolic intervention can prevent the senescence of ISC due to H3K36me3 deficiency. Our findings reveal a crucial role for H3K36me3 in maintaining the epigenetic landscape that orchestrates FAO and determines intestinal stem cell functions, emphasizing the role of FAO as a key modulator between H3K36me3 and ISC aging, suggesting that metabolic intervention may help mitigate age-related ISC dysfunction.

BACKGROUND: Dengue may affect the heart, manifesting as asymptomatic electrocardiographic (ECG) abnormalities to fulminant myocarditis. The role of routine ECG monitoring in dengue management remains unclear. We, therefore, studied ECG changes in patients with non-severe dengue. METHODS: In this observational study, we performed and interpreted 12-lead ECGs (admission, discharge and follow up) on hospitalised Vietnamese adults with confirmed dengue and measured the PR, QRS, and Fridericia-corrected QT intervals (QTcF). RESULTS: A total of 330 ECGs were performed in 136 patients (64 females; median age, 24 years; range, 16–72 years). Of these, 132 patients had WHO-defined non-severe dengue and 4 had severe dengue. Elevated CK-MB fractions (> 25 IU/L) were detected in 64 of 134 patients (47.8%). The most common ECG abnormality was sinus bradycardia (< 60 beats/min), with 54 episodes occurring in 39 patients (28.7%), most frequently on illness day 10, 9/34 (26.5%) patients. The mean QTcF increased during the first 10 illness days in parallel with a decrease in mean heart rate; only 3 females (461–501 ms) and 1 male (460 ms) had sex-specific QTcF prolongation (≥ 460 ms female, ≥ 450 ms male). Female sex and illness day were significant predictors of QTcF variability over time. Pathological T-wave inversion in the inferior (II, III, aVF) and lateral chest (V5, V6) leads were observed in 12 patients (8.8%), and were mostly transient. CONCLUSIONS: ECG changes associated with nonsevere dengue in predominantly young Vietnamese adults without comorbidities were mostly mild, non-specific, and transient with limited impact on the QTcF interval. Routine ECGs in non-severe dengue in our setting did not provide clinically actionable information.

BACKGROUND: The interaction between tumour-associated neutrophils (TANs) and angiogenesis plays a crucial role in tumour progression. However, the specific regulatory mechanisms by which different TANs populations influence angiogenesis in colorectal cancer (CRC) remain poorly understood. METHODS: The study integrates our own dataset with publicly available single-cell RNA sequencing (scRNA-seq) data to analyze the infiltration of various single-cell types in colorectal cancer (CRC). Using in vitro experiments, immunofluorescence, and animal models treated with anti-VEGFR antibodies, we investigated the role of tumor-associated neutrophils (TANs) in CRC and angiogenesis. We conducted RAB31 knockdown cell line, RNA sequencing, Western blotting, quantitative PCR, ELISA, tube formation assays, CCK8, and clonogenic assays, combined with in vivo experiments, to elucidate the impact of RAB31 in the tumor microenvironment (TME) and the mechanism by which tumor-derived RAB1 regulates TANs recruitment. RESULTS: Single-cell analysis revealed a significant infiltration of myeloid cells, particularly TANs, in CRC tumour tissue compared to normal tissue. RAB31 was found to be involved in malignant cell pathways, promoting CRC progression. Depletion of TANs substantially suppressed tumour growth and angiogenesis, while VEGFR blockade inhibited tumour growth and altered TANs infiltration in response to angiogenesis status. In vitro, knockdown of RAB31 expression inhibited tumour cell proliferation, and CXCL2 protein secretion was significantly reduced in shRab31-transfected tumour cells. In vivo, tumour tissue from shRab31 models showed decreased angiogenesis and reduced Neutrophil Extracellular Traps (NET) production, alongside reduced TANs infiltration. Moreover, CXCR4 expression was significantly elevated in TANs, and tumour cells influenced TANs through the CXCL2–CXCR4 axis. RAB31 regulated the PI3K–Akt signalling pathway, which is linked to neutrophil recruitment in CRC. CONCLUSIONS: Our findings suggest that tumour-derived RAB31 regulates the CXCL2–CXCR4 signalling pathway in CRC. RAB31 promotes CXCL2 secretion through the PI3K-Akt signalling pathway, facilitating TANs recruitment and angiogenesis, thereby driving tumour growth. This work enhances our understanding of the complex role of TANs and RAB31 in CRC progression and provides new insights into potential therapeutic strategies targeting the tumour microenvironment.

<h4>Background & aims</h4>Helicobacter pylori (H pylori) eradication significantly reduces gastric cancer risk but offers limited benefit for patients with advanced atrophic gastritis. After eradication, a persistent inflammatory-cancer-transformation microenvironment (ICTM) drives gastric carcinogenesis, warranting further investigation.<h4>Methods</h4>We constructed a single-cell atlas of 22 gastric antral mucosae across disease stages, including normal controls, H pylori-infected non-atrophic gastritis, H pylori-infected chronic atrophic gastritis, and post-eradication chronic atrophic gastritis (stratified by pathological improvement). Key findings were validated via multiplex immunofluorescence and primary cancer-associated fibroblast (CAF) experiments.<h4>Results</h4>H pylori infection profoundly alters the ICTM within immune and stromal cells during and after eradication. Following eradication, epithelial cells exhibit distinct trajectories after eradication, with the pathological improvement group showing increased MUC5AC expression and the non-improvement group displaying a malignant trajectory. Immunomodulatory CAFs (iCAFs) critically promote epithelial malignancy and immunosuppression. CCN2⁺ iCAF1 and C3⁺ iCAF3 are enriched in infected tissues and persist in post-eradication patients with CAG. H pylori infection reprograms iCAFs, leading to extracellular matrix reorganization and senescence with senescence-associated secretory phenotype. This reprogramming promotes epithelial malignant transformation and stemness. Infection-driven reprogramming of the iCAF-epithelial niche is accompanied by an immunosuppressive state after eradication.<h4>Conclusions</h4>H pylori infection induces persistent ICTM after eradication. Infection-driven iCAF differentiation contributes to epithelial malignant transformation and a potential immunosuppressive microenvironment, linking to gastric carcinogenesis. Our findings underscore the critical role of ICTM transformation, highlighting the need to improve ICTM for post-H pylori eradication therapies, and indicate that specific iCAF subtypes represent promising intervention targets.

<h4>Background</h4>Due to the overlapping clinical features of neurodegenerative dementia (NDD)-including Alzheimer's disease (AD), frontotemporal dementia (FTD), dementia with Lewy bodies (DLB), and progressive supranuclear palsy (PSP), accurate diagnosis remains challenging in early stages. Multiple dementias can be caused by short tandem repeat (STR) expansions. However, systematic investigation of known pathogenic STRs in large dementia cohorts remains lacking.<h4>Methods</h4>We used ExpansionHunter (EH) to assess 22 neurodegenerative disease-associated STRs in whole-genome sequencing (WGS) data from 950 patients with AD, 222 patients with FTD, 165 patients with DLB, 231 patients with PSP, and 1522 cognitively normal controls. Repeat primed-polymerase chain reaction (RP-PCR) was performed to validate EH calls that exceeded the intermediate thresholds of STR. We also attempted to use ExpansionHunter Denovo (EHDn) to detect C9orf72 expansions missed by EH.<h4>Findings</h4>EHDn improved the detection rate of C9orf72 expansions. After sample quality control, 33 PCR-validated pathogenic expansions were identified in nine genes (C9orf72, ATXN8OS, NOTCH2NLC, HTT, FMR1, DMPK, AR, CACNA1A, and PPP2R2B) among 1559 patients with NDD, accounting for 2.12% of cases. Burden-based logistic regression analyses demonstrated that the presence of pathogenic STR expansions was significantly associated with NDD status (OR = 3.57, p = 4.70 × 10^-2). Additionally, intermediate-length TBP alleles showed a nominal enrichment in PSP compared with controls (2.61% vs 0.54%; OR = 6.25, p = 2.00 × 10^-2).<h4>Interpretation</h4>Our findings provide evidence for the clinical pleiotropy of STRs in NDD and their involvement in NDD pathogenesis.<h4>Funding</h4>This study was supported by the National Natural Science Foundation of China(U22A20300, 82502244, 823714434, 82071216), the National Key R&D Program of China(2023YFC3603700), STI2030-Major Projects(2021ZD0201803), Outstanding Youth Fund of Hunan Provincial Natural Science Foundation(2024JJ2097), Youth Fund of Hunan Provincial Natural Science Foundation(2025JJ60696), Hunan Health Commission Grant(20232460), Postdoctoral Fellowship Program of CPSF(GZC20233185), China Postdoctoral Science Foundation (2025M772318), and the Scientific Research Program of FuRong Laboratory (2024PT5108).

Reprogramming of spinal astrocytes into motor neurons holds great promise for spinal cord injury in regeneration medicine. Here we identified a set of four transcription factors-achaete-scute complex homolog-like 1, myelin transcription factor 1 like, POU class 3 homeobox 2, and ISL LIM homeobox 1-collectively referred to as the 4 F cocktail. The 4 F cocktail reprograms rat and human reactive astrocytes into motoneuron-like cells. The reprogrammed cells display neuronal morphology and stain positive for microtubule-associated protein 2 (MAP2), a neuronal-specific marker, and choline acetyltransferase, which is a known motor neuron-specific marker. Early in the process of astrocytic reprogramming mediated by the 4 F cocktail, quantitative real-time reverse transcription polymerase chain reaction revealed that the expression of the astrocytic gene glial fibrillary acidic protein was inhibited. Additionally, the expression of the neural progenitor cell markers SRY-box transcription factor 2 and neural cell adhesion molecule 1 was upregulated within 5 days, while the motor neuron progenitor marker oligodendrocyte transcription factor 2 was activated within 7 days. Furthermore, the expression of the neural genes Map2, synapsin I, tubulin β-3, and neurofilament heavy, as well as the motor neural genes ISL LIM homeobox 1, motor neuron and pancreas homeobox 1, LIM homeobox 1, LIM homeobox 3, and NK6 homeobox 1, and survival of motor neuron 1 increased in the reprogrammed cells after induction with the 4 F cocktail. More importantly, 4 F cocktail-reprogrammed motoneuron-like cells can release acetylcholine and exhibited vulnerability to glutamate-induced excitotoxicity. Our findings demonstrate that the 4 F cocktail first reprograms spinal astrocytes into a neural progenitor-like and motor neuron progenitor-like intermediate plastic state, which then gives rise to motoneuron-like cells.

<h4>Aims</h4>This study aimed to assess the nutritional status of COVID-19 patients within the first 24 h of Intensive Care Unit (ICU) admission and to determine its association with the risk of mechanical ventilation and mortality.<h4>Methods</h4>This retrospective study analyzed electronic medical records from 342 COVID-19 patients admitted to Hanoi Medical University Hospital, Vietnam, between September 2021 and April 2022. The Global Leadership Initiative on Malnutrition (GLIM) criteria were applied to assess the risk of malnutrition.<h4>Results</h4>According to the GLIM criteria, 77.8% of patients had moderate or severe malnutrition upon ICU admission. Compared with patients without malnutrition, those who were malnourished were significantly older (<i>p</i> < 0.001) and had lower BMI, hemoglobin, and red blood cell counts (<i>p</i> < 0.05). Univariate logistic regression analysis identified several factors associated with respiratory therapy and mortality among ICU COVID-19 patients, including age, vaccination status, nutritional status, and BMI. In multivariate logistic regression analysis, malnutrition remained a significant predictor of clinical outcomes. The risk of requiring mechanical ventilation and death were 3.55-fold and 2.03-fold higher, respectively, in malnourished patients.<h4>Conclusion</h4>The study revealed a high prevalence of malnutrition among ICU patients with COVID-19. Furthermore, malnutrition was independently associated with an increased risk of mechanical ventilation and mortality. These findings underscored the importance of early nutritional screening and intervention as integral components of routine management for critically ill COVID-19 patients.

This review examines the design and synthesis of peptide-based supramolecular cages, highlighting the versatility and functional diversity achievable by incorporating small peptides as structural components. Inspired by natural supramolecular architectures, synthetic peptide cages offer unique advantages, including tunable chirality, structural predictability, biocompatibility, and ease of functionalisation. The discussion focuses on two principal strategies. The first involves cages in which peptides constitute the primary structural framework, with cage geometry dictated either by intrinsic backbone conformations or by externally imposed directional interactions such as metal coordination. The second covers hybrid systems in which peptides play a functional rather than framework-determining role and are integrated with rigid aromatic or synthetic scaffolds that define the overall architecture. These approaches enable precise control over cage geometry, cavity characteristics, and dynamic behaviour, facilitating applications in biosensing, targeted drug delivery, molecular separation, and environmental remediation. By bridging principles from natural assembly and synthetic supramolecular chemistry, peptide cages represent a powerful platform for developing next-generation functional materials.

Cold autoimmune hemolytic anemia (cAIHA) is a rare form of anemia characterized by antibody-mediated red blood cell destruction at low temperatures (i.e., below 98.6°F or 37°C). Patients with severe cAIHA exhibit hemoglobin levels under 8 g/dL. Affected patients present with hepatosplenomegaly, hypotension, tachycardia, and an increased risk of death with a 5-year survival rate of 63.5%. Treatment with immunosuppressants and lifestyle modifications can improve cAIHA symptoms. However, the presence of a concurrent disease process can worsen clinical outcomes. In our report, we discuss the clinical management of acute decompensated cirrhosis secondary to heterozygous hemochromatosis complicated by cAIHA. Hemochromatosis is characterized by increased iron deposition that results in organ dysfunction. On admission, the patient presented with anemia, weight loss, and ascites. Although heterozygous hemochromatosis is typically asymptomatic, the presence of cAIHA appears to have contributed to the patient's liver dysfunction. Requiring different treatment regimens, the case underscores the difficulty of managing multiple rare hematological conditions.

The pervasive contamination of water resources by heavy metal ions poses serious threats to environmental safety and human health, creating an urgent demand for simple, sensitive, and selective sensing platforms. Herein, a robust polyurethane foam (PUF)-supported UiO-66-NH₂-calcein composite is developed as a solid-state fluorescent sensor for the selective detection of Cu²⁺ ions in aqueous media. The immobilization of calcein within the porous UiO-66-NH₂ framework and its subsequent integration into a PUF matrix provides a stable, highly accessible, and reusable fluorescence platform. Structural and spectroscopic characterization confirms the successful formation and homogeneous distribution of the composite components. Fluorescence screening against a series of metal ions demonstrates that, although several ions can interact with the sensor, a markedly enhanced and selective fluorescence response toward Cu²⁺ is achieved under mildly acidic conditions (pH 5). The sensor exhibits a linear fluorescence response toward Cu²⁺ over the micromolar concentration range with a low detection limit of 0.011 µM. The composite also exhibits mechanical integrity and operational reusability, with no significant signal loss. The foam-based architecture enables rapid mass transfer, improved light penetration, and convenient handling, making the platform well-suited for portable and on-site water analysis. This work presents an effective strategy for constructing MOF-polymer fluorescent composites for selective Cu²⁺ sensing and practical environmental monitoring. The PUF/UIO-66-NH-calcein nanocomposite exhibits remarkable fluorescence sensitivity toward multiple metal ions at pH 7, achieving ultra-low limits of detection of 0.0013 µM for As³⁺, 0.0049 µM for Cd²⁺, 0.009 µM for Zn²⁺, 0.0049 µM for Cu²⁺, and 0.0081 µM for Ca²⁺, while maintaining stable performance under aqueous conditions.

<h4>Background</h4>Vibrio vulnificus is a halophilic marine bacterium capable of causing rapidly progressive septic shock and necrotizing fasciitis, particularly in immunocompromised individuals. Mortality rates remain high due to the fulminant nature of the infection and diagnostic challenges.<h4>Case presentation</h4>A 70-year-old female with poorly controlled type 2 diabetes presented with fever and septic shock 48 hours after ingesting raw seafood. The clinical course was characterized by rapid deterioration, severe coagulopathy, and the development of extensive necrotizing fasciitis in the right upper limb. Vibrio vulnificus infection was confirmed via next-generation sequencing (NGS) on hospital day 6. Despite broad-spectrum antibiotic therapy and fluid resuscitation, surgical intervention was delayed until day 9 following multidisciplinary consultation. Intraoperative findings revealed extensive "dishwater" necrosis. The patient suffered from recurrent sepsis, graft failure, and deep vein thrombosis. Due to the rapid progression and delayed source control, the outcome was unfavorable, leading to discharge against medical advice.<h4>Conclusion</h4>This case underscores the high mortality risk associated with Vibrio vulnificus infection in immunocompromised individuals, particularly those with diabetes mellitus. Observations from this patient highlight that antibiotic therapy alone is often insufficient for necrotizing soft tissue infections. It is emphasized that in these high-risk populations, early recognition must trigger immediate, aggressive surgical debridement alongside antimicrobial therapy. Delays in surgical source control, even while awaiting molecular confirmation, can irreversibly compromise patient survival.

Whitlockite (WH), a natural magnesium-containing calcium phosphate second in abundance only to hydroxyapatite (HAp) in the inorganic phase of bone tissue, has attracted great attention in bone repair recently. Previous studies have shown that porous structures play a pivotal role in bone defect repair using biomaterials. However, the traditional fabrication process of porous ceramic through high-temperature sintering frequently results in the dehydration and breakdown of WH. To overcome this challenge, we develop an innovative fabrication of porous whitlockite ceramic <i>via in-situ</i> phase transformation from calcium phosphate ceramic. Furthermore, we perform molecular dynamics (MD) and determine intermediates to delve into a comprehensive exploration of the underpinning mechanism of the transformation. <i>In-vitro</i> cellular responses reveal the superior cell proliferation, adhesion, spreading, migration, and osteogenic and angiogenic differentiation of porous WH ceramic. Mechanistically, our results reveal that the Mg²⁺-mediated regulation of ATP levels activates the AMPK-FoxO signaling axis, which enhances osteogenesis through autophagy. Intramuscular implantation evaluation confirms the satisfactory osteoinduction, and further rat mandibular defect implantation reveals significant bone regeneration in comparison to calcium phosphate ceramic. Together, the porous WH ceramic encompasses enhanced potential for the regenerative amelioration of bone defects.

Innate-like T cells (ILTCs) link innate immune responses with adaptive immune functions. This group includes invariant natural killer T (iNKT) cells, mucosa-associated invariant T (MAIT) cells, and γδ T cells. ILTCs detect transformed or stressed cells via non-classical antigen presentation pathways. For example, iNKT cells recognize CD1d-presented glycolipids, MAIT cells respond to MR1-presented metabolites from riboflavin pathways, and γδ T cells sense phosphoantigens through butyrophilin-dependent mechanisms and stress ligands. These features support early tumor control and shape downstream immunity by promoting dendritic cell activation, NK cell function, and priming of tumor-reactive CD8⁺ T cells. In established tumors, ILTC activity is frequently suppressed. Reduced antigen presentation, inhibitory cytokines, hypoxia, and metabolic constraints, including lactate accumulation and kynurenine production, limit effector responses and promote hyporesponsive states. Transcriptional regulators such as TOX, NR4A family members, and BATF are associated with these programs. This review discusses ILTC roles in tumor surveillance, immune escape, and therapeutic strategies to restore their function.

Mitochondrial dysfunction lies at the core of numerous cardiac pathologies, yet restoring mitochondrial health remains a therapeutic frontier. In recent years, extracellular vesicles (EVs) have emerged as nature's delivery nanocarriers, capable of transporting a wide array of biomolecules, including mitochondrial-associated microRNAs (mito-miRs). These miRNAs regulate bioenergetics, redox homeostasis, and apoptotic signaling-making them prime candidates for non-cellular mitochondrial therapy. This review explores the evolving landscape of mitochondrial miRNA encapsulation within EVs, focusing on their potential to restore mitochondrial transcriptional and metabolic programs governing ATP synthesis and redox balance, enhance cellular energy output, and mitigate oxidative stress. We integrate insights from stem cell biology, RNA epigenetics, systems cardiology, and bioengineering, offering a unifying framework for therapeutic applications across ischemic heart disease, heart failure, and chemotherapy-induced cardiomyopathy. An integrative narrative synthesis of recent peer-reviewed literature was performed across major biomedical databases, prioritizing mechanistic studies linking EV-mediated mito-miR delivery to cardiomyocyte mitochondrial function. By harmonizing multi-omic signaling, vesicle engineering, and mitochondrial medicine, this review seeks to guide future research toward targeted, customizable, and scalable bioenergetic interventions-unlocking a next-generation path for cardiovascular regeneration.

The <i>KMT2A</i> gene, located at chromosome band 11q23, encodes a lysine methyltransferase essential for hematopoietic gene regulation. While <i>KMT2A</i> rearrangements are common in acute myeloid leukemia (AML) and B-cell acute lymphoblastic leukemia (B-ALL), <i>KMT2A</i> amplification is rare, occurring in ~1% of AML cases and even less frequently in B-ALL. Given its rarity, understanding <i>KMT2A</i> amplification in B-ALL is crucial for improving diagnostics and therapy. We report two adult B-ALL cases with <i>KMT2A</i> amplification. Patient 1, a 58-year-old male, had <i>KMT2A</i> amplification (6~18 copies in 68.5% of bone marrow cells), a complex karyotype, and a pathogenic <i>TP53</i> variant (c.524G>A, p.Arg175His). He underwent induction chemotherapy but passed away after two months due to complications. Patient 2, a 66-year-old female, had <i>KMT2A</i> amplification (8~11 copies in 87.5% of peripheral blood cells) and <i>CRLF2</i> rearrangement, representing the first reported case of <i>de novo</i> Ph-like B-ALL with <i>KMT2A</i> amplification in an adult. She deteriorated rapidly and died within four days. In addition to these two cases from our cohort, we review nine published cases with <i>KMT2A</i> amplification in B-ALL, which showed frequent <i>TP53</i> alterations, emphasizing the clinical and genetic characteristics of this aggressive leukemia subtype. These cases highlight the high-risk nature of <i>KMT2A</i>-amplified B-ALL, particularly in older adults, where prognosis is poor and linked to <i>TP53</i> variants or <i>CRLF2</i> rearrangement. Our review underscores the need for genetic profiling to improve risk stratification and treatment. Given the limited documented cases, further research is essential to develop better therapeutic strategies for <i>KMT2A</i>-amplified B-ALL.

<h4>Background</h4>Mastocytosis is a rare clonal hematological neoplasm, characterized by cutaneous manifestations in children and categorized as: maculopapular cutaneous mastocytosis (MPCM), diffuse cutaneous mastocytosis (DCM) and mastocytoma. Systemic mastocytosis (SM) typically occurs in adults with c-<i>KIT</i> D816V mutation. Additional genetic mutations (<i>TET2</i>, <i>NRAS</i>, <i>SF3B1</i>, <i>ASXL1</i>, etc.) have been detected using Next-Generation Sequencing (NGS) in the adult population while limited information is available in the pediatric setting.<h4>Methods</h4>36 patients (pts) with pediatric mastocytosis diagnosed between 1997 and 2021 were included. Peripheral blood samples were collected to detect c-<i>KIT</i> D816V mutation, using both RT-PCR and ddPCR techniques, and to investigate other molecular mutations using NGS panel for rare and myeloid genes.<h4>Results</h4>Median age of lesion onset was 4.7 months (range birth-17.8 years). 58% of the cohort underwent cutaneous biopsy after a median 3.77 months from lesion onset (range 2.49 months-11.6 years). 20 (55%) were classified as MPCM, 10 (28%) as DCM and 6 (17%) as mastocytoma. Median tryptase value at the onset was 5 ng/mL: MPCM (range 1.2-141 ng/mL) vs. DCM (range 2.71-19.4 ng/mL) vs. mastocytoma (range 3.8-7.3 ng/mL). Two MPCM pts developed indolent SM (ISM) after 10 and 20 years from the onset of disease. RT-PCR identified c-<i>KIT</i> D816V mutation in 4 pts (2 MPCM, 1 DCM, 1 ISM). NGS revealed the precedent mutation in 3 pts, c-<i>KIT</i> D816Y and c-<i>KIT</i> Y553C in 2 pts. An additional 10 myeloid gene mutations were detected by NGS: 5 already known (<i>ASXL1</i> G1397S; <i>JAK2</i> L393V; c-<i>KIT</i> D816Y; <i>LNK</i> E208Q; <i>TET2</i> Y867H) and 5 not previously described (<i>ETV6</i> A215P; c-<i>KIT</i> Y553C; <i>NFE2</i> I291T; <i>SH2B3</i> G382D; <i>SH2B3</i> L438V). A single mutation was found in 7 pts (3 MPCM, 3 DCM, 1 ISM), while two or more mutations in 3 DCM pts. Overall, 9/36 pts (5 DCM, 3 MPCM, 1 mastocytoma) presented spontaneous complete regression of cutaneous lesions after a median time of 25 months (range 17 months-25 years).<h4>Conclusion</h4>c-<i>KIT</i> mutations resulted in 35% of the children tested. The RT-PCR technique resulted more sensitive in finding c-<i>KIT</i> D816V, while NGS in detecting other mutations whose prognostic roles require further investigation.

<h4>Background</h4>Although immune dysregulation is implicated in both autoimmune diseases and cancer, comparative pathogenesis and immune response mechanisms between systemic lupus erythematosus (SLE) and colorectal cancer (CRC) remain elusive. This study identifies common molecular biomarkers and pathogenic pathways shared between SLE and CRC via multi-omics analysis.<h4>Methods</h4>Integrated datasets including SLE (GSE61635, GSE50772, GSE142016) and CRC cohorts (GSE39582, GSE17536, E-MTAB-8107, COAD, READ), were analyzed. Differential expression analysis identified shared genes, and machine learning screened four hub prognostic genes. A risk model based on these genes was constructed to evaluate SLE screening and CRC prognosis. Multi-omics approaches explored mutational profiles, immune infiltration, drug sensitivity, and signaling pathways. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), staining were performed to confirmed the expression levels of the hub genes. The impact of DNASE1L3 knockdown in the human monocyte cell line THP-1 on phagocytosis of apoptotic intestinal epithelial cell line NCM460 was evaluated using RT-qPCR and flow cytometry.<h4>Results</h4>We identified 58 shared differentially expressed genes (DEGs) between SLE and CRC, enriched in apoptosis, oxidative phosphorylation, and immune infiltration. Machine learning highlighted four hub genes (<i>DNASE1L3</i>, <i>PTPN14</i>, <i>SELENBP1</i>, <i>ECRG4</i>), forming a robust predictive model for SLE occurrence and CRC prognosis. Shared immune infiltration patterns and small-molecule drug candidates were observed. Single-cell and spatial transcriptomic analyses revealed DNASE1L3 predominantly in myeloid cells. Cellular experiments revealed that reduced DNASE1L3 levels significantly compromised macrophage efferocytosis of apoptotic cells, accompanied by a decrease in M2 macrophage proportion, an increase in M1 macrophage proportion, diminished LOX enzyme activity, and elevated levels of interleukin-1 β and tumor necrosis factor-α.<h4>Conclusion</h4>SLE and CRC exhibit overlapping DEGs, immune profiles, and signaling pathways. The model based on the shared genes-<i>DNASE1L3</i>, <i>PTPN14</i>, <i>SELENBP1</i>, and <i>ECRG4</i>-offers novel insights for precise intervention in both diseases.

Comb and skin pigmentation are key external traits in domestic chickens, closely correlated and serving as important visual cues for consumers. The Anyi Gray chicken, an indigenous breed from Jiangxi Province, China, exhibits two stable phenotypes: red comb with white skin (RAY), and purple-gray comb with purple-gray skin (PAY). Although skin color is completely linked to comb color, the genetic basis of this coordination remains unclear. We performed whole-genome resequencing (∼ 10 × depth) on RAY and PAY individuals, followed by a case-control genome-wide association study (GWAS). A highly significant variant, g.10818413 T > C, located in the promoter region of EDN3, was identified (P < 4.065343e-09). Transcription factor binding site prediction revealed that this mutation disrupts an NFAT5 binding site. Genotyping of 97 Anyi Gray chickens showed complete concordance between genotype and phenotype: all PAY individuals carried the TC genotype, while all RAY individuals had the TT genotype; no CC genotypes were detected. Validation in an F1 hybrid population (n = 61) and in 140 individuals from other Jiangxi native breeds confirmed the absence of the CC genotype. Functional assays demonstrated that NFAT5 negatively regulates EDN3 transcription. Dual-luciferase reporter assays confirmed NFAT5 binding to the EDN3 promoter and its suppressive effect. Tissue expression analysis showed significantly higher EDN3 expression in the comb, skin, and chest muscle of PAY individuals compared to RAY individuals, reinforcing the phenotypic linkage. These findings indicate that the EDN3 g.10818413 T > C mutation reduces NFAT5 binding affinity, cis-regulating EDN3 transcription and driving the coordinated pigmentation of the comb and skin. This study reveals a shared regulatory mechanism underlying the phenotypic correlation between comb and skin pigmentation in Anyi Gray chickens.

Visceral adipose tissue (VAT)-derived extracellular vesicles (EVs) have emerged as key mediators of inter-organ communication, yet their role in nicotine-induced atherosclerosis remains poorly defined. Here, we demonstrate that nicotine markedly enhances secretion of VAT-EVs and that these EVs are preferentially internalized by macrophages within atherosclerotic plaques, thereby accelerating lesion progression. Functionally, nicotine-induced VAT-EVs promote macrophage inflammation, oxidative stress, and foam cell formation. High-throughput profiling identified miR-210-3p as a dominant pro-atherogenic cargo within VAT-EVs, and its inhibition significantly attenuated nicotine-induced atherosclerosis <i>in vivo</i>. Mechanistically, miR-210-3p directly targets Krüppel-like factor 7 (KLF7), amplifying macrophage inflammatory responses and promoting plaque progression. Collectively, these findings uncover a previously unrecognized role of adipose-derived EVs in smoking-related vascular injury and highlight EV-derived miR-210-3p as a promising therapeutic target in nicotine-associated atherosclerosis.

c-Jun, a core component of the activating protein-1 (AP-1) transcription factor complex, regulates cellular processes including proliferation, differentiation, survival, apoptosis, and oncogenesis. c-Jun functions by dimerizing to bind DNA and modulates the expression of genes such as Bcl-2, cyclin D1, and pro-inflammatory cytokines, enabling context-dependent transcriptional control. Its role in neurodegenerative diseases has gained attention due to its regulation of oxidative stress, inflammation, and apoptosis. In Parkinson's disease, Alzheimer's disease, and Huntington's disease, dysregulated c-Jun expression accelerates dopaminergic neuron loss via oxidative damage, contributes to amyloid-β-induced synaptic toxicity, and mediates neuronal apoptosis and inflammation, respectively. Despite its degenerative role, c-Jun also promotes axonal regeneration and stress adaptation, revealing a dual function that depends on context and stimulus severity. This paradox underscores its ability to promote survival under mild stress and apoptosis under chronic damage. Emerging therapeutic strategies targeting c-Jun-via small-molecule inhibitors (e.g., SP600125), RNA interference, or modulation of upstream c-Jun N-terminal kinase (JNK)-are being explored. However, challenges remain in achieving specificity, as c-Jun's ubiquitous expression raises concerns about off-target effects. This review highlights recent advances in understanding c-Jun's complex role in neurodegeneration and its therapeutic potential, emphasizing its value as both a mechanistic regulator and a target for preserving neuronal integrity in neurodegenerative diseases.

<h4>Objective</h4>Cerebral meningiomas are the most common primary intracranial tumors in adults. Treatment of symptomatic tumors is normally surgical; tumors not suitable for surgery can be irradiated. While there is good data on the effectiveness of acute therapy, little is known about the effects of long-term team-based multidisciplinary inpatient neurological rehabilitation. We analyzed the outcome of these patients undergoing neurological rehabilitation.<h4>Methods</h4>We performed a retrospective analysis of patients with cerebral meningioma who underwent specialized rehabilitation. We analyzed routine demographic and clinical data; the outcome was measured with the Barthel Index (BI) in patients with a BI of ≤90 on admission.<h4>Results</h4>We analyzed 151 patients. Neuropsychological deficits were evident in 93 patients, and 9% had speech disorders. BI increased from 66.8 to 75.2%. Examination of factors influencing treatment success revealed that the number of secondary diagnoses had an influence on the average increase in the BI. No correlation was found for the other independent variables, including age, sex, tumor localization, stage, resection (complete or incomplete), complications, and length of stay.<h4>Conclusions</h4>Even given the limitations of our analysis, rehabilitation appears to be effective in these patients. However, further investigations with a matched control group would be desirable to verify our hypothesis. Furthermore, studies regarding optimal intensity, timing, long-term outcome, and modality of rehabilitation are necessary.

Low-intensity pulsed ultrasound (LIPUS) has emerged as a versatile, non-invasive physical modality with growing potential in regenerative medicine and neural repair. Advances in ultrasound physics and biomedical engineering have enabled precise spatiotemporal control of acoustic stimulation, positioning therapeutic ultrasound as an alternative to conventional pharmacological and surgical interventions that often suffer from limited targeting and substantial side effects. Unlike high-intensity focused ultrasound, which relies primarily on thermal ablation, LIPUS operates within a low-energy, non-thermal regime and modulates cellular behavior through mechanical cues, mechano-transduction, and downstream biological responses. Accumulating evidence demonstrates that LIPUS regulates calcium dynamics, cytoskeletal remodeling, neurotrophic factor expression, inflammation, myelination, and local vascular remodeling, thereby promoting functional recovery in both peripheral and central nerve injury models. Moreover, the integration of LIPUS with biomaterials, including piezoelectric scaffolds and acoustically responsive drug delivery systems, has expanded its functionality from direct stimulation to on-demand electrical signaling and controlled therapeutic release. Despite these advances, challenges remain regarding parameter standardization, mechanistic consistency, and clinical translation. In this review, we summarize the systems, parameters, and biological mechanisms underlying LIPUS, discuss its applications in peripheral and central nerve injury repair, and highlight emerging strategies and translational barriers toward intelligent, multimodal, and personalized ultrasound-based therapies.

Loss of ovarian hormones following menopause or ovariectomy is associated with increased anxiety, cognitive impairment, and dysregulation of hypothalamic neuroendocrine pathways. MicroRNAs (miRNAs) and tRNA-derived fragments (tRFs) are emerging classes of small non-coding RNAs that act as post-transcriptional regulators of stress, inflammation, and synaptic function; however, their coordinated involvement in estradiol-mediated hypothalamic regulation remains poorly understood. In this study, adult female mice were assigned to control, estradiol-treated, ovariectomized (OVX), or OVX plus estradiol groups. Anxiety- and cognition-related behaviors were assessed using the open field, Y-maze, and elevated plus maze tests. Circulating estradiol levels and hypothalamic gonadotropin-releasing hormone (GnRH) expression were quantified by ELISA. Hypothalamic mRNA, miRNA, and tRF expression profiles were analyzed by RNA sequencing, followed by differential expression analysis, functional enrichment, integrative network construction, and quantitative real-time PCR validation. Ovariectomy induced anxiety-like behaviors, impaired working memory, reduced estradiol levels, and increased hypothalamic GnRH expression, all of which were reversed by estradiol treatment. Transcriptomic analysis identified 376 differentially expressed miRNAs, 182 differentially expressed tRFs, and 439 differentially expressed mRNAs, enriched in pathways related to stress responses, neuroendocrine regulation, synaptic signaling, metabolic homeostasis, and neuroinflammation. Integrated miRNA-mRNA and tRF-mRNA network analyses revealed several estradiol-responsive miRNAs (including miR-200a-5p, miR-182/183-5p, miR-381-3p, miR-148a-3p, and miR-10 family members) predicting key hub genes such as <i>Gcg</i>, <i>Wnt4</i>, <i>Prkacb</i>, <i>Sgk1</i>, <i>Fpr2</i>, and <i>Aldoa</i>, and key tRFs like tRFdb-1003, tRFdb-1013, tRFdb-1026, tRFdb-3001a and tRFdb-5020a, targeting hub genes such as <i>Wnt4</i>, <i>Prkacb</i>, <i>Sh3rf2</i>, <i>Hpse</i>, <i>Cxcr2</i> and <i>Zbtb16</i> respectively. Collectively, these findings demonstrate that estradiol ameliorates OVX-induced behavioral and endocrine dysfunction by reorganizing hypothalamic miRNA- and tRF-mediated regulatory networks involved in stress adaptation, synaptic homeostasis, and neuroimmune signaling.

<h4>Background</h4>Early diagnosis of familial hypercholesterolemia (FH) is crucial to improve long-term outcomes. FH diagnosis relies on elevated low-density lipoprotein cholesterol (LDL-C) levels, familial clinical characteristics, and identification of pathogenic variants in FH-related genes. Secondary factors, such as overweight and obesity, are known to influence lipid profiles in the general population. More recently, polygenic risk scores based on single-nucleotide polymorphisms (SNPs) have been proposed as additional determinants of LDL-C levels.<h4>Methods</h4>We enrolled 214 pediatric subjects with LDL-C levels ≥95th percentile (after 6 months of dietary intervention) and with at least one parent with LDL-C levels ≥ 95th percentile. All participants underwent biochemical and auxological assessment and genetic testing for FH. In a subgroup of 60 subjects, LDL-C polygenic scores based on 6- and 12-SNPs were calculated.<h4>Results</h4>Pathogenic variants confirming heterozygous FH were identified in 190 subjects (variant-positive, V+); 17 were variant-negative (V-), yielding a mutation detection rate of 91.8%. An additional seven patients carrying variants of uncertain significance were excluded from the primary analysis. LDL-C was modestly higher in V+ than V- subjects using both Friedewald (212 vs. 188 mg/dL; <i>p</i> = 0.035) and Martin-Hopkins formulas (208 vs. 187 mg/dL; <i>p</i> = 0.041), while the other main clinical and laboratory parameters were similar. In V+, LDL-C was higher in subjects with null variants, compared to those with defective variants. Body mass index (BMI SDS) was inversely correlated with HDL-C (<i>p</i> < 0.001), and obesity (BMI z-score > 2 SDS) was associated with lower HDL-C and higher LDL-C, non-HDL-C, and ApoB. With regard to the polygenic scores, 12- and 6-SNP scores showed overlap between V+ and V-, and published cut-offs did not discriminate lipid severity in our population; however, in V+ subjects, the 12-SNP score acted as a phenotype modifier, being independently associated with higher LDL-C and non-HDL-C levels after adjustment for age, sex, and BMI SDS.<h4>Conclusions</h4>In children selected by LDL-C ≥ 95th percentile, together with autosomal dominant familial hypercholesterolemia, genetic confirmation of FH is achieved in the vast majority of cases. Variant type (null vs. defective), BMI, and polygenic background contribute to phenotypic heterogeneity, supporting the need to address other factors alongside genetic diagnosis. Further validation is needed before polygenic scores can be implemented in routine clinical practice.

<b>Background</b>: The peptidyl-prolyl cis-trans isomerase PIN1 regulates multiple oncogenic and tumor-suppressive pathways and is frequently overexpressed in human cancers. Although pharmacological inhibition of PIN1 has shown antitumor potential, existing PIN1-targeting degraders lack systematic structure-activity relationship (SAR) analyses and display inconsistent cellular efficacy, leaving the therapeutic relevance of PIN1 degradation unclear. <b>Methods</b>: Two series of PIN1-targeting PROTACs were designed using the covalent inhibitor sulfopin as the PIN1 binder and ligands for either cereblon (CRBN) or von Hippel-Lindau (VHL). Systematic SAR studies focused on linker structure and jointing atom composition. PIN1 degradation was assessed by Western blotting in multiple cancer cell lines, and further investigated through a series of computational and mechanistic experiments. Antitumor efficacy and safety were evaluated in an MCF-7 xenograft mouse model with preliminary pharmacokinetic analysis. <b>Results</b>: SAR analysis revealed that short, linear linkers and reduced hydrogen bond donor content markedly enhanced PIN1 degradation, whereas VHL-recruiting PROTACs showed inferior cellular activity. These studies identified <b>PC2</b>, a CRBN-recruiting PROTAC, as a lead compound. <b>PC2</b> selectively induced ubiquitin-proteasome-dependent PIN1 degradation with minimal global proteomic or transcriptomic perturbation. Despite modest antiproliferative effects in vitro, <b>PC2</b> significantly suppressed tumor growth in vivo without observable toxicity and achieved effective intratumoral PIN1 degradation. <b>Conclusions</b>: This study defines SAR-guided design principles for PIN1-targeting PROTACs and demonstrates that selective PIN1 degradation can produce robust antitumor activity in vivo. <b>PC2</b> represents the first PIN1 degrader validated in animal models and supports targeted PIN1 degradation as a viable anticancer strategy.

<h4>Aim and background</h4>Cytochrome P450 (CYP450) enzymes are the primary hepatic Phase I oxidative biotransformation system for many drugs and xenobiotics; variability in CYP capacity is therefore a key determinant of metabolic reserve. Reserve varies with developmental ontogeny, genotype, and acquired suppression (e.g., cytokine-mediated phenoconversion). Early infancy represents a developmental window in which clearance capacity and redox/energetic buffering may be comparatively constrained. We introduce a hypothesis-generating Three-Axis Convergence Framework (TCF) modeling interacting effects of (i) developmental/genetic reserve limits, (ii) immune-cytokine modulation of metabolism, and (iii) exposure/disposition context, and translate this synthesis into a structured postmortem interpretive tool integrated into the primary medicolegal autopsy, using an enhanced analytic panel applied selectively based on case context and specimen validity/QC to support mechanistic characterization of infant deaths remaining unexplained (often SUID/SIDS).<h4>Methods</h4>A structured narrative synthesis was conducted spanning developmental pharmacology, pharmacogenetics, immunology, redox biology, neuropathology, and toxicology. Routine medicolegal postmortem practices used in SUID investigations were reviewed to identify measurement gaps that may limit mechanistic resolution in unexplained cases. The synthesis was formalized into five analytic domains: CYP450 capacity, immune/cytokine load, redox balance/energetics, neurochemical integrity, and xenobiotic/metal burden.<h4>Results</h4>The Metabolic Vulnerability Index (MVI) operationalizes the TCF as a five-domain ordinal scoring system (0-15). Domain 1 is anchored by hepatic CYP protein abundance (a more postmortem-stable proxy than CYP activity assays, which are generally constrained by rapid functional decay and QC limitations), normalized to adult reference and interpreted against age-matched developmental expectations. Domain-combination lookup tables route users to 14 mechanistically defined archetypes and specify modifier/exposure-context documentation. Appendices define an operational postmortem workflow, specimen validity rules, analytic QC constraints, detection limits, and a worked example. A Cytokine-Metabolic Suppression Profile (CMSP) is presented as an interpretive coherence summary and does not modify MVI scoring or certification.<h4>Conclusion</h4>The MVI provides a structured framework for describing multidomain physiologic constraints in unexplained infant deaths alongside standard forensic practice. In this way, the TCF, MVI, and CMSP together offer a disciplined response to long-standing mechanistic uncertainty in early life-by enabling systematic measurement, coherent interpretation, and transparent identification of evidence gaps, rather than asserting new causes.

<h4>Background</h4>The clinical relevance of DNA methylation in head and neck cancer (HNC) is increasingly recognized, yet controversies persist regarding its prognostic significance. We conducted a systematic review and meta-analysis adhering to the PRISMA 2020 guidelines to synthesize evidence on this association.<h4>Methods</h4>A comprehensive literature search was performed across five databases (PubMed, Web of Science Core Collection, Cochrane Library, Embase, and Scopus) from inception to October 31, 2025. Eligible studies included original clinical investigations (prospective/retrospective cohort, case-control) reporting prognostic outcomes [overall survival (OS), disease-free survival (DFS)] stratified by DNA methylation status in histologically confirmed HNC patients. Pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated using random-effects models.<h4>Results</h4>Nineteen studies involving 3,251 patients were included. High DNA methylation was significantly associated with poor prognosis in HNC: OS (HR =1.50, 95% CI: 1.19-1.90, P=0.001) and DFS (HR =2.44, 95% CI: 1.74-3.47, P<0.001). Significant heterogeneity was observed for OS (I²=71%, P<0.001) and moderate heterogeneity for DFS (I²=42%, P=0.10), partially attributed to regional differences and variability in detection methods. Confounding adjustment status confirmed the robustness of the results.<h4>Conclusions</h4>DNA methylation plays a pivotal role in HNC carcinogenesis and progression, emerging as a robust and promising prognostic biomarker. However, its predictive value for treatment response remains unproven, requiring further investigation in treatment-stratified cohorts.

<h4>Background</h4>Ovarian cancer (OV) is the leading cause of mortality among gynecological malignancies, often diagnosed at an advanced stage and prone to recurrence after treatment. In order to improve the prognosis, there is an urgent clinical need to identify novel strategies for early intervention and prognosis prediction. Sphingolipids are both important components of cell membranes and closely related to cell signaling. Key enzymes and intermediates of sphingolipid metabolism have critical roles in regulating biological processes such as proliferation and apoptosis of cancer cells, and some of the anticancer drugs targeting sphingolipid metabolism have already entered into clinical trials. However, the prognostic value of sphingolipid metabolism-related genes (SRGs) in OV remains unclear. This study aims to systematically evaluate the prognostic significance of SRGs in OV and construct a prognostic risk model to improve survival prediction.<h4>Methods</h4>In this study, we integrated transcriptomic profiles and corresponding clinical data of OV patients from the Cancer Genome Atlas (TCGA; https://portal.gdc.cancer.gov/) and the Gene Expression Omnibus (GEO; https://www.ncbi.nlm.nih.gov/) databases. Through univariate and multivariate Cox regression analyses, we identified five SRGs to construct a prognostic signature of OV. Based on the signature-derived risk scores, all samples were stratified into high- and low-risk groups. To further evaluate the signature's clinical utility, we comprehensively assessed its associations with immune microenvironment, immunotherapy response and chemotherapy sensitivity. Finally, <i>in vitro</i> experiments were performed to validate the functional role of the key gene <i>CERK</i> in the model.<h4>Results</h4>Patients stratified according to risk scores exhibited statistically significant differences in survival outcomes. The robustness and predictive accuracy of this signature were consistently validated in both internal and external cohorts. Comprehensive analysis of the immune microenvironment and immunotherapy response revealed that patients in the low-risk group were more likely to derive clinical benefit from immunotherapy. Moreover, drug sensitivity analysis indicated that the low-risk group was more responsive to olaparib, whereas the high-risk group showed increased sensitivity to Topotecan. Critically, functional experiments demonstrated that CERK knockout in this model significantly suppressed proliferation, migration, and invasion capacities in SKOV3 and ES-2 cell lines.<h4>Conclusions</h4>We developed a sphingolipid metabolism-related prognostic signature (SRPS), which demonstrated robust performance in predicting survival outcomes and guiding personalized therapeutic strategies.

<h4>Background</h4>Oral lichen planus (OLP) and oral lichenoid lesions (OLL) are potentially malignant disorders that share clinical and histopathological features. Oral epithelial dysplasia (OED) is a key indicator of malignant transformation risk in these disorders. This study aimed to investigate possible associations between the severity of OED and sociodemographic variables, hepatitis C infection, and clinical aspects in patients with OLP and OLL.<h4>Material and methods</h4>The sample comprised 65 biopsies from 56 participants with OLP or OLL, classified according to the criteria of the American Academy of Oral and Maxillofacial Pathology (excluding the absence of OED as a diagnostic criterion for OLP). Data were retrieved from electronic medical records, and photographs taken at the time of biopsy were independently reviewed by two stomatologists and one oral pathologist to classify the clinical patterns. The presence and severity of OED were assessed according to the 2017 World Health Organization system and evaluated independently by two oral pathologists.<h4>Results</h4>A statistically significant association was observed between OED severity and patient age (p=0.004). No significant associations were found with sex, skin color, harmful habits, hepatitis C history, clinical pattern, or anatomical site.<h4>Conclusions</h4>Older patients were more likely to present moderate/severe OED, highlighting the importance of age as a risk factor for progression in OLP and OLL.

<h4>Introduction</h4>Cholangiocarcinoma (CCA) is an aggressive malignancy with poor prognosis and limited treatment options. Molecular heterogeneity and the tumor immune microenvironment play crucial roles in CCA progression and therapeutic response, but comprehensive stratification remains insufficient for guiding treatment decisions.<h4>Methods</h4>This study performed integrated multi-omics profiling as well as immunohistochemical validation on 424 CCA patients. Multidimensional data, including genomic, epigenomic, transcriptomic, and immune profiling, were analyzed to identify clinically relevant molecular subtypes.<h4>Results</h4>Four distinct molecular subtypes of CCA were identified, each exhibiting unique clinical and immunological features: C1 (proliferation): driven by <i>TP53/KRAS</i> mutations and CpG island methylator phenotype (CIMP+) hypermethylation, showing T helper 17 cells infiltration and poor outcomes; C2 (Immune-Suppressed Macro_LYVE1 [ISM_LYVE1]): stroma-rich with LYVE1⁺ macrophages and epithelial-mesenchymal transition (EMT) activation; C3 (immune-activated Macro_C1QC (IAM_C1QC)): enriched in C1QC⁺ macrophages, CD8⁺ T-cells, and metabolic pathways, highly responsive to immune checkpoint blockade (ICB; 75% overall response rate); C4 (immune exclusion): <i>FGFR2</i>-altered and <i>IDH1</i>-mutant, with an immunologically cold phenotype.<h4>Conclusion</h4>We validated ATP2B1 as a novel prognostic biomarker and developed a 160-gene classifier for subtype prediction. The C3 subtype's exceptional ICB response, independent of conventional biomarkers (PD-L1/microsatellite instability/tumor mutational burden), highlights the clinical utility of this classification system in guiding precision immunotherapy for CCA.

<h4>Summary</h4> Nucleosomes with their associated modifications underlie genome organisation and regulation. Replication and transcription require nucleosome disassembly to access the DNA template. How this is orchestrated without jeopardizing chromatin function remains unknown. Here, we reveal a general, global requirement of the histone chaperone FACT in mammalian replication, transcription and chromatin maintenance. Upon acute FACT depletion, replisome and RNA polymerase progression is halted genome-wide and chromatin structure in their wake collapses with reduced nucleosome occupancy, irregular spacing and intermediate assemblies. Chromatin states deteriorate as modified histones are lost due to a lack of histone recycling. Chromatin fiber disorder further manifests in the 3D genome, triggering active genes to coalesce in aberrant microcompartments. This establishes a unifying mechanistic basis for mammalian chromatin replication and transcription, with FACT mediating nucleosome disruption and re-assembly and thereby guarding against spurious chromatin aggregation. Nucleosome organization can therefore dynamically regulate genome architecture.

<h4>Background</h4>Hypothalamic kisspeptin neurons are master regulators of reproduction in mammals, including humans and rodents, via direct stimulation of gonadotropin-releasing hormone (GnRH) neurons. Therefore, they play a central role in puberty onset and gonadal function in mammals. Arcuate kisspeptin neurons express both neurokinin B (NKB) and dynorphin A (Dyn). Thus, these neurons are also referred to as KNDy neurons, and accumulating evidence suggests that KNDy neurons are responsible for tonic GnRH/gonadotropin release and the consequent folliculogenesis/steroidogenesis. Notably, mutations in genes encoding kisspeptin (KISS1), kisspeptin receptor (GPR54), or NKB (TAC3) in humans, or deletion of these genes in rodents, cause hypogonadotropic hypogonadism. Therefore, cellular models of KNDy neurons are useful for elucidating the pathogenesis of reproductive disorders and for developing novel therapies for these disorders. Here, we established a method to differentiate KNDy neurons from mouse embryonic stem cells (mESCs).<h4>Methods</h4>mESCs were incubated in culture medium containing agents that enhance Sonic Hedgehog signaling, a ventralizing signal, to induce the differentiation of ventral hypothalamic organoids. Kisspeptin-expressing cells were generated by adding a Notch inhibitor, dispersing the aggregates, and transferring them into a two-dimensional culture system. Kisspeptin-, NKB-, and Dyn-expressing cells were identified by immunohistochemistry, and kisspeptin secretion into the culture medium was quantified by enzyme-linked immunosorbent assay.<h4>Results</h4>Immunoreactivity for kisspeptin, NKB, and Dyn was detected in differentiated organoids derived from mESCs. Furthermore, kisspeptin secretion was evident in culture supernatants of differentiated KNDy neurons.<h4>Conclusions</h4>This is the first report demonstrating the differentiation of KNDy neurons from pluripotent stem cells, which can be applied to create cellular models for various diseases caused by KNDy peptide deficiency and may contribute to the development of novel therapeutic approaches and understanding cellular mechanisms regulating mammalian reproduction.

Human T-cell leukemia virus type 1 (HTLV-1) causes adult T-cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy, and its viral transactivator Tax is central to disrupting host transcriptional control. Although individual interactions between Tax and host factors have been investigated, their global network and functional impact remain unclear. In this study, we performed proteomic analyses of Tax-associated complexes in HTLV-1-infected T-cells and mapped interactions between host factors and Tax. In addition to known nuclear factor kappa B (NF-κB) regulators, we identified the transcription factor interferon regulatory factor 4 (IRF4) as a novel interactor. Co-immunoprecipitation experiments confirmed Tax-IRF4 binding, and chromatin profiling revealed that Tax reprograms genome-wide IRF4 occupancy. In infected T-cells, IRF4 was enriched in super-enhancer (SE) regions, where Tax, IRF4, and NF-κB factors colocalized at ATL-related loci, and this was accompanied by histone acetylation changes and transcriptional activation. Functionally, Tax and IRF4 co-expression cooperatively restructured chromatin accessibility and induced aberrant gene expression programs. Moreover, ATL-associated IRF4 mutants (K59R, L70V, and S114N) partially reproduced Tax-driven chromatin remodeling and transcriptional reprogramming, suggesting that convergent mechanisms drive leukemogenesis. Collectively, these findings demonstrate that Tax exploits IRF4 to establish a novel regulatory mechanism. This interaction drives widespread epigenomic reprogramming in HTLV-1-infected T-cells. Overall, our study underscores the central role of Tax in IRF4 exploitation to induce infection-specific epigenomic changes, which offers novel insights into ATL pathogenesis and therapeutic strategies.

Increasing consumer demand for high-quality pork has led the industry to focus on attributes like color, marbling, and tenderness. Since genetics plays a crucial role in such traits, this study explored the relationship between specific genetic variants and key pork quality parameters. Six candidate single nucleotide polymorphisms (SNPs) were analyzed in pigs from commercial farms in northeastern Argentina: RYR1 1843 C > T, RN (200R > Q/199I > V), CAST 638 S > A, CAST 76872G > A, SOX6A (A42812066G > A), and SOX6B (B43023574G > C) using polymerase chain reaction-restriction fragment length polymorphism analysis. Meat quality traits— pH, color, drip, thawing and cooking losses, marbling, moisture, and shear force—were measured and statistically analyzed. The RYR1-T allele was associated with faster pH decline, higher L*, CL and reduced tenderness (p < 0.05), suggesting susceptibility to pale, soft, exudative meat. Considering the major effect of RYR1, the influence of other variants was examined within RYR1 backgrounds. RN carriers showed effects color, drip, thawing and cooking losses and shear force (p < 0.05). CAST polymorphisms influenced pH decline and water retention (p < 0.05), while SOX6 SNPs were associated with pH, color, marbling, and water-holding capacity (p < 0.05). This study presents the initial characterization of these genetic markers within Argentinean herds, providing breeders with actionable data for genetic selection and advancing the field of porcine genetics.

<h4>Background</h4>Chronic kidney disease (CKD) affects approximately 14% of the UK population and is associated with significant exercise intolerance, partly due to skeletal muscle dysfunction. While exercise is a potential therapeutic strategy, the molecular response of skeletal muscle to exercise in CKD remains poorly understood. This study aimed to characterise transcriptomic changes in skeletal muscle 24 hours after aerobic (AE) or combined aerobic and resistance exercise (CE) in non-dialysis CKD.<h4>Methods</h4>This study utilised muscle biopsies from participants in the ExTRA CKD trial with stage 3b-4 CKD stages 3b-4 (AE: 24 (15-32) ml/min/1.73m2; CE: 25 (19-31) ml/min/1.73m2). Participants (n = 4 per group) were randomised to 12 weeks of thrice-weekly AE or CE. Vastus lateralis skeletal muscle biopsies were collected at baseline and 24h after the first bout of exercise. RNA was extracted for Bulk RNA sequencing. Bulk RNA sequencing was performed, and differentially expressed genes (DEGs) were identified between baseline and post-exercise samples, followed by pathway enrichment analysis.<h4>Results</h4>Following AE, 1480 genes were upregulated and 1554 downregulated. CE resulted in 556 upregulated and 115 downregulated genes. The most upregulated gene after AE was CHI3L1 (log₂FC 10.7), followed by SAA2 and PTX3, all associated with inflammation. After CE, SFN (log₂FC 6.8) and MT1A were among the most highly upregulated. Enrichment analysis showed strong activation of inflammatory and cellular senescence pathways, and downregulation of mitochondrial function-related processes, particularly after AE.<h4>Conclusion</h4>Both AE and CE triggered robust inflammatory gene expression responses in CKD skeletal muscle, which may be indicative of early repair processes. Unexpectedly, mitochondrial-related pathways were downregulated at 24h post exercise. In the absence of earlier post exercise timepoints, it is not possible to determine whether these findings reflect impaired mitochondrial adaptation, or instead represent a recovery phase return of mitochondrial gene expression levels to baseline. These results highlight mitochondrial dysfunction may be a potential barrier to effective exercise adaptation and a possible therapeutic target in this population.

<h4>Background</h4>Amnestic mild cognitive impairment (aMCI) represents a transitional stage between normal aging and Alzheimer disease (AD), where early intervention is critical for preserving cognition and delaying or preventing progression to dementia. Due to the limited availability of curative pharmacological treatments, there is growing interest in traditional and indigenous medicinal interventions, such as Bacopa monnieri (Linn.) or Brahmi, a widely used Ayurvedic nootropic herb. Bacopa consumption is known to enhance cognitive performance in healthy individuals and is associated with alterations in pathways related to branched-chain and aromatic amino acid biosynthesis. These pathways have been implicated in MCI due to AD pathophysiology. Hence, the efficacy of Bacopa in aMCI and the mediating metabolic changes need to be systematically evaluated through clinical trials.<h4>Objective</h4>The aim of this study is to assess the effects of B. monnieri on cognitive function and plasma metabolites in individuals with aMCI (early or prodromal AD).<h4>Methods</h4>This study will employ a double-blind, randomized, placebo-controlled trial design, with 76 participants (38 per group) of aMCI diagnosed using the clinical National Institute on Aging-Alzheimer's Association 2011 criteria in a tertiary care setting in India. Participants will receive either B. monnieri (Linn.) (300 mg standardized plant extract) or a matched placebo daily for 12 weeks. Comprehensive cognitive assessments (0, 12, and 24 wk) and untargeted plasma metabolomic profiling (0 and 12 wk) will be conducted to evaluate both cognitive changes and alterations in plasma metabolites. MetaboAnalyst 6.0 (with integrated features like Kyoto Encyclopedia of Genes and Genomes and the Human Metabolome Database) will be used for a comprehensive statistical and functional analysis pertaining to the metabolomics. The primary outcome will be a change in the composite z score of memory between the 2 groups at 12 weeks. The secondary outcomes will include alterations in metabolites and pathways, and adverse events at 12 weeks, and cognitive performance at 12 and 24 weeks.<h4>Results</h4>The study was funded in October 2023, and the first participant was enrolled in April 2024. As of November 2025, a total of 60 participants have been recruited, with a mean (SD) age of 62.2 (8.1) years. The cohort predominantly comprises men (53/60, 88.3%), individuals with at least a high school education (47/60, 78.3%), and participants at elevated risk for cognitive decline. Specifically, 39 of 60 participants (65%) have diabetes, 29 (48.3%) have hypertension, and 13 (21%) report a positive family history of dementia. Data collection will conclude in June 2026, after which data analysis will begin and be completed by September 2026, with primary findings targeted for publication in spring 2027.<h4>Conclusions</h4>This protocol investigates the efficacy of B. monnieri (Linn.) in improving cognitive function and altering the blood metabolites in patients with aMCI. If effective, this intervention could provide an accessible and cost-effective approach to manage early AD in resource-limited settings.

The GAIA-CLL13 trial showed that venetoclax-obinutuzumab (Ven-O) based regimens, with or without ibrutinib, offer superior efficacy compared to chemoimmunotherapy (CIT) with regards to progression free survival (PFS) in fit, treatment-naïve (TN) CLL patients. However, their higher costs warrant a cost-effectiveness evaluation. This study assessed the cost-effectiveness of Ven-O versus venetoclax with rituximab (Ven-R), venetoclax-obinutuzumab-ibrutinib (Ven-O-I), and CIT in fit, TN CLL patients without TP53 aberrations across the Netherlands, Norway, and Denmark. A state-transition Markov model including later line treatment was applied to estimate costs, life years (LYs), and quality-adjusted life years (QALYs) over a 38-year horizon. Results were sensitive to long-term OS assumptions. In the base-case analysis, extrapolating OS and PFS for each treatment separately, all venetoclax-based treatments appeared cost-effective compared to CIT in all three countries. ICERs for Ven-R, Ven-O, and Ven-O-I versus CIT were €35 840, €32 513, €30 331 for the Netherlands, €39 881, €38 099, €26 381 for Norway, and €34 010, €37 804, €33 215 for Denmark. In the sensitivity analyses, however, cost-effectiveness was lost when only allowing separate OS and PFS extrapolations for statistically significant differences at 60-month follow-up. Furthermore, cost-effectiveness results were sensitive to varying assumptions about national willingness-to-pay (WTP) thresholds, IGHV-status, and time horizon. In conclusion, venetoclax-based treatments may be considered a cost-effective treatment option for fit, TN CLL patients without TP53 aberrations in the Netherlands, Norway, and Denmark, but the pricing process for targeted agents should take the uncertainty about cost-effectiveness into account when negotiating pricing of medication. Longer follow-up data is needed to address the uncertainties.

Patient-derived brain organoids have emerged as a powerful model for investigating the mechanisms underlying neurological and psychiatric disorders. They provide novel insights into autism spectrum disorder (ASD), a heterogeneous neurodevelopmental condition whose underlying mechanisms remain poorly understood. Recent advancements in generating electrophysiological functional 3D brain organoids enable the study of molecular and network-level neuronal activity. Here, we aimed to characterize the neurophysiological underpinnings of ASD by comparing electrophysiological properties of brain organoids derived from eleven individuals diagnosed with autism spectrum disorder, 10 with monogenic syndromic ASD across five genetic subtypes, and 1 with idiopathic ASD, to organoids derived from 4 neurotypical control individuals. We identified distinct differences in baseline activity (resting state) and evoked responses (synaptic plasticity and network dynamics) across ASD subgroups. To comprehensively assess these differences, we applied dimensionality reduction (principal component analysis, PCA) to integrate multiple electrophysiological features into a unified framework. Our findings reveal subtype-specific neurophysiological alterations in ASD brain organoids, offering mechanistic insights into ASD heterogeneity and potential applications for early diagnostics, drug screening, and therapeutic development.

In atopic dermatitis (AD), skin barrier and immune dysfunction result in chronic tissue inflammation, yet our understanding of the tissue ecosystem remains incomplete. Here, we generate a multi-modal census of 280,518 cells from whole skin tissue samples from 17 adults, including 11 AD patients, integrating it with 430,186 cell profiles from four previous studies into a comprehensive human skin cell atlas. Reconstruction of keratinocyte differentiation revealed disrupted cornification in AD associated with signals from an immune and stromal multicellular community - comprising MMP12⁺ and migratory dendritic cells (DCs), cycling innate lymphoid cells (ILC), natural killer cells, inflammatory CCL19⁺ IL4I1⁺ fibroblasts, and clonally expanded IL13⁺IL22⁺IL26⁺ T cells connected by intercellular feedback loops predicted to impact community assembly. Subsets from this community, along with disrupted cornified keratinocytes, were enriched in GWAS, suggesting that dysfunction in this communication network may initiate AD. Our work highlights disease-associated cell subsets and interactions in chronic skin inflammation.

The existence of human hippocampal neurogenesis has long been disputed^1-12 and its relevance in cognition remains unknown. Recent studies have established the presence of proliferating progenitors and immature neurons and a reduction in the latter in Alzheimer's disease (AD)^11,13. However, their origin and the molecular networks that regulate neurogenesis and function are poorly understood. Here we studied human post-mortem hippocampi obtained from different cohorts: young adults with intact memory, aged adults with no cognitive impairments, aged adults with extraordinary memory capacity (SuperAgers)^14,15, adults with preclinical intermediate pathology or adults with AD. Using multiomic single-cell sequencing (single-nucleus RNA sequencing and single-nuclei assay for transposase-accessible chromatin with sequencing), we analysed the profiles of 355,997 nuclei isolated from the hippocampus samples and identified neural stem cells, neuroblasts and immature granule neurons. Dysregulated neurogenesis was largely associated with changes in chromatin accessibility. Analyses of transcription factors and target gene signatures that distinguished each of the groups revealed early alterations in chromatin accessibility of neurogenic cells from individuals with preclinical AD, and such changes were even more evident in samples from individuals with AD. We identified a distinct profile of neurogenesis in SuperAgers that may reflect a 'resilience signature'. Finally, alterations in the profile of astrocytes and CA1 neurons govern cognitive function in the ageing hippocampus. Together, our study points to a multiomic molecular signature of the hippocampus that distinguishes cognitive resilience and deterioration with ageing.

BACKGROUND: Immune tolerance breakdown at the maternal-fetal interface is implicated in unexplained recurrent pregnancy loss (URPL), but the interplay between T cell hyperactivation and dendritic cells (DCs)-mediated signaling remains poorly defined. METHODS: First-trimester decidual tissues from 5 healthy controls and 6 URPL patients underwent single-cell RNA sequencing (scRNA-seq, 10× Genomics). Computational analyses included clustering (Seurat), trajectory inference (scTour), intercellular communication (CellChat) and metabolic pathway enrichment (Gene Ontology and scMetabolism). Flow cytometry was performed from 11 patients and 11 healthy controls. Spatial validation was performed via multiplex immunohistochemistry and immunohistochemistry on 12 additional controls and 12 URPL cases. Statistical significance was assessed using Student’s t-test. RESULTS: URPL decidua exhibited marked CD3+ T cells and MX1+effector T (Tem) cells infiltration and activation. Flow cytometry analysis confirmed a significant decidua-specific upregulation of T cell activation markers CD25 and CD69 specifically on the MX1+Tem subset in URPL patients compared to controls. MX1+Tem cell subset demonstrated interferon hyperactivation, proliferative hyperactivity and lipid-biased immunometabolism. Pseudotemporal analysis positioned MX1+ Tem cells between classical Tem and exhausted T cell states, suggesting progressive differentiation. CellChat identified DCs as key regulators of MX1+ Tem expansion via aberrant ICOSL signaling, validated by spatial co-localization of ICOSL+ DCs and MX1+ Tem cells in URPL tissues. CONCLUSION: Our findings demonstrate that the aberrant activation and proliferation of MX1+Tem cells as a key immunological feature associated with URPL patients.

BACKGROUND: Dietary approaches to stop hypertension (DASH) diet as a balanced dietary pattern is designed to help manage hypertension. The scientific evidence hypothesized that following the DASH diet may lead to reductions in cardiometabolic risk. We aimed to design a clinical trial to assess the impact of the DASH diet on cardiometabolic and atherogenic indices in individuals with metabolic syndrome (MetS). METHODS: Sixty participants diagnosed with MetS were randomly divided into two groups: the intervention group (adhering to DASH diet) or the control group (following a common healthy diet). Key outcomes of the present study including cardiometabolic index (CMI) as our primary outcome, and atherogenic index of plasma (AIP), atherogenic coefficient (AC) and castelli risk index II (CRI-II) as secondary outcomes were evaluated at the baseline and at the end of the study. RESULTS: While one participant excluded from the trial, 59 subjects completed the study. Baseline values of CMI (p = 0.33), AIP (p = 0.27), AC (p = 0.44) and CRI-II (p = 0.17) showed no significant difference between the intervention group and the control group. After adjusting for confounding factors, the intervention group demonstrated a significant decrease compared to the control group in CMI (−0.38±0.29 vs. −0.02±0.47; p = 0.001), AIP (−0.04±0.04 vs. 0.01 ± 0.07; p = 0.001), AC (−0.31±0.54 vs. 0.01 ± 0.72; p = 0.04) and CRI-II (−0.30±0.37 vs. −0.03±0.39; p = 0.009). CONCLUSIONS: Adherence to the DASH diet appears to be effective in reducing CMI and some atherogenic indices. Further well-designed clinical trials with longer intervention durations are needed. TRIAL REGISTRATION: The trial was registered on 21 October 2022 at Iranian Registry of Clinical Trials (IRCT20180201038585N12, https://irct.behdasht.gov.ir/trial/66161 ).

BACKGROUND: Identification of genetic variations associated with early-onset high myopia (eoHM) provides a genetic basis for risk assessment and prevention of this disease. METHODS: Whole-exome sequencing (WES) was performed on 41 probands with eoHM with or without other abnormalities. RESULTS: Sixteen high myopia-associated variants identified in 13 probands involved 13 genes comprising 11 autosomal dominant and 2 X-linked genes. The frequency of variants in SNRNP200, ARR3, and COL2A1 was 13%, which was slightly greater than that of other genes. A total of 46% were associated with inherited retinal diseases documented in the RetNet database. According to the relevant guidelines and standards, 9.7%(4/41) of the probands had suspected genetic pathogenic variations through combined clinical-genetic assessment (ID2,3,8,11). Interestingly, we found that the genetic diagnosis rate was significantly correlated with the specific clinical phenotypic characteristics of the patients. The diagnosis rate of patients with ultrahigh myopia was significantly greater than that of patients with high myopia. CONCLUSIONS: Genetic analysis identified the pathogenic factors of many cases of eoHM, revealing a strong association between eoHM candidate genes and hereditary retinal diseases. EoHM is the predominant clinical presentation in most hereditary ocular diseases.

BACKGROUND: Bronchiectasis (BE), Chronic Obstructive Pulmonary Disease (COPD) and Primary Immunodeficiency (PID) are distinct respiratory conditions that share features such as chronic inflammation, airway obstruction, and recurrent infections. Despite distinct underlying causes, disease manifestations often overlap clinically, complicating diagnosis and treatment. We aimed to characterise shared and disease-specific immune mechanisms in BE, COPD, and PID using a multi-omics approach across blood and sputum. METHODS: Adults with BE, COPD, PID, and healthy controls underwent serum and sputum protein quantification, sputum proteomics, and transcriptomic profiling of sputum and blood. Neutrophil fractions were included as covariates in transcriptomic analyses. RESULTS: Distinct clinical profiles were observed across disease groups. Serum analyses showed changes in inflammatory and extracellular matrix markers, particularly in BE and COPD. Sputum from BE patients showed elevated inflammatory markers and pathogen burden. Proteomic profiling demonstrated that BE and COPD shared neutrophil-driven inflammatory pathways, while PID exhibited a unique humoral immune signature. Transcriptomic analyses further confirmed neutrophil abundance as a major driver of gene expression variation across disease cohorts. After adjusting for neutrophil fractions, disease-specific differentially expressed genes (DEG) were identified, including OLFML2B, FZD2, and STAB1 in BE, and GPR15, METTL7B, and RNASE1 in COPD. PID samples showed no significant DEGs but enrichment of interferon signalling pathways. CD177 and DEFA3 were overexpressed beyond variation driven by neutrophil abundance, highlighting association with key inflammatory drivers in BE and COPD. CONCLUSIONS: This study uses an integrative multi-omics analysis to improve the understanding of disease heterogeneity and distinct inflammatory pathways in BE, COPD and PID.

West Nile virus (WNV) and Japanese encephalitis virus (JEV) are major mosquito-borne pathogens that cause severe neurological illnesses in humans, including encephalitis and meningitis. Neurons are the primary target of WNV and JEV infections in the brain. However, the pathogenic mechanisms and host immune response associated with WNV and JEV infections in the primary neurons are not fully characterized. Herein, using high-throughput RNA sequencing (RNA-Seq), we performed a comparative transcriptomic analysis of the primary mouse neurons following infection with WNV and JEV. We identified differentially expressed host genes (DEGs) that were commonly induced in neurons by both viruses, as well as those uniquely regulated in a virus-specific manner. We observed pronounced changes in gene expression profiles that indicate activation of a robust antiviral response accompanied by signatures of neuronal dysfunction. Notably, interferon-stimulated genes such as Irf7, Isg15, and Ifnb1 were markedly upregulated in WNV- and JEV-infected neurons. Additionally, we observed the significant upregulation of proinflammatory chemokine and cytokine genes such as Il6, Cxcl2 and Cxcl10 which was further validated by Luminex assay. Ingenuity Pathway Analysis (IPA) revealed the activation of several cell death pathways including pyroptosis and necroptosis. We also showed the significant dysregulation in pattern recognition receptors and neurotransmitter receptors such as glutamatergic and acetylcholine receptors. This dysregulation extended to the synaptic signaling and reduced synaptogenesis in infected neurons. Quantitative RT-PCR confirmed the upregulation of key cytokines like Il6 and Ifna, further establishing the validity of the RNA-Seq approach. Overall, this study provides insights into the pathological changes in the neurons following WNV and JEV infections.

Nerve regeneration and axonal growth can be significantly enhanced by electrical stimulation (ES) using conductive polymers. But in addition to ES, nerve cell function and how they react to substrate conductivity are also essential factors in nerve repair. As a result, ES, in conjunction with appropriate materials (such as tissue scaffolds), has been used to treat nerve damage and has demonstrated significant promise for peripheral nerve regeneration. In this work, electrospinning was used to create conductive composite scaffolds made of polyglycolic acid@polycaprolactone/aniline pentamer (PGCL/AP). We later verified that PC12 cells cultured on these scaffolds exhibited improved migration, proliferation, and vitality upon electrical stimulation. Furthermore, In the established 10-mm peripheral nerve injury model, the PAP + ES group achieved outcomes comparable to the autograft group in the assessment of functional and morphological recovery, encompassing SFI, myelination rate, and gastrocnemius muscle evaluation.

Myopia represents a refractive anomaly characterized by impaired vision resulting from a misfocused image in front of the fovea. Although numerous genes linked to high myopia (HM) have been identified, the exact etiology and pathogenesis mechanisms of HM remain predominantly obscure. In a prior investigation, a mutation in the <i>P4HA2</i> gene was identified in association with HM. To illuminate the potential mechanisms of action of <i>P4HA2</i> in HM, we established a <i>P4HA2</i>-knockout mouse line (<i>P4ha2</i> ^-/-) and a P4HA2-knockout HEK293 cell line for this study. <i>P4ha2</i> ^-/- mice exhibited compromised visual acuity and altered light transmission pathways as evidenced by multiple biometric assessments. Furthermore, we observed a time-dependent disruption in the arrangement of collagen fibrils in the sclera and cornea of the <i>P4ha2</i> ^-/- mice, attributed to diminished thermal stability due to decreased collagen hydroxylation. Our findings also revealed elevated fibronectin levels and reduced Collagen I expression in the sclera and cornea of the <i>P4ha2</i> ^-/- mice, as well as in P4HA2-knockout HEK293 cells, suggesting an imbalance in extracellular matrix (ECM) components that could further perturb light transmission pathways, which induced HM-associated refractive error. In summary, <i>P4HA2</i> contributes significantly to the pathogenesis and progressive deterioration of refractive error by accelerating collagen degeneration via reduced collagen hydroxylation.

Campylobacter is a major cause of foodborne illnesses worldwide; however, comparative data on its occurrence and antimicrobial resistance (AMR) in retail meat across Asian regions remain limited. Therefore, this study investigated the prevalence, quantitative burden, and genomic and phenotypic AMR characteristics of Campylobacter in retail chicken and pork from Vietnam and Japan. Multiplex real-time polymerase chain reaction without enrichment successfully detected Campylobacter and revealed country-specific patterns, with Vietnamese chicken showing a substantially higher contamination rate (43.9%) with a predominance of C. coli (20.7%; n = 82) and Japanese chicken exhibiting a lower contamination rate (33.3%) with a predominance of C. jejuni (47.6%; n = 21). Notably, pork contamination was rare (2.2%; n = 46). Culture-based isolation recovered five C. jejuni isolates and one C. coli isolate from Japan and three C. coli isolates from Vietnam, using a membrane filter method necessary to obtain Vietnamese strains because of heavy Escherichia coli overgrowth. Whole-genome sequencing identified widespread quinolone resistance-associated gyrA mutations and a multidrug-resistant C. coli genotype in Vietnamese isolates harboring chromosomal determinants conferring resistance to multiple antimicrobial classes. Phenotypic susceptibility testing revealed universal resistance to fluoroquinolones and tetracyclines, along with carbapenem resistance, in several Japanese isolates and cephalosporin resistance in C. coli isolates from both countries. Overall, our findings indicate a markedly high contamination rate and AMR burden in Vietnamese retail chicken meat and underscore the need for stringent Campylobacter surveillance and effective risk-mitigation strategies in high-prevalence regions.

<h4>Background</h4>Selective serotonin reuptake inhibitors (SSRIs) are a recommended first line medication for the treatment of major depressive disorder, due to higher tolerability and lower risk of adverse effects than other antidepressants. The mechanisms by which SSRIs reduce depressive symptoms are not well understood, but are hypothesised to include direct effects on serotonin signalling and synaptic remodelling, and indirect effects on inflammation. Indirect or off-target effects may be detectable in blood and can be investigated using methylome- and transcriptome-wide approaches.<h4>Methods</h4>The Staged Treatment in Early Psychosis (STEP) clinical trial included a 6-month long randomised, placebo-controlled trial of the SSRI fluoxetine in a cohort of young people at ultra-high risk for psychosis. A methylome-wide association study (MWAS; N_{Total (before/after/both)} = 104 (52/52/44), N_{SSRI (before/after/both)} = 45 (21/24/18)) and differential expression analysis were performed on longitudinal blood samples collected at the start and end of the 6 months to identify changes in DNA-methylation and gene expression associated with medium-term SSRI exposure.<h4>Results</h4>Four methylation CpGs (cg26253898, cg09719563, cg22216017, cg26017656) were significantly associated with SSRI exposure (FDR < 0.1, 2 CpGs at FDR < 0.05) and annotated to genes involved in glucose metabolism, synaptic remodelling and inflammation (GCG, COL23A1, PEG10, SGCE, MFGE8). Gene-set enrichment analyses of genes annotated to the top 100 CpGs identified significant tissue-specific enrichments in artery, adipose and spleen tissues, and in the 'postsynaptic density' GO term. No genes were differentially expressed, including genes annotated to the significant methylation CpGs.<h4>Conclusions</h4>Medium-term SSRI use during the STEP trial was associated with changes in DNA-methylation that may partially explain the potential antidepressant mechanisms and adverse effects of SSRIs, however replication in other cohorts is necessary to establish if these changes are generalisable to SSRI use more broadly.

Mitochondrial DNA copy number (mtDNA-CN) is a metric of mitochondrial function that has been associated with a variety of diseases including cardiovascular disease and all-cause mortality. To investigate genes and pathways affected by mtDNA-CN variation, we perturbed HEK 293T cells with ethidium bromide to deplete mtDNA. Using RNASeq and methylation microarrays, we evaluated transcriptomic and methylomic changes in treated cell lines. We observed an 8-fold decrease in mtDNA-CN and compensatory shifts in mitochondrial transcription to support mtDNA replication. Nuclear transcriptomic and methylomic analysis highlighted changes in metabolic pathways, including oxidative phosphorylation and canonical glycolysis. Longitudinal analyses revealed that the identified genes and pathways have different response timing, with nuclear response lagging behind mitochondrial response. These findings further elucidate the mechanisms behind mtDNA maintenance and responses to cellular energetics as well as mitochondrial-nuclear crosstalk dynamics.

The World Health Organization has classified five species of <i>Candida</i> as pathogens of critical concern on its recent 'Fungal Pathogen Priority List'. <i>Candida albicans</i> is the most common species aetiologically associated with human and animal mucosa; however, we know almost nothing about the environmental survival and the transfer pathways of <i>C. albicans</i> in natural environments. Here, we have isolated <i>C. albicans</i> from the faeces of newly arrived migratory geese in two agricultural fields in Scotland (the United Kingdom). All 14 confirmed isolates were characterised in terms of their pathogenicity (in a <i>Galleria</i> larval infection model) and resistance to four classes of antifungal drugs. Multilocus sequence typing (MLST) was used to assess the relatedness of these isolates with <i>C. albicans</i> isolated from the faeces of sheep grazing in the same field. Finally, mesocosms were used to test whether <i>C. albicans</i> in geese faeces could persist and remain viable when incorporated into agricultural soils. All <i>C. albicans</i> isolates were virulent, and exhibited resistance to at least one of the four classes of antifungal drugs against which they were screened. Maximum likelihood phylogenetic analysis of MLST sequences revealed that <i>C. albicans</i> isolated from goose and sheep faeces were relatively conserved, although they did cluster independently on separate clades. <i>C. albicans</i> remained viable in common agricultural soils, and after 60 days, the concentrations of all three representative <i>C. albicans</i> isolates had only declined by one log. The unrestricted movement of migratory birds provides a significant opportunity for the widespread dissemination of pathogens. Although <i>C. albicans</i> is not generally considered to be zoonotic, its introduction into new environments and subsequent persistence in agricultural systems has the potential to facilitate transmission between animal species or humans. Therefore, it is critical that more emphasis be put on monitoring animal migration and the potential for cross-boundary movement of pathogens, particularly with climate change widening the geographic range for pathogen persistence.

Iron is an essential micro component and is involved in numerous critical cellular processes and in energy production. While its roles in oxygen transport and in oxidative phosphorylation are well documented, it remains to be elucidated, whether iron modulates specific cellular processes in different organs. Iron deficiency has been found to lead to a decrease in the capacity of osteoclasts to dissolve amorphous calcium phosphate. Furthermore, levels of iron in the cellular environment led to significant changes in the levels of transcripts encoding iron transport proteins. Within the present study, the uptake of iron by osteoclasts and the kinetics of intracellular transport were analyzed. For this purpose, M-CSF (Macrophage-Colony Stimulating Factor) dependent non-adherent osteoclast progenitor cells were differentiated to osteoclasts in media containing M-CSF and RANKL (Receptor Activator of NF-κB Ligand). For the upregulation of iron transport capacity, media were supplemented with Deferoxamine, an iron complexor, rendering the cultures virtually iron-free. To analyze iron uptake by osteoclast like cells, holo-transferrin, loaded with ⁵⁵Fe was added to the cells and iron uptake was quantitated in whole cell lysates and in fractionated cells. The data demonstrates that Deferoxamine-treated osteoclasts absorb higher quantities of iron as compared to untreated control cells. By density gradient centrifugation, cell associated iron can be separated into two major pools. Pool I represents non-transferrin associated iron in cytoplasmic fractions, while pool II contains transferrin/ transferrin receptor associated iron. Within 4 h of incubation in iron-deficient medium, pool I disappears, as does transferrin. Pool II iron and transferrin receptor, however, remain detectable. Furthermore, iron peaks did not associate with ferritin nor with mitochondria, demonstrating that these two mechanisms of iron storage did not become activated during the course of the study. The data thus demonstrates that iron uptake by osteoclasts can be modulated by exogenous iron and that cell associated iron forms either a labile iron pool of free iron that is lost within a short period of time or a vesicular pool of non-transferrin bound iron that remains stable over the experimental period. No further trafficking of iron into ferritin particles or mitochondria was detected.

<h4>Background</h4>Mass-forming (MF) type is the most common, accounting for 57.1-83.6% of intrahepatic cholangiocarcinoma (ICC), with a poor prognosis. Transarterial chemoembolization (TACE) can induce necrosis of tumor cells, induce the release of tumor antigens, enhance the immune response of tumor-specific CD8⁺ T cells, and regulate the proliferation of Treg cells. However, real-world data directly comparing TACE combined with programmed cell death protein-1 (PD-1) versus programmed death ligand-1 (PD-L1) inhibitors in MF-ICC are lacking. Therefore, we aimed to evaluate the efficacy and safety between the different immune checkpoint inhibitors (ICIs) (PD-1/PD-L1 inhibitors) in MF-ICC, and to explore prognosis-related clinical factors and preliminary immune mechanisms underlying this combined therapy.<h4>Methods</h4>A total of 50 patients with MF-ICC who underwent TACE combined with ICIs at Beijing Friendship Hospital and Beijing Ditan Hospital from May 2020 to December 2024 were retrospectively enrolled. Least absolute shrinkage and selection operator (LASSO) regression was used to screen the risk factors of overall survival (OS). Survival was estimated using the Kaplan-Meier method and compared by the log-rank test. Univariable Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for the association between treatment regimen and survival. In parallel, we analyzed dynamic changes in immune cells before and after ICI treatment using the single-cell RNA sequencing dataset GSE208205 and further validated these findings by flow cytometry.<h4>Results</h4>The median overall survival (mOS) and median progression-free survival (mPFS) for TACE-PD-L1 followed by therapy were 18 and 13 months, which were significantly longer than those with TACE-PD-1 sequential therapy (mOS: 12 months, HR: 0.42, 95% CI: 0.17-1.03, P=0.047; mPFS: 8 months, HR: 0.29, 95% CI: 0.12-0.73, P=0.006). In exploratory multivariable analysis, pre-treatment monocyte-to-lymphocyte ratio (MLR), Child-Pugh classification, total bilirubin (TBIL), and alanine aminotransferase (ALT) emerged as potential prognosis-related factors for OS. Single-cell analysis showed that CD4⁺ T and CD8⁺ T cells were markedly increased after treatment, while circulating tumor cells and vascular endothelial cells were decreased. This was further validated by the flow cytometry. Moreover, regardless of treatment status, ICC patients in the PD-L1 groups exhibited higher levels of CD4⁺ and CD8⁺ T cells compared to the PD-1 group, whereas B cells were lower in the PD-L1 group than in the PD-1 group.<h4>Conclusions</h4>TACE combined with PD-L1 inhibitors was associated with longer survival than TACE combined with PD-1 inhibitors in patients with mass-forming ICC. The immune system, particularly lymphocytes, plays a critical role in the efficacy of combination therapy. In addition, several baseline inflammation- and liver function-related factors (MLR, Child-Pugh class, TBIL, ALT) were associated with OS in exploratory analyses.

Cardiovascular disease (CVD) remains the leading cause of worldwide morbidity and mortality. Studies have found that there is a significant genetic component contributing to CVD development. Advances in genome sequencing technologies have revolutionized the identification of disease-causing variants in the human genome. With the development of whole genome sequencing (WGS), the understanding of these variants has been deepened as it enables comprehensive detection of many variants in the genome including structural variants (SVs). SVs are large genomic variants that are present in the genome of an organism and play a significant role in disease. Numerous techniques are being used to detect SVs with varying accuracy levels. Due to the limited number of focused research studies on SVs and CVD, there is a rich opportunity for further investigation with the aim of utilizing SV data in disease diagnosis and treatment plans. Emerging evidence highlights the role of SVs in CVD and the importance of adopting WGS approaches to unravel the genetic architecture of CVD. Moreover, integrating SV data with population scale epidemiology and advanced risk prediction models would enhance CVD prevention by enabling more personalized treatment strategies. This review aims to describe the different types of SVs and their involvement in CVD development and then to discuss WGS-based SV detection methods and future clinical implementations. We also report an overview of the SVs identified across various CVD types and different bioinformatics tools that can be used to detect SVs in WGS data.

MicroRNAs (miRNAs) are ~19-25-nt post-transcriptional regulators whose dysregulation promotes hallmark cancer traits and therapy resistance. This review synthesizes translational principles for developing miRNA therapeutics in oncology, integrating miRNA biology and target engagement with delivery design and clinical experience. We summarize key determinants that shape efficacy and safety, including sequence and chemistry choices, biodistribution and intracellular delivery, dosing strategy, and biomarker-informed patient selection. We compare the main therapeutic modalities, miRNA mimics and inhibitors, and evaluate leading delivery approaches relevant to cancer, including lipid-based systems, polymer-based carriers and conjugates, and extracellular vesicle-inspired platforms, highlighting trade-offs in stability, specificity, immune activation, and tumor exposure. Early clinical programs such as MRX34, TargomiR/MesomiR-1, and cobomarsen, together with experience from non-oncology indications, illustrate both opportunities and practical constraints on tolerability and regimen optimization. We conclude with pragmatic priorities for the field, including standardized analytics for isoforms and target engagement, PK/PD- and biomarker-guided dose selection, and rational combination strategies to safely integrate miRNA-based interventions into precision oncology.

To meet the miniaturized cooling demands of high-heat-flux electronic devices, metal foams-featuring high specific surface area and multiscale porous structures-are considered promising candidates for enhancing flow boiling evaporation. However, pore density (PPI) and grooved geometry (channel aspect ratio, AR) jointly regulate vapor-liquid distribution, rewetting, and flow resistance, thereby constraining overall performance. Here, flow boiling experiments were conducted on nickel and copper foams with pore densities of 100, 500, and 1000 PPI and AR values of 0.7, 1.0, and 1.3. Heat transfer coefficient (HTC), wall superheat (ΔT), and pressure drop (Δp) were systematically evaluated, complemented by transient two-phase simulations revealing vapor fraction, temperature, and pressure drop distributions. A pronounced non-monotonic pore-density dependence is observed: 500 PPI achieves an optimal balance between heat-transfer enhancement and flow resistance, whereas 100 PPI suffers from vapor accumulation and temperature non-uniformity, and 1000 PPI is penalized by excessive permeability resistance and pore-scale confinement. An optimal AR of 1.0 promotes efficient vapor venting and stable rewetting. Under the optimal configuration (500 PPI, AR =1.0), a limiting heat flux of 348.6 W/cm², corresponding to the HTC of 55.4 kW/(m² · K), and a limiting HTC of 130.3 kW/(m² · K) are achieved, providing quantitative design guidelines for metal-foam two-phase evaporators.

Metazoa evolved regulatory networks to balance the expression of their sex chromosomes. In Drosophila , males have a single gene-rich X chromosome, whereas females have two. Balanced X/autosome expression is essential for viability, and in male flies is achieved by activation of genes on the X through the male-specific-lethal (MSL) dosage compensation complex (DCC). This ribonucleoprotein assembly contains long, non-coding roX RNAs. To dissect the functional requirements of roX in a cell-based system, we deleted the roX2 gene in male S2 cells and selected two independent lines lacking detectable roX RNA. In the absence of roX , the remaining MSL protein complex was unable to associate with known or newly identified binding sites and thus failed to activate transcription. Surprisingly, the X/autosome expression ratio appeared nevertheless compensated. Cytogenetic and genomic analyses revealed that both roX -deficient cell populations had acquired additional X chromosomes. Apparently, chromosome gains due to mis-segregation made up for the loss of DCC-mediated dosage compensation. Interestingly, ectopic expression of full-length roX2 , but not of shortened derivatives, fully restored DCC binding and normalized the karyotype. These findings illustrate that X chromosome dosage compensation is critical for viability even in cultured cells and provide a striking example of rapid evolution under stringent selection.

The ubiquitin-proteasome system (UPS) represents an evolutionarily conserved machinery governing proteostasis through spatiotemporal regulation of protein degradation. While spermatogenesis involves multilayered regulatory mechanisms spanning translation to dynamic post-translational modifications (PTMs), the identity of UPS-associated E3 ligases orchestrating germ cell-specific protein turnover remains elusive. Here, we identify a testis-specific E3 ubiquitin ligase complex comprising elongin B/C, Cullin-2 (CUL2), RING-box protein-1 (RBX1), and SOCS box protein ASB9, designated ECS^ASB9. Genetic ablation of ECS^ASB9 in mice via ubiquitous Asb9 knockout (KO) or spermatid-specific elongin B/C conditional KO disrupts spermiogenesis and compromises fertility. Mechanistic studies reveal that ECS^ASB9 engages tubulin beta 4 A (TUBB4A) through substrate recognition, catalyzing K48-linked polyubiquitination at lysine 379 (K379) to promote proteasomal degradation. Notably, Tubb4a^K379R knock-in (KI) mice phenocopy the spermiogenesis defects observed upon ECS^ASB9 deficiency. Clinically, we identify three hemizygous missense variants in X-linked ASB9 among Chinese males with idiopathic infertility. Male mice bearing orthologous ASB9 variant exhibit oligoasthenoteratozoospermia (OAT) and subfertility, mirroring human phenotypes. Taken together, our findings establish ECS^ASB9 as an important regulator of spermatogenic proteostasis and provide mechanistic insights into UPS-mediated tissue-specific degradation, while implicating ASB9 variants in male infertility pathogenesis.

The dysfunctional reconstitution of the intestinal barrier is pivotal in driving the initiation of inflammatory pathogenesis in Crohn's disease (CD), although the exact pathophysiology underlying this phenomenon has yet to be definitively characterised. This study aimed to investigate the role of the histone methyltransferase mixed lineage leukaemia 1 (MLL1) in the development of CD-like colitis and to elucidate the mechanism by which MLL1 promotes epithelial cell differentiation. Colonic tissue specimens from CD patients and TNBS-induced murine models were analysed to assess MLL1 expression dynamics. The functional impact of MLL1 on murine colitis modelling CD was systematically investigated through clinical symptom scoring, histopathological profiling and quantitative evaluation of intestinal barrier integrity. The role of MLL1 in promoting epithelial cell differentiation and repairing the intestinal barrier was investigated through immunofluorescence and western blotting. Additionally, potential mechanisms underlying the reparative effects of MLL1 on intestinal barrier function were explored. MLL1 expression was upregulated in colonic tissues from CD patients and TNBS-induced murine colitis models. In contrast, MLL1 suppression in the TNBS cohort attenuated mucosal inflammation and downregulated pro-inflammatory cytokine production (IL-1β, IL-6, TNF-α) within the colonic mucosa. Additionally, reduced MLL1 expression increased the differentiation capacity of intestinal epithelial cells, including goblet cells, absorptive cells and tuft cells, and promoted barrier function restoration in injured colons and lipopolysaccharide-stimulated colonic organoids. MLL1 downregulation activated the Gata4/Bmp4 signalling pathway, which may contribute to the reparative effects of MLL1 on intestinal barrier integrity. Downregulating MLL1 expression promotes intestinal epithelial cell differentiation by activating the Gata4/Bmp4 pathway. These findings elucidate a pathophysiological mechanism wherein MLL1 suppression potentiates intestinal barrier restoration, thereby attenuating colitis severity in murine models. The observed therapeutic efficacy positions MLL1 inhibition presents a novel strategy for CD management.

Inflammation is a major driver of preterm birth, a common pregnancy disorder and the leading cause of childhood death. T regulatory (Treg) cells are essential mediators of maternal fetal tolerance and are critical for constraining uterine inflammation. In some tissue settings, loss of Foxp3 expression can cause instability in Treg cell lineage commitment, elevated production of proinflammatory cytokines and compromised suppressive function. Whether preterm birth susceptibility is associated with loss of lineage fidelity and adoption of proinflammatory phenotypes in Treg cells is unknown. In this study, we investigated the lineage stability of Treg cells in vivo in pregnant mice using a Foxp3 fate-mapping system and models of preterm birth induced by late-gestation inflammatory challenge with lipopolysaccharide (LPS) or interleukin-1β (IL-1β). Ex-Foxp3-expressing (ex-Foxp3) cells were observed in the uterus-draining lymph nodes (udLNs) in non-pregnant mice and in similar abundance across normal gestation, and a proportion expressed proinflammatory cytokines IFNγ and/or IL-17A. Bulk RNA-sequencing of sorted Treg and ex-Foxp3 cells from late-gestation udLNs revealed substantial loss of the Treg cell lineage program in ex-Foxp3 cells, characterized by reversal in expression of canonical Treg cell genes and pathways. Late gestation LPS or IL-1β administration to induce preterm birth did not expand the ex-Foxp3 cell population in the uDLNs or uterine decidua. We conclude that uterine Treg cells exhibit a high level of lineage stability in pregnancy regardless of proinflammatory challenge. Whether there is any biological or pathophysiological significance of ex-Foxp3 cells in gestational tissues remains to be defined.

PURPOSE: Rare cancer patients face dire prognoses due to high malignancy and scarce treatment options, underscoring an imperative for personalized strategies. This study aims to evaluate the potential of precision medicine to extend survival in rare cancer patients by leveraging personalized tumor organoids (PTOs) as a discovery platform to comprehensively explore and generate new treatment opportunities. METHODS: We performed whole-journey clinical mapping in a neuroendocrine cervical cancer (NECC) patient with chemo-resistant metastases. Longitudinal PTOs from three recurrent lesions and autologous tumor-infiltrating lymphocytes (TILs) were established to correlate clinical responses and discover new therapies. A co-culture system integrating PTOs and autologous TILs was developed to replicate clinical responses and optimize TIL therapy through rebuilding tumor antigen microenvironment. RESULTS: PTOs recapitulated tumor evolution and resistance imprints, enabling identification of a highly effective targeted therapy candidate post-chemotherapy failure. The co-culture system faithfully mirrored clinical immunotherapy responses, validating its potential as a companion diagnostic. Utilizing PTOs to rebuild the autologous tumor antigen microenvironment during TIL production generated a personalized, optimized TIL population. These optimized TILs exhibited enhanced anti-tumor ability and their administration correlated with clinical tumor regression in the patient. Generalizability of this PTO-based optimization strategy was validated across both patients and cancer types. CONCLUSION: This study establishes that survival for rare cancer patients hinges on integrated, personalized companion diagnostics and therapy optimization. By bridging iterative tumor models with clinical interventions, we provide a scalable conceptual framework to overcome therapeutic nihilism and unlock precision-driven survival benefits.

Engineered virus-like particles (eVLPs) have emerged as a promising class of delivery systems for genome editing agents. By combining the efficient cellular entry of viral vectors with the safety advantages of nonviral platforms, eVLPs enable transient delivery of ribonucleoproteins such as Cas9, base editors, and prime editors. Successive design strategies, from rational engineering to directed optimization, have progressively improved particle assembly, cargo stability, and editing efficiency. In parallel, pseudotyping approaches have expanded the versatility of eVLPs for cell-type-specific targeting. Recent preclinical studies highlight their potential across diverse applications, ranging from monogenic disease models to complex disorders, and support their advancement toward clinical translation. This review summarizes the structural and production principles of eVLPs, surveys key advances in their development, and discusses therapeutic opportunities and future challenges for their application in genome editing.

Integrin-targeted therapies are under investigation for HIV-associated neuroinflammation, yet their effect on CNS antiviral immunity remains undefined. We examined the role of α4 integrin in T cell-mediated neuroimmune surveillance using SIV-infected macaques with α4 blockade and T cell-specific α4-deficient mice. In macaques, α4 blockade preserved CD4+ Th1 cell access to the brain parenchyma but impaired CD8 effector recruitment, disrupting antiviral control. Despite stable cerebrospinal fluid viral loads, hippocampal SIV RNA increased under blockade. Single-cell analyses revealed α4 enrichment in CD8 effector memory (Tem) cells; blockade reduced inferred CD8+ Tem-monocyte interactions and heightened innate immune activation in the hippocampus. Microscopy demonstrated persistent SIV-induced microglial simplification despite treatment. Th1 CD4 effectors correlated positively with gray matter viral RNA, whereas α4β7+ CD8+ T cells correlated inversely, implicating impaired CD8+ Tem recruitment in elevated parenchymal viral burden. In mice, α4 proved dispensable for CD4 trafficking to inflamed brain but essential for CD8 effector access across CNS compartments and for both subsets to reach skull marrow. These findings establish that α4 integrin governs CD8-mediated neuroimmune surveillance through coordinated cellular positioning, with blockade enabling viral seeding while disrupting spatially organized antiviral defense.

<h4>Objective</h4>To determine whether hysterectomy for benign diseases is associated with the risk of dementia in middle-aged women.<h4>Methods</h4>We conducted a retrospective cohort study using data from the Korean National Health Insurance Service database (2002-2020). Women aged 40-59 years who underwent hysterectomy for benign indications (n=16,818) were propensity score-matched (1:1) to controls who had not received hysterectomy. Subjects were followed up until the diagnosis of dementia, death, or the end of the study period (2020). Dementia (all types), Alzheimer's disease (AD), and vascular dementia (VaD) were identified by International Classification of Diseases, 10th revision codes. Cox proportional hazards models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for dementia associated with hysterectomy.<h4>Results</h4>Median follow-up was 11.4 years. Dementia occurred in 302 (1.8%) women who did not undergo hysterectomy and 257 (1.5%) women who underwent hysterectomy (P=0.061). Cox analysis revealed that hysterectomy was not significantly associated with all-cause dementia (HR, 0.865; 95% CI, 0.724-1.033), with a non-significant trend towards reduced risk. Subgroup analysis also failed to identify any significant association; AD (HR, 0.696; 95% CI, 0.463-1.048) and VaD (HR, 0.625; 95% CI, 0.284-1.377) were not significantly associated with an increased hazard.<h4>Conclusion</h4>In this large Korean cohort, hysterectomy for benign diseases in women aged 40-59 years was not associated with a significant change in the subsequent risk of dementia. Collectively, our results indicate that hysterectomy was not associated with an increased incidence of dementia, including in analyses stratified by adnexal surgery.

The primitive gut tube of mammals initially forms as a simple cylinder consisting of the endoderm-derived, pseudostratified epithelium and the mesoderm-derived surrounding mesenchyme. During mid-gestation, a dramatic transformation occurs in which the epithelium is both restructured into its final cuboidal form and simultaneously folded and refolded to create intestinal villi and intervillus regions. Here, we show that the mesenchymal winged helix transcription factor Foxl1, itself induced by epithelial hedgehog signaling, controls villification by activating BMP and PDGFRα and the planar cell polarity factor Fat4 in epithelial-adjacent telocyte progenitors either directly or indirectly. In the absence of Foxl1-dependent mesenchymal signaling, villus formation and the separation of epithelial cells into mitotic intervillus and postmitotic villus are delayed, and the differentiation of secretory progenitors temporarily blocked. Thus, Foxl1 orchestrates key events during the epithelial transition of the fetal mammalian gut.

The mosquito fat body plays key roles in metabolism and immunity, yet its cellular diversity and specialization remain poorly understood. This study analyzed 97,650 nuclei from the female Anopheles gambiae abdominal body wall at single-nucleus resolution, identifying seven major cell types. Fat body trophocytes are most abundant ( ~ 85%), with five subpopulations: basal (T1, T2), metabolically enriched (T3), immune-responsive (T4), and a vitellogenic group (T5) found only in blood-fed females. Sessile hemocytes comprise 7.4% of cells and expression of lipid biosynthesis enzymes increase in oenocytes (1.1%) after immune priming. T4 trophocytes consistently express immune genes, while various cell types respond to bacterial infection. Blood feeding induces extensive transcriptomic changes, notably upregulating vitellogenin and DNA replication genes, indicating trophocyte endoreplication and metabolic shifts. Vitellogenin mRNA was expressed in the first layer of trophocytes facing the hemolymph with apical subcellular localization. This high-resolution atlas reveals specialized trophocyte roles in mosquito immunity and reproduction.

In most eukaryotic cells, euchromatin is localized in the nuclear interior, whereas heterochromatin is enriched at the nuclear envelope (NE). This conventional chromatin organization is established by heterochromatin tethering to the NE; however, its importance for cellular homeostasis is largely unexplored. One tether is constituted by the lamin B receptor (LBR) in mammals, but the enigmatic nature of other redundant tethers has hampered functional analyses. Here we demonstrate that downregulation of abundant, ubiquitous NE proteins can induce the global detachment of heterochromatin from the NE and its repositioning to the nuclear interior. We identify LBR and lamina-associated polypeptide 2 (LAP2) as key factors for peripheral heterochromatin positioning in differentiated and pluripotent mammalian cells. Their long-term loss leads to changes in three-dimensional chromatin organization and a reduction in repressive epigenetic marks, especially H3K27me3. These changes are associated with massive deregulation of gene expression, activation of antiviral innate immunity, and defects in cell fate determination.

The pathogenic mechanisms leading to neurodegeneration in Huntington disease (HD) are not fully understood but involve accumulation of toxic mRNA and protein products in the brain. Recent studies described an unconventional autophagic pathway involving DNA and RNA degradation through DNautophagy and RNautophagy that is regulated by the lysosomal protein SID1 transmembrane family member 2 (SIDT2). Interestingly, SIDT2 has been shown to bind to the expanded CAG repeat in the mutant huntingtin (mHTT) transcript and lower mHTT in vitro. The aim of the present study was to determine whether SIDT2 levels are altered in HD and whether manipulation of SIDT2-mediated RNautophagy can alter HD pathology. We demonstrate a significant reduction of SIDT2 protein levels in the striatum and in the lateral hypothalamic area in postmortem HD brains compared to control cases without effects on SIDT2 mRNA levels. In frontal cortical postmortem HD tissue, we show a CAG-repeat-length-dependent increase in the frequency of SIDT2-immunoreactive intranuclear inclusions. In postmortem tissue of an HD case with Vonsattel grade 0, we demonstrate SIDT2- and mHTT-immunoreactive inclusions not only in the frontal cortex, but also in the striatum and the lateral hypothalamic area. In the R6/2 mouse model of HD, we show that SIDT2 inclusions form at later stages than mHTT inclusions. Overexpression of SIDT2 using adeno-associated viral vectors injected into the hypothalamus of R6/2 mice led to a reduction of mHTT inclusions in the lateral hypothalamic area. Similarly, in a neuronal cell model, overexpression of SIDT2 reduced soluble and insoluble mHTT exon 1 protein levels. Taken together, our results reveal novel pathology in clinical HD cases and in experimental models, characterized by the accumulation of SIDT2-immunoreactive inclusions, while demonstrating the efficacy of overexpressing SIDT2 for lowering detrimental mHTT species. Targeting SIDT2-mediated RNautophagy may offer a potential strategy to ameliorate the molecular pathology in HD.

Pyogenic liver abscess is a rare and under-recognized complication of gangrenous or perforated appendicitis in children. Management typically requires a combination of antibiotic therapy and abscess drainage. Although uncommon in the pediatric population, pyogenic liver abscess is more frequently observed in the elderly people. With an aging population, it is increasingly important for young physicians to recognize its clinical presentation and treatment strategies. This report describes an 11-year-old boy admitted with fever, impaired consciousness, nausea, and vomiting-initially raising concern for meningitis. A CT scan revealed a hepatic abscess and findings consistent with acute appendicitis. Following multidisciplinary consultation, the patient underwent percutaneous drainage of the liver abscess and an appendectomy. His postoperative course was uneventful, and he was discharged with a liver drain in place. Given the limited reliability of history and physical examination in pediatric patients with septic shock-especially in emergency settings-clinicians must maintain a broad differential diagnosis.

Microautophagy (MI-autophagy) is an umbrella term for intracellular degradative pathways that entail the invagination or protrusion of the limiting membranes of endolysosomal compartments, that is, late endosomes and mammalian lysosomes or yeast and plant vacuoles, followed by pinching-off of the membrane into the lumen of the organelle. During these processes, the material specifically and nonspecifically targeted for degradation is sequestered within the invaginating or protuberating membrane. In contrast to macroautophagy, the molecular mechanisms underlying MI-autophagy are largely unknown due to their diversity and complexity in location, regulation and molecular machinery requirements. Here, we review recent progress in the field of MI-autophagy, describing the molecular basis and functions of the MI-autophagic pathways reported to date in eukaryotic cells, from yeast to mammalian and plant cells.

In the last few years, gene therapy, holding the promise for long-term disease correction through a one-time treatment, has transitioned from experimental research to approved medicine. Several gene therapies are now available for congenital blood disorders, notably for hemophilia and hemoglobinopathies. In this review, we discuss each of the six therapies that now have regulatory approval for treatment in the United States: Roctavian (valoctocogene roxaparvovec) for hemophilia A, Beqvez (fidanacogene elaparvovec) and Hemgenix (etranacogene dezaparvovec) for hemophilia B, Lyfgenia (lovotibeglogene autotemcel) for sickle cell disease, Zynteglo (betibeglogene autotemcel) for β-thalassemia, and Casgevy (exagamglogene autotemcel) for either sickle cell disease or β-thalassemia. The underlying principles for these treatments vary and include both in vivo and ex vivo methods, lentiviral and adeno-associated viral (AAV) vectors, and gene silencing by the CRISPR-Cas9 gene editing system. Consequently, they pose correspondingly disparate risks and benefits when compared to other treatment modalities and to each other. Although long-term effects are not yet entirely understood, given the novelty of these therapies, knowledge on patient outcomes is continuously increasing. Overall, results are very encouraging, often freeing patients from the need for coagulation factor or red blood cell (RBC) infusions, albeit that for some of these diseases there is room for further improvement in terms of safety and therapeutic durability, which may be achieved with next-generation gene therapy products. However, improvements are needed to address issues with durability of results, side effects, and accessibility of these therapies.

Neurodegenerative diseases are traditionally viewed as age-associated conditions, characterized by distinct biochemical, cellular, and clinical features. However, emerging evidence suggests that their origins may trace back to much earlier stages of life. In this review, we synthesize insights from molecular genetics, developmental neurobiology, and systems neuroscience to examine the hypothesis that selective neuronal vulnerability can arise from developmental misprogramming. We explore how early-life processes-ranging from neurogenesis to synaptic maturation and circuit formation-can imprint long-lasting susceptibilities that manifest as degeneration decades later. Crucially, we highlight that many neurological disorders share early developmental commonalities that may predispose individuals to neurodegenerative vulnerability later in life. This is most apparent in familial forms of these diseases but may also emerge through embryonic or perinatal interactions with environmental or polygenic risk factors. Furthermore, we emphasize the importance of human-specific developmental features, which not only advance our understanding of brain formation but also reveal unique vulnerabilities to neurodegenerative diseases-insights that are increasingly accessible through advances in 3D organoid modeling. Together, these perspectives support a conceptual reframing of neurodegeneration as a late-onset neurodevelopmental disorder. This shift opens promising avenues for early diagnosis, prevention, and precision therapeutics, redirecting focus from late-stage intervention to fostering developmental resilience.

The dynamic lipid modification known as protein palmitoylation is essential for modulating protein activity and subcellular distribution. This process is increasingly recognized as a pivotal molecular mechanism governing the balance between cellular survival and death. This paper explores the molecular regulation of palmitoylation within diverse pathways of regulated cell death, for instance, in necroptosis, ferroptosis, pyroptosis, and apoptosis. The core findings indicate that by controlling the stability, membrane anchoring, and interactions of key signaling proteins, palmitoylation can precisely regulate the ultimate fate of the cell. Additionally, the dysregulation of palmitoylation is closely linked to the pathogenesis of major human diseases, including cancer, neurodegenerative disorders, and inflammatory diseases. For instance, the process can display a dual role in tumor progression, acting to either promote or inhibit it. Concurrently, it is essential for the inflammatory signaling in pyroptosis and for mounting a cellular defense against ferroptosis. A deeper understanding of these regulatory networks provides highly promising therapeutic targets for disease intervention. Targeting the activity of specific palmitoylation-related enzymes has emerged as an innovative strategy for developing novel therapies for a range of diseases, demonstrating significant clinical translational potential.

<h4>Introduction</h4>Bovine viral diarrhea virus (BVDV) can induce diarrhea and mucosal tissue damage, and in severe cases, may result in fatal. Targeted adjuvants capable of enhancing intestinal IgA antibody responses represent an effective strategy for preventing bovine viral diarrhea.<h4>Methods</h4>In this study, an <i>Alhagi honey</i> polysaccharide Pickering emulsion (AHPPE) adjuvant was developed by incorporating <i>Alhagi</i> honey polysaccharide (AHP) and retinoic acid (RA) with an aluminium-based adjuvant. Mice were subsequently immunized via intramuscular injection to evaluate the adjuvants' immunostimulatory potential.<h4>Results</h4>The results demonstrate that the particle size of AHPPE is 2133 nm, with excellent dispersion and stability. The encapsulation efficiencies for BVDV and AHP were 66.8% and 73.2%, respectively. AHPPE demonstrated recruitment of antigen-presenting cells at the injection site and activated IgA cells in the duodenum, jejunum, and ileum, inducing to increased IgA expression across multiple intestinal segments. Furthermore, AHPPE significantly induced serum IgG production and elevated levels of cytokines, including IL-4, IL-10, IL-17, IFN-γ, and TNF-α (<i>P</i><0.05). Through sequencing analysis, it was found that IgA production may be induced via Intestinal immune network for IgA production, thereby mediating intestinal mucosal immune responses.<h4>Conclusion</h4>Collectively, these findings indicate that AHPPE adjuvant administered via injection can simultaneously induce effective systemic immune and intestinal mucosal immunity. Therefore, as a novel intestinal-targeted adjuvant, AHPPE shows potential to enhance the specific intestinal mucosal immunity and systemic immunity of BVDV vaccine.

Although a range of glomerular diseases profoundly affect glomerulus-associated cells, a comprehensive understanding of their molecular alterations is still lacking. Here, we performed in-depth analysis of glomerular data from mouse models of primary and secondary glomerulopathies and constructed a multi-disease cellular landscape of glomerular cells. We identified a putative subset of proliferative glomerular endothelial cells(gECs) that highly expresses genetic susceptibility genes associated with multiple glomerular diseases. Podocytes exhibited shared injury-associated cell types across different disease models. A podocyte subset highly expressing <i>Endou</i>, <i>Cd200</i>, <i>Lgmn</i>, <i>Il18</i>, <i>Dmpk</i>, and <i>Spon2</i> was predominantly derived from ob/ob mice, whereas another podocyte subset with high expression of <i>Selenbp1</i>, <i>Lpar1</i>, <i>S100a8</i>, <i>S100a9</i>, and <i>Sult1a1</i> was mainly observed in adriamycin-induced mice. Mesangial cells shared common injury-related alterations across diseases (high expression of <i>Cxcl1</i>, <i>egr1</i>, <i>hspa1b</i>, <i>socs3</i> and <i>dnajb1</i>), while ob/ob mice exhibited a distinct mesangial cell subset (high expression of <i>aldh1a2, thbs1</i> and <i>fbln5</i>). In contrast, the gECs displayed similar molecular changes across different diseases without giving rise to disease-specific subtypes. Intercellular ligand-receptor analysis underpins the recruitment of immune cells by injured mesangial cells and podocytes via specific engagement of pairs such as CXCL and MIF, respectively. Our study systematically elucidates the molecular alterations of glomerulus-associated cells across various diseases, providing a foundation and strategic insights for future targeted therapies tailored to specific glomerular disease contexts.

Autoinflammatory diseases are characterized by inappropriate activation of innate immunity resulting in excessive or persistent inflammation in the absence of infection. γδ T cells possess innate-like properties, including rapid responsiveness to stress-induced self-molecules, phosphoantigens, and inflammasome-derived cytokines, while retaining adaptive effector functions. Neutrophils and macrophages are well-established drivers of autoinflammatory disease, but increasing evidence implicates γδ T cells as key intermediaries by linking innate immune activation to tissue-specific inflammatory pathology. Here, we review evidence that in both monogenic and multifactorial autoinflammatory diseases-including, for example, familial Mediterranean fever, hyper-immunoglobulin (Ig) D syndrome, gout, Behçet's disease, Still's disease, atherosclerosis, and neurodegenerative disorders-γδ T cells display altered frequencies, activation states, cytokine polarization, and tissue recruitment. In inflammasome-driven diseases, skewing of γδ T cells toward interleukin (IL)-17 production has been observed, often accompanied by reduced interferon (IFN)γ secretion, thereby amplifying neutrophilic inflammation and tissue damage. In other diseases, e.g., Behcet's disease, IFNγ and tumor necrosis factor (TNF)α producton predominate. Transcriptomic and tissue-based analyses support the accumulation and functional specialization of γδ T cells at sites of sterile inflammation. Collectively, these findings position γδ T cells as central amplifiers and modulators of inappropriate innate immune activation in the context of autoinflammatory diseases. Improved understanding of γδ T cell subset-specific regulation may inform novel therapeutic strategies targeting autoinflammatory diseases.

Cleft palate only (CPO) is a multifactorial craniofacial malformation with significant genetic and epigenetic contributions. Among these, microRNAs (miRNAs) have emerged as key regulators of palate development, although their alterations in CPO remain incompletely characterized. In this study, we performed a comprehensive miRNA expression analysis on palatal tissues from an Italian cohort of non-syndromic CPO patients, compared with a human embryonic palatal mesenchymal (HEPM) cell line. Using the NanoString^® nCounter^® platform for miRNA profiling, we identified significant deregulation of several miRNAs, notably the upregulation of miR-205-5p and miR-200c-3p and the downregulation of miR-125a-5p in CPO tissues. Based on these expression changes, a functional analysis was carried out to identify potential target genes. Validation in primary cell cultures derived from patient tissues confirmed these expression patterns. Functional analyses and target predictions implicated PAX9, a key transcription factor essential for palatogenesis, as a probable target of miR-205-5p, while miR-125a-5p was associated with the regulation of PRTG and PRSS35-genes involved in neural crest cell biology and extracellular matrix remodeling, respectively. Although modulation of certain predicted targets of miR-200c-3p was observed, in vitro inhibition experiments did not show significant changes in gene expression, highlighting the complexity of miRNA regulatory networks and the need for further studies to unravel these interactions. These findings identify miRNA alterations associated with CPO tissue and fibroblasts, highlighting novel candidate pathways for further mechanistic and therapeutic investigation.

Phytochemicals, including flavonoids, stilbenoids, alkaloids, terpenoids, and structurally related synthetic small molecules, exhibit a broad spectrum of beneficial pharmacological effects. These effects stem not only from interactions with specific protein targets but also from their capacity to modify the physical properties of biological membranes. A key membrane property influenced by these plant-derived compounds is the electrical potential drop at the membrane-water interface, which plays a crucial role in numerous cellular processes. Changes in membrane potential impact the function of embedded proteins and ion channels, thereby modulating cell signaling, transport, and pharmacological responses. This review compiles data on how diverse plant and synthetic small molecules alter membrane physical characteristics, particularly the dipole component of the boundary potential in lipid bilayers primarily composed of phosphatidylcholine, a predominant membrane lipid in mammals and fungi. In-depth analysis of structure-activity relationships in this context elucidates how various structural modifications affect the compounds' ability to shift membrane electrical potential. Understanding these relationships can pinpoint molecular features that drive membrane interactions and facilitate the discovery and design of more potent dipole-modifying agents with therapeutic potential.

Age-defined type 1 diabetes (T1D) endotypes, T1DE1 and T1DE2, are characterized by reproducible differences in pancreatic immunopathology and clinical course. In particular, these endotypes differ in the extent and composition of lymphocytic insulitis and in the extent of loss of insulin-producing β cell mass, at diagnosis. However, blood-based biomarkers that may distinguish these endotypes and inform the underlying immune–islet biology axis at diagnosis remain limited. Here, we characterized the clinical features and profiled circulating microRNAs (miRNAs) in plasma from two independent INNODIA cohorts of individuals with newly diagnosed stage 3 T1D (discovery, n=115; replication, n=147), stratified into age-defined endotypes (T1DE1, <7 years; T1DE2, ≥13 years; and intermediate T1DInt, 7–12 years). Differential-expression and age-adjusted models were coupled to orthogonal ddPCR validation. Putative miRNAs cellular sources were inferred using reference miRNA expression atlases. Biological context was explored via correlations of miRNAs with whole-blood transcriptomics. Clinically, T1DE1 was associated with lower β-cell function and higher first-year C-peptide decline, alongside distinct islet autoantibody patterns, consistent with an immunologically aggressive endotype. Small RNA-seq analysis and ddPCR validation identified a reproducible signature in which miR-150-5p, a B-and T-lymphocyte related miRNA, and miR-375-3p, a β cell enriched molecule, were consistently increased in T1DE1 compared with T1DE2 across both cohorts. MiR-150-5p retained robust association with T1DE1 even after age adjustment, and neither miRNA was associated with age in non-T1D pediatric datasets, supporting T1D endotype specificity. The increased circulating miR-150-5p signal was not explained by differences in peripheral blood B-or T-cell frequencies in high-parameter flow-cytometry subsets, and its levels correlated inversely with whole-blood expression of the immune-associated miR-150-5p target genes MPPE1 and RABGAP1L. Finally, applying a rule-based combined classifier (miR-150-5p and miR-375-3p “high”) achieved re-stratification of T1D individuals, including those in the intermediate age group, into two miRNA-defined groups with distinct β cell functional trajectories. Collectively, these data suggest circulating miR-150-5p and miR-375-3p as non-invasive biomarkers linked to endotype-associated biology at T1D diagnosis, with potential utility for endotype-centered stratification and trial enrichment.

Redox balance is crucial for maintaining normal physiological functions. Its disruption by oxidative stress can trigger or exacerbate a series of pathological cascades, ultimately contributing to various chronic diseases, particularly inflammatory disorders. Inhibiting oxidative stress and its associated pathological cascades may alleviate these diseases, a process often linked to the activation of nuclear factor erythroid 2-related factor 2 (Nrf2). Initially characterized as a redox-sensitive transcription factor, Nrf2 is now recognized as a pivotal regulator of an extensive network of antioxidant genes, effectively counteracting oxidative stress and its detrimental effects. Consequently, advances in understanding Nrf2 activators and their regulatory mechanisms have accelerated the development of Nrf2-targeted therapies, demonstrating significant potential for preventing and treating chronic inflammation diseases. Many natural phytochemicals, particularly flavonoids, have been identified as Nrf2 activators that can ameliorate inflammatory responses. Furthermore, therapy with mesenchymal stromal/stem cells (MSCs) is a highly researched treatment approach with the potential to confer immunomodulatory, anti-inflammatory, anti-apoptotic and antimicrobial effects. Owing to their superior safety profile compared to conventional therapeutics, MSCs are gaining prominence as sustainable long-term treatment options, although their precise molecular mechanisms remain to be fully elucidated. This review focuses on the activation mechanisms of Nrf2 and its clinical and preclinical inducers, with particular emphasis on the mechanistic insights and therapeutic applications of natural flavonoids and MSCs in the prevention or treatment of inflammatory diseases. More importantly, it summarizes the profound role of flavonoid-MSCs combinatorial therapy in the intervention of inflammatory diseases, pointing out novel therapeutic strategies and future prospects for modulating the Nrf2 signaling pathway in the treatment of inflammatory disorders.

The neurotoxicity of emerging contaminants, polyhalogenated carbazoles (PHCZs), is elusive. In this study, we investigated the potential toxicity of 13 prevalent PHCZs utilizing a network toxicology approach, which revealed shared molecular targets associated with Parkinson's disease (PD). Molecular docking simulations assessed the binding affinities of these PHCZs for eight key PD-related targets, identifying monoamine oxidase B (MAOB) as a critical target. Among the dihalogenated PHCZs, 2,7-dibromocarbazole (2,7-BCZ) exhibited the highest binding affinity to MAOB. Comparative molecular docking and dynamics simulations suggest that the inhibition of MAOB activity by 2,7-BCZ is a potential initiating event in PHCZ-induced neurological disorders. <i>In vivo</i> experiments confirmed that 2,7-BCZ exposure highly correlates with α-synuclein aggregation, a hallmark of PD pathology. Transcriptomic sequencing of 2,7-BCZ-exposed SH-SY5Y cells, combined with analysis of public PD microarray data, identified shared transcriptional alterations in genes including <i>CLSTN2</i>, <i>CBLN1</i>, <i>AGTR1</i>, <i>DLK1</i>, and <i>DDC</i>. By integrating pathways from PD-related targets of PHCZs, differentially expressed genes in 2,7-BCZ-exposed cells, and public PD data sets, we further elucidated key biological pathways through which 2,7-BCZ may contribute to PD pathogenesis, particularly dopaminergic synapse function and neurodevelopmental regulation. Collectively, this study not only highlights the potential role of PHCZs in PD, elucidating the potential biological mechanisms by which PHCZs may exacerbate PD, but also exemplifies an innovative, animal-sparing approach using New Approach Methodologies (NAMs) to assess environmental pollutants' risks in neurodegenerative adverse outcomes.

Huntington's disease (HD) is a neurodegenerative disorder caused by the expansion of CAG repeats in the gene encoding huntingtin. Since accumulation of mutant huntingtin (mHtt) leads to dysfunction of numerous cellular pathways and toxicity, reducing levels of the mutant protein represents a key therapeutic objective in HD. We found that ubiquitination of mHtt by E3 ubiquitin ligase Nedd4 promotes clearance of the mutant protein. Knockdown of Nedd4 increased toxicity of mHtt in mouse primary neurons and in a fly model of HD, suggesting the protective role of Nedd4. Importantly, levels of Nedd4 were decreased in mHtt-expressing neurons through impaired mTORC1 activity, suggesting a feedback loop of mHtt accumulation and Nedd4 reduction that leads to accumulation and, ultimately, toxicity of mHtt. These findings suggest that restoring Nedd4 activity may offer a novel therapeutic opportunity for HD.

Gold nanoparticles (AuNPs) have attracted significant attention due to their broad range of applications in the health and environmental fields and the fact that gold is an inert metal with low physiological toxicity, making it a favorable candidate for biomedical use. In line with the growing interest in green nanotechnology, plant-based nanoparticle synthesis has emerged as a sustainable, low-cost, and environmentally friendly alternative to conventional methods. Among these, AuNPs synthesized using plant extracts have been extensively studied, particularly concerning their antimicrobial properties. In this sense, this study aimed to perform a bibliometric review of the antimicrobial potential of plant-based AuNPs focusing on articles published between 2014 and 2025. Searches were conducted at the Web of Science, PubMed, and Scopus databases, totaling 190 eligible studies. India is the leading country in terms of number of publications (36.84%), with leaves as the most employed plant part (42.6%). The reported AuNPs synthesized from various plant parts, including seeds, fruits, as well as leaves, exhibited significant antimicrobial activities. Although some AuNPs, such as those synthesized from seeds, were more effective against Gram-positive bacteria, others, such as those from fruits, showed greater activity against Gram-negative strains, and several leaf-based ones demonstrated broad-spectrum efficacy against both groups. These antimicrobial performance variations are primarily attributed to the presence of specific organic compounds like flavonoids, terpenoids, and phenolics unique to each plant species used in the synthesis process. Escherichia coli and Staphylococcus aureus were the most frequently tested bacterial strains, with main AuNPs effects comprising damage to their thiol and peptidoglycan cell walls. One hundred thirty three (Emmanuel et al. in Microb Pathog 113:295-302, 2017) articles expanded their evaluations to include fungi, protozoa and cytotoxic effects, with many reports encouraging results against antibiotic-resistant bacterial strains, reinforcing the potential of plant-based AuNPs as innovative antimicrobial therapy alternatives.

Early spring sowing of maize in semi-arid, wind-eroded regions is increasingly threatened by cold snaps due to climate change.These events, often coupled with uneven soil moisture distribution,compromise seedling emergence and early development. Identifying critical temperature and moisture thresholds is essential to ensure successful germination in these vulnerable environments.A factorial experiment was conducted in a controlled environment using maize seeds (Zea mays L.) exposed to diurnal temperature cycles.Treatments included five minimum temperatures (0,2,4,6,8°C), three chilling durations (2,4,6 hours),and four soil moisture levels (60,70,80,90% field capacity). Key germination metrics,including final germination rate, weighted germination time,synchrony,delay days,and seedling dry matter at day 30,were measured and analyzed using three-way ANOVA and Pearson correlations. Temperatures below 6°C significantly delayed germination and reduced final germination rates,particularly under low moisture conditions.Moisture levels ≥80% effectively mitigated chilling effects at moderate temperatures(4 ~ 6°C).Extended chilling durations further suppressed germination.The strongest interaction was observed between minimum temperature and soil moisture.Seedling dry matter accumulation was also significantly affected by all three factors and their interactions.Soil moisture serves as a critical buffer against chilling stress during maize germination. This study provides quantitative benchmarks for temperature and moisture combinations that optimize early maize emergence under extreme spring weather, offering practical insights for precision moisture management in semi-arid agriculture.

Major depressive disorder (MDD), bipolar disorder (BPD), and attention-deficit hyperactivity disorder (ADHD) are prevalent, highly heritable psychiatric disorders with significant degrees of genetic overlap. Using open-sourced summary statistics from the Psychiatric Genomics Consortium and 1000 Genomes European reference panel, we fit a latent factor model (F1) capturing the shared genetic liability across MDD, BPD, and ADHD. Multivariate GWAS identified 350 linkage disequilibrium-independent loci, 105 of which have not been previously reported by the contributing univariate GWASs. Univariate, bivariate, and trivariate mixture models elucidated both shared and trait-specific polygenicity across the disorders. Gene-level analysis with Hi-C coupled MAGMA identified a total of 2936 novel dopaminergic associations across the constituent disorders that went undetected in univariate analyses. Among the top genes associated with F1, protein tyrosine phosphatase receptor type D emerged as a promising candidate. Cell typing and brain tissue enrichment for F1 further implicated the cerebellum and cholinergic neurons. These findings demonstrate how multivariate approaches can elucidate shared biological mechanisms, providing new etiological insights into individual disorders and implicating therapeutic targets for the treatment of psychiatric comorbidity.

Catechol-modified polymers, such as DOPA-functionalized systems, have recently gained significant interest for a variety of biomedical applications, particularly in their role as antibacterial adjuvants due to their oxidative activity and ability to generate reactive oxygen species (ROS). Current catechol-functionalized polymers, however, often suffer from a restricted number of catechol groups, limited biocompatibility and solubility, and low stability due to the rapid oxidation under physiological conditions. In this study, we developed a water-soluble, biocompatible DOPA-modified biodynamer (DOPA-BD), leveraging the principles of constitutional dynamic chemistry (CDC). DOPA-BD was synthesized via polycondensation of DOPA-hydrazide and the hexaethylene glycol-conjugated carbazole dialdehyde (CA-HG), forming dynamic imine and acylhydrazone linkages between the monomers. As a result of its dynamic covalent backbone, DOPA-BD exhibits biodegradability and undergoes pH-responsive degradation under mildly acidic conditions typically found at infection sites, leading to a more than 3-fold increase in DOPA-hydrazide release compared to physiological pH. Interestingly, driven by CDC, DOPA-BD folds into a nanorod structure with a hydrodynamic diameter of ∼7.8 nm, surrounded by HG chains that offer water solubility and biocompatibility. Moreover, the incorporation of the DOPA-derivative in each repeating unit yields a polymer with exceptionally high catechol content, which remains stable and resistant to oxidation for 72 h in physiological buffer conditions. Regarding its antibacterial applicability, DOPA-BD demonstrated synergistic antibacterial activity with Azithromycin (AZM) against AZM-resistant <i>E. coli</i>, enhancing the antibiotic's efficacy by 4-fold. Our study indicates that DOPA-BD induces ROS production in the respective bacterial strain, suggesting ROS generation as one of the possible mechanisms contributing to the observed synergy. Overall, DOPA-BD represents a promising alternative strategy to potentiate antibacterial activity against resistant strains, holding strong potential for future antibacterial applications.

Sactipeptides are a class of natural product peptides with remarkable antibiotic properties that are defined by the presence of thioaminoketals in their structure. Recently, we reported the first total synthesis of a sactipeptide in our synthesis of enteropeptin A. The key to our synthesis involved the use of a dithiophosphoric acid catalyzed Markovnikov hydrothiolation of dehydroamino acids. With this reaction, thioaminoketals found in sactipeptides can be prepared directly from a dehydroamino acid and a cysteine residue. This article summarizes our initial approach toward enteropeptin synthesis and the evolution of our strategy that ultimately enabled the synthesis of these peptide natural products. Our first strategy involved late-stage Markovnikov hydrothiolation of an 8-mer peptide containing a dehydroamino acid and a cysteine residue that was unsuccessful. The second strategy involved an annulation reaction between a methyl ester of a dehydroamino acid and a cysteine with an unprotected amine that forged the central thiomorpholine ring albeit in low yield. The third strategy involved a divergent synthesis of the enteropeptins by early stage formation of the thiomorpholine ring by Markovnikov hydrothiolation followed by amidative coupling of the N- and C-terminal peptide fragments. This modular strategy enabled the unified synthesis of the enteropeptin sactipeptides.

Neuronal maturation involves a tightly regulated cessation of growth and acquisition of polarity, ultimately leading to synapse formation. While essential for circuit stability, maturation marks the loss of regenerative capacity after central nervous system (CNS) injury. The molecular programs coupling maturation to regenerative decline remain incompletely understood. Here, we show that the transcriptional and epigenetic signatures enabling axon growth in dorsal root ganglion (DRG) neurons are lost as they transition from immature, non-polarized cells to mature, pseudo-unipolar neurons. We identify the transcriptional co-regulator CITED2 as a key epigenetic switch, active in immature and regenerating DRG neurons but silent after non-regenerative spinal cord injury (SCI). Cited2 overexpression reactivates growth programs, enhancing regeneration in vivo after SCI. Mechanistically, CITED2 reinstates developmental epigenetic and transcriptional profiles, decoupling maturation from regenerative failure. Pharmacogenomic screening identified CITED2 as a target of the clinically approved HDAC inhibitor Panobinostat, which promoted axonal growth, sprouting, and functional recovery post-injury. These findings position CITED2 as a key regulator of sensory neuron plasticity and a novel therapeutic target for CNS repair.

BACKGROUND: The inner dynein arm (IDA) plays a crucial role in regulating ciliary and flagellar beating; however, the molecular mechanisms underlying IDA regulation remain largely unclear. METHODS: Whole-exome sequencing (WES) was performed to identify candidate pathogenic variants in an infertile man with asthenoteratozoospermia. A knockout mouse model was generated using CRISPR/Cas9 to investigate the pathophysiological effects of the identified variant. Phenotypic characterization included semen parameter analysis (e.g., sperm concentration, motility), morphological assessments (electron microscopy, immunostaining, histology), and mechanistic studies (e.g., scRNA-seq, proteomics, co-immunoprecipitation, mass spectrometry, computational prediction). RESULTS: A homozygous missense variant in DNAH3 was identified in the patient. DNAH3, an IDA component, is predominantly expressed in post-meiotic cells, specifically within the sperm flagella. Notably, Dnah3 knockout (Dnah3−/−) mice exhibited abnormal flagellar morphology and male infertility. Immunofluorescence and transmission electron microscopy revealed that the absence of IDAs in Dnah3−/− male mice. Additionally, scRNA-seq analysis indicated that Dnah3 deficiency in elongating spermatids significantly alters the expression of genes related to sperm motility. Combined comparative proteomics and co-immunoprecipitation analyses demonstrated that DNAH3 forms a complex with DNALI1. Furthermore, DNAH3-associated male infertility in human and mice was resolved by intracytoplasmic sperm injection (ICSI). CONCLUSIONS: Our findings suggest that the interaction between DNAH3 and DNALI1 is critical for flagellum assembly. DNAH3 is a candidate gene for the genetic diagnostic of severe asthenoteratozoospermia and primary male infertility.

Alzheimer’s Disease represents the most significant form of neurodegenerative disease worldwide with progressive dementia and synaptic dysfunction. Though the accumulation of β-amyloid and hyperphosphorylated tau protein is the most observed pathological feature of AD, the emergence of ribosomal dysfunction and oxidative stress has recently gained interest. In this study, we conducted a comprehensive multi-omics investigation, which included transcriptomic, proteomic, and lipidomic analyses, on cortical region from 5xFAD mice, a transgenic model of AD. Gene and protein expression analysis demonstrated ribosomal pathways were largely affected in the cortex. Histological and immunohistochemistry imaging showed increased amyloid-β and tau pathology leading to extensive cortical neurodegeneration. RNAseq analysis revealed increased oxidative RNA damage, indicating a potential mechanism of ribosomal stress. Elevated expression of RPL11, RPL6, and other large ribosomal subunit proteins was observed, consistent with impaired protein synthesis. This dysregulation may contribute to neurodegenerative processes in AD. Among the large subunit ribosomal proteins, Rpl29 were downregulated at the gene expression level in AD, although its protein expression revealed a statistically insignificant rise. Comparison between the transcriptome and proteome demonstrated evidence of impaired translation, suggesting failed translational control. Lipidomic analyses revealed alterations in the levels of phospholipids, sphingolipids and lipid mediators in AD that is closely linked to the alterations in the neuroinflammatory pathways at the transcriptomic and proteomics levels. Multi-omics integration demonstrates that ribosomal dysregulation, oxidative stress, and protein homeostasis are affected, leading to neuronal damage in AD. According to this study, ribosomal malfunction plays a significant role in the pathophysiology of AD and serve as a potential target for therapeutic interventions.

<h4>Background</h4>Skewed myelopoiesis in the bone marrow has been identified as a key driver of multiple sclerosis (MS) progression. Interestingly, SARS-CoV-2 infection, which has a severe impact on MS patients, can also induce similar skewed myelopoiesis. This shared phenotype raises the question of whether a common mechanism underlies the skewed myelopoiesis in both diseases. Previous studies in mice have demonstrated that the dysregulation of endogenous retroelements (EREs) in HSPCs leads to skewed myelopoiesis. Building on this, we sought to determine whether ERE dysregulation contributes to the skewed myelopoiesis observed in MS and after COVID-19, which remains challenging.<h4>Methods</h4>We undertook a joint investigation of two public single-cell/nuclei cohorts respectively representing MS and following COVID-19. Both cohorts were processed through an identical bioinformatic pipeline to ensure comparable assessment of gene and ERE expression.<h4>Results</h4>We observed enhanced myelopoiesis in the bone marrow of MS patients compared to healthy controls, along with downregulation of the ERE repressor H3.3 and concomitant EREs overexpression. Notably, a similar epigenetic and transcript feature was found in post-COVID-19 individuals.<h4>Conclusion</h4>The H3.3^low/ERE^high signature may not only explain the common skewed myelopoiesis in MS and post-COVID-19 conditions, but also provide a mechanistic link between infection and the innate immune reprogramming that drives MS progression. This offers a novel therapeutic insight for MS.

Metformin is an antidiabetic drug that has been tested widely as an anti-cancer agent. However, data from clinical trials have been mixed. Evidence for metformin's efficacy in HER2+ breast cancer exists. Hence, we evaluated whether WBP2, a HER2-coamplified gene, can regulate the response of HER2+ breast cancer to metformin. Identification of biomarkers for predicting metformin response has implications in repurposing metformin for precision oncology. The effect of WBP2 on breast cancer response to metformin was studied using in vitro and mouse models. The mechanism of WBP2 on metformin-induced AMPK activation was elucidated, and its co-expression with p-AMPK was examined in clinical specimens using IHC. RNA-seq analyses were performed to elucidate WBP2's mechanism in energy metabolism. WBP2 inhibited the metformin response of HER2+ breast cancer in vitro and in vivo. These effects were concomitant with WBP2-mediated repression of metformin-induced AMPK activation and mTOR inhibition in HER2+ breast cancer cells, a lower AMP:ATP ratio state, and enhanced glycolytic capacity and mitochondria respiration. Analysis of HER2-positive breast cancer samples supports the negative correlation between WBP2 expression and activated AMPK observed in vitro. RNA-seq analysis revealed the potential mechanism of WBP2 in regulating ATP production processes and preferential effect of WBP2 on metformin response in HER2+ breast cancer. This study reported a novel role of WBP2 in cancer metabolism and energetics that contributes new insights into the molecular etiology of cancer. WBP2 may be a biomarker for patient stratification, paving the way towards repurposing metformin for precision oncology.

The recognition of microRNAs as components of animal-source foods (ASFs) with epigenetic characteristics and regulation has spurred research in an interesting direction, particularly in understanding their microRNAs (miRNAs) fraction. Thus, a constant supply of them through food intake, with equally conserved targets, may facilitate their accumulation in tissues rich in their targets. Here, we consider the potentially dominant miRNAs in animal-source foods (ASFs) documented in the literature, identified through a frequency-weighted ordinal recurrence approach. <i>let</i>-7d-5p, miR-101-3p, and miR-133b consistently showed dominant rankings in a product-specific manner in lean meat. In meat fat, let-7i-5p, miR-30c-5p, and miR-23a-3p were highly ranked. Among various types of meat offal, miR-145-5p, miR-92-5p, and miR-24-3p emerged as the predominant miRNAs. Similarly, in dairy products, miR-200a-3p, miR-200c-3p, miR-223-3p, miR-25-3p, miR-29a-3p, and miR-29b-3p were recurrently dominant, whereas miR-17-5p, miR-184, miR-30e-5p, and miR-92b-3p showed a comparable prevalence in seafood. Even though bioinformatic approaches suggest miRNAs from raw ASFs showed major enrichment of processes and pathways culminating in epithelial barrier integrity modulation, such putative functions tend to be equally enriched by predicted targets of the miRNAs in processed products. Product-specific highly ranked miRNAs from food categories stipulate possible preferential enrichment in contexts of cell-cell adhesion, cytoskeletal dynamics, and inflammatory control by meat (lean, fat, offal), immune homeostasis by dairy, and neural signalling by seafood, providing hypotheses for future functional studies. However, a limited understanding of their stability during gastrointestinal transit may present a more immediate limitation to their potential translational applicability.

Pancreatic ductal adenocarcinoma (PDAC) shows substantial heterogeneity in cysteine dependence and ferroptosis sensitivity. We identify two PDAC subtypes distinguished by EMT status: mesenchymal-like cells are highly cysteine-dependent and rapidly undergo ferroptosis upon cystine deprivation or system xc^- inhibition, whereas epithelial-type cells are ferroptosis-resistant. Selenium supplementation protects cells from erastin-induced ferroptosis, and this protection persists even when intracellular glutathione (GSH) is depleted, supporting an additional GPX4-independent protective mechanism. Sepp1 knockdown does not alter sensitivity, indicating that selenium's protective effect is independent of Sepp1. Instead, epithelial-type cells rely on both cytosolic and mitochondrial thioredoxin reductases (TrxR1 and TrxR2) to maintain ferroptosis resistance. Chemical inhibition of thioredoxin reductases abolishes selenium-mediated protection and sensitizes epithelial cells to ferroptosis inducers, while dual genetic suppression of TrxR1 and TrxR2 similarly restores ferroptosis sensitivity. These findings uncover a selenium-thioredoxin redox axis that functions independently of GPX4 and contributes ferroptosis resistance in epithelial-type PDAC cells. Co-targeting cysteine metabolism and thioredoxin reductases may therefore represent a rational strategy to overcome ferroptosis resistance in some PDAC subtypes.

Spinal cord injury (SCI) is a devastating neurological disorder that can result in permanent disability and reduced quality of life, characterized by heterogeneous injury mechanisms and limited tools for accurate early diagnosis and prognostic stratification. The clinical course of SCI is driven not only by the initial mechanical insult but also by complex secondary injury cascades involving neuroinflammation, axonal degeneration, demyelination, and maladaptive repair responses. Current diagnostic and prognostic approaches, which rely largely on neurological examination and imaging, provide limited insight into these dynamic molecular processes. In this context, extracellular vesicles (EVs) have emerged as a biologically compelling source of biomarkers for SCI. EVs are released by neurons, glial cells, endothelial cells, and immune cells and carry molecular cargo that reflects cellular stress, injury severity, and endogenous repair activity. Increasing evidence indicates that EV-associated proteins and regulatory microRNAs (miRNAs) encode injury-specific signatures related to neuronal and glial damage, inflammatory signaling, metabolic stress, and functional recovery potential. In this review, we summarize the current knowledge on EV biology in SCI and discuss emerging evidence supporting EV-derived proteins and miRNAs as promising tools for refining diagnosis and prognosis. Our aim is not only to consolidate established findings but also to highlight EV-based molecular signatures as a developing framework for precision biomarker discovery in SCI.

<h4>Background</h4>Liver metastasis accounts for most deaths in colorectal cancer patients, but we still lack a clear understanding of how this devastating process unfolds at the molecular level. Identifying these mechanisms could lead to better ways to predict and treat metastatic disease.<h4>Methods</h4>We performed proteomic analysis on 38 tissue samples, including benign adenomas, primary colorectal tumors, and liver metastases, to map the molecular changes that drive cancer spread to the liver.<h4>Results</h4>We found striking differences in protein expression patterns across tissue types. Primary tumors showed massive protein changes compared to normal tissue (nearly 1,900 alterations), but liver metastases were surprisingly selective in their modifications (just 89 unique changes from primary tumors). This specificity indicates that liver colonization depends on precise molecular adjustments rather than random cellular disruption. The proteins involved in metastasis fell into three main categories: tissue restructuring to support tumor growth, mechanisms to avoid immune detection, and blood clotting factors that help cancer cells establish themselves in the liver. We validated three biomarkers with clinical potential: HDHD3 shows promise for detecting existing metastases, while OLFM4 and ZNF34 can help identify primary tumors at high risk for spreading.<h4>Conclusions</h4>Our work reveals that colorectal cancer liver metastasis involves targeted molecular changes rather than widespread cellular chaos. The biomarkers we identified could improve early detection of metastatic risk and guide more personalized treatment decisions for patients.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is the most common subtype of non-small cell lung cancer (NSCLC) and has high mortality rates. However, practical strategies for guiding clinical therapies for LUAD are still lacking. This study aimed to analyze the expression profiles and mutation features of 20 m6A (N6-methyladenosine) regulators in LUAD patients. It also systematically explored the biological roles of these m6A methylation regulators and their links to tumor immunity in LUAD, ultimately providing a theoretical basis for clinical treatment approaches.<h4>Methods</h4>RNA sequencing data for 20 m6A methylation regulators and clinical information for LUAD patients were sourced from the Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) databases. The relationship between insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) and immune cell infiltration in LUAD was analyzed using CIBERSORT. The "GSVA" R package (version 1.38.2) was employed to perform Gene Set Variation Analysis (GSVA). The protein-protein interaction (PPI) network of these m6A-related genes was built using the STRING database. The Tumor Immune Dysfunction and Exclusion (TIDE) algorithm was applied to predict clinical responses to immune checkpoint inhibitors, while the oncoPredict R package evaluated chemotherapeutic responses. We collected clinical specimens to validate Kaplan-Meier survival analysis and used immunohistochemistry to differentiate between high- and low-expression groups.<h4>Results</h4>Sixteen m6A modification regulators showed significant abnormal expression in LUAD tissues. Univariate Cox and Least Absolute Shrinkage and Selection Operator (LASSO) logistic regression analyses revealed that IGF2BP1 was the only independent predictor of LUAD after adjustment for common clinical markers. The mutation rates of m6A modification regulators in LUAD were below 10%. Further studies demonstrated that IGF2BP1 expression was strongly associated with immune infiltration, immune checkpoint expression, the effectiveness of immunotherapy in LUAD patients, and the incidence, progression, metastasis, and treatment resistance of lung adenocarcinoma. Additionally, patients with high IGF2BP1 expression had worse prognoses. We developed a nomogram combining IGF2BP1 expression with five other predictive risk factors. The ROC and calibration curves showed that the nomogram was well-calibrated and effectively distinguished between high- and low-expression LUAD patients. The results from the clinical validation cohort were consistent with these previous analyses.<h4>Conclusions</h4>Our findings suggest that the m6A modification influences the tumor microenvironment and that IGF2BP1 acts as an independent predictor of immunotherapy response in LUAD. It may serve as an innovative biomarker for LUAD prognosis and tumor immunity status.

Once relegated to the background of striatal circuitry, parvalbumin-expressing interneurons are now emerging as central players in health and disease. Acting as true gatekeepers, striatal PV interneurons are well-described for their role in synchronizing striatal output and balancing excitation and inhibition to sustain coordinated motor and cognitive functions. In this review, we highlight recent advances in understanding their developmental origins, molecular identity, physiological properties, and their roles in striatal function. Furthermore, we examine converging evidence implicating PV interneurons in Huntington's disease and Autism Spectrum Disorder, where their structural, molecular, and functional alterations position them at the intersection of neurodegenerative and psychiatric research.

<h4>Background</h4>Direct Oral Anticoagulants (DOACs) and antidepressants are often co-administered, but their combined effect on bleeding risk in real-world settings is poorly quantified. This study aimed to evaluate this risk using pharmacovigilance data.<h4>Methods</h4>Data spanning quarter 3 (Q3) 2010 to quarter 1 (Q1) 2025 were analyzed from the Food and Drug Administration Adverse Event Reporting System (FAERS) database, comparing bleeding adverse events reported for DOACs monotherapy versus DOACs combined with antidepressants. Bleeding signals were assessed using the Reporting Odds Ratio (ROR).<h4>Results</h4>The proportion of bleeding events was 31.00% (64,165/207,000) in the DOACs monotherapy group versus 57.92% (307/530) in the DOACs-antidepressant combination therapy group. Concomitant use of DOACs with antidepressants was associated with a significant increase in overall bleeding risk (reporting odds ratio [ROR] = 1.45, 95% confidence interval [CI]: 1.29-1.63). Selective serotonin reuptake inhibitors (SSRIs) presented the highest class risk (ROR 1.78, 95% CI 1.54-2.04), with apixaban plus paroxetine showing the strongest signal (ROR 14.12, 95% CI 7.62-26.15). Nervous system bleeding was also elevated (ROR 1.86, 95% CI 1.44-2.40). Notably, mirtazapine significantly increased nervous system bleeding risk (ROR 9.83, 95% CI 4.92-19.67) despite its non-SSRI mechanism.<h4>Conclusion</h4>Co-administration of antidepressants and DOACs significantly elevates bleeding risk, especially for the nervous system. Clinicians must exercise heightened caution, particularly with SSRIs and when using mirtazapine, and further validation studies are needed.

<h4>Introduction</h4>Cerebral Palsy (CP) is characterized by permanent, non-degenerative motor function deficits with increasing evidence of genetic contributions. Although prenatal and perinatal risk factors are well recognized, the underlying etiopathology remains incompletely understood. This study aimed to improve diagnostic accuracy and elucidate the genetic architecture of CP and CP-like phenotypes through systematic genomic analyses.<h4>Methods</h4>Patients with clinically confirmed CP or CP-like presentations were recruited, and biological samples were stored in the ACU-Biobank. Whole-exome and whole-genome sequencing data were analyzed using a validated in-house pipeline incorporating comprehensive variant filtering, prioritization, and re-phenotyping.<h4>Results</h4>Pathogenic or likely pathogenic variants were identified in 36.4% (24/66) of patients, while variants of uncertain significance (VUS) were detected in 25.8% (17/66). Identified variants involved genes such as <i>SPAST</i>, <i>KIF1A</i>, <i>PLA2G6</i>, <i>CTNNB1</i>, <i>L1CAM</i>, and <i>SYNGAP1</i>. These results demonstrate a substantial contribution of rare monogenic variants to CP and CP-like phenotypes, reflecting extensive genetic heterogeneity.<h4>Discussion</h4>Our findings support the increasing evidence that genetic factors contribute significantly to CP etiology and emphasize the importance of integrating genomic testing into clinical evaluation. The systematic use of exome and genome sequencing improves diagnostic yield and enables genotype-informed classification, aiding targeted management and genetic counseling for affected individuals.

Chimeric antigen receptor (CAR)-engineered immune cells have revolutionized cancer immunotherapy, expanding from the established success of CAR-T cells to a diverse array of cellular platforms. While seven Food and Drug Administration-approved CAR-T cell products demonstrate unprecedented efficacy in hematologic malignancies, significant limitations persist, including severe inflammatory toxicities, resistance in solid tumors, and manufacturing barriers. These challenges have catalyzed extensive research to extend CAR engineering into alternative effector cell types, such as unconventional T cell subsets, natural killer (NK) cells, macrophages, neutrophils, and dendritic cells, as well as non-immune platforms. Each cell type exhibits distinct antitumor mechanisms, persistence profiles, safety characteristics, and manufacturing requirements, positioning them to address complementary therapeutic needs. This review provides a comprehensive overview of diverse CAR-engineered cellular platforms, encompassing their biological properties, advantages, sourcing strategies, and manufacturing processes, alongside current clinical progress and optimization approaches. Beyond oncology, these platforms have demonstrated significant potential in treating autoimmune diseases, infections, cardiac fibrosis, and senescence-associated disorders. By leveraging distinct immune and non-immune cell types to mediate cytotoxicity or suppress pathogenic cells, CAR technology provides versatile therapeutic avenues across varied disease contexts. Through synthesis of recent advances in CAR platform diversity, this review identifies opportunities for targeted optimization and explores future directions for broadening CAR-based therapeutic applications.

Beyond their lipid-lowering effects, statins exert pleiotropic properties, including anti-inflammatory, antioxidant, and antifibrotic actions, which may confer therapeutic benefits in patients with metabolic dysfunction-associated steatotic liver disease (MASLD). This systematic review and meta-analysis aimed to evaluate the impact of statin therapy on clinical outcomes in adults with MASLD. A comprehensive literature search was conducted across PubMed, Embase, Cochrane CENTRAL, Web of Science, and Scopus from inception through January 10, 2026. Observational studies and randomized controlled trials comparing statin therapy with placebo or no treatment in adults with MASLD were eligible for inclusion. Eight observational studies met the inclusion criteria and were included in the quantitative synthesis. The primary outcomes were hepatocellular carcinoma incidence, all-cause mortality, and liver-related mortality. Statin use was associated with a significantly lower risk of hepatocellular carcinoma (pooled risk ratio (RR) 0.56, 95% CI 0.45-0.71), all-cause mortality (pooled RR 0.81, 95% CI 0.72-0.91), and liver-related mortality (pooled RR 0.54, 95% CI 0.37-0.79). Moderate to substantial heterogeneity was observed across studies. These findings suggest that statin therapy may confer meaningful hepatic and survival benefits in patients with MASLD, supporting their proactive use, particularly given their concomitant cardiovascular advantages. Nonetheless, the observational nature of the included studies underscores the need for large-scale randomized controlled trials to establish causality and define optimal treatment strategies.

Hepatocellular carcinoma (HCC) is characterized by substantial heterogeneity and immune tolerance, and its therapeutic efficacy is profoundly influenced by the immune microenvironment. Ferroptosis, an iron-dependent form of regulated cell death, is closely linked to tumor immune regulation. We conducted an integrative multi-omics analysis to systematically delineate the composition and function of the ferroptosis-immunity network in HCC. Using TCGA data, we identified ferroptosis-related prognostic genes. By integrating these with immune features via weighted gene co-expression network analysis (WGCNA), we defined 55 ferroptosis-immune microenvironment-related genes (FIMRGs). Single-cell transcriptomic analysis showed that, among immune cell types, macrophages exhibited the highest ferroptosis-immunity activity. Spatial transcriptomics revealed that macrophages with high ferroptosis signaling (FehighMac) preferentially infiltrated tumor nests. These macrophages engaged in robust interactions with T cells via ligand-receptor axes such as ICAM1-ITGAX/ITGB2, TNF-TNFR, and LGALS9-CD45, potentially promoting T-cell exhaustion and shaping an immunosuppressive microenvironment. We identified CAPG-positive macrophages (CAPG + Mac) as the key driver of this process. Characterized by suppressed ferroptosis, enhanced glutathione metabolism, and upregulated immune checkpoints, CAPG + Mac appear to foster an immune-tolerant microenvironment. A prognostic model constructed from CAPG + Mac signature genes effectively stratified HCC patients into high- and low-risk groups and demonstrated stable predictive performance in external validation. This study reveals that CAPG + Mac putatively modulate ferroptosis signaling to influence immune homeostasis, highlighting them as a key target for HCC progression and immunotherapy responsiveness.

Adolescent major depressive disorder (MDD) involves complex and heterogeneous alterations in brain functional organization. A possible explanation is the disruption of individualized brain maturation during adolescence, which typically transitions from lower-order sensory to higher-order association regions. To test this hypothesis, we analyzed resting-state functional MRI data using advanced matrix decomposition techniques to extract organizational gradients, temporal dynamics, and information flow. Clustering sensory-association gradient features in our main sample (N_MDD = 302 and N_Controls = 207; ages 11-17) revealed two subtypes. Subtype 1 showed bottom‑up (sensory‑to‑association) flow, reduced synergy but heightened redundancy in sensory cortices, and a highly modular yet globally inefficient network-patterns consistent with aberrant sensory processing and impaired integration. Subtype 2 instead displayed a top‑down (association‑to‑sensory) flow, reduced synergy in association areas, elevated redundancy in sensory regions, and network efficiency between subtype 1 and healthy controls, suggesting compensatory higher‑order engagement. Subtypes also differed in age-related changes, clinical profiles, and neurobiological processing. These patterns were largely replicated in an independent sample (N_MDD = 73 and N_Controls = 28). Overall, the sensory-association axis distinguishes MDD subtypes, integrating functional heterogeneity, cortical dynamics, developmental trajectories, and genetic influences, offering new insights for pathophysiology and precision psychiatry.

BACKGROUND: T/myeloid mixed-phenotype acute leukemias (MPAL-T/M) is a type of rare and high-risk acute leukemia that carries both T- and myeloid- lineage markers. The diagnosis of MPAL requires integration of clinical, immunophenotypic, and genetic information. Optical Genome Mapping (OGM) technology is a particularly powerful tool to profile genome-wide variants and locate complex chromosomal rearrangements that may guide diagnosis and risk stratification in MPAL-T/M. CASE PRESENTATION: We report two MPAL-T/M cases with comprehensive clinical, immunophenotype, and genetic results. A shared chromosomal rearrangement between chromosomes 14q32 and 16q24, t(14;16)(q32;q24), was detected in both leukemias. The t(14;16) has breakpoints 500-900 kb downstream to the BCL11B gene on chromosome 14, and within or near LINC01081 on chromosome 16, juxtaposing BCL11B enhancer to a novel gene on chromosome 16. In addition, both leukemias were positive for WT1 mutations and negative for FLT3-ITD. CONCLUSION: These cases represent the first report of recurrent translocation, t(14;16), in MPAL-T/M. The t(14;16) with these breakpoints is consistent with the juxtaposition of BCL11B enhancer to a novel gene on chromosome 16, similar to BCL11B::TLX3 in T-ALL. This report highlights the clinical relevance of OGM in identifying critical gene rearrangement that creates enhancer hijacking in the diagnosis of MPAL-T/M. These discoveries, along with further chromatin topology and gene expression studies, may provide a new window into the underlying biology of these aggressive leukemias.

<h4>Objective</h4>Glucose tolerance (GT) is a major effector for adipose tissue (AT) remodeling in obesity, yet its molecular mechanisms remain incompletely defined. We hypothesized that the biophysical and molecular profiles of AT-derived small extracellular vesicles (sEV^AT) change in response to glucose availability and differ by GT status.<h4>Methods</h4>sEV^AT were isolated from plasma of individuals with normal GT (NGT) and impaired GT (IGT) (n = 5/group) at fasting (0 h) and 1 h post glucose challenge during oral glucose tolerance test (OGTT). sEV^AT were characterized for size, concentration, surface expression of insulin receptor-α (INSRα), proteome, and insulin signaling-related miRNAs. C2C12 myotubes were treated with sEV^AT for 48 h, followed by quantification of 84 insulin signaling-related genes.<h4>Result</h4>The size and concentration of sEV^AT did not differ between groups. At fasting, INSRα expression on sEV^AT was comparable; however, groups exhibited opposite directional changes at 1-h OGTT. LC-MS/MS identified significant proteomic differences between NGT and IGT sEV^AT. miR-27a-5p and miR-145a-5p levels in sEV^AT also differed significantly by GT status. Notably, treatment with sEV^AT (IGT-0 h) significantly downregulated insulin signaling-related genes in myotubes.<h4>Conclusions</h4>Distinct molecular signatures in sEV^AT offer a unique insight into AT dysfunction during IGT and offer novel diagnostic and therapeutic targets.

Hereditary hemochromatosis is a common inherited disorder of iron metabolism. It is classically associated with hepatic, cardiac, and endocrine complications. Neurologic manifestations, particularly ischemic stroke, are less commonly recognized and remain underrepresented in the literature. Our case focuses on a 54-year-old man with hereditary hemochromatosis who developed a subacute left pontine infarction in the absence of an identifiable cardioembolic source or large-vessel disease. Extensive evaluation failed to reveal a clear traditional etiology, prompting consideration of nontraditional risk factors. Given the patient's history of iron overload, a hypercoagulable and microvascular mechanism was considered. Iron overload has been associated with endothelial dysfunction, oxidative stress, and prothrombotic changes, which may predispose a patient to ischemic events. Posterior circulation structures, supplied by small perforating vessels, may be particularly vulnerable to microvascular injury. This case highlights the potential role of hereditary hemochromatosis as an underrecognized contributor to ischemic stroke. Awareness of iron overload-related hypercoagulability may broaden the differential diagnosis in patients with cryptogenic or posterior circulation strokes. Further studies are needed to better define the relationship between iron metabolism and cerebrovascular risk.

Prostate cancer lineage plasticity is associated with changes in DNA methylation and enhancer of zeste homolog 2 (EZH2) activity. How these epigenetic programs functionally interact to modulate transcriptional reprogramming in neuroendocrine prostate cancer (NEPC) is not well understood. In this study, we demonstrate that hypomethylated regions of DNA preferentially accumulate the repressive mark, H3K27me3. We established an NEPC mouse model with deletion of Ezh2 in the background of Pten and Rb1 loss plus human MYCN overexpression. Deletion or pharmacological inhibition of EZH2 in NEPC murine or patient-derived models leads to a genome-wide rewiring of DNA methylation, characterized by hypomethylation and upregulation of neuroendocrine-lineage genes along with hypermethylation and repression of polycomb repressive complex 2 (PRC2) targets. On the other hand, deletion of DNA methyltransferase 1 (DNMT1) results in significant changes in H3K27me3 distribution, particularly affecting bivalent promoters bearing both H3K27me3 and active H3K4me3 marks. In NEPC models, neuroendocrine-lineage genes are repressed upon DNMT1 deletion associated with increased H3K27me3. Conversely, in prostate adenocarcinoma models, DNMT1 deletion leads to de-repression of neuroendocrine lineage genes with a loss of H3K27me3 marks. Our findings reveal a functional interplay between two repressive epigenetic machineries that mediates lineage plasticity in prostate cancer.

An abnormal amplification of the CAG trinucleotide repeat in the huntingtin (HTT) gene causes Huntington’s disease (HD), a devastating neurological illness. The striatum and cortex are primarily affected by the gradual neuronal dysfunction and loss caused by the mutant huntingtin (mHTT) protein. Although there have been notable improvements in symptomatic management, curative treatments remain unavailable. A promising treatment option that offers specifically designed interventions to decrease or eradicate mutant HTT expression is gene therapy. RNA-targeted treatments (antisense oligonucleotides, RNA interference), DNA editing methods (Clustered Regularly Interspaced Short Palindromic Repeats/caspase 9, zinc finger nucleases, Transcription Activator-like Effector Nuclease), and advanced delivery systems (viral and non-viral vectors, lipid nanoparticles, exosomes) are just a few of the many approaches that are presently being researched. AMT-130 and tominersen clinical trials provide crucial information about the viability, security, and effectiveness of gene therapy for HD. It is hoped that developments in genome editing and delivery methods would make gene therapy a viable treatment for HD. However, challenges remain, including immunological responses, blood-brain barrier penetration, and off-target consequences. This review delves into the latest developments in HD gene therapy, highlighting new approaches, obstacles, and potential future paths to a permanent cure.

BACKGROUND: The uterus is a pivotal organ for mammalian reproduction, directly determining reproductive success by orchestrating embryo implantation, placental development, fetal nourishment, and parturition. However, the molecular mechanisms regulating high fecundity in the uterus across different stages of the estrous cycle remain unclear. This study aimed to elucidate the genetic regulation of goat fecundity through integrated proteomic and transcriptomic analyses of uterine tissues. RESULTS: Twenty healthy female Yunshang black goats (2–3 years old, body weight 52.22 ± 0.43 kg) were stratified into high- and low-fecundity groups during the follicular (FH and FL, n = 5 per group) and luteal (LH and LL, n = 5 per group) phases. Using data-independent acquisition (DIA) mass spectrometry, we quantified 4,455 proteins. Weighted gene co-expression network analysis (WGCNA) identified the lightcyan module as highly correlated with high fecundity in the luteal phase, with hub proteins including IDH2, PSAT1, MDH1, UBQLN1, RPLP1, SEC23B, RAD23B, PSPH, and MTHFD2. Additionally, 125 and 183 differentially abundant proteins (DAPs) were detected in the FH vs. FL and LH vs. LL comparisons, respectively. Biological adhesion processes, closely linked to transport activity, were implicated in uterine function, with key proteins such as PKP4, COL4A1, LPCAT4, ARRB1, SERPINA5, CDK9, and HDAC7 identified as potentially critical for embryo–uterine communication. Integrated transcriptomic and proteomic analyses revealed significant upregulation of the relaxin signaling pathway during the follicular phase, which may promote uterine tissue remodeling, extracellular matrix degradation, and angiogenesis, thereby facilitating preparation for embryo implantation. During the luteal phase, enhanced activity of the terpenoid backbone biosynthesis pathway was observed, which may indicate an increase in the production of cholesterol and steroid hormone precursors, potentially supporting metabolic demands and pre-receptive steroid signaling in the uterus. CONCLUSION: Our findings demonstrate that specific protein co-expression modules and hub proteins are closely associated with high fecundity in goats. The stage-specific activation of key pathways, including relaxin signaling and terpenoid backbone biosynthesis, may reflect dynamic molecular reprogramming of the uterus during the estrous cycle. This study provides insights into the molecular mechanisms underlying uterine prolificacy in Yunshang Black goats. However, these findings are based on proteomic and transcriptomic data, and further experimental validation is needed to confirm the causal relationships.

Spatial cellular context is crucial in shaping intratumor heterogeneity. However, understanding how each tumor establishes its unique spatial landscape and what factors drive the landscape for tumor fitness remains significantly challenging. Here, we analyze over 2 million cells from 50 tumor biospecimens using spatial single-cell imaging and single-cell RNA sequencing. We develop a deep learning-based strategy to spatially map tumor cell states and their surrounding environmental architecture, and find that different tumor cell states can be organized into distinct clusters, or "villages," each supported by unique microenvironments. Notably, tumor cell villages exhibit village-specific molecular co-dependencies between tumor cells and their microenvironment and are associated with patient outcomes. Perturbation of molecular co-dependencies via random spatial shuffling of the microenvironment results in destabilization of the corresponding villages. We validate our findings using single-cell, spatial, and bulk transcriptome data from 740 liver cancer patients. This study provides insights into understanding tumor spatial landscape and its impact on tumor aggressiveness.

Chondrosarcoma (CS) is the second most common bone sarcoma with a cartilage matrix. Angiogenesis and integrated stress response (ISR) exert a vital influence on the development of CS. This research aimed to conduct a comprehensive analysis to pinpoint angiogenesis and ISR-related potential biomarkers in CS and to elucidate their potential molecular mechanisms. CS data were from GEO. Potential biomarkers were identified and confirmed using differential expression analysis, WGCNA, and expression assessment. Moreover, enrichment analysis was employed to examine relevant pathways. Molecular regulatory network, compound prediction, and molecular docking analyses further explored the key regulatory roles of potential biomarkers in CS. GSE184118 was used to determine key cells and perform pseudo-time and cell communication analyses. Finally, RT-qPCR was used to confirm potential biomarker expression levels. Overall, three potential biomarkers (HSPA8, LMNA and SERPINH1) were determined, and their expression trends were consistent across the GSE30835 and GSE22855 datasets. Potential biomarkers were significantly enriched in the pathways like "medicus variant mutation caused aberrant HTT to 26S proteasome mediated protein degradation" in CS. Moreover, 9 transcription factors (TFs) (like STAT1), 69 key microRNAs (miRNAs) (like hsa-miR-361-3p), and 78 long non-coding RNAs (lncRNAs) (like NEAT1) were found to have relationships with potential biomarkers, and potential biomarkers had stable binding affinity with adenosine diphosphate (ADP) and lonafarnib. Moreover, pseudo-time analysis demonstrated a notable correlation between potential biomarkers' expression and differentiation status of key cells (stromal cells (excluding leucocytes)), and cell communication revealed the strong interactions between stromal cells and chondroid clusters 1. Importantly, RT-qPCR confirmed higher expression of HSPA8, LMNA and SERPINH1 in CS patients. The findings suggested that HSPA8, LMNA and SERPINH1 might offer novel insights for the development of targeted therapies for CS associated with angiogenesis and ISR.

Lgr5+ stem cells play crucial roles in maintaining intestinal epithelial cell homeostasis. However, the cellular heterogeneity and underlying regulatory programs of Lgr5+ small intestinal stem cells (ISCs) remain elusive. In this study, we profiled gene expression and chromatin accessibility of Lgr5+ ISCs at single-cell resolution to gain a deeper understanding of the lineage specification and early fate determining mechanisms. Our analysis identified a total of 6 subsets of Lgr5+ cell populations, which exhibited heterogeneity in gene expression and chromatin structure. We found that early fate-determining processes diverged the absorptive and secretory lineages within Lgr5+ cells. We further constructed gene regulatory networks controlling lineage determination and identified Foxa3 as a key transcription factor that regulates the differentiation of the intestinal secretory precursor. In vitro knockdown of Foxa3 disrupted the differentiation of Paneth cells by modulating Peroxisome-Proliferator-Activated Receptors (PPARs). Further Foxa3-targeted CUT&Tag sequencing analysis also verified that Foxa3 predominantly drives Paneth cell differentiation in the small intestine by regulating the expression of core genes in the PPAR signaling pathway. These results provide a comprehensive reference map for advancing our understanding of intestinal epithelial development and related diseases.

Inflammation plays a vital role in the initiation and progression of hepatocellular carcinoma (HCC). Whether the inflammatory state of HCC peripheral liver tissue also affects tumor progression remains unclear. The medical data of HCC patients at Nanjing Drum Tower Hospital was retrospectively reviewed. Patients were divided into two groups on different inflammation grades in the peritumoral liver tissue: G 0–1 and G ≥ 2. As a preliminary exploration, RNA-seq was conducted on a limited cohort of six HCC tissues (three per group). Gene Set Enrichment Analysis (GSEA) was conducted to detect differential activation of Hallmarks pathways. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analyses were conducted on differentially expressed genes between these two groups to elucidate enriched pathways and biological processes. Integrated machine learning algorithms and multivariate Cox regression analysis were used for prognostic gene screening and model establishment. 163 patients were included in the G 0–1 group and 207 patients in the G ≥ 2 group. More significant liver fibrosis, as well as severe coagulation and hepatic function impairment, were related to the G ≥ 2 group. Inflammatory genes, hepatic fibrosis-related genes, and Hallmarks pathways INFLAMMATORY_RESPONSE were significantly upregulated in the G ≥ 2 group. Conversely, coagulation factors, complement molecules, fibrinolysis-related molecules, hepatic metabolism-related genes, and Hallmarks pathways COAGULATION and BILE_ACID_METABOLISM were markedly upregulated in the G 0–1 group. KEGG and GO enrichment analyses yielded results consistent with above findings. The established model based on seven inflammatory-related genes performed well in prognostic prediction and risk stratification across multiple datasets. HCC patients with G ≥ 2 had an inflammatory and fibrotic phenotype with a worse prognosis, while patients with G 0–1 retained better coagulation and liver metabolic functions.

Bone metastases represent a major cause of morbidity in advanced cancers, yet the neural regulation of metastatic growth within bone remains largely unexplored. The skeletal system is richly innervated by sensory and sympathetic nerve fibers that influence bone remodeling, hematopoiesis, and immune surveillance. Emerging evidence suggests that disseminated tumor cells exploit these neural circuits to create a growth-permissive microenvironment. Tumor-secreted neurotrophic factors can induce nerve sprouting, while sympathetic activation via β-adrenergic receptors promotes osteoclastogenesis, immunosuppression, and tumor proliferation. Neuropeptides such as substance P and calcitonin gene-related peptide exert dual effects on bone cells and infiltrating immune populations, further shaping the metastatic niche. The interplay between neural signals, osteolytic activity, and immune modulation positions the neuro-bone axis as a critical but underappreciated driver of metastatic progression. In this review, we synthesize current evidence on the anatomy and function of bone innervation, tumor-induced neural remodeling, and neuro-immune-osteoclast interactions. We highlight preclinical and clinical data supporting neuromodulatory strategies, including β-blockers, neurotrophin inhibitors, and targeted nerve ablation, as potential adjuncts to standard bone metastasis therapies. Finally, we identify key knowledge gaps, including the need for spatial and functional mapping of nerve-tumor interfaces and for integrating neuroimaging into bone metastasis detection. By framing the neuro-bone axis as a therapeutic target, we aim to catalyze interdisciplinary research that bridges oncology, neuroscience, and bone biology, with the goal of disrupting neural support for metastatic growth.

Gene delivery/administration and, in particular, small interfering RNA (siRNA) delivery represent a therapeutic challenge, though very effective carriers have yet to be identified. Cyclodextrins (CDs) are cyclic oligosaccharides with unique host-guest inclusion capabilities, widely recognized in the pharmaceutical field for their ability to enhance drug solubility and bioavailability. Their excellent biocompatibility and chemical versatility make them powerful building blocks for the design of supramolecular nanovectors (NVs). Thanks to their facility of functionalization, CDs are highly versatile and have found numerous applications across various fields. In this context, CD-based NVs are currently explored as non-viral agents to transport and release siRNA. Recent studies suggest that self-assembled NVs based on CDs can improve the transfection and safety of siRNA delivery. This review provides a comprehensive overview of the most recent advances in the design of NVs based on CDs and their use for siRNA delivery, discussing the role played by structural differences and chemical functionalization in the context of encapsulation and release.

Mutations in TFRC, which encodes transferrin receptor 1 (TfR1/CD71), have emerged as a rare but significant cause of inborn error of immunity (IEI). TfR1 is a crucial membrane glycoprotein responsible for receptor-mediated iron uptake, playing fundamental roles in erythropoiesis and immune function, and linking impaired iron homeostasis to profound immune dysfunction. Notably, TFRC deficiency provides a biologically instructive human model demonstrating that iron uptake via TfR1 is a non-redundant metabolic and signaling checkpoint required for antigen receptor–driven immune activation, clonal expansion, and immune homeostasis. This review summarizes the molecular mechanisms of TfR1, including its interactions with transferrin, and explores how pathogenic TFRC mutations impair iron uptake, affect immune cells, lymphocyte activation, and clonal diversity. We also examine TfR1 in the context of immunodeficiency, linking its roles in iron metabolism and immune regulation to clinical outcomes. By integrating current knowledge on TfR1 biology, molecular mechanisms with patients’ clinical presentation, observations, variants, and therapeutic approaches, this review highlights knowledge gaps and unresolved questions, providing a framework for future research toward elucidating TfR1 mechanisms, improving patient diagnosis, and ultimately enabling the development of targeted therapies for TfR1-mediated immunodeficiency.

Although irreversible electroporation (IRE) is an approved ablation therapy for liver cancer, the border of the IRE-induced inflammatory margin (IM) remains poorly defined, posing a challenge for investigating IRE-specific immune and metabolic alterations within this area. To address this, we employed a male C57BL/6 orthotopic liver cancer model and integrated spatial transcriptomics, spatial metabolomics, single-cell RNA sequencing, and cytometry by time-of-flight (CyTOF). This multiomics approach enabled precise spatial mapping of the IM and revealed a pronounced infiltration of Ly6C^lowPD-L1^hi that phenotypically resemble lipid-associated macrophages (LAMs). Further analysis uncovered profound lipid metabolic reprogramming within the IM, including biosynthesis of unsaturated fatty acids, arachidonic acid and sphingolipid metabolism, which appears to sustain the immunosuppressive phenotype of LAMs. Collectively, our study uncovers an IRE-specific immunosuppressive microenvironment in the IM and propose potential metabolic targets that might be effectively combined with IRE to improve therapeutic outcomes.

Cell death is a fundamental process that maintains tissue homeostasis and shapes the tumour microenvironment. Cancer cells often evade or reprogram cell death pathways, which leads to malignancy and therapy resistance. On the other hand, the cell death of non-malignant cells significantly influences the adverse effects of anticancer therapy, such as the onset of mucositis or enteritis. Emerging evidence identifies both the tumour and gut microbiomes as critical modulators of these processes. In turn, cell death reshapes the microbial ecosystem by altering nutrient landscapes and immune signalling. Although both cell death and the microbiome are well-studied in cancer, their intersection remains underexplored. This review article addresses this gap by summarising how microbes and microbial metabolites modulate cancer cell death pathways and shape responses to anticancer therapy. We integrated current knowledge on this complex interplay, focusing on key metabolic mediators that can influence cell fate decisions. Understanding this bidirectional crosstalk offers new opportunities to "preserve the good and eliminate the bad" within the tumour-microbiome axis, paving the way for precision microbiome-informed cancer therapies.

Poultry is a major nutritional source providing food for large human populations. Infectious diseases threaten the productivity of poultry flocks and diminish animal welfare. Recent advances in genome editing have significantly contributed to our understanding of various physiological aspects and have helped elucidate the interaction between the chicken host and pathogens. Several chicken lines were generated, including those with Type I and Type III interferon receptor knockouts, those lacking specific T cell populations, and those missing contributing factors to V(D)J recombination, such as the recombination-activating gene 1 (RAG1). In addition, researchers achieved resistance to the avian influenza virus (AIV) by targeting acidic nuclear phosphoproteins. Finally, reinstating retinoic acid-inducible gene I (RIG-I) and RING finger protein 135 (RNF135) in the chicken revealed new insights into their evolutionary role, particularly during host-pathogen interactions with AIV. This review provides an update about recent achievements in genome editing of chickens, particularly in immunology and disease resistance.

<h4>Background</h4>Proteomic signatures may enhance the prediction of cardiometabolic diseases for targeted prevention. We evaluated whether a proteomic risk score (ProtRS) improves the prediction of incident hypertension.<h4>Methods</h4>Within the UK Biobank Proteomics data set, participants at risk for incident hypertension were randomly split into derivation (n=25 158) and test (n=10 781) sets. A ProtRS was trained in the derivation cohort using least absolute shrinkage and selection operator penalized Cox regression and evaluated in the test set with sequential adjustment for demographics, clinical risk factors, and a polygenic risk score (PRS). External replication was performed in the Asklepios study (n=793).<h4>Results</h4>In UK Biobank Proteomics (2312 events), each 1-SD higher ProtRS was associated with incident hypertension after covariate adjustment (hazard ratio, 1.68 [95% CI, 1.59-1.78]; <i>P</i><0.001). Adding the protein score to demographics, clinical variables, and PRS improved model fit (global χ² from 1733 to 2048, <i>P</i><0.001), discrimination (C-index, 0.745-0.765), net reclassification improvement (19.0% [95% CI, 15.9-22.0]), and integrated discrimination improvement (2.9% [95% CI, 2.4-3.8]). In Asklepios (221 events), the ProtRS was associated with an increased risk of hypertension (hazard ratio, 1.32 [95% CI, 1.13-1.55]; <i>P</i><0.001) after covariate adjustment, and improved global χ² (from 120.4 to 132.4; <i>P</i><0.001), C-index (from 0.723 to 0.736) and discrimination (net reclassification improvement, 13.1% [95% CI, 1.2%-24.1%]). Pathway analysis highlighted neutrophil degranulation, insulin-like growth factor, PI3K signaling, and extracellular matrix remodeling.<h4>Conclusions</h4>A ProtRS improves prediction of incident hypertension beyond demographics, clinical, and genetic information in UK Biobank Proteomics, with supportive external replication. Proteomic information may enhance individualized hypertension risk stratification and provide biologic insights into disease development.

<h4>Background</h4>Pulmonary function is linked to cardiovascular disease risk; however, the underlying mechanisms remain unclear. We aimed to identify protein biomarkers associated with pulmonary function and examine their impact on incident chronic obstructive pulmonary disease, coronary heart disease, heart failure, and all-cause mortality.<h4>Methods</h4>Data from White and Black Americans in the Atherosclerosis Risk in Communities study (visit 2: N=11 354, mean age=57 years; visit 5: N=3517, mean age=75 years), a prospective cohort, were analyzed. Linear regression assessed associations between protein levels and pulmonary function measures, including forced expiratory volume in 1 second and forced vital capacity. The impact of the identified proteins on incident chronic obstructive pulmonary disease, coronary heart disease, heart failure, and mortality was estimated using logistic regression and Cox proportional hazards models. Pathway enrichment and Mendelian randomization explored underlying biological functions and causal effects.<h4>Results</h4>Of 4766 proteins analyzed, 364 were cross-sectionally associated with forced expiratory volume in 1 second (and forced vital capacity (false discovery rate<0.05). Ninety-four and 270 proteins had concordant positive and negative effects, respectively. Five pathways related to pulmonary and cardiac function were enriched. Of the 364 proteins, 112 were linked to all 4 outcomes, where 86 were associated with increased risk (odds ratio/hazard ratio [OR/HR], 1.05-1.42) and 26 with reduced risk (OR/HR, 0.69-0.96). Six proteins (STAT3 [signal transducer and activator of transcription 3], MIC-1 [growth differentiation factor 15], apoA-II [apolipoprotein A-II], TPST1 [protein-tyrosine sulfotransferase 1], integrin a1b1 [integrin alpha-I: beta-1 complex], and BLC [C-X-C motif chemokine 13]) showed potential inverse causal effects on with forced expiratory volume in 1 second and forced vital capacity, and integrin a1b1 demonstrated consistent inverse associations with chronic obstructive pulmonary disease, coronary heart disease, and heart failure risks.<h4>Conclusions</h4>Proteins associated with pulmonary function may influence CVD risk. Six proteins, including integrin a1b1, represent promising targets for future interventions.

Accurate identification of protein-ligand binding sites is an essential step in structure-based drug discovery. Herein, we present SwinSite, a deep learning framework that leverages a hybrid architecture combining 3D convolutional neural networks and hierarchical vision transformer modules to predict ligand binding sites based on a 3D structure of a target protein. SwinSite encodes spatial information by voxelizing a protein structure into 3D grids centered around surface residues, allowing for a detailed spatial representation of the protein's surface environment. By combining local feature extraction with hierarchical self-attention via shifted windows, SwinSite effectively captures both fine-grained geometric features and long-range dependencies. Evaluations on multiple benchmark data sets demonstrate that SwinSite outperforms existing CNN- and GNN-based ligand binding site detection methods consistently, highlighting its robustness and generalization ability.

Mouse intestinal organoids are ideal models for investigating intestinal development and diseases. The full potential of these models hinges on the ability to precisely engineer their genome, yet traditional methods for CRISPR-based editing in 3D cultures often surfer from low efficiency, high cytotoxicity, and inconsistent post-editing differentiation, which limits their applications. Here, we developed an electroporation approach mediated by ribonucleoprotein (RNP)-CRISPR that achieves over 90% gene editing efficiency in mouse intestinal organoids. Using this optimized method, we generated APC-knockout organoids that exhibit Wnt pathway hyperactivation, demonstrated by R-spondin1-independent growth, increased nuclear β-catenin, and enhanced proliferation. Our method addresses a critical technical gap in murine organoid research, offering a scalable platform for intestinal disease modeling.

<h4>Purpose</h4>The relationship between metabolic dysfunction-associated steatotic liver disease (MASLD) and acromegaly is unclear due to the complex metabolic effects of growth hormone (GH). GH stimulates gluconeogenesis, glycogenolysis, and lipolysis, increasing free fatty acids, a risk factor for MASLD, but may also protect against hepatic fat accumulation. In this study, we aimed to evaluate the impact of acromegaly and GH levels on liver steatosis and fibrosis using non-invasive FibroScan imaging.<h4>Methods</h4>This cross-sectional study included 58 patients with acromegaly, 61 age- and sex-matched metabolically comparable controls, and 82 healthy controls. Hepatic steatosis and fibrosis were assessed via controlled attenuation parameter (CAP) and liver stiffness measurement (LSM) by vibration-controlled transient elastography. Moderate to severe steatosis was defined as CAP > 260 dB/m, and significant fibrosis as LSM ≥ 8.0 kPa.<h4>Results</h4>Both acromegaly patients and metabolically healthy individuals exhibited lower CAP (241.8 ± 50.0 and 231.7 ± 51.9 dB/m) and LSM (4.7 ± 1.4 and 4.8 ± 1.8 kPa) scores than metabolically matched controls (CAP 281.7 ± 61.2 dB/m; LSM 5.5 ± 1.8 kPa; CAP p < 0.001; LSM p = 0.012). Moderate-to-severe steatosis was observed in 36.2% of acromegaly patients, 27.7% of healthy individuals, and 68.8% of metabolically matched controls (p < 0.001), while significant liver fibrosis occurred in 5.2%, 7.3%, and 14.7%, respectively (p = 0.154). GH levels were lower in acromegaly patients with MASLD (p < 0.001) compared to those without. CAP correlated negatively with GH and positively with BMI, waist circumference, and triglycerides, whereas LSM correlated positively with age, BMI, and triglycerides.<h4>Conclusions</h4>Hepatic steatosis was less frequent in acromegaly than in metabolically comparable individuals, and GH was inversely associated with hepatic steatosis scores, possibly suggesting a protective role of GH independent of metabolic comorbidities.

Heat hormesis describes the beneficial adaptations resulting from transient exposure to mild heat stress, which enhances stress resilience and promotes healthy aging. While heat hormesis is widely observed, much remains to be learned about its molecular basis. This study bridges a critical knowledge gap through a comprehensive multiomic analysis, providing key insights into the transcriptomic and chromatin accessibility landscapes throughout a heat hormesis regimen in Caenorhabditis elegans. We uncover highly dynamic, dose-dependent molecular responses to heat stress and reveal that while most initial molecular changes induced by mild stress revert to baseline, key differences emerge in response to subsequent heat shock challenge that likely contribute to physiological benefits. We further demonstrate that heat hormesis extends life span specifically in wild-type animals, but not in germline-less mutants, likely due to transient disruption of germline activities during mild heat exposure, which appears sufficient to trigger pro-longevity mechanisms. This finding points to tissue-specific responses in mediating the physiological outcomes of heat hormesis. Importantly, we identify several highly conserved regulators of heat hormesis that likely orchestrate gene expression to enhance stress resilience. Among these regulators, some (MARS-1/MARS1, SNPC-4/SNAPc, FOS-1/c-Fos) are broadly required for heat-hormesis-induced benefits, whereas others (ELT-2/GATA4, DPY-27/SMC4) are uniquely important in specific genetic backgrounds. This study advances our understanding of stress resilience mechanisms, points to multiple new avenues for future investigations, and provides a molecular framework for promoting healthy aging through strategic mid-life stress management.

Transcriptional programs in renal cell carcinoma (RCC) have been linked to tumor heterogeneity and clinical outcomes, but analogous efforts to define chromatin programs shaping disease biology have been limited. Here, we generated single-cell ATAC-seq profiles from patients with RCC and integrated them with three previously published datasets to identify chromatin programs in tumor cells. We identified an interferon response program enriched in <i>BAP1</i>-mutant tumors, and, in bulk ATAC-seq cohorts with linked clinical data, this program was associated with poor prognosis. Mechanistic analyses in isogenic models suggested that <i>BAP1</i> loss induces a tumor-intrinsic interferon response, with dysregulated endogenous retroviruses as a potential upstream trigger. We further characterized the <i>BAP1</i> mutation-associated tumor microenvironment across single-cell, bulk, and multiplex immunofluorescence data, identifying features of both inflammation and immune evasion. Together, our findings nominate tumor-intrinsic interferon signaling as a candidate driver of <i>BAP1</i>-associated aggressiveness in RCC and highlight immune evasion pathways as potential therapeutic targets.

Ustilago maydis infection in maize causes hypertrophic leaf tumors; however, the underlying mechanisms driving this excessive cell growth are unknown. In this study, we identify Hap1 (hypertrophy-associated protein 1) as an effector and virulence factor that regulates mesophyll cell hypertrophy. Using CRISPR-Cas9 mutagenesis, we demonstrate that Hap1 contributes to endoreduplication and starch accumulation in infected tissues. Transcriptomics revealed Hap1-dependent upregulation of starch biosynthesis and cell cycle genes, as well as suppression of plant defense. This links Hap1 to metabolic and cell cycle reprogramming, and immune suppression. To identify the target of Hap1 that drives metabolic reprogramming, we investigated its interaction with ZmSnRK1α in maize. We found that Hap1 interferes with the phosphorylation of SnRK1 substrates and that two Hap1-interacting effectors, Hip1 and Hip2, enhance its protein stability. We conclude that Hap1 contributes to the reprogramming of maize metabolism and cell cycle, as well as mesophyll cell hypertrophy, by modulating the SnRK1 signaling pathway to regulate starch biosynthesis and host defense responses.

Bone marrow fibrosis is the most extensive matrix remodeling of the microenvironment and can include de novo formation of bone (osteosclerosis). Spatiotemporal information on the contribution of distinct bone marrow niche populations to this process is incomplete. We demonstrate that fibrosis-inducing hematopoietic cells cause profibrotic reprogramming of perivascular CXCL12-abundant reticular (CAR) progenitor cells, resulting in loss of their hematopoiesis-support and upregulation of osteogenic and pro-apoptotic programs. In turn, peritrabecular osteolineage cells (OLCs) are activated in an injury-specific, Wnt-dependent manner, comparable to skeletal repair. OLCs fuel bone marrow fibrosis through their expansion and skewed differentiation, resulting in osteosclerosis and expansion of Ly6a⁺ fibroblasts. NCAM1 expression marks peritrabecular OLCs and their expansion into the central marrow is specific for fibrosis in mice and patients. Peritrabecular stromal β-catenin expression is linked to fibrosis in patients, and inhibition of Wnt signaling reduces bone marrow fibrosis and osteosclerosis, possibly being a clinically relevant therapeutic target.

Exosomes are a type of extracellular vesicles with a diameter of 30-150 nm, formed within multivesicular bodies of late endosomes, and released into the extracellular environment before being taken up by recipient cells. Exosomes transfer bioactive molecules, such as proteins and various RNAs, between cells and thereby influence recipient cell behavior. Among these molecules, long non-coding RNAs (lncRNAs) are not only selectively packaged into exosomes but have also been reported to participate in exosome biogenesis and secretion. Furthermore, they can contribute to cancer development by regulating gene expression through epigenetic mechanisms such as miRNA sponging, protein interactions, and methylation. These mechanisms appear to drive tumorigenesis by altering the tumor microenvironment, including macrophage polarization, immune evasion and suppression, angiogenesis, and lymphangiogenesis-mediated metastasis. In addition, exosomal lncRNAs can be readily detected in serum, highlighting their potential as biomarkers for liquid biopsy-based diagnosis. Finally, exosomal lncRNAs have been reported to contribute to drug resistance in various cancers. Therefore, therapeutic strategies targeting exosomal lncRNAs are expected to improve clinical outcomes in cancer patients.

<b>Background</b>: Energy metabolism progressively deteriorates from a healthy state to non-alcoholic fatty liver disease (NAFLD), and circulating lipopolysaccharide (LPS) may contribute to this process. However, previous studies have analyzed healthy individuals and NAFLD patients together, leaving stage-specific associations unclear. Whether LPS and its surrogate marker, lipopolysaccharide-binding protein (LBP), show similar relationships during NAFLD development also remains unknown. This study evaluated the associations between plasma LPS and LBP concentrations with clinical parameters in healthy individuals and NAFLD patients. <b>Methods</b>: We conducted a cross-sectional study of 31 healthy individuals (median age [IQR]: 31 (26-43) years) and 31 NAFLD patients (59 (54-70) years). Plasma LPS and LBP concentrations and clinical parameters were measured. Correlations were assessed using Spearman's rank analysis, followed by multivariate regression adjusting for age, sex, and BMI. <b>Results</b>: Plasma LPS and LBP concentrations were significantly higher in NAFLD patients compared to healthy individuals. Additionally, in the univariate regression analysis for all study participants, plasma LPS concentrations were correlated with obesity, blood pressure, liver function, lipid metabolism, and glucose metabolism. Plasma LBP concentrations were also correlated with age, obesity, blood pressure, liver function, lipid metabolism, glucose metabolism, and inflammatory cytokines. In healthy individuals, LPS correlated positively with triglycerides (TG), remaining significant after adjustment and exclusion of participants with any clinical test values outside the normal range. This association was not observed in NAFLD patients. Plasma LBP did not correlate with TG in either group; however, it was inversely associated with hepatic fat fraction in NAFLD patients, although this association was attenuated after adjusting for alanine aminotransferase. <b>Conclusions</b>: Plasma LPS correlates with TG even in clinically healthy individuals, suggesting LPS may influence lipid metabolism before NAFLD onset.

Huntington's disease is a progressive neurological disorder marked by motor, cognitive, and psychiatric symptoms. Currently, there are no definitive diagnostic tools or effective treatments to halt or reverse the disease. In recent years, surface-engineered nanosystems have emerged as innovative therapeutic platforms, offering significant promise in overcoming the limitations of traditional approaches. These nano systems, including liposomes, dendrimers, polymeric nanoparticles, and solid lipid nanoparticles, offer significant potential by targeting and modulating intricate biochemical pathways involved in the progression of Huntington's disease. Their defining advantage lies in the ability to selectively deliver therapeutic agents to specific regions of the brain with high precision. Through the use of various nanoscale carriers, these particles can successfully traverse the protective barrier between the blood and brain tissue, enabling the direct delivery of treatment agents to the regions affected by Huntington's disease. This targeted approach not only enhances the therapeutic efficacy but also minimizes unwanted systemic side effects. This review highlights recent advancements in nanosystem development, addressing previous challenges and setbacks in the field, particularly in overcoming the blood-brain barrier and improving treatment delivery. The review further explores the evolving mechanisms of nanosystem delivery and their functional impact in experimental models of Huntington's disease. While the primary focus remains on therapeutic applications, we also briefly discuss recent developments in nanoparticle-based diagnostics. Although several challenges, particularly regarding comprehensive safety assessments and the current absence of nanoparticles approved by the United States Food and Drug Administration for Huntington's disease, this review underscores the transformative potential of nanosystems for future therapeutic applications.

<h4>Background</h4>The aim of this study was to define the relationships between mean corpuscular volume (MCV) and 12 clinical and laboratory variables in <i>HFE</i> p.C282Y (rs1800562)/p.H63D (rs1799945) compound heterozygotes.<h4>Methods</h4>We retrospectively studied self-reported non-Hispanic white adult compound heterozygotes with transferrin saturation (TS) > 50% and serum ferritin (SF) > 300 µg/L (men) or TS > 45% and SF > 200 µg/L (women) who participated in primary care-based screening. In post-screening evaluations, we excluded participants with anemia, pregnancy, or medication use that increases MCV. We defined heavy alcohol intake as > 28 g/day in men and > 14 g/day in women. We determined associations of MCV with the following clinical and laboratory variables: age, sex, body mass index (BMI), diabetes, daily intakes of heme, non-heme, and supplemental iron, daily intakes of alcohol, swollen or tender second/third metacarpophalangeal (MCP) joints, reports of therapeutic phlebotomy, TS, and SF.<h4>Results</h4>There were 74 participants (37 men, 37 women) of mean age 59 ± 12 (SD) years. Mean screening TS and SF were 65±13% and 529 ± 169 µg/L (men) and 59 ± 14% and 376 ± 195 µg/L (women). Post-screening values did not differ significantly. Mean MCV was 95.7 ± 4.0 fL. There was a negative correlation of MCV with BMI (P = 0.0488) and positive correlations of MCV with age (P = 0.0098), daily heme iron intake (P = 0.0333), and daily alcohol intake (P = 0.0113). Mean MCVs of 19 participants with and 55 without heavy alcohol intake were 97.8 ± 3.8 fL and 95.0 ± 3.9 fL, respectively; P = 0.0074). Linear regression on MCV confirmed positive associations with age (P = 0.0064) and daily alcohol intake (P = 0.0151). MCV was not significantly associated with sex, diabetes, daily intakes of non-heme and supplemental iron, swollen or tender second/third MCP joints, reports of therapeutic phlebotomy, TS, or SF.<h4>Conclusion</h4>MCV in <i>HFE</i> p.C282Y/p.H63D compound heterozygotes with high iron phenotypes is positively associated with age and daily alcohol intake, after adjustment for other variables.

Maternal exposure to the pyrethroid pesticide deltamethrin (DM), even below the No Observed Adverse Effect Level (NOAEL), has been increasingly associated with models of neurodevelopmental disorders (NDDs) such as attention-deficit hyperactivity disorder (ADHD) and autism-spectrum disorders (ASD). While recent studies have documented the impact of maternal DM exposure on offspring hippocampal circuits, its effects on the nucleus accumbens (NAc)-a key region implicated in NDDs-remain poorly understood. The NAc is composed of over 80% medium spiny neurons (MSNs), which serve as its sole output and rely on specialized local inhibitory interneurons to regulate firing patterns and synaptic integration. Here, we report subtype-specific dysfunction in parvalbumin-positive (PV+) inhibitory interneurons in the NAc of offspring following maternal DM exposure in mice. Whole-cell patch-clamp recordings from genetically labeled PV+ interneurons revealed that high input-resistance (IR) PV interneurons exhibited unique vulnerability. DM exposure significantly reduced maximum firing frequency and lowered action potential threshold, while low IR PV interneurons remained unaffected. Additionally, the probability of more frequent excitatory synaptic inputs onto high IR PV interneurons were significantly reduced. Mapping our data onto publicly available patch-seq data from a Mini-Atlas revealed that high IR PV interneurons express high levels of Shisa8, an accessory protein regulating AMPA receptor kinetics, and low levels of Grin2A, an NMDA receptor component, suggesting that specific local synaptic circuit disruption in DM-sensitive neurons. Thus, maternal DM exposure selectively impairs a subtype of inhibitory interneurons with unique physiological properties, potentially disrupting synaptic integration in the reward circuit and contributing to NDD-like phenotypes.

Radial glia are a specialized population of neural progenitor cells that persist throughout embryogenesis and into adulthood. Throughout this period, radial glia reside in a highly dynamic microenvironment that influences various biological decisions that govern typical cortical development. Subsequently, radial glia must fine-tune their responses to numerous environmental cues throughout development. The establishment of the cortical vasculature coincides with neurogenesis and dramatically alters the radial glia microenvironment by increasing oxygen and metabolite delivery. In addition, a synergistic spatial relationship between radial glia and endothelial cells regulates multiple aspects of radial glial biology. Here, we discuss crosstalk between radial glia and the cortical vasculature/endothelial cells throughout development, including the influence of extrinsic angiogenic processes and our growing understanding of the intricate spatial relationships between radial glia and endothelial cells.

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease that leads to progressive joint destruction, extra-articular manifestations, disability, and increased mortality. Early detection, particularly in seronegative patients, remains challenging because current diagnostic criteria based on joint involvement, serology, and acute-phase reactants may fail to identify disease at its earliest stages. Epigenetic mechanisms, including DNA and RNA methylation, histone modifications, and non-coding RNAs (ncRNAs), provide a dynamic interface between genetic predisposition and environmental triggers in RA pathogenesis. Peripheral blood (plasma, serum, and cellular fractions) is an accessible, minimally invasive source for monitoring systemic molecular alterations over time. To capture the latest evidence, we performed a structured literature search using curated keywords covering RA, epigenetic mechanisms, DNA and RNA methylation, m6A, histone modifications, miRNAs, lncRNAs, circRNAs, and blood-based fractions (peripheral blood mononuclear cells (PBMCs), plasma, serum, whole blood). Emerging data indicate that blood-based epigenetic signatures not only reflect disease activity but also hold promise as prognostic biomarkers, predictors of treatment response, and tools for personalized therapeutic strategies. In this review, we synthesize current knowledge on blood-based epigenetic alterations in RA, focusing on DNA methylation, histone modifications, and multiple classes of ncRNAs, including less widely studied species such as piRNAs, snoRNAs, Y-RNAs, snRNAs, and tRNA-derived fragments, with an emphasis on studies published between 2020 and 2025. We highlight the translational potential of multilayered epigenetic signatures as innovative diagnostic and prognostic tools that could advance early detection and guide precision-medicine approaches in RA.

This review synthesizes the current state of knowledge on medical-coating materials. Fundamental theories are examined with respect to material taxonomy, physicochemical properties, biocompatibility profiles, and antimicrobial mechanisms. Clinical applications are reviewed for implants, wound dressings, catheters, and endovascular stents. Technological advances are highlighted in nanotechnology, surface-modification strategies, and the development of stimulus-responsive "smart" coatings. The analysis traces the field's evolution, noting its growing market and clinical adoption, while identifying persistent challenges in long-term biosafety and translation. Future directions are projected toward synergistic modification for multifunctionality, patient-specific designs, and next-generation intelligent systems. By integrating contemporary research, this review aims to inform and guide the future development and clinical application of advanced coating technologies.

<h4>Introduction</h4>Temporal lobe epilepsy is the most prevalent form of drug-resistant focal epilepsy and is frequently associated with neuronal cell loss and astrogliosis in the hippocampus, i.e. hippocampal sclerosis (HS).<h4>Methods</h4>In this study, we performed mass spectrometry-based proteomic profiling of microdissected hippocampal, neocortical, and white matter tissue obtained from TLE patients and respective control samples.<h4>Results</h4>In hippocampal TLE tissue, we observed significant upregulation of proteins involved in complement system activation, extracellular matrix (ECM) organization, and astrocyte reactivity, indicative of active inflammatory remodeling within the sclerotic hippocampus. Conversely, synaptic proteins, including glutamate and gamma-aminobutyric acid (GABA) receptors, along with other regulators of synaptic structure and function, were markedly downregulated. Interestingly, in neocortical and white matter regions from the same TLE patients, immune- and ECM-related proteins were downregulated or unchanged, whereas synaptic proteins were preserved or upregulated.<h4>Discussion</h4>These region-specific molecular signatures suggest that inflammatory-driven ECM remodeling is spatially restricted to the epileptogenic hippocampus, where it may contribute to synaptic destabilization and network dysfunction. Together, our findings support the hypothesis that inflammatory ECM remodeling in the hippocampus plays a central role in epileptogenesis in TLE. In contrast, the neocortical and white matter regions may undergo compensatory adaptions. The convergence of immune and ECM-related alterations on synaptic structures highlights a potential pathophysiological axis in epilepsy and points to novel molecular targets for therapeutic intervention.

Public speaking is a fundamental form of communication across a wide range of domains; however, the neural mechanisms underlying audience engagement during different speeches remain poorly understood. In particular, it is unclear which functional brain networks support the dynamic fluctuations of audience engagement and what neurobiological processes underlie these effects. In this study, we used naturalistic fMRI combined with intersubject correlation (ISC) analysis to examine how carefully selected and matched speeches, with varying levels of audience engagement, influence neural activity. Our results revealed that the more engaging speech elicited significantly greater interbrain neural synchronization, as indexed by ISC, across a broad range of brain regions. Notably, these engagement-related effects were most prominent in networks associated with language processing and theory of mind, highlighting their critical roles in facilitating shared audience experiences during compelling public communication. A sliding-window analysis further revealed substantial temporal fluctuations in interbrain synchronization throughout the speech. Additionally, neurobiological annotation analyses identified strong associations between engagement-related ISC effects and molecular pathways involved in trans-synaptic signaling, suggesting that intrabrain neuronal communication may contribute to modulating interbrain synchronization. By integrating naturalistic fMRI with ISC analyses, this study offers a promising framework for investigating dynamic neural synchronization among audience members. These findings have broad implications for fields such as education and leadership development, where a deeper understanding of the neural basis of audience engagement could inform strategies to enhance public speaking and communication effectiveness.

<h4>Background</h4>Gemcitabine plus cisplatin (GC) has been the global standard for advanced biliary tract cancer (BTC). The triplet regimen gemcitabine-cisplatin-S-1 (GCS) demonstrated superiority in the MITSUBA trial, but its real-world effectiveness remains unclear. We compared survival outcomes of GCS versus GC, focusing on conversion surgery (CS).<h4>Methods</h4>We retrospectively analyzed 542 patients with unresectable BTC treated between 2017 and 2024 at 19 Japanese institutions. Patients received GC (<i>n</i> = 310) or GCS (<i>n</i> = 232). Survival was evaluated using multivariable Cox regression, 90-day landmark analysis, and propensity score matching (PSM) to adjust for baseline imbalances.<h4>Results</h4>Patients treated with GCS achieved greater tumor shrinkage (median -23.0% vs. -10.0%, <i>p</i> = 0.014) and a higher CS rate (16.4% vs. 4.5%, <i>p</i> < 0.001) than GC. Median progression-free survival was 8.6 versus 5.4 months (<i>p</i> = 0.002), and median overall survival (OS) was 17.2 versus 11.6 months (<i>p</i> = 0.006). In multivariable analysis, GCS was associated with a lower risk of death (HR 0.80, 95% CI 0.65-0.98, <i>p</i> = 0.035), with consistent results after PSM. Fifty-two patients underwent CS, with comparable perioperative safety and R0 resection rates between regimens. Patients who underwent CS achieved markedly longer OS; in the overall cohort, median OS was 31.0 months in the GCS with CS group and not reached in the GC with CS group (<i>p</i> = 0.131).<h4>Conclusions</h4>Treatment with GCS was associated with longer survival compared with GC in unresectable BTC, alongside a higher rate of conversion surgery, which was associated with favorable long-term outcomes.

<title>Abstract</title> <p>Targeted protein degradation (TPD) is a therapeutic strategy to remove disease-causing proteins by routing them to the ubiquitin-proteasome, autophagy, or lysosme machineries. For instance, proteolysis-targeting chimeras (PROTACs) are synthetic hetero-bifunctional small molecules that simultaneously bind the target and an E3 ubiquitin ligase to drive ubiquitination and degradation by the proteasome. Despite considerable success, designing such molecules is challenging and the number of currently addressable ubiquitin E3 ligases is limited. Here we demonstrate hetero-bifunctional de novo designed proteins as alternatives for TPD to access more targets and ligases. First, we develop a stable and highly adaptable helix-turn-helix scaffold for presenting different binding sites. Next, we use computational protein design to incorporate and embellish hot-spot- binding sites to target BCL-xL, plus short linear motifs (SLiMs) for KLHL20 ligase recruitment. The resulting mono- and bi-functionalised proteins bind the targets in vitro, and the latter degrade BCL-xL in cells leading to apoptosis.</p>

The rising prevalence of hepatocellular carcinoma (HCC) in the last decade is mostly attributable to the growing epidemic of metabolic dysfunction-associated steatotic liver diseases (MASLD). However, the transition from steatosis to steatohepatitis (MASH) and ultimately to HCC is not fully understood. As an executioner protein of necroptosis, the mixed-lineage kinase domain-like protein (MLKL) has been proposed to contribute to MASH and HCC development. To investigate its role in disease progression, mice whose liver parenchymal cells (LPCs) no longer expressed MLKL (Mlkl^LPC-KO) were compared to their control counterparts (Mlkl^fl/fl) using an experimental model combining diabetes induction and a high-fat high-sugar diet (HFHSD) for 4, 8, or 12 weeks. Notably, HFHSD failed to induce detectable hepatic necroptosis in Mlkl^fl/fl mice, with no phosphorylated MLKL observed by Western blot. Both genotypes displayed similar steatosis and mild fibrosis, consistent with comparable MASH severity, and this condition progressed to HCC. Interestingly, the incidence of liver tumors in Mlkl^LPC-KO mice was significantly reduced, which was associated with a delay in the onset of systemic and hepatic inflammation. At the early stage of the disease (4th week), the absence of MLKL in LPCs appeared to confer a protective effect on the liver, reducing metabolic stress, as reflected by a lower ceramide-to- sphingomyelin ratio, along with oxidative stress and DNA damage. Altogether, our data suggest that MLKL in LPCs contributes to HCC initiation in the context of MASH, potentially involving its described non-canonical role within mitochondria, promoting oxidative stress, a cancer hallmark. This study provides new insights into evaluating the therapeutic potential of targeting MLKL, as its inhibition in LPCs may represent an effective strategy for treating MASH-related HCC.

Bacillus amyloliquefaciens SC06 (BaSC06) has emerged as a promising probiotic for improving animal gut health and immuno-protection. However, the underlying molecular mechanisms remain incompletely understood. In this study, we employed porcine intestinal organoids as an ex vivo model and induced oxidative stress using diquat to systematically evaluate the protective effects of BaSC06. We evaluated antioxidant capacity, apoptosis-related markers, intestinal stem cells differentiation markers, and associated signaling pathways. Pretreatment with BaSC06 significantly alleviated oxidative damage by reducing intracellular reactive oxygen species levels and resultant apoptosis. Cellularly, BaSC06 promoted intestinal stem cells proliferation and favored differentiation toward Paneth cells while suppressing differentiation into other epithelial lineages. Mechanistically, these effects were mediated by the activation of the Wnt signaling pathway, and it was further confirmed by using a specific Wnt inhibitor. Overall, our findings uncover a probiotic-Wnt axis through which BaSC06 confers protection against oxidative stress, involving modulation of intestinal stem cells fate and enhancement of epithelial barrier function.

The microbiological quality of corneal tissue for transplantation must be monitored in a validated procedure complying with the European Pharmacopoeia (Ph. Eur.). An automated culture system is used to detect test strains in the presence of the medium. The Ph. Eur. mentioned C. acnes ATCC 11827 strain has allegedly been isolated from skin. This strain is known to display variable growth in routine matrix validation. As the Ph. Eur. also opens the use of clinical isolates, this study analyzed the use of clinical isolates for matrix validation.We compared the time-to-detection (TTD) in absence and presence of cornea organ culture medium (cocm) of 24 C. acnes strains, preferring clinical isolates, in an automated culture system.In the first step clinical strains (n=23) were examined, thus 4 slow-growing (TTD 305,1-336 h), 5 intermediate-fast-growing (TTD 250,1-305 h), 5 fast-growing strains (TTD 164,3-250 h) and nine non-blood-culture-detectable strains (TTD>336 h) were identified. Seven strains showed a reproducible growth signal in cocm-absence. In presence of cocm, three strains (CA02, CA10, CA22) showed a reproducible growth signal, while the reference strain was not blood-culture-detectable (bcd) in four of five experiments. The strain with the preferred growth characteristics (DSM 117854) was tested according to the Ph. Eur. conditions and showed bcd growth in all measurements (n=5) in presence of medium, in contrast to the reference strain.The use of a clinical isolate of C. acnes could improve matrix validation of cocm quality control testing and thus the care of cornea-transplant-patients. C. acnes DSM 117854 provides an isolate that promises more reliable results in matrix validation.

Oral cancer, a common malignancy of the head and neck, remains therapeutically challenging largely due to the frequent development of chemotherapy resistance. Autophagy, a key cellular stress-response pathway, contributes to oral cancer cell survival and drug tolerance under therapeutic pressure. Emerging evidence indicates that targeting the initiation phase of autophagy—particularly when combined with conventional agents such as cisplatin—may help reverse resistance and improve treatment efficacy. In this review, we summarize the molecular mechanisms governing autophagy initiation, highlight key actionable targets, and discuss the synergistic effects observed when autophagy inhibitors are paired with chemotherapeutic drugs. We place particular emphasis on newly developed initiation-stage inhibitors, bioactive natural products, and nanotechnology-enabled formulations that show potential for overcoming chemoresistance in oral cancer. By integrating recent advances, this article aims to provide a mechanistic framework and translational considerations to support the development of more precise therapeutic strategies for chemotherapy-resistant oral cancer.

Exosomal circular RNAs (circRNAs) have emerged as key regulators of tumor apoptosis, with significant implications for cancer development and treatment response. These persistent, covalently closed-loop RNAs are preferentially packed into exosomes, which are nanoscale extracellular vesicles that allow cells to communicate within the tumor microenvironment. This study investigates the molecular mechanisms underlying exosomal circRNA synthesis, selective cargo sorting, and their modulatory effects on apoptotic signaling pathways in various malignancies, including hepatocellular carcinoma, lung, gastric, pancreatic, and colorectal cancers. Exosomal circRNAs regulate apoptosis primarily through microRNA sponging, interaction with RNA-binding proteins, and the encoding of functional peptides, which influence tumor cell survival, treatment resistance, and metastatic potential. We highlight recent advances in the translational potential of exosomal circRNAs as diagnostic biomarkers and therapeutic targets for overcoming apoptosis evasion in malignancies. Targeting exosomal circRNA-mediated apoptotic networks is a promising approach to precision oncology and better clinical outcomes. This review highlights the critical need for more mechanistic and clinical investigations to utilize exosome biology in cancer therapy innovation.

Unraveling the molecular mechanisms underlying long-range mutational effects on function is a challenging prospect. In this study, we delineate the consequences of a mutation located at a site distal to the functional region in a promiscuous acyl-CoA binding domain. A W86A mutation at a distal C-terminal segment induces two-stage noncooperative unfolding in an otherwise cooperatively unfolding protein, with the mutant exhibiting properties of a molten-globule with large equilibrium fluctuations at 310 K. Under the same conditions, NMR reveals a fourfold increase of a slow-exchanging alternate conformation in the W86A mutant, relative to the wild-type. These manifest as altered dynamic and thermodynamic coupling patterns promoting significant conformational excursions of helix 1, thus stabilizing an intermediate involving unfolded or undocked helix 1. The nonnative interactions mediated by helix 1, in turn, partially occlude the ligand-binding pocket influencing function in a chain-length-dependent manner. Further, the two-stage unfolding switches to a single transition in the W86A mutant in the presence of the ligand while simultaneously suppressing the population of the alternate conformation. Our results provide insights into how mutations could trigger long-range effects through modulation of the native ensemble properties, and the uniquely interconnected nature of the underlying molecular mechanisms.

Hydroxyl radical protein footprinting (HRPF) is a well-established technique used to analyze the binding sites and conformational alterations of proteins. Inline LC-flash oxidation can efficiently capture and characterize structural changes between dynamic protein-ligand complexes as they elute from the LC column. The current work focuses on the development of multidose inline SEC-FOX (size-exclusion chromatography-flash oxidation) for the quantitative topographical analysis of the dynamic protein-ligand complex of human antithrombin III (ATIII) and unfractionated heparin (UFH). ATIII alone and the ATIII-UFH complex were separated by an inline SEC system and flash oxidized at multiple radical doses, followed by proteolytic digestion and LC-MS/MS analysis. A total of 21 oxidized peptides, including three glycopeptides with two major glycoforms each, were reproducibly detected. Six peptides including one glycopeptide (48-53, 58-70, 189-197 A2G2S2 glycoform, 108-114, 333-348, and 360-370) were significantly protected from oxidation upon UFH binding, and one peptide (242-257) showed a significant increase in oxidation upon UFH binding. Residue-level analysis showed that three amino acids (M49, E113, and F329) were significantly protected from modification in the ATIII-UFH complex compared to ATIII alone. Two of these three amino acids are proximal to the known pentasaccharide binding site, while the third is distal and suggests a potential extended interaction of the full-length UFH with ATIII, which was well supported by computational modeling data. We interpret these findings in the context of previously published biophysical data and present a computational model of UFH binding consistent with our findings.

Huntington's disease (HD) is a fatal neurodegenerative disorder caused by an expanded CAG repeat in the <i>HTT</i> gene, producing mutant huntingtin (mHTT) that misfolds into β-sheet-rich aggregates and drives neuronal loss. Current HTT-lowering strategies face challenges, including invasive delivery and nonselective suppression of wild-type HTT. Here, we report the development and synthesis of proteolysis-targeting chimeras (PROTACs) to selectively degrade aggregated mHTT. The lead compound, PROTAC <b>2'</b>, consists of a (pyridylvinyl)aniline aggregate-binding ligand linked via polyethylene glycol spacers to pomalidomide, an E3 ligase recruiter for cereblon. PROTAC <b>2'</b> selectively degraded mHTT aggregates without affecting wild-type huntingtin and significantly reduced mHTT-induced cytotoxicity in the cell model. LC-MS/MS analysis confirmed the blood-brain barrier (BBB) penetration ability of PROTAC <b>2'</b> following subcutaneous administration. In an R6/2 HD mouse model, continuous PROTAC <b>2'</b> delivery via osmotic pumps improved body weight, motor coordination, and survival, correlating with reduced mHTT aggregation and neuroinflammation in the brain. These results highlight the therapeutic potential of aggregate-selective degradation as a disease-modifying strategy for HD, providing a promising alternative to conventional HTT-lowering approaches and supporting the broader potential of PROTAC-based therapeutics for neurodegenerative proteinopathies.

The arcuate nucleus of the hypothalamus plays a central role in sensing and integrating nutritional, hormonal, and neural signals that regulate feeding, energy homeostasis, growth, and reproduction, all of which show pronounced sex differences. However, the cellular mechanisms underlying these responses remain poorly understood. We performed snRNA-seq of the mediobasal hypothalamus, focusing on the arcuate nucleus, in female and male mice under different nutritional states. Analysis of 42 cell types revealed that Agrp neurons were most sensitive to nutritional changes, dopaminergic neurons showed strong sex-specific differences, and KNDy neurons were highly responsive to both sex and nutrition. Pomc neurons displayed moderate nutritional sensitivity. Most glial populations were stable, although microglia and oligodendrocytes showed moderate variation. Cell-cell communication analysis identified neurotrophic factor signaling as a key pathway regulated by sex and nutrition. This study represents a major effort to comprehensively characterize sex-specific differences in arcuate nucleus response across nutritional conditions.

This study presents the development of a novel adsorbent based on hydroxyapatite (CaHAp) grafted with sodium benzoate (SB) at varying proportions (5%, 10%, 15%) via a double decomposition method. Structural and chemical characterizations (XRD, FTIR, SEM, TGA/DTA) confirmed the successful integration of SB, with noticeable improvements in crystallinity (+ 10%), porosity, and thermal stability compared to pristine CaHAp. Adsorption experiments using methylene blue (MB⁺) as a model pollutant showed that the CaHAp-(SB)15 composite achieved an adsorption efficiency of 90%. The adsorption process followed Langmuir isotherm (R² > 0.96) and pseudo-second-order kinetics (R² > 0.99), indicating monolayer chemisorption as the dominant mechanism. Increasing SB content correlated with a significant rise in adsorption capacity from 5.41 to 9.42 mg·g⁻¹, demonstrating the role of –CO2⁻ groups in enhancing electrostatic interactions with cationic MB⁺. DFT simulations supported the experimental findings, showing favorable interactions between SB and CaHAp at both molecular and atomic levels, particularly through non-covalent forces such as van der Waals and electrostatic interactions. This combined theoretical–experimental approach highlights CaHAp-(SB)15 as a promising, low-cost, and eco-friendly adsorbent for dye-contaminated wastewater, offering high efficiency, reproducibility, and environmental compatibility.

Body mass index (BMI) changes throughout life with age-varying genetic contributions. We use a random regression model to investigate the genetic contribution to BMI trajectories from ages one to 18 years in 6,291 ALSPAC participants with 65,930 repeated BMI measurements. Here we show the estimated SNP-based heritability of BMI at 9.5 years is 28.4% (SE = 4.8%), and 23.8% (SE = 4.2%) for rate of change in BMI from one to 18 years. The genetic correlations between early childhood and adolescence are low (genetic correlation between two and 17 years is 0.108 [SE = 0.146]). We find that the first principal component of the trajectory, explaining 89% of genetic variation, captures effects which increase in magnitude from early childhood to adolescence and then plateau. A second axis explaining 9% of the genetic variance has opposite effects on BMI between early and later ages. Our findings demonstrate the value of RRMs to reveal age-specific genetic influences on BMI across development.

Epstein-Barr virus (EBV) infects approximately 90-95% of the global population^1,2 and persists in B cells as a lifelong infection³. Previous EBV infection is associated with autoimmune and neoplastic disease⁴. Still, the biological basis of host control during EBV persistence remains unclear. Here we report the identification of non-genetic and genetic factors that are associated with EBV control during persistent infection. Using blood-based genome sequence data from 486,315 UK Biobank and 336,123 All of Us participants, we identified short-read pairs mapping to the EBV genome in 16.2% and 21.8% of individuals, respectively. EBV read detection (EBVread⁺) reflects increased viral load in blood cells, as shown by orthogonal measurements, and was associated with HIV infection, immunosuppressive drug intake and current smoking. Genome-wide analyses of EBVread⁺ identified strong associations at the major histocompatibility complex (MHC), including 54 independent human leukocyte antigen (HLA) alleles of MHC classes I and II, and at 27 genomic regions outside MHC. Epistasis with distinct HLA alleles of MHC class I was observed at the ERAP2 locus. Analysis of individuals with EBV-associated diseases⁴ revealed a higher polygenic burden of EBVread⁺ for HLA alleles at MHC class I in multiple sclerosis (driven by HLA-A*02:01) and at MHC class II in rheumatoid arthritis. Phenome-wide analyses identified a polygenic overlap of EBVread⁺ with inflammatory bowel disease, hypothyroidism and type 1 diabetes. Our study establishes by-products of human genome sequencing as a surrogate marker of EBV viral load. This will facilitate investigation and treatment for EBV and other persistent viral infections.

Cell-free DNA analysis via methylation and fragmentation profiling has advanced minimally invasive cancer detection; however, broader application has been limited by small cohorts and inconsistent data processing. Here we collated 1,074 cfMeDIP-seq profiles across 9 studies, comprising cancer samples from 11 cancer types, carriers of Li-Fraumeni syndrome and healthy controls. We developed a uniform computational workflow to mitigate technical and biological confounders across cohorts. This analysis identified 14,202 pancancer differentially methylated regions for cancer detection, along with cancer-specific markers for subtype monitoring. Fragmentomic profiling revealed distinguishing differences in 5' end motifs, fragment lengths and nucleosome footprints across cancers. Integrating methylome and fragmentome features enhanced cancer detection and classification. Validation in 220 independent samples, including 3 cancer types absent from the primary dataset, confirmed the robustness of our findings. Altogether, this work provides a pancancer cell-free DNA resource of 1,294 samples to support future methylome and fragmentome studies.

BACKGROUND: The developmental phase is a pivotal biological period for the maturation of the gut microbiota and the establishment of lifelong metabolic health. During these period, dietary patterns that induce dysbiosis, such as the high-fat, low-fiber “cafeteria diet,” disrupt the production of key metabolites in the gut-metabolite axis, including short chain fatty acids (SCFAs) and indole-3-propionic acid (IPA). This study employs a multi-omics approach to examine the impact of cafeteria diet exposure during the developmental period (days 21–56) in 21-day-old male Wistar rats on microbiota composition, SCFA, and IPA levels, and to assess the extent to which concurrent probiotic administration can mitigate these disruptions. RESULTS: The cafeteria diet led to a marked reduction in alpha diversity indices (Shannon p = 0.021; Simpson p = 0.034) and altered the Firmicutes/Bacteroidetes ratio (p = 0.015). Beta diversity analysis indicated a distinct separation between groups (PERMANOVA p = 0.002). Metabolite analysis revealed significant reductions in acetic acid (p = 0.004), isobutyric acid (p = 0.094), butyric acid (p = 0.0014), valeric acid (p = 0.0001), heptanoic acid (p = 0.0125), and IPA (p = 0.002), whereas probiotic administration largely restored these levels. At the species level, cafeteria diet markedly increased Segatella copri, while probiotic intervention partially restored beneficial taxa such as Faecalibacterium prausnitzii and butyrate-producing genera (Anaerostipes hadrus, Intestinimonas butyriciproducens, Blautia wexlerae, and Flintibacter sp. KGMB00164), as evidenced primarily by shotgun metagenomics. Correlation analysis further revealed strong positive associations between butyrate and F. prausnitzii (ρ = 0.65, p = 0.003) and between IPA and B. longum (ρ = 0.68, p = 0.002). Collectively, these results highlight the protective role of probiotic intervention against diet-induced dysbiosis by reinforcing microbiota metabolite interactions. CONCLUSIONS: By integrating metagenomic and metabolomic analyses, this multi-omics study demonstrates that exposure to a high-fat cafeteria diet during the developmental period disrupts microbiota composition and metabolite production, whereas concurrent probiotic administration largely prevent these effects, serving a protective role in the gut-metabolite axis. The study underscores the potential of early-life probiotic intervention, supports SCFA and IPA production, as a critical strategy to optimize microbiota-metabolite interactions and promote long-term gut and systemic health.

BACKGROUND: The tumor microenvironment of oral squamous cell carcinoma (OSCC) is shaped by complex metabolite-mediated cell-cell communication (mCCC), the functional role of which remains incompletely understood. This study aimed to identify key mCCC pathways in OSCC and elucidate the mechanisms by which tumor cells respond to these metabolic signals. METHODS: Single-cell RNA sequencing data from the Gene Expression Omnibus (GEO) database were analyzed using the Metabolite-mediated cell communication modeling by single-cell transcriptome algorithm to identify mCCC events. High-dimensional weighted gene co-expression network analysis (hdWGCNA), in combination with the Mime algorithm, was employed to construct prognostic models and screen for core genes. Enhancer regions were identified by analyzing histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing data from the GEO database. Chromatin immunoprecipitation-quantitative PCR was performed to validate H3K27ac and transcription factor enrichment at these enhancers. M2 macrophages were cultured under glutamine-deprived conditions to generate conditioned medium. OSCC cells treated with L-methionine-DL-sulfoximine were cultured in M2 macrophage-derived conditioned medium, and cell proliferation and invasion were assessed using Cell Counting Kit-8 and Transwell assays. RESULTS: Glutamine secretion from M2 macrophages to tumor cells, mediated by SLC38A5, was identified as the core mCCC pathway upregulated in metastatic OSCC lesions. Knockdown of SLC38A5 significantly inhibited the uptake of M2 macrophage-derived glutamine by OSCC cells, thereby suppressing their proliferation and invasion. CNIH4 was identified as a key effector mediating the response of OSCC cells to M2 macrophage-derived glutamine. Mechanistically, the transcriptional activation of CNIH4 was driven by the direct binding of the transcription factor FOXM1 to its enhancer. Notably, M2 macrophage-derived glutamine upregulated the activity of the CNIH4 enhancer and promoted FOXM1 recruitment in a coordinated manner. Overexpression of CNIH4 rescued the impaired proliferation and invasion induced by FOXM1 knockdown, an effect dependent on the uptake of M2 macrophage-derived glutamine. CONCLUSIONS: In the OSCC microenvironment, glutamine derived from M2 macrophages is taken up by tumor cells through SLC38A5. This process enhances FOXM1 binding to the CNIH4 enhancer, thereby activating CNIH4 expression and promoting OSCC cell proliferation and invasion. Targeting the glutamine/SLC38A5/FOXM1/CNIH4 axis may offer a rational strategy for mCCC-based precision therapy.

Microvillus inclusion disease (MVID) is a congenital diarrheal disorder, caused by inactivating mutations in myosin Vb (MYO5B). MYO5B-deficient mice and cell lines have demonstrated the importance of MYO5B in brush border development; however, the previous models lacked specificity to test intestinal stem cell functions. In the present study, we investigated the effects of progressive MYO5B deficiency originating in intestinal crypt cells utilizing mouse models. Our transcriptomic and multiplex immunostaining datasets demonstrate that MYO5B is critical for intestinal stem cell function. MYO5B-deficient crypts acquire a hyperproliferative phenotype with incomplete cell differentiation in vivo and an elevated organoid formation rate compared to control crypts. An evident disruption in mitochondrial structure and fatty acid metabolism likely underlies these crypt phenotypes. Consistent with mouse models, MVID patient biopsies demonstrate abnormal expansion of the proliferative zone along with villus blunting. These data reveal the direct role of MYO5B in intestinal epithelial progenitor cell functions.

MicroRNAs (miRNAs) play critical roles in ocular surface diseases. In our recent study, through microarray analysis, miR-138-5p was identified as one of the most prominent miRNAs expressed in aniridia patients derived primary limbal epithelial cells (pLECs). Although miR-138-5p expression was significantly elevated in aniridia samples, its specific role in the pathogenesis of aniridia-associated keratopathy (AAK) remained unclear. Therefore, the present study focused on exploring the potential functional impact of upregulated miR-138-5p on limbal epithelial stem cell maintenance and function. Cultured pLECs of healthy human donors isolated from corneoscleral rims were transfected with miR-138-5p mimics. The gene and protein expression level of direct target genes of miR138-5p were evaluated. Additionally, cell proliferation, apoptosis, and cell cycle progression was assessed. Overexpression of miR-138-5p significantly downregulated the expression of several target genes, including Cyclin D (CCND1 and CCND3), Hypoxia-Inducible Factor 1-alpha (HIF1A), Forkhead Box C1 (FOXC1), Caspase 3 (CASP3), and Fos-like 1 (FOSL1). Protein levels of Cyclin D1 were also significantly reduced. Additionally, miR-138-5p transfection markedly inhibited cell proliferation and cell cycle progression. Our results demonstrate that miR138-5p can regulate CCND1 expression, and that its overexpression inhibits limbal epithelial cell proliferation and induces cell cycle arrest, suggesting that miR-138-5p acts as a negative regulator in limbal epithelial cells. Therefore, identifying strategies to suppress miR-138-5p expression at the ocular surface in congenital aniridia could have therapeutic potential for slowing the progression of AAK. Nonetheless, further in vivo studies are needed to fully elucidate the mechanistic link between miR-138-5p and AAK pathogenesis.

<i>KMT2A</i>-rearranged B-cell acute lymphoblastic leukemia (<i>KMT2A</i>r B-ALL) exhibits significant heterogeneity in age of onset, developmental origins, and clinical outcomes. The interplay of individual factors influencing early treatment response within this high-risk molecular subtype remains poorly elucidated. To identify determinants of early treatment response to induction chemotherapy, we analyzed 465 <i>KMT2A</i>r B-ALL cases spanning a wide age range (1 month to 89 years) by integrating transcriptomic and genomic profiling with functional drug response and measurable residual disease (MRD) kinetics. We observed a strong inverse correlation between MRD clearance with advancing age (P = 2.1E-04), proximity to early B-cell-precursor developmental state (low maturity score, P = 1.3E-03), and <i>AFF1</i> as fusion partner (P = 7.0E-04). A multivariable analysis confirmed the strong impact of maturity (P = 0.02) and <i>KMT2A</i> fusion partner (P = 0.03) on MRD clearance, supporting the concept that the cell's developmental state defines therapy response. Gene expression analysis identified cellular traits that relate to MRD clearance (e.g., chromatin organization, immune modulation, and proliferation). This gene expression classifier grouped cases not only by MRD clearance but also by ex vivo sensitivity to induction therapy drugs. Notably, good responders to ex vivo induction drugs were characterized by a higher maturity score (P = 1.8E-03), whereas for less mature <i>KMT2A</i>r B-ALL cases, response profiles suggested higher Venetoclax sensitivity. Our study provides an integrative framework linking developmental phenotype, fusion partner, and MRD kinetics across the full age spectrum of <i>KMT2A</i>r B-ALL. These insights may support future risk-adapted strategies and therapeutic targeting, particularly in immature <i>KMT2A</i>r B-ALL.

Microfluidics-based preparation methods for cell-laden hydrogel microspheres are well-suited for large-scale comparative analysis of single or few cells. However, in existing studies, the preparation of cell-laden hydrogel microspheres and the cell culture process are typically separated, requiring the fabricated microspheres to be eluted and transferred from the preparation device to cell culture dishes or plates for cultivation. This transfer process can easily compromise sterility, while conventional cell culture methods consume more reagents and cause microsphere stacking, hindering single-cell observation and analysis. To address these issues, this paper presents an integrated microfluidic chip that sequentially enables droplet generation with cell encapsulation, gel droplet solidification, hydrogel microsphere trapping, and microsphere-based cell culture and analysis, facilitating the cultivation and observation of single or small numbers of cells. Integrating cell-laden microsphere preparation and 3D cell culture within a sealed chip structure reduces contamination risks associated with cell transfer, enables automation of multiple cell analysis workflows, and minimizes reagent and sample consumption. Using polydimethylsiloxane (PDMS) with good gas permeability and processability as the chip material, biocompatible fluorinated oil was selected as the oil phase for microsphere preparation. A mild sodium alginate-calcium ion gelation system was employed, where calcium ions were released under acidic conditions after droplet generation to trigger solidification, yielding uniform hydrogel microspheres. Under optimized conditions, the single-cell encapsulation efficiency for test samples of human myeloid leukemia cells (K562) was 33.8% ± 1.8%, with a size uniformity coefficient of variation (CV) reaching 3.85%. Cells encapsulated within hydrogel microspheres were cultured in 286 on-chip independent cell culture chambers, achieving >95% viability after 24 h.

Rhabdomyosarcoma is a small round-cell soft tissue tumor that occurs mainly in pediatric and adolescent/young adult (AYA) patients but also rarely in adults. Multidisciplinary treatments including multidrug therapy and local therapy (surgery and/or radiation) are the current standard of care, and treatment strategies are determined according to the estimated risk based on the patient's age, site of onset, and histologic type, as well as the disease stage. New treatment developments in recent years have been based on risk; lower cumulative doses of alkylating agents to reduce late toxicity for low-risk patients are being studied, and long-term maintenance therapy or the addition of new drugs inhibitors to standard multidisciplinary therapy for intermediate- to high-risk patients have been investigated. For high-risk and metastatic patients, novel molecular targeted drug candidates are being evaluated. The target candidates for rhabdomyosarcoma have included the RAS-signaling pathway, ALK, NTRK, FGFR, and MSI-High. In addition, fusion genes (e.g., <i>PAX3/7-FOXO1</i>), which play an important role in diagnostic and prognostic factors, are also being investigated as potential therapeutic targets as their underlying backgrounds are gradually becoming clear. This review summarizes the overall picture of the development of novel therapies for rhabdomyosarcoma and discusses the direction that should be taken in the future.