Gene Literature Dashboard

<h4>Introduction</h4>Lung adenocarcinoma (LUAD) is one of the leading causes of cancer-related deaths worldwide. While NCAPD2 has been implicated in promoting tumorigenesis across various cancer types, its specific role in LUAD remains underexplored. This study aims to elucidate the molecular mechanisms by which NCAPD2 contributes to LUAD progression, with a focus on its involvement in the AKTMDM2/E2F1 positive feedback loop.<h4>Materials and methods</h4>NCAPD2 expression in LUAD and normal tissues was analyzed using Western blotting and immunohistochemistry (IHC). Functional assays, including colony formation, wound healing, Transwell assays, and in vivo mouse models were conducted to evaluate the impact of NCAPD2 on LUAD cell proliferation, invasion, and metastasis. RNA sequencing and protein interaction experiments were used to investigate the role of NCAPD2 in the PI3K/AKT/MDM2 pathway and its interaction with E2F1.<h4>Results</h4>This study first identified that NCAPD2 expression is significantly upregulated in LUAD tissues, particularly in higher pathological stages. NCAPD2 overexpression promoted LUAD cell proliferation and metastasis, while its knockdown inhibited tumor growth and invasion. Mechanistically, NCAPD2 activated the PI3K/Akt pathway, facilitating the interaction between MDM2 and E2F1, reducing E2F1 ubiquitination, and increasing its expression. Furthermore, E2F1 enhanced NCAPD2 transcription, forming a positive feedback loop that drives LUAD progression.<h4>Conclusion</h4>This study reveals a novel role of NCAPD2 in promoting LUAD progression through the AKT/MDM2/E2F1 positive feedback loop. These findings provide new insights into the molecular pathogenesis of LUAD and suggest NCAPD2 as a potential therapeutic target for improving patient outcomes.

Neuroimaging techniques offer valuable insights for understanding pharmacological treatment effects in neuropsychiatric disorders. Here, we present a novel approach that simultaneously assesses hemodynamic and neurometabolic brain responses to psychotropic drugs using interleaved pharmacological magnetic resonance imaging (phMRI) and pharmacological magnetic resonance spectroscopy (phMRS). This method was tested using a double-dose, placebo-controlled, randomized, crossover design using S-ketamine as the pharmacological agent. We acquired 7 Tesla phMRI and phMRS data to evaluate time- and dose-dependent effects of S-ketamine in 32 healthy controls. S-ketamine elicited robust phMRI responses in the dorsofrontal, cingulate, and insular cortices, which correlated with glutamate and opioid receptor maps and subjective dissociation scores. These hemodynamic changes were paralleled by increases in glutamate and lactate, especially at higher doses. Furthermore, accuracy in predicting received condition (placebo, a low, or a high S-ketamine dose) increased when combining both techniques. Here, we show for the first time that concurrent phMRI and phMRS assessments provide important complementary insights into the functional brain response to pharmacological interventions.

Chronic visceral pain is a key symptom of irritable bowel syndrome. Modulation of voltage-gated calcium and potassium channels by G protein-coupled receptors plays a key role in dampening nociceptive transmission. Both baclofen and the analgesic peptide α-conotoxin Vc1.1 activate GABA_B receptors (GABA_BR), resulting in inhibition of Ca_V2.2 and Ca_V2.3 calcium channels to reduce colonic nociception. Recent studies have also shown that GABA_BR activation potentiates G-protein-coupled inwardly rectifying potassium (GIRK)-1/2 channels in mammalian sensory afferent neurons. In this study, we investigated the expression of these ion channel targets in rodent and human dorsal root ganglion (DRG) neurons, including those innervating the colon. We examined how Ca_V2.2 and GIRK channel antagonists, as well as a GIRK channel activator, influence the passive and active electrical properties of adult mouse DRG neurons. We also assessed the effects of α-conotoxin Vc1.1 on neuronal excitability in the presence of the selective Ca_V2.2 antagonist ω-conotoxin CVIE and the GIRK channel activator ML297. We further evaluated the impact of the GIRK channel antagonist tertiapin-Q on excitability in mouse colonic DRGs and afferents and explored the role of hyperpolarization-activated cyclic nucleotide-gated (HCN) channels. Our findings demonstrate that both Ca_V2.2 inhibition and GIRK channel potentiation reduce excitability in mouse DRGs, likely mediating the antinociceptive effects of Vc1.1 and baclofen observed in vivo. However, GIRK channel potentiation appears to play only a limited role in modulating excitability in colon-innervating DRGs and colonic afferents. These findings suggest that neurons innervating different body regions use distinct mechanisms to regulate excitability and nociceptive signalling.

Votoplam is a novel, orally bioavailable, small molecule HTT gene splicing modifier that is being developed for the treatment of Huntington's disease. This was a single dose, open-label, two-period, crossover food effect study that evaluated the effect of high- and low-fat meals on 20 mg votoplam in healthy participants. There was a washout of 21 days between the two periods. Twenty-six participants completed the study. There were minimal changes in the bioavailability and pharmacokinetics of votoplam following administration of a single dose of 20 mg votoplam when taken with low-fat and high-fat meals relative to fasted condition. The mean C_max, AUC_0-last, and AUC_0-inf for votoplam following administration of a single dose of 20 mg votoplam were 1.4-fold, 1.2-fold, and 1.1-fold higher, respectively, in high-fat fed conditions, and were 1.3-fold, 1.1-fold, and 1.1-fold higher, respectively, in the low-fat fed conditions, when compared to fasted conditions. There were no relevant safety findings in any of the treatment groups. Votoplam can be administered with or without food in patients.

Pregnancy- and lactation-associated osteoporosis (PLO) describes a fragility fracture presentation around pregnancy/lactation. Presentation often includes multiple vertebral fractures, but can also involve hip, sacral/pelvic, or other fractures. Substantial bone structural deficits and low bone formation rate have been documented. Most have no known secondary cause. Many have a history of childhood fracture and/or family history of osteoporosis. These characteristics, together with early onset and disease severity, lead to the hypothesis that genetic factors may contribute to PLO. We enrolled 110 women with PLO (mean #fractures = 6, vertebral fractures in 88 %) in an exome sequencing (ES) study. Analyses identified rare (<1 % allele frequency in gnomAD) predicted deleterious variants (RPDV) in 33/110 (30 %) women. All were heterozygous; two participants had multiple RPDV. No RPDV in COL1A1/COL1A2 were identified. 28/110 (25 %) had RPDV in genes related to WNT signaling, critical to bone formation: LRP5 (n = 19), LRP6 (n = 6), WNT1 (n = 2) or WNT1&LRP5 (n = 1). Seven had RPDV related to renal/calcium handling (SLC34A1, SLC34A3, SLC9A3), or other osteoporosis mechanisms (PLS3 (n = 3), HGD (n = 1)). Those with RPDV did not differ from those without in terms of BMD, fracture characteristics, and most clinical characteristics. Among 110 PLO women, exome sequencing analyses identified a potential genetic osteoporosis contribution in 30 %, suggesting that many genetic contributors to PLO have yet to be elucidated. The finding of variants related to WNT signaling in 25 % of the cohort is consistent with the predominantly low bone formation phenotype of PLO and may have implications for prognosis and treatment response.

Endometriosis is a hormone-dependent disease, in the pathophysiology of which sex hormones (androgens, estrogens, etc.) are involved. The level of bioactive androgens/estrogens (in the free state) in the organism largely depends on sex hormone-binding globulin (SHBG), which binds/transports a significant portion of the androgens/estrogens of the body and, due to this, changes the amount of these hormones in a free state (bioactive), which may be important in the development of endometriosis. The study was devoted to identifying the link between the genetic determinants (single nucleotide polymorphisms [SNPs]) of SHBG (according to predating genome-wide associative studies [GWAS]) and the risk of endometriosis in the Caucasian women of Russia. The study was accomplished on a total sample of 1368 women (395 endometriosis; 973 endometriosis free [controls]). Nine loci with an impact on SHBG level in predating GWAS have been examined. The search for associations of these loci with endometriosis was carried out: both their independent effects and interlocus interactions with an in silico interpretation of the functionality/pathways in which endometriosis-related loci and strongly linked SNPs were involved have been evaluated. Polymorphic locus rs440837 (A > G) <i>ZBTB10</i> correlated with endometriosis development (recessive genetic model): the SHBG-raising genotype GG rs440837 (A > G) <i>ZBTB10</i> serves as a risk factor for the disease formation; its presence in the genotype almost doubles the risk of endometriosis (OR = 1.91; 95%CI = 1.13-2.98; p_perm = 0.024; power = 81.13%). The SHBG-impacts of 7 SNPs from 9 analyzed loci such as rs17496332 (A > G) <i>PRMT6</i>, rs780093 (C > T) <i>GCKR</i>, rs10454142 (T > C) <i>PPP1R21</i>, rs3779195 (T > A) <i>BAIAP2L1</i>, rs440837 (A > G) <i>ZBTB10</i>, rs7910927 (G > T) <i>JMJD1C</i>, and rs8023580 (T > C) <i>NR2F2</i> interacting with each other have been endometriosis-associated. Endometriosis-causal SNP rs440837 (A > G) <i>ZBTB10</i> and 5 proxy SNPs determine the DNA interaction in the region of 3 genes (<i>RP11-48B3.3</i>, <i>RP11-48B3.4</i>, <i>ZBTB10</i>) with 22 transcription factors and, due to this, affect the processes of development of the endocrine system, gene transcription regulation, TGF-beta signaling pathway, regulation of cell proliferation/differentiation, etc. In conclusion, the results of this study showed the endometriosis risk effect of the SHBG-impact polymorphic variants.

<h4>Background</h4>Calcium is essential for bone health, but its intake remains inadequate in populations with limited dairy consumption. Fish bones, abundant in calcium and phosphorus, represent a largely untapped natural resource for dietary calcium supplementation. The application of high-energy ball milling (HEM) presents a promising strategy to enhance calcium bioavailability by producing particles at the nanoscale.<h4>Methods</h4>This study used HEM to convert fish bones from Euthynnus affinis (tuna) and Lutjanus sp. (snapper) into nanoparticles. The resulting powder was analyzed for particle size, morphology, crystallinity, thermal, flowability, hygroscopicity, mineral composition, and proximate constituents.<h4>Results</h4>The HEM process successfully reduced the particle size of snapper and tuna bone powder to 729.83 and 847.93 nm, respectively, without altering their crystallinity. Morphological analysis revealed irregularly shaped particles, while thermal analysis indicated an exothermic decomposition pattern. No hygroscopic properties were observed in any of the samples. The nanonization process preserved the intrinsic mineral composition of the fish bones. Calcium/Phosphorus ratios remained >1.8. Elemental analysis confirmed high levels of calcium, phosphorus, and essential minerals such as potassium, magnesium, and sodium, while the protein and ash contents met nutritional standards.<h4>Conclusions</h4>Fish bone-derived nanoparticles produced via HEM exhibit advantageous physicochemical characteristics for nutraceutical applications and represent a promising source of bioavailable calcium to support bone health.

This study introduces an innovative hybrid approach combining salting-out and adsorption for the highly efficient removal of crystal violet (CV) dye from aqueous solutions. The method leverages high-ionic-strength brine discharge from the Complex of El-Outaya (CEO, ENASEL, Biskra, Algeria) and micro-mesoporous biochar derived from calves' horn cores (BHC-800). Results demonstrate that both undiluted and diluted brine significantly enhance CV removal, while BHC-800, with a surface area of 258 m² g^-1, exhibits a maximum Langmuir adsorption capacity of 106.1 mg g^-1 (at 20 °C ± 2). Thermodynamic analysis confirms a spontaneous (Δ<i>G°</i> < 0) and exothermic (Δ<i>H°</i> = -0.86 kJ mol^-1) process, with increased interfacial disorder (Δ<i>S°</i> = 93.53 J mol^-1 K^-1). The synergistic effect of salting-out and adsorption achieved ~99.8% removal of CV at an initial concentration of 1000 mg L^-1. Furthermore, BHC-800 exhibited excellent reusability, maintaining high adsorption efficiency over multiple cycles. Economic assessment revealed operational costs of 0.45-0.89 US$ m^-3 for 60% brine discharge. Biochar production costs were 0.076-0.18 US$ kg^-1, translating to 7.5-17.2 (10^-4 US$) per gram of CV removed. This dual strategy not only offers an eco-friendly and cost-effective solution for dye-laden water but also promotes the valorization of saline effluents and animal byproducts, addressing critical environmental challenges in industrial wastewater treatment.

Skeletal muscle is the largest organ system in mammals. To investigate the differences in energy metabolism across various skeletal muscles in Yili horses, this study examined muscle fiber type distributions through immunohistochemical staining of muscles, including the splenius, triceps brachii, longissimus dorsi, and gluteus medius. The splenius and gluteus medius muscles, which exhibited the greatest differences in the proportion of slow-twitch fiber area, were selected for further comparison of differential metabolites and transcriptomic expression profiles between slow-twitch and fast-twitch fibers. A total of 27 energy metabolism-related differential metabolites, including pyruvate and lactate, were identified, along with 432 differentially expressed genes, such as PFKM and ALDOA. Additionally, 164 differentially expressed miRNAs, including miR-499 and miR-24-3p, were detected. We found highlighted differences in LDHA expression between the gluteus medius and splenius muscles, which may influence the conversion of fast and slow muscle fibers by modulating the glycolysis/gluconeogenesis pathway. The miRNA-mRNA targeting relationships established here warrant further validation. These findings provide valuable insights into the molecular mechanisms underlying energy metabolism differences in Yili horses.

<h4>Background</h4>Melanoma is a highly heterogeneous neoplasia in which transcriptional profile encodes much of the biological diversity that determines tumor progression and therapeutic response. To refine its molecular stratification and profiles characterization, we conducted an in silico transcriptomic analysis.<h4>Methods</h4>Public microarray datasets from the GEO and ArrayExpress were examined, and the E-MTAB-6697 expression dataset was selected. We used a K-Means clustering algorithm to stratify 194 tumor samples into expression-driven subgroups and analyzed each one to define their transcriptional and biological profiles. Differential expression analysis between identified clusters and controls was performed. Additionally, we applied Weighted-Gene correlation analysis to identify coordinated expression hubs in the tumor dataset and tested the resulting modules for correlation with the identified clusters.<h4>Results</h4>Unsupervised clustering of melanoma transcriptomic profiles identified three distinct molecular subtypes characterized by divergent biological programs. While all clusters shared the dysregulation of pathways involved in epidermal differentiation, immune response, and lipid metabolism, they diverged in proliferation, phenotypic plasticity, metabolic adaptation, and apoptotic regulation. Cluster A was characterized by enrichment in DNA replication, repair, and mitochondrial metabolism modules, suggesting a proliferative yet genomically stable state. Cluster B showed enrichment in immune and cytokine signaling pathways alongside reduced proliferative activity, consistent with a quiescent or transitional phenotype. Cluster C displayed coordinated enrichment in cell-cycle, DNA-maintenance, and neuroectodermal reprogramming pathways, indicating a highly plastic and proliferative subtype. Despite these molecular distinctions, all clusters retained an "immunologically hot" profile (IPS 7-8), indicating potential responsiveness to immunotherapy.<h4>Conclusions</h4>These findings provide an overview of the functional characteristics of melanoma heterogeneity and identify biological processes that could be targeted by drugs for the development of tailored therapies for each subtype. Nevertheless, future studies in independent clinically annotated cohorts would be required.

Sodium-glucose co-transporter 2 (SGLT2) inhibitors, initially developed to treat type 2 diabetes mellitus, work by blocking SGLT2 proteins in the proximal convoluted tubules of the nephrons, thereby preventing glucose reabsorption into the bloodstream. Subsequently, its mechanism reveals additional cardiovascular and renal benefits, necessitating further investigation into its effects on heart failure. This condition affects millions and leads to diminished quality of life, frequent hospitalizations, and high mortality rates. This review aims to evaluate several large randomized controlled trials (RCTs) investigating the favourable outcomes of SGLT2 inhibitors on heart failure and its related variables, including hospitalizations secondary to the condition, mortality rate, kidney function, and left ventricular ejection fraction (LVEF). Nine RCTs were reviewed systematically and meta-analyzed to assess the effectiveness of SGLT2 inhibitors. A pooled risk ratio was calculated under a random-effects model, along with heterogeneity analysis, to summarize the overall effect across studies. The studies analyzed two primary clinical outcomes of mortality and rehospitalization due to heart failure. SGLT2 inhibitors were associated with a 20% relative reduction in mortality (risk ratio (RR) = 0.80) and a 24% relative risk reduction of rehospitalization due to heart failure (RR = 0.76). These findings support the cardioprotective effects of SGLT2 inhibitors, as well as significant reductions in hospitalization and mortality. The efficacy of SGLT2 inhibitors was observed across diverse demographic and clinical subgroups, supporting their role as a valuable therapeutic option for managing heart failure.

As the global population ages, the prevalence of neurodegenerative diseases (NDDs)including Alzheimer's disease, Parkinson's disease, Huntington's disease, Multisystem Atrophy (multiple system atrophy), and amyotrophic lateral sclerosiscontinues to rise, largely driven by environmental, metabolic, and lifestyle risk factors. Advances in computational technologies, particularly machine learning (ML) and deep learning, are reshaping research in this field. This review summarizes the major features of these diseases and emphasizes the role of ML in drug discovery, virtual screening, drug repurposing, and drug combination optimization. Representative approaches include support vector machines for classification, convolutional neural networks|convolutional neural network for imaging analysis, recurrent neural networks for temporal biomedical data, and transformers for multimodal integration. These methods highlight the potential of computational strategies to improve therapeutic development. In addition, the review underscores the substantial incidence rates and socioeconomic burden of these conditions, which have made them focal points for algorithmic innovation. With research evolving rapidly, the development of AI-driven approaches is expected to enable more effective, targeted interventions and improve patient outcomes. This Perspective provides a concise overview of current progress and identifies promising future directions in the fight against NDDs.

Electrochemical immunosensors are emerging as promising tools for cancer detection due to their simplicity, portability, and high sensitivity. Colorectal cancer (CRC), the third most common cancer in the United States, remains challenging to diagnose early, as the standard method, colonoscopy, is invasive and often avoided. To address this gap, a graphene-gold-based immunosensor was developed for the early detection of CRC by targeting colon cancer-secreted protein-2 (CCSP-2), a biomarker overexpressed in CRC patients. The sensor was fabricated by attaching graphene oxide (GO) to a gold (Au) electrode using 4-aminothiophenol (4-ATP) as a linker, followed by immobilization of CCSP-2 antibodies (Anti-CCSP-2) and blocking with bovine serum albumin (BSA). Characterization of the immunosensor using linear sweep voltammetry (LSV), Raman spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) confirmed successful modifications. The attachment of graphene oxide contributed to an enhancement in current response, likely due to partial reduction and improved electron transfer at the modified surface. The sensor demonstrated a good linear response (<i>R</i> ² = 0.979) to CCSP-2 antigen (CCSP-2) concentrations ranging from 1 ng/μL to 100 ng/μL, with a limit of detection (LOD) of 0.17 ng/μL and a sensitivity of 0.031 (ng/μL)^-1. Selectivity was validated using CRC cell extracts (CACO-2) and human kidney extracts (HEK), showing a more significant signal for CACO-2. These findings suggest that the developed immunosensor is a reliable and sensitive platform for CCSP-2 detection, with the potential for adaptation as a point-of-care device for early CRC screening.

Collagen type I, a key component of the extracellular matrix, underpins cellular organization and regeneration but remains challenging to process into scalable, reproducible, and biologically faithful 3D materials. Although several recent platforms have achieved high-throughput collagen microgel fabrication, most rely on chemical modification or rigid process parameters that compromise native collagen structure or limit biological versatility. A scalable yet biologically unmodified system remains lacking. Here, we developed a pressure-tunable microfluidic platform that enables the rapid, high-throughput generation of collagen microbeads with controlled size, fibrillar architecture, and mechanical properties for versatile downstream biofabrication. By adjusting collagen concentrations (0.25-3 mg mL^-1) and thermal self-assembly, we produced monodisperse microbeads (45-1000 μm) at rates exceeding 10,000 beads min^-1 (~166 Hz) while preserving the native triple-helical structure. The microbeads exhibited tunable stiffness (0.6-2.8 kPa) and pore structures confirmed by scanning electron microscopy, enabling systematic modulation of the 3D microenvironment for cell culture and bioprinting. When incorporated into alginate-gelatin bioinks, the collagen microbeads enhanced rheological stability, shear-thinning, and print fidelity, supporting sustained cell growth that reached a 13-fold increase in proliferation over 21 days; independently, cell-laden collagen microbeads functioned as self-contained microbioreactors enabling multicellular spheroid formation and continuous EV secretion. This modular and naturally derived, unmodified collagen microbead technology unites tunability, throughput, and functional scalability, establishing a versatile platform for tissue modeling, drug testing, and therapeutic EV biomanufacturing.

Major depressive disorder (MDD) is a multifaceted psychiatric illness with profound global consequences. To illuminate its molecular underpinnings, we employed a genome-driven integrative systems neuroimmunology approach to analyze transcriptomic profiles from 3114 individuals (1877 MDD patients and 1237 controls). This analysis uncovered coordinated neuroimmune transcriptomic shifts, marked by altered expression of genes involved in innate immune regulation, suppression of inflammatory responses, and pathways related to sensory perception, visual system development, and synaptic signaling. These alterations were consistently observed in both peripheral blood leukocytes (PBLs) and brain regions implicated in MDD. Among 31 genes jointly dysregulated in blood and brain, four stood out as robust predictors of MDD in PBLs: NEGR1, PPP6C, SORCS3, and PAX6. Of these, PAX6, a gene previously linked to MDD by GWAS, also exhibited differential expression in the amygdala and was functionally enriched in pathways governing immune modulation, vesicle trafficking, and neurodevelopmental processes, such as neuronal fate determination. In contrast, NEGR1, PPP6C, and SORCS3 showed no significant changes in brain expression, suggesting a predominantly peripheral role. Importantly, these transcriptomic insights were reinforced in a murine model of chronic stress, where immunophenotyping revealed elevated PAX6 expression in peripheral myeloid cells. Together, these findings reveal a shared neuroimmune signature across the brain and immune system in MDD, highlighting PAX6 as a promising mechanistic link and potential biomarker for this disorder.

A signature of 16 serum proteins that were previously profiled using the aptamer-based Somascan technology highlighted the roles of the e2 allele of APOE in lipid regulation via apolipoprotein B (APOB) and apolipoprotein E (APOE) and in inflammation. Here, the serum protein signature of APOE is validated and expanded using a combination of mass-spectrometry, ELISA, Luminex, blood transcriptomics, and antibody-based Olink serum proteomics. Some of the findings were replicated in the UK Biobank using antibody-based Olink serum proteomics. This analysis replicated the association between APOB and the e2 allele of APOE, detected a new, robust pattern of association between APOE genotypes and the serum level of APOE, and discovered new associations between APOE genotypes and the complex of apolipoproteins APOC1, APOC2, APOC3, APOC4, APOE, APOF, and APOL1. In addition, 13 new proteins correlated with APOE genotypes. This extended signature includes granule proteins CAMP, CTSG, DEFA3, and MPO secreted from neutrophils and points to olfactomedin 4 (OLFM4) as a new target for the prevention of Alzheimer's disease.

Humans are chronically exposed to mixtures of environmental contaminants. Exposure to endocrine-disrupting chemicals (EDCs) contributes to increased health impairment observed globally. This study aimed to evaluate the endocrine-disruptive and oxidative stress potential of a human-relevant, complex chemical mixture in vitro. By testing chemical class subgroup mixtures, the identity of toxicological drivers and mixture additivity could be investigated. A 50-component mixture was compiled based on Swedish human blood concentrations (xHBC), consisting of six subgroup mixtures: polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (PCB mixture), brominated flame retardants (BFR mixture), per- and polyfluoroalkyl substances (PFAS mixture), pesticide mixture, synthetic phenolic contaminants (phenol mixture), and phthalate mixture. These were tested in four chemically activated luciferase gene expression (CALUX) assays: dioxin responsive (DR-), estrogen receptor α (ERα-), androgen receptor. (AR-), and nuclear factor erythroid 2-related factor 2 (Nrf2)-CALUX, along with an adipocyte cell assay. The total mixture caused significant agonistic activity in DR- and ER-, and antagonistic activity in AR-CALUX at 0.1-15 xHBC, depending on the assay. Mixture additivity was assessed in ERα-, DR-, and anti-AR-CALUX using subgroup mixtures and the concentration addition (CA) model. The total mixture followed the CA model in ERα-, anti-AR- and DR-CALUX. The toxicological drivers of these activities were mainly the PCB and phenol mixture. A significant increase in differentiated adipocytes was observed at 100 xHBC. These results raise concerns regarding potential health effects on the endocrine system. The additive effects at human-relevant concentrations observed in this study motivate considering mixtures in regulatory contexts to protect the well-being of future generations.

Alzheimer's disease (AD) still lacks a conclusive treatment, largely due to an incomplete understanding of the molecular mechanisms involved. To enhance our knowledge of AD pathogenesis and identify potential therapeutic targets, this study integrates differential gene expression analysis, pathway enrichment, hub gene discovery, protein-protein interaction (PPI) clustering, and transcription factor/protein kinase regulation into a single, cohesive pipeline. This comprehensive systems-level approach moves beyond single-gene analyses to offer a broader, mechanistically focused insight into AD biology. Using RNA-seq data from the CA1 region of the hippocampus-a subregion selectively affected in early AD-we identified 1,104 differentially expressed genes (DEGs). Among the enriched pathways, "7-alpha-hydroxycholesterol" was upregulated, while "vacuolar organization" was downregulated in AD samples. Furthermore, five novel hub genes (MRPS7, RPL5, GFM1, RAD51, and ASPM) were identified within the PPI network. The first three-MRPS7, RPL5, and GFM1-along with ACO2 and MT-ATP6, are potentially linked to hereditary forms of AD due to their roles in mitochondrial function. We also discovered four collaborative clusters within the network that notably associated with the "inflammatory response", "7-alpha-hydroxycholesterol", "Mitochondrial dysfunction" and "Oxidative phosphorylation" pathways, making them promising candidates for therapeutic and diagnostic investigation due their behavioral information members. Additionally, we identified ten transcription factors (GATA2, CHD1, THRA, IRF7, ZBTB48, POLE4, ZNF219, SLC2A4RG, NR1D1, and RXRA) and one protein kinase (PRKCZ) as potential regulatory elements in AD. This study broadens our understanding of Alzheimer's disease by identifying five candidate hub genes, two functional PPI clusters, two signaling pathways, and eleven regulatory proteins, thereby laying the groundwork for future therapeutic and diagnostic developments in molecular AD research.

Thyroid hormone (TH) is crucial to maintaining nervous system's normal function and is essential for formation of dendrites, axons, and myelin sheaths of neurons, which form the basis for transmitting messages between neurons, and the deficiency of TH affects their development, which in turn affects nervous system's normal operation. Recent articles have found that TH deficiency or abnormal levels not only affect mood and cognitive function but are also closely related to the development of a series of neurodegenerative disorders, like Alzheimer's, Parkinson's and Huntington's diseases, along with multiple sclerosis. The pathologic mechanisms of such diseases are complex, and it is difficult to confirm clinical diagnosis, and there exists a shortage of valid therapies, which can greatly reduce patients' life quality once they develop. Therefore, a deep comprehension of TH mechanism in neurodegenerative disorders and exploring factors affecting its relationship with neurodegenerative disorders are of great clinical and research value. This article reviews the research progress in this field so far, analyzes the functional changes of TH in neurological pathological states, and explores its potential in clinical applications, aiming to provide new orientation for neurodegenerative disorders' future diagnosis and therapy, and thus for developing more valid therapeutic strategies.

BACKGROUND: Establishing a comprehensive characterization of the regulatory landscapes of cattle tissues facilitates a better understanding of the biological mechanisms responsible for condition-dependent phenotypes that drive tissue-specific gene regulation, developmental processes, and responses to environmental or physiological cues. This can be achieved through the characterization of gene expression, transcript usage and open chromatin accessibility. While much of this work has been done in human and biomedical model species, there is a lack of research in cattle that jointly characterizes chromatin accessibility and transcript usage alongside gene expression to define tissue-specific regulatory landscapes in cattle. Samples of prefrontal cortex (PFC), liver (L), and gracilis skeletal muscle (SM) were collected from four 3-month-old Angus steers and subjected to ATAC-seq and RNA-seq analyses. Differential gene expression (DGE) and transcript usage (DTU) were quantified across the tissues and evaluated in PFC-L, PFC-SM, and L-SM pairwise comparisons. RESULTS: In total, 4,840, 4,151, and 3,663 genes were identified as differentially expressed, while 209, 236, and 154 genes exhibited differential transcript usage in PFC-L, PFC-SM, and L-SM comparisons, respectively. The genes CLTA, SLC25A3 and P2RX5 were highlighted for DTU between tissues, and further comparison of predicted protein sequences identified structural differences between CLTA and P2RX5 isoforms suggesting tissue-specific roles. Differential chromatin accessibility analysis revealed 2,576, 1,820, and 2,960 differential peaks in promoter regions across the same pairwise tissue comparisons. Functional enrichment revealed that PFC regulatory genes were associated with synaptic formation, myelination, and neurodevelopment, L genes with homeostatic regulation and lipid metabolism, and SM genes with muscle differentiation, myonuclear addition, and hypertrophy. These patterns aligned with ATF, HNF, and MEF2 motif enrichment in PFC, L, and SM promoter peaks, respectively. Furthermore, the integration of ATAC-seq and RNA-seq results revealed 62 genes in PFC, 93 genes in L, and 59 genes in SM as potential tissue-specific regulatory genes. CONCLUSIONS: These findings provide insight into the complex regulatory mechanisms governing biological processes in the PFC, L, and SM of young Angus steers. This work integrates chromatin accessibility, transcript usage, and gene expression across multiple physiologically important cattle tissues, providing a comprehensive view of tissue-specific regulatory landscapes and generating a resource to support future studies in cattle genomics.

Circular RNAs (circRNAs) are a unique class of endogenous, single-stranded non-coding RNAs characterized by a covalently closed circular structure. They exhibit tissue-specific expression patterns and have been implicated in the pathogenesis of a wide range of diseases. In the brain, circRNAs are abundantly expressed and evolutionarily conserved, where they play critical roles in neurodevelopment, neuronal differentiation, and synaptic plasticity. Recent studies have revealed that circRNAs are broadly expressed across eukaryotic species and display distinct expression profiles associated with various pathological conditions, particularly neuropsychiatric disorders. However, despite the identification of thousands of circRNAs through high-throughput sequencing technologies, only a small subset has been functionally characterized. In this review, we underscore the emerging significance of circRNAs in neurological and psychiatric disorders. Their exceptional stability and diverse modes of action suggest that circRNAs could serve as key nodes within future precision medicine frameworks. Nevertheless, progress toward clinical translation is currently limited by incomplete mechanistic understanding, small study cohorts, and persistent challenges in therapeutic development. To overcome these barriers, more systematic, large-scale, and mechanism-driven investigations, coupled with innovative strategies and technologies, will be required to fully realize the clinical potential of circRNAs.

In contact with saliva, tooth enamel is covered by biomolecules forming an initial biofilm. Microorganisms attach to the initial biofilm and form the bacterial biofilm, which can provoke diseases. Therefore, dental biofilms are the focus of preventive research. Enamel consists mainly of hydroxyapatite (HAP). Yet, the composition of dental apatite differs between individuals and influences enamel properties. Standardized surfaces might therefore be useful for biofilm research. Synthetic enamel-like HAP pellets perfectly meet the criteria for such well-defined samples. However, systematic investigations of synthetic HAP on oral biofilm formation have never been performed, especially not in comparison to enamel. Therefore, we systematically compared the in situ biofilm formation on synthetic HAP and enamel to investigate the suitability of HAP as a substitute for natural enamel in biofilm formation studies. We observed no differences in formation kinetics, microstructure and subject-specificity of the initial biofilm on both materials. Furthermore, at the proteome level the development of the biofilm on HAP follows the formation patterns observed for enamel. Formation kinetics and morphology of the bacterial biofilm were also subject-dependent and not distinguishable between the two materials. However, the bacterial viability on HAP was higher than on enamel. For bacterial biofilm viability studies, synthetic HAP may therefore even be the preferred substrate as it is more beneficial for identifying antimicrobial agents. In summary, the results prove synthetic HAP as perfect substrate for dental biofilm studies.

Neuroendocrine prostate cancer (NEPC) is a highly aggressive and lethal subtype of prostate cancer (PCa) that often emerges in response to androgen receptor pathway inhibitors (ARPIs), which are widely used in treating metastatic castration-resistant and hormone-sensitive prostate cancer. The incidence of NEPC is increasing, yet effective therapeutic strategies remain limited due to an incomplete understanding of its molecular drivers. Through transcriptomic analyses of human prostate tumor samples, we identified the histone lysine demethylase KDM4A as uniquely overexpressed in human and mouse NEPC compared to prostate adenocarcinoma. Functional validation demonstrated that KDM4A is a key regulator of NEPC progression and a promising therapeutic target, as knockdown or knockout of KDM4A suppresses NEPC cell proliferation in vitro and tumor growth in vivo. Mechanistically, we found that KDM4A directly regulates the transcription of the oncogene MYC, which we show is essential for NEPC cell growth through knockdown and inhibitor studies. Importantly, pharmacologic inhibition of KDM4A using QC6352, a potent pan-KDM4 inhibitor, significantly reduced NEPC cell proliferation in vitro and tumor growth in vivo, providing proof-of-concept for therapeutic targeting. Collectively, our findings establish KDM4A as a critical epigenetic driver of NEPC through MYC regulation and demonstrate its therapeutic potential for this lethal disease that currently lacks effective treatments.

The High Mobility Group Nucleosome-binding (HMGN) proteins are small, abundant nuclear proteins that directly bind nucleosomes and form a major component of chromatin. HMGN proteins localize to enhancers and actively transcribed genes across the genome; however, their roles in regulating chromatin structure and transcription remain poorly understood. Although it is well established that HMGN proteins bind to the H2A-H2B acidic patch on nucleosomes, other potential nucleosome targeting mechanisms, including histone post-translational modifications and histone variants, remain unclear. To investigate the nucleosomal binding preferences and function of HMGN proteins, we engineered mouse embryonic stem cells (mESCs) lacking HMGN1 and/or HMGN2 (Hmgn1^-/- mESCs, Hmgn2^-/- mESCs, and Hmgn1^-/-Hmgn2^-/- mESCs) and profiled gene expression and localization of architectural proteins. In the absence of these HMGN proteins, ∼1000 genes were differentially expressed, including cell identity genes, with most genes being downregulated. Nucleosome binding assays revealed preferential binding of HMGN1 and HMGN2 proteins to nucleosomes with acetylated H3 tail residues and nucleosomes containing the histone variant H2A.Z. In addition, in vitro acetylation assays demonstrated that binding of HMGN1 and HMGN2 to nucleosomes reduces p300-mediated acetylation of H3K18, H3K23, and H3K27 residues. An epiproteomic mass spectrometry analysis of histone tail modifications revealed that Hmgn1^-/-Hmgn2^-/- mESCs have increased steady-state levels of H3K27me2 and H3K27me3, but not H3 tail acetylation, relative to WT cells. Together, these findings show that HMGN proteins function as both sensors and modulators of the histone post-translational modification landscape inside cells, playing a critical role in the dynamic balance between active and repressive chromatin states.

<h4>Background & aims</h4>The quiescence-activation transition of cancer stem cells regulated by environmental stimuli has been shown to potentially contribute to cancer maintenance and regrowth after therapy. However, little is known about how the neoplastic niche couples with neighboring signals to control the activation of quiescent rectal cancer stem cells during radiotherapy.<h4>Methods</h4>Lineage-tracing experiments were performed usingHopx^CreERT2;Rosa^tdTomato mice and organoids to visualize the dynamics and radioresistance ofHopx-expressing cells in vivo. BrdU pulse-chase assays and cell cycle analysis were used to identify whether Hopx⁺ stem cells were label-retaining cells (LRCs). The paracrine pro-survival effect of radiotherapy-induced dying cancer cells on neighboring Hopx⁺ quiescent stem cells was analyzed in the context of both apoptosis and necroptosis blockade. Human rectal cancer organoids and patient-derived xenografts (PDXs) were used to assess the radiotherapy-enhanced efficacy of Hopx targeting.<h4>Results</h4>Lineage tracing experiments revealed that Hopx⁺ quiescent stem cell subpopulation exhibited an enhanced regeneration, which functionally drove the recurrence of rectal cancer after irradiation. Mechanistically, iron released from radiotherapy-induced tumor cell death triggered a Stat3-dependent pro-survival program in neighbor-surviving Hopx⁺ quiescent stem cells. Interestingly, we demonstrated activated Hopx⁺ cancer stem cells antagonized ferroptosis that should be caused by iron-overload via the inhibition of de novo lipid synthesis.<h4>Conclusions</h4>Collectively, quiescent cancer stem cells could establish a new dependency on anti-apoptotic programs in their dying neighbors. This study highlights targeting and regulating Hopx⁺ quiescent stem cells could be a promising therapeutic approach to overcome the refractoriness of human rectal cancer.

The Raf kinase inhibitor protein (RKIP) functions as both a metastasis suppressor and immune enhancer, exerting its influence over several key oncogenic signaling pathways, including the MAPK, NF-κB, and PI3K pathways. Recent studies have highlighted a potential interplay between RKIP and hypoxia-inducible factors (HIFs), particularly in the hypoxic tumor microenvironment (TME). Hypoxia is known to reprogram cellular metabolism, enhance angiogenesis, and facilitate immune escape. Through analysis of cross-talk signaling pathways between RKIP and HIFs, we establish the presence of a dysregulated RKIP-hypoxia axis in cancer. Notably, many cancers simultaneously express low levels of RKIP and high levels of HIFs an expression pattern that strongly correlates with the emergence of immune evasion mechanisms. Herein, we report on the mechanisms by which this dysregulated axis mediates immune evasion. These include the molecular regulations of RKIP and HIFs expressions, and the low expression of RKIP and high expression of HIFs in several cancers. We report on the mechanisms underlying immune evasion by the RKIP-hypoxia axis by examining various factors intimately involved in immune evasion, such as the upregulation of PD-L1, matrix metalloproteinases (MMPs), anti-apoptotic molecules, CD47, and the enhanced frequencies of regulatory T cells (Tregs), myeloid-derived suppressor cells (MDSCs), tumor-associated macrophage (TAM) polarization, and decreased antigen presentation. Thus, hypoxia-induced repression of RKIP establishes a feedforward loop that sustains immune evasion and tumor aggressiveness. Therapeutically, we propose that targeting the RKIP-hypoxia axis offers a new strategy to restore immune surveillance and counteract tumor progression. We present various means to target the inhibition of hypoxia as well as the induction of RKIP. Elucidating the molecular crosstalk between RKIP and hypoxic stress responses opens a new paradigm for strategies that enhance the efficacy of immunotherapies and overcome tumor resistance.

The challenges associated with solubility and bioavailability of porphyrinoid-type photosensitizers in photodynamic therapy require solutions that are based on modern drug carriers, including polymeric nanoparticles. With that in mind this review discusses poly(lactic-<i>co</i>-glycolic acid, PLGA)-based polymeric nanoparticles encapsulating selected well-known photosensitizers, such as protoporphyrin IX, tetrahydroxyphenylporphyrin, chlorin e6, and tetracarboxyphenylporphyrin, with a view to the physicochemical and biological properties. Also discussed are their potential medical applications towards photodynamic and sonodynamic therapy. PLGA-based nanoparticles, encapsulating photosensitizers, were analysed in terms of particle size, surface charge, morphology, loading efficiency, release kinetics, and stability. Moreover, the cellular uptake and subcellular localisation of carriers were considered in correlation to polymer composition and surface functionalisation. Special attention was given to how PEGylation, lipid-hybrid coatings, or the incorporation of additional therapeutic or imaging agents has modulated both the physicochemical properties and biological activities of photosensitizers. The comparative assessment of different porphyrinoid-based photosensitizers highlighted how hydrophobicity, amphiphilicity, and molecular structure have an influence on encapsulation efficiency and therapeutic outcomes. Furthermore, issues such as the premature release of photosensitizers, along with limited bioavailability, and limited penetration through biological barriers were addressed as well as some proposed mitigation strategies. Overall, this review highlights the versatility of PLGA nanoparticles as a powerful platform for photosensitizer delivery, with promising implications for advancing polymer-based nanomedicine and improving the efficacy of photodynamic therapy.

Postmenopausal osteoporosis (PMOP) arises from estrogen deficiency, which disrupts bone remodeling by shifting the balance toward bone resorption over osteogenesis. Glycolytic regulation has emerged as a critical mechanism governing osteoclast differentiation and resorptive activity. Blocking lactate transport through monocarboxylate transporters (MCTs) suppresses glycolysis, thereby attenuating these processes and highlighting MCT inhibition as a potential therapeutic target. The MCT inhibitor AZD3965 blocks lactate transport, thereby downregulating NF-κB/MAPK signaling, increasing intracellular lactate levels, and ultimately suppressing osteoclast formation and bone resorption in vitro. To achieve targeted delivery and reduce off-target effects, a bone-targeted, reactive oxygen species (ROS)-responsive nanocarrier (PH/DPA@A) was engineered by integrating a bone-affinitive DSPE-PEG-Asp₈ (DPA) ligand with a ROS-cleavable phenylboronic acid pinacol ester-hyaluronic acid (PH) shell to encapsulate AZD3965. The nanoparticles exhibited a mean diameter of ∼179 nm, well-defined ROS-triggered drug release kinetics, and high in vivo bone-targeting efficiency. In vitro, PH/DPA@A inhibited osteoclast formation and resorptive activity at levels comparable to free AZD3965, indicating preserved pharmacological potency. In ovariectomized (OVX) mice, systemic PH/DPA@A administration increased femoral bone mineral density and improved trabecular number, thickness, and connectivity, as confirmed by micro-computed tomography. These findings demonstrate that the bone-targeting, ROS-responsive design enables efficient in vivo delivery and metabolic modulation in osteoporotic bone, supporting PH/DPA@A as a multifunctional nanoplatform with translational potential for postmenopausal osteoporosis therapy.

<b>Objective</b>: Neuronal growth regulator 1 (<i>Negr1</i>) is a GPI-anchored neuronal cell adhesion molecule of the IgLON superfamily associated with multiple psychiatric disorders. This study aimed to investigate behavioral and molecular adaptations to social isolation (SI) stress in <i>Negr1</i>-deficient (<i>Negr1^-/-</i>) mice. <b>Methods</b>: Male and female <i>Negr1^-/-</i> and wild-type (Wt) mice (n = 10 per group) were exposed to two weeks of SI or group housing (Ctl). Behavioral assays assessed exploratory and anxiety-like behavior. Gene expression analyses in the prefrontal cortex and hippocampus were performed using RT-qPCR, focusing on GABAergic, neurotrophic, and IgLON family genes. <b>Results</b>: SI-induced weight loss in <i>Negr1^-/-</i> mice compared to Wt was evident in both sexes but more pronounced in males. Behaviorally, SI Wt males showed stress-induced hyperactivity compared to Ctl Wt, whereas SI <i>Negr1^-/-</i> males exhibited blunted exploratory behavior relative to SI Wt in the open field test (OFT). <i>Negr1^-/-</i> females showed reduced exploration in the elevated plus maze (EPM), suggesting increased anxiety. Hippocampal <i>Pvalb</i> was downregulated in SI <i>Negr1^-/-</i> mice of both sexes compared to Wts, with a stronger decrease in males, indicating heightened male vulnerability in GABAergic interneuron function. In males, SI reduced hippocampal <i>Bdnf</i> in both genotypes, whereas <i>Ntrk2</i> (TrkB) upregulation occurred only in <i>Negr1^-/-</i> mice, suggesting a genotype-specific compensatory response. Hippocampal expression of <i>Fgfr2</i> and IgLON members (<i>Ntm1a/1b</i>, <i>Lsamp1a/1b</i>) was increased in SI <i>Negr1^-/-</i> males compared to SI Wt, with minimal changes in females. <b>Conclusions</b>: <i>Negr1</i> deficiency leads to sex-specific behavioral and molecular adaptations to social isolation stress, highlighting the role of <i>Negr1</i> in modulating neurotrophic and GABAergic signaling pathways under adverse environmental conditions.

Drug-induced liver injury (DILI) is an uncommon but potentially serious adverse reaction that can arise from the use of various antidepressants. Vortioxetine, widely used for the treatment of major depressive disorder due to its favorable efficacy and tolerability profile, has been infrequently associated with hepatic toxicity. We describe the case of a 59-year-old female who developed acute hepatitis with cholestatic features after long-term vortioxetine therapy. Extensive evaluation ruled out viral, autoimmune, and metabolic causes. Liver function improved after the discontinuation of the drug but worsened upon rechallenge, necessitating permanent withdrawal of the drug. A subsequent episode of hepatic failure after exposure to venlafaxine suggested a class-related susceptibility to selective serotonin reuptake inhibitor (SSRI)-induced hepatotoxicity. The patient recovered following cessation of antidepressants and treatment with N-acetylcysteine. This report underscores the importance of considering DILI in patients treated with vortioxetine or other serotonergic antidepressants.

Live cells in tissue are plastic, phenotypically dynamic, and modify their function in response to genetic and environmental perturbations. To unleash the power of live-cell imaging to identify phenotype-genotype-function coupling over time, we report the development of a standardized Shape-Appearance-Motion (SAM) “phenome” and SAM-Phenotype-Observation-Tool (SPOT), that act as an image-“transcriptome” and image-“transcriptome analyzer” respectively, and provide unbiased and comprehensive description of morpho-dynamic phenotypes without prior knowledge. We developed and applied SAM-SPOT to our simulated organoids database with known ground-truth and >1.6 million mouse and human organoid instances with defined genetic and chemical perturbations. SAM-SPOT can effectively and robustly characterize 3D morpho-dynamics from 2D projection videos. Combined with single-cell RNA sequencing, SAM-SPOT revealed that altered WNT signaling, but not mutant RAS or p53, predisposes intestinal organoids to irregular morphogenesis. SAM-SPOT advances biomedical discovery by empowering live-cell imaging to identify phenotype-genotype-function relationships through large-scale and cost-effective label-free live-cell imaging.

Following the publication of the above paper, it was drawn to the Editor's attention by an interested reader that the middle and right‑hand protein blots shown for the RIPK4 data in Fig. 2B (relating to the PANC‑1‑Rsh1 and PANC‑1‑Rsh2 experiments) were strikingly similar to western blot data shown in Fig. 3B for the RAF‑1 data (and the same PANC‑1‑Rsh1 and PANC‑1‑Rsh2 experiments), albeit the bands were presented with different exposures/a change in contrast, also with apparent horizontal flipping and vertical resizing. Upon contacting the authors, they realized that errors had been made during the assembly of the experimental images presented in Fig. 3B. These errors were likely to have resulted from oversights made during the process of data consolidation and figure assembly; specifically, this led to the inadvertent use of incorrect images for the RAF‑1 western blot results in both the PANC‑1 cell line (as was correctly identified by the interested reader on PubPeer) and in the Capan‑1 cell line (which the authors identified themselves upon performing their own subsequent review). The authors were also able to present photos of the raw, unedited versions of the gels to the Editorial Office. A revised version of Fig. 3, now showing the correct data for the RAF‑1 blots for both the PANC‑1 and Capan‑1 cell lines, as specified above, is shown on the next page. The authors confirm that the errors made in assembling Fig. 3 did not have a major impact on the conclusions reported in the above article, and they thank the Editor of <i>International Journal of Oncology</i> for allowing them the opportunity to publish a Corrigendum. Furthermore, all the authors agree to the publication of this Corrigendum, and apologize to the readers for any inconvenience caused. [International Journal of Oncology 52: 1105‑1116, 2018; DOI: 10.3892/ijo.2018.4269].

Genetic variants influencing gene expression have been extensively studied at the transcriptional level. How these variants affect downstream processes remains unclear. We quantitated ribosome occupancy in prefrontal cortex samples from the BrainGVEX cohort and integrated these data with transcriptomic and proteomic profiles from the same individuals. Through cis-QTL mapping, we identified genetic variants associated with transcript level (eQTLs), ribosome occupancy (rQTLs), and protein level (pQTLs). Notably, only 34% of eQTLs have their effects propagated to the protein levels, suggesting widespread post-transcriptional attenuation. Using both a gene-based approach and a variant-based approach we identified omics-specific QTLs that associated with brain disorder GWAS signals and found the majority of them to be driven predominantly by transcriptional regulation. Consistently, using a TWAS approach, we identified 74 SCZ risk genes across the three omics layers, 52 were discovered using transcriptome with 68% showing limited impact on protein expression. Our findings indicated that many disease-associated variants act through regulatory mechanisms that do not lead to an observable impact on the protein level.

Leukodystrophies are a diverse group of genetic disorders affecting the central nervous system white matter. Since their initial identification over a century ago, significant advancements have been made in understanding their genetic and clinical profiles. Yet, disease modifying therapies are limited, despite significant clinical impact characterized by progressive neurological decline leading to severe disability and early mortality. This underscores the need for advanced disease models to facilitate the understanding of disease mechanisms and the development of early therapeutic interventions. Stem cells have emerged as a transformative tool in leukodystrophy research, enabling the generation of patient-specific cells otherwise inaccessible for study. We have conducted the first scoping review of stem cell-based disease modeling in leukodystrophies, highlighting recent developments, challenges, and future directions in leveraging these models to enhance our understanding and aid in the development of therapies for these debilitating disorders.

Lung development generates a complex tree-like architecture through proximal-distal patterning and branching morphogenesis. However, the gene regulatory programs governing embryonic lung development remain poorly understood. Here, we present a comprehensive single-cell multi-omics atlas of mouse embryonic lungs, integrating gene expression and chromatin accessibility profiles. Through systematic analysis, we identify 13 distinct cell types and map cis-regulatory elements, peak-to-gene linkages, and transcription factors underlying lung development. Leveraging this multi-modal dataset, we uncover lineage-determining transcription factors driving cell differentiation, including the Activated Protein-1 complex. We further delineate gene regulatory networks involving diverse transcription regulators, including CCCTC-binding factor (CTCF). Using the Ctcf conditional knockout mouse, coupled with histological and multi-omics analyses, we demonstrate that CTCF orchestrates lung progenitor maintenance and branching morphogenesis by modulating both gene expression and chromatin accessibility. Thus, our study provides a multi-omics resource and mechanistic insights for transcriptional regulation of lung morphogenesis.

The human CYFIP1 gene is linked to Autism Spectrum Disorder (ASD) and Schizophrenia (SCZ), both associated with brain connectivity defects and corpus callosum abnormalities. Previous studies demonstrated that Cyfip1-heterozygous mice exhibit diminished bilateral functional connectivity and callosal defects-resembling observations in ASD and SCZ patients. Here, we demonstrate that CYFIP1 is crucial for cortical axonal development and identify insufficient calcium uptake as the pivotal mechanism. In vivo, Cyfip1 heterozygosity delays callosal axon growth and arborization. Additionally, Cyfip1-deficient cortical neurons and axons have reduced intracellular calcium, along with impaired mitochondria morphology, activity, and motility. Mechanistically, CYFIP1 binds and stabilises the mRNA of specific voltage-gated calcium channel subunits, explaining the decreased calcium concentration in Cyfip1^+/- cells. Notably, elevating intracellular calcium rescues delayed axonal growth and mitochondrial defects in Cyfip1-deficient neurons. These findings highlight that, by regulating mRNA metabolism, CYFIP1 ensures proper callosal development, offering insights into brain connectivity disruptions underlaying neurodevelopmental disorders.

In the treatment of ovarian cancer, PARP inhibitors such as olaparib have shown promise, yet resistance in advanced cases remains a significant challenge. Our study identified SLC7A5 as a key gene associated with olaparib resistance through differential gene expression and prognostic analysis. High SLC7A5 expression was found to correlate with poor prognosis. In vivo and in vitro experiments revealed that SLC7A5 enhances olaparib resistance by stabilizing ACLY and promoting de novo lipid synthesis via the ERBB2 axis, independent of leucine. Mechanistically, SLC7A5 upregulates ERBB2 transcription through ELK1, and ERBB2 competes with CUL3 to prevent ACLY degradation. These findings suggest that targeting SLC7A5 may reverse olaparib resistance, offering new strategies for combination therapies and improving clinical outcomes in ovarian cancer treatment.

Extracellular vesicles (EVs) are an essential signalling entity in human plasma implicated in health and disease. Still, their core protein and lipid componentry, which lie at the centre of EV form and function, remain poorly defined. Here we performed high-resolution density gradient fractionation of over 140 human plasma samples to isolate circulating EVs, and systematically constructed their quantitative proteome (4,500 proteins) and lipidome (829 lipids) landscapes using mass spectrometry. We identified a highly conserved panel of 182 proteins (including ADAM10, STEAP23 and STX7) and 52 lipids (including PS, PIPs, Hex2Cer and PAs), providing a deep survey of hallmark molecular features and biological pathways characteristic to circulating EVs. We also mapped the surfaceome diversity, identifying 151 proteins on the EV surface. We further established a set of 42 proteins and 114 lipids features that served as hallmark features of non-EV particles in plasma. We submit ADAM10 and PS(36:1) as conserved EV biological markers that precisely differentiate between EV and non-EV particles. Our findings, which can be explored via an open-source Shiny web tool ( evmap.shinyapps.io/evmap/ ), will serve as a valuable repository to the research community for a clearer understanding of circulating EV biology.

Brugada syndrome (BrS) is an inherited cardiac arrhythmic disorder associated with an increased risk of malignant ventricular arrhythmia and sudden death. Mendelian randomisation implicated N-palmitoyl glycine (PalGly) in BrS risk and identified BrS-associated proteins (DCC, CR1, CTSB, NAAA, DEFB1, EPHA1, IGF1/IGFBP3/ALS, and LTA), for which molecular docking further predicted moderate binding affinities with PalGly. PalGly enhanced calcium sparks in ventricular cardiomyocytes without affecting Na_v1.5 or K_v4.3/KChiP2 but activated TRPC5 (EC₅₀ 104 nM), as confirmed by patch-clamp. TRPM4, a channel mediating sodium influx at negative potentials and reported to link to BrS when mutated, was directly inhibited by PalGly (IC₅₀ = 7 nM). Functionally, PalGly shortened APD in cardiomyocytes and QT in male rabbit hearts, whereas ML204 (TRPC5 inhibitor) further shortened APD in isolated cardiomyocytes. Transcriptomic and lipidomic analyses further indicated immune pathway suppression. Our study underscores the involvement of PalGly, TRPC5, and inflammation-related proteins in the pathophysiology of BrS.

<h4>Background</h4>The Varroa destructor represents one of the most significant global threats to honeybee colonies. Genome-wide association studies (GWAS) play a crucial role in identifying genetic markers associated with Varroa resistance in honeybees. Six scenarios of genome-wide association studies (additive, dominance, and epistatic for each case‒control and quantitative approach) were implemented to analyze SNP markers associated with hygienic behavior in worker bees.<h4>Results</h4>In additive GWAS (case‒control), 10 SNPs were identified, whereas additive GWAS (quantitative) revealed 23 SNPs. For the dominance GWAS, 2 SNPs were found in the case‒control study, and 13 SNPs were found in the quantitative analysis. Moreover, the epistatic GWAS (case‒control) identified 10 paired SNPs, and the epistasis GWAS (quantitative) revealed 13 paired SNPs associated with uncapping behavior. These scenarios, along with some overlapping findings, highlighted unique SNPs. The results demonstrate that the type of data definition and genetic model used significantly influence the identification of SNPs associated with the uncapping trait. Additionally, combining multiple methods enhances SNP detection and provides deeper insights into the genetic architecture of Varroa resistance. The identified SNPs were distributed across the genome and presented distinct features from the perspective of minor allele frequency across different models. Furthermore, enrichment analysis revealed that genes related to these SNPs are predominantly involved in pathways related to the stress response and immunity.<h4>Conclusions</h4>Together, our research provides new insights into the genetics underlying varroa resistance in honeybees. By leveraging genomic data and different GWAS models, this study contributes significantly to the understanding of the genetic architecture of varroa resistance.

BACKGROUND: Ferroptosis, a type of cell death driven by iron metabolism, has been implicated in polycystic ovary syndrome (PCOS), although the evidence remains conflicting. This study aimed to comprehensively investigate the relationship between PCOS and ferroptosis-related iron metabolism, with a particular focus on its contribution to ferroptosis. METHODS: We performed a systematic review and meta-analysis including clinical studies from PubMed, Embase, and Web of Science up to November 2024 to analyse and compare the levels of serum ferritin, iron, hepcidin, and transferrin saturation levels between women with PCOS and the controls. Additionally, a case-control study on serum ferritin levels and placental expression of ferroptosis-related genes was conducted in 16 pregnant women with a prepregnancy diagnosis of PCOS and 21 controls, at Women’s Hospital, School of Medicine, Zhejiang University, China, in 2022. RESULTS: Twenty-two studies were included, and significant differences in serum ferritin levels were found between women with PCOS and controls (standardized mean difference (SMD): 0.86; 95% confidence intervals (CI): 0.40 to 1.32; P < 0.01). Moreover, serum iron (SMD: 0.46; 95% CI: 0.04 to 0.87; P = 0.03) and transferrin saturation (mean difference (MD): 4.85; 95% CI: 2.21 to 7.49; P < 0.01) levels were significantly greater in women with PCOS, whereas serum hepcidin levels were lower (SMD: -2.09; 95% CI: -3.30 to -0.87; P < 0.01). A case-control study revealed significantly elevated serum ferritin levels in the third trimester and notably upregulated expression of placental ferroptosis-related genes (ACSL4, TP53, GPX4, SLC7A11, and FTH1) at full term in the PCOS group compared with the control group. CONCLUSION: Our findings highlight the involvement of ferroptosis-related dysregulation of iron metabolism in the pathophysiology of PCOS and suggest that altered placental ferroptosis is potentially linked to adverse pregnancy outcomes. Clinical health care providers should be more careful about decision-making concerning iron supplements for women with PCOS, especially pregnant individuals. Further work is needed to explore the regulatory mechanisms underlying iron metabolism in women with PCOS. CLINICAL TRIAL NUMBER: Not applicable.

Mycobacterium tuberculosis (Mtb) is an intracellular pathogen responsible for tuberculosis (TB), a disease that remains the leading infectious cause of death worldwide. Several host cell proteins have been reported to play a significant role in regulating Mtb infection and replication. Among these, TRIM proteins are a large family of E3 ubiquitin ligases that have been described as being involved in host defence against Mtb, by controlling two important cellular processes: autophagy and the inflammatory response. Autophagy is a highly conserved cellular process deputed to the degradation of damaged or unnecessary components, such as organelles and protein aggregates. Autophagy is induced by a variety of stress stimuli, including nutrient deprivation, hypoxia, infection, oxidative stress and DNA damage, to maintain cellular homeostasis and survival. Autophagy plays a crucial role in host defense against infections by selectively targeting and delivering intracellular pathogens, like bacteria, viruses, and parasites to the lysosomes for degradation. Inflammation is a biological coordinated cellular process involving the secretion of cytokines and chemokines by innate immune cells, that can be triggered by various factors, including damaged cells, toxins and pathogens. Pathogen-Associated Molecular Patterns (PAMPs), found on microbes, are recognized by pattern recognition receptors (PRRs) on immune cells, which trigger signaling pathways leading to pro-inflammatory responses and the elimination of the pathogen. Danger-Associated Molecular Patterns (DAMPs) are host-derived molecules released from damaged cells that activate the same PRRs to initiate or perpetuate inflammation. In this review, we discuss the emerging role played by TRIM proteins in regulating autophagy and pro-inflammatory response during Mtb infection, pointing out the crucial role of TRIM in the complex crosstalk between these processes.Clinical trial number Not applicable.

<h4>Background</h4>Red blood cells (RBCs), essential for oxygen transport and carbon dioxide removal, are pivotal for maintaining systemic metabolic homeostasis. However, global blood shortages and limitations in current transfusion practices underscore the urgent need for alternative sources, such as in vitro-generated RBCs. Among these, induced pluripotent stem cell (iPSC)-derived RBCs have gained attention for their potential patient-specific, pathogen-free, and immunologically compatible solutions. Yet, conventional assays of RBC function provide only a partial view of the complex molecular programs that govern erythropoiesis and maturation. These gaps motivate the integration of multi-omics platforms to comprehensively profile the developmental and functional states of in vitro-generated RBCs.<h4>Main body</h4>iPSC-derived RBCs hold broad translational promise, yet recapitulating erythropoiesis in vitro remains difficult given the niche's hypoxia, extracellular cues, and multilayered regulation. We review erythroid development from primitive to definitive programs, the roles of HSCs and erythroblastic islands, and key pathways (e.g. EPO, GATA1, KLF1, TGF-β), with emphasis on transcriptional networks and chromatin remodeling that drive maturation. Integrating multi-omics provides a quantitative view of erythropoietic state transitions, explicitly linking gene-regulatory programs with metabolic and proteomic remodeling. By clarifying the mechanisms behind incomplete enucleation and hemoglobin switching, this systems-level perspective guides optimization toward scalable iPSC-derived RBC manufacture. We outline how transcriptomic, epigenomic, proteomic, and metabolomic readouts together map the regulatory landscape, and we discuss translational applications-transfusion medicine, rare blood type management, disease modeling, and drug delivery-alongside safety, ethical, and regulatory considerations.<h4>Conclusion</h4>This review underscores both current advances and persistent challenges in characterizing in vitro-generated RBCs, and it proposes a translational framework that integrates multi-omics data into their development and evaluation. By explicitly linking molecular regulation with process control and quality metrics, this approach provides a roadmap to optimize functional performance and ensure clinical readiness of in vitro-generated RBCs.

<h4>Background</h4>Spinal cord injury (SCI) initiates a cascade of secondary pathological processes significantly impairs functional recovery and leads to complications such as neuropathic pain. Arachidonic acid (AA) and its metabolites play crucial roles through various metabolic pathways. These pathways contribute to neuroinflammation, tissue damage, and functional impairment following SCI.<h4>Aim</h4>We aim to provide a comprehensive outline on AA metabolic pathways, enzymes and signaling cascades in spinal cord injury pathophysiology, in order to develop new perspectives in treatment and diagnosis.<h4>Key scientific concepts</h4>We present a comprehensive review on AA metabolism and pathways in the different pathology process of SCI, such as neuronal loss, demyelination, neuropathic pain and neuroinflammation with different cell types involved. We also discussed the complex interactions of AA metabolites pose challenges for the development of effective therapies. Finally, we outlooked future research directions aimed at optimizing AA-targeted treatments and expanding their clinical applicability to enhance recovery in patients with SCI.

Ataxin-2 (Atx2) is a general RNA-binding protein involved in processes such as RNA processing and metabolism in cells. Atx2 is also a polyglutamine (polyQ) tract-containing protein; its abnormal expansion can lead to protein aggregation associated with neurodegenerative diseases. Previous studies have shown that the C-terminal intrinsically disordered regions (c-IDRs) of Atx2 participate in its condensation and aggregation processes. To elucidate the role of polyQ expansion in biomolecular condensation and aggregation, we studied the N-terminal fragments of Atx2 (namely, Atx2-N317 and Atx2-N81) that preserve a polyQ tract and compared their molecular behaviors in cells to those of the full-length Atx2. We found that the molecular mobility of the N-terminal fragments decreases with the increasing length of polyQ, indicating that polyQ expansion promotes a gradual phase transition to an irreversible and insoluble state. Moreover, the molecular state and mobility of Atx2-N317 are not distinct from those of Atx2-N81, regardless of the presence of other domains, demonstrating that the polyQ tract is a direct and sufficient element for protein condensation and aggregation, while the Like Sm (LSm) and LSm-associated (LSmAD) domains and their interactions with RNA are not necessary for these processes. This result is also validated through the in vitro investigation of Atx2-N81 with different polyQ expansions. This study reveals that polyQ expansion controls the biomolecular condensation and aggregation of the N-terminal fragments of Atx2 and is thus thought to modulate the dynamic behaviors of the full-length protein as well, which is implicated in the pathological accumulation of Atx2 in cells.

This study aimed to collaboratively investigate the mechanism of variations in intramuscular fat (IMF) content in Wandong cattle using transcriptomics and metabolomics techniques. Longissimus dorsi (LD) muscle samples were collected from thirteen free-range Wandong cattle in Fengyang County, Anhui Province, China. From this initial cohort, eight animals closely matched in age and body weight were selected. Based on IMF content measured by Soxhlet extraction, these eight cattle were divided into two groups: the high-IMF (HF, n = 4) and low-IMF (LF, n = 4) groups. Subsequent analyses were performed on integrated datasets comprising the transcriptome, metabolome, and fatty acid profile. The results revealed a significant increase in IMF in the HF group compared to the LF group (<i>p</i> < 0.05). Specifically, α-linolenic acid (C18:3n3) and γ-linolenic acid (C18:3n6) were significantly more abundant in the LF group compared to the HF group (<i>p</i> < 0.05), whereas oleic acid (C18:1n9c) and cis-9-palmitoleic acid (C16:1) predominated in the HF group. However, saturated fatty acids (SFAs), such as myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), and Margaric acid (C17:0), did not show significant differences (<i>p</i> > 0.05). A total of 9164 differentially expressed genes (DEGs) were identified via transcriptome analysis, with 2202 genes upregulated and 6962 genes downregulated in the HF group compared to the LF group. The expression profiles exhibited a distinct pattern, characterized by the upregulation of genes such as FABP1, SREBF1, and LIPE, while genes including SCD, PPARGC1A, and LEP were downregulated. GO enrichment analysis demonstrated that the majority of DEGs were predominantly abundant across 25 distinct functional categories distributed across the three primary ontologies. KEGG pathway analysis further identified 341 significantly enriched signaling pathways in the HF group (<i>p</i> < 0.05), predominantly involving metabolic pathways, FoxO, AMPK, and PPAR signaling pathways. Untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics analysis revealed 404 differential accumulated metabolites (DAMs), with 187 in positive ion mode and 217 in negative ion mode (<i>p</i> < 0.05). These DAMs were notably enriched in pathways such as glycerophospholipid metabolism, terpene and steroid biosynthesis, fatty acid degradation, and fatty acid metabolism. Notably, C16:1, C18:1n9c, arachidonic acid (peroxide free) (C20:4n6), oleoyl-L-carnitine, and linoleoyl-carnitine were identified as key players in lipid metabolism. Integrating transcriptomics with metabolomics data unveiled significant associations between DAMs linked to lipid metabolism and DEGs. Specifically, C18:1n9c exhibited a positive correlation with LPIN3, while C16:1 showed negative associations with PPAP2B, PPAP2A, CDS2, HADHA, LPL, HSD17B12, ELOVL5, ACSL1, and ACOX1, and positive correlations with PLA2G15, CDIPT, AGPSBG1, and GPD1. In summary, the variation in IMF content in Wandong cattle is co-regulated by key genes (SREBF1, ACSL1, SCD) via the AMPK, PPAR, and FoxO signaling pathways, coupled with alterations in specific fatty acid metabolites such as C18:1n9c, C16:1, and C20:4n6. These findings provide critical molecular insights for the genetic selection and breeding of Wandong cattle, which are renowned for their superior meat quality.

Ferroptosis, recently proposed as a novel type of cell death, is characterized by unique characteristics and recognition functions. It is involved in diverse physiological processes and in the onset and progression of various diseases and is characterized by reactions between reactive oxygen species (ROS) and iron-dependent lipid peroxidation. This process is finely regulated by a variety of metabolic pathways. Ferroptosis fundamentally differs from conventional cell death mechanisms such as apoptosis, necrosis, and autophagy. In recent years, research on ferroptosis in the field of ophthalmology has gradually emerged, and a large amount of evidence has shown that it is closely related to the occurrence and development of ophthalmic diseases such as age-related macular degeneration (AMD), diabetic retinopathy (DR), retinal ischemia-reperfusion injury (RIRI), retinitis pigmentosa, dry eye disease, cataracts, and glaucoma. This paper provides a comprehensive review of the latest advancements in ferroptosis within ophthalmological research and systematically describes the molecular mechanisms and pathophysiological significance of ferroptosis in the pathogenesis and progression of ophthalmic diseases. Exploring the mechanisms of ferroptosis holds promise for the delivery of novel molecular targets and therapeutic approaches to prevent and treat ophthalmic diseases. Additionally, its clinical translational and application are anticipated to surmount current therapeutic limitations and emerge as a significant direction for breakthroughs in the precision medicine era.

UNC13s are presynaptic proteins essential for neurotransmitter release at chemical synapses. In this study, we present eleven patients from nine families with severe neurodevelopmental impairments, who carry rare, biallelic <i>UNC13C</i> single-nucleotide variants (SNVs). Six missense variants, each identified in compound heterozygosity in one of three of these patients, were introduced into the <i>Drosophila melanogaster</i> ortholog <i>unc13</i> using a previously established CRISPR/Cas9-based method for rapid and scarless genomic modifications, hypothesising that they underlie the observed clinical manifestations. However, none of the introduced mutations influenced Mendelian ratios, negative geotaxis, or lifespan of the fruit flies. Interestingly, two variants located outside the gene regions encoding known UNC13C domains caused a decreased ethanol sensitivity in <i>Drosophila</i>, while the Thr1729Met substitution within the C₁ domain resulted in increased ethanol sensitivity. Molecular dynamics simulations of the latter mutant gene product suggested that the altered protein conformation enhances exposure of the ethanol-binding site, thereby increasing sensitivity to ethanol. These findings reinforce previous evidence highlighting the critical role of the C₁ domain in ethanol sensitivity. Given the involvement of the C₁ domain in synaptic plasticity this result might implicate an influence of the Thr1729Met on synaptic function.

This study aimed to determine coagulation changes in bitches with pyometra based on a series of coagulation tests and thromboelastography (TEG), and to assess the incidence of disseminated intravascular coagulation (DIC). Eighteen bitches with pyometra and thirty-four control bitches were examined. Haematological, biochemical, and following haemostasis parameters were measured, including: prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin activity (AT), and levels of fibrinogen (FBG), d-dimers (DD), tissue factor (TF), plasminogen (PLG), tissue plasminogen activator inhibitor 1 (TPAI-1), and thromboelastography. DIC was considered present if three or more of these parameters were abnormal: platelet count (<153 × 10⁹/l), PT (>8.1 s), aPTT (>25.5 s), FBG (<0.6 g/l), DD (>0.2 mg/l), and AT (<107%). Significant differences were found in the PT, aPTT, FBG, DD, TPAI-1, clotting time, α-angle, and maximal amplitude. According to our scoring system, two patients were DIC positive. The study found alterations in several coagulation tests and hypercoagulable TEG tracings in bitches with pyometra, which point to excessive activation of coagulation, delayed fibrinolysis, and the presence of DIC. No patient bled abnormally, which may suggest that DIC is not overt in the majority of pyometra patients.

To date, a variety of covalent cysteine-reactive chemical probes have been reported. The most common ones include maleimide for bioconjugation and iodoacetamide alkyne (IAA) for chemoproteomics analyses. Their applicability, however, is limited due to, e.g., the hydrolytic lability of maleimide adducts and the slow reaction kinetics as well as the inability to record the entirety of the cysteinome with IAA. This is compounded by the missing potential to fine-tune their reactivity tailored to advanced applications. To generate a high-reactivity, cysteine-selective chemical probe with broad utility, we have performed an in-depth investigation into the acrylophenone scaffold. The aryl group connected to the vinyl ketone chemotype can be readily substituted, which provides the potential to fine-tune the reactivity and install a bioorthogonal handle. We took advantage of this feature by modifying acrylophenone-alkyne (APA) with two ortho chlorine groups to generate <i>ortho</i>-dichloroacrylophenone-alkyne (CAPA), which increased the stability of the probe and the yield of its cysteine adducts. To showcase the reactivity, we performed reaction rate analyses with model reagents. The selectivity was demonstrated by specifically labeling cysteine residues within two peptides under physiological conditions. To investigate its utility toward bioconjugation reactions, we performed the stoichiometric labeling of two proteins. Remarkably, CAPA was successfully implemented as a high-reactivity cysteine-selective chemical probe into activity-based protein profiling (ABPP) experiments using both in-gel fluorescence (in-gel ABPP) and mass spectrometry analyses. The chemoproteomics workflow, named isoDTB-ABPP, allowed us to highlight that CAPA provides a complementary approach to IAA in expanding the coverage of the cysteinome.

<b>Background/Objectives:</b> Atrazine (ATZ) is a widely used herbicide, and most studies of its reproductive toxicity have been conducted <i>in vivo</i> using animal models, where ATZ disrupts redox homeostasis, leading to male reproductive dysfunction. However, its molecular mechanisms of action in human spermatogenic cells remain poorly understood. Huntington's disease (HD), an autosomal dominant disorder caused by abnormal CAG repeat expansion in the <i>HTT</i> gene, exhibits heightened oxidative stress sensitivity and mitochondrial dysfunction, which may further impair reproductive function. This study investigated ATZ effects on human spermatogenesis using an <i>in vitro</i> spermatogenesis (IVS) model derived from human induced pluripotent stem cells (hiPSCs), focusing on Nrf2-mediated oxidative responses and apoptotic regulation during spermatogonial stem cell-like cell (SSCLC) differentiation in wild-type (WT) and HD hiPSC lines. <b>Methods:</b> Two WT and two HD hiPSC lines carrying 44 (HD1) and 180 (HD2) CAG repeats were treated with ATZ (0, 0.01, 1, or 10 μM) for 30 days, followed by differentiation into SSCLCs for 15 days under continuous exposure. Expression of pluripotency (<i>OCT4</i>, <i>SOX2</i>), oxidative stress (<i>NFE2L2</i>, <i>SOD1</i>, <i>GPX1</i>, <i>NQO1</i>), cell cycle (<i>CDK1</i>), apoptosis (<i>BCL2</i>, <i>BAX</i>, <i>CASP3</i>, <i>CASP9</i>, <i>FAS</i>, <i>FASLG</i>), and spermatogenic markers (<i>DAZL</i>, <i>ZBTB16</i>, <i>GFRA1</i>, <i>PIWIL2</i>) were assessed by immunocytochemistry and qRT-PCR. <b>Results</b>: Long-term ATZ exposure affected pluripotency markers in hiPSCs and SSCLC differentiation in a cell line-dependent manner. WT cells exhibited early differentiation suppression without significant apoptosis. HD1 cells were highly sensitive: low ATZ doses (0.01-1 μM) partially activated intrinsic and extrinsic apoptotic pathways, whereas high-dose ATZ (10 μM) reduced Nrf2-target and spermatogenic gene expression, strongly impairing SSCLC maturation. HD2 cells showed pronounced oxidative stress with robust Nrf2-driven antioxidant responses and <i>BCL2</i> that supported differentiation at low doses. However, excessive oxidative or proliferative signaling, including <i>CDK1</i> upregulation at high ATZ concentrations, disrupted redox balance and SSCLC differentiation in HD2 cells. <b>Conclusions</b>: ATZ exerts dose- and genotype-dependent effects on IVS through coordinated regulation of oxidative stress and apoptosis. These findings highlight the interplay between Nrf2-mediated antioxidant defenses, apoptotic signaling, and genetic background in shaping spermatogenic outcomes, providing mechanistic insight into ATZ-induced reproductive toxicity in a human-relevant <i>in vitro</i> spermatogenesis model.

<b>Background:</b> Systemic inflammatory factors may be altered by periodontitis and/or COVID-19, potentially increasing the risk of adverse pregnancy outcomes, a relationship that remains unclear. <b>Objective:</b> This study aimed to identify associations between periodontitis and COVID-19 during pregnancy, evaluating the influence of dental care on obstetric variables through set theory and probability. <b>Methods:</b> A quantitative, cross-sectional, and correlational study was conducted in two phases. The first phase analyzed 156 medical records from 5 institutions, including gynecological and periodontal data; the second phase examined 104 records from a single institution selected for data completeness (2020-2021). Descriptive statistics, bivariate analysis, chi-square tests, and odds ratios were applied. Set operations (union, intersection) and relative probabilities were calculated using R and Excel. Sets represented dental care, dental disease, COVID-19 diagnosis, gestational age, neonatal weight, and complications. <b>Results:</b> In Phase 1, 37% of pregnant women were COVID-19-positive, 44% vaccinated, 51.9% underwent cesarean section, and 5.12% had periodontitis. In Phase 2, 76 pregnant women did not receive dental care, while 28 did; among them, 6 were COVID-19-positive. Mean neonatal weight ranged from 2336 g (dental care) to 2271 g (no dental care). COVID-19-positive pregnant women showed fewer complications and a higher proportion of normal-weight neonates. Gingivitis was the most frequent periodontal condition (75%). No statistically significant differences were observed between the analyzed sets. Conclusions: no direct relationship was found between periodontitis and neonatal weight in COVID-19-positive cases. Dental care did not influence maternal-fetal outcomes. The methodology provides an innovative framework for clinical analysis through mathematical abstraction.

Carcass weight (CW) is a major determinant of beef yield and market value in Korea, yet the genetic basis of this trait remains largely unexplored in cattle from Jeju Island. In this study, we performed a genome-wide association study (GWAS) using both a mixed linear model (MLM) and the FarmCPU approach, followed by pathway and network analyses to identify loci and biological functions underlying CW variation. A total of 256 Jeju cattle (92 Jeju Black and 164 Jeju Black × Hanwoo crossbreds) were initially sampled. One crossbred sample failed genotyping, leaving 255 animals (92 Jeju Black and 163 crossbreds) for analysis. Animals were genotyped using the Illumina BovineSNP50 v3 BeadChip, and 39,055 high-quality single nucleotide polymorphisms (SNPs) were retained after quality control. The MLM analysis detected no genome-wide significant associations, whereas the FarmCPU analysis identified six significant loci on <i>Bos taurus</i> chromosomes 3, 5, 6, 10, and 13, each explaining 2.55-9.58% of the phenotypic variance. Candidate genes located near these loci included <i>EIF2B3</i>, <i>HECTD3</i>, <i>SOX5</i>, <i>KLF6</i>, <i>PHACTR3</i>, and two uncharacterized protein-coding genes. Functional enrichment analysis identified biologically relevant pathways including lysine degradation, tryptophan metabolism, glycerolipid metabolism, fatty acid biosynthesis, extracellular matrix-receptor interaction, and signaling cascades such as PI3K-Akt and Rap1, although most pathways were not statistically significant after FDR correction. Protein-protein interaction (PPI) network analysis using STRING highlighted modules of signaling, extracellular matrix, and metabolic genes. These clusters suggest that coordinated interactions among these pathways contribute to carcass growth and development. These findings provide new insights into the molecular basis of CW in Jeju Black and Hanwoo × Jeju Black crossbred cattle and identify candidate genes and pathways that may be useful for genomic selection and the sustainable improvement of Jeju Black cattle populations.

C-reactive protein (CRP) is a well-known acute phase reactant and putative biomarker for advancing and chronically established inflammation. Its biological activity across its multiple isoforms plays various roles in the initiation, potentiation, and resolution of inflammation. Its molecular signaling within the tissue microenvironment regulates cell-cell communication across cell types (e.g., epithelial cells, endothelial cells, fibroblasts, adipocytes, and immune cells) and affects the development of conditions such as cancer that are subject, at least in part, to inflammatory signaling. Considering the dynamic nature of CRP in modulating disease progression, and the growing evidence of the context-dependent direct molecular activity of CRP on regulating intra- and inter-cellular signaling, it is critical to further understand how this integral molecule alters cell signaling pathways. Although the ability of CRP to directly interact with some extracellular matrix proteins involved with inflammation and disease has been reported as early as the mid-1980s, recent advances in unbiased proteomics have revealed a broader interactome of protein-protein interactions (PPIs) involving CRP. The present study evaluates the CRP PPIs identified to date and explores the potential novel regulatory capacity of CRP on multiple key cellular functions in metabolism and cell-cell signaling, offering an updated framework of the possible biological activities of CRP relevant to tumorigenic processes.

<h4>Background</h4>Ovarian cancer is a rare cancer, it has the worst prognosis and the highest mortality rate, especially in high-grade serous ovarian cancer (HGSOC). High-throughput data generation is developed and provides an opportunity to investigate molecular pathways involved in cancer progression. The purpose of this study is to explore the role of main genes linked to the immune system and immune microenvironment in HGSOC using bioinformatics approaches to introduce promising biomarkers.<h4>Materials and methods</h4>After a systematic search was performed in the Gene Expression Omnibus database, high-throughput sequencing, two RNAseq datasets were selected. The integrated dataset was used to obtain differentially expressed (DE) genes. The immune infiltrate abundances related to DE genes were determined. The pathway, gene-set, and hallmark analyses were performed for the extraction of the immune system-related genes. Then, genes that were included in an integrated network, with acceptable AUC for immunotherapy response, were selected, and survival, gene expression, and mutation analyses were done.<h4>Results</h4>The <i>NCF1, MAP4K1</i>, <i>CIITA, CD3E</i>, <i>PTPN22</i>, <i>SAMHD1</i>, <i>LCP2</i>, <i>TSHR, DOCK2</i>, <i>FANCD2</i>, <i>GNAI1</i>, <i>GNG12</i>, <i>ZBP1</i>, <i>TXK</i>, <i>STAT4</i>, and <i>RASGRP2</i> genes offer as key genes linked the immune system. <i>MAP4K1</i> and <i>TXK</i> genes have targeted compounds.<h4>Conclusions</h4>The current study recognized the most relevant genes involved in the immune system in high-grade serous ovarian cancer. The findings of this study provided a holistic understanding of the tumor microenvironment that can be used for introducing therapeutic targets.

Amongst genome-wide association studies (GWAS) across diverse populations, allelic effect heterogeneity may arise due to differences in genetic ancestry and environmental exposures. This heterogeneity impacts the refinement of sets of potential causal variants underlying genetic associations through statistical fine-mapping. We introduce two multi-ancestry fine-mapping methods, MR-MEGAfm and env-MR-MEGAfm, allowing for multiple causal variants in a genomic region. Both methods integrate GWAS summary statistics and differing linkage disequilibrium from multiple cohorts; env-MR-MEGAfm additionally incorporates summary-level environmental covariates. Through simulations, we show that, when allelic heterogeneity is correlated with environmental exposures and ancestry, env-MR-MEGAfm yields improved resolution over MR-MEGAfm and similar resolution to SuSiEx. In twelve sex-stratified African GWAS of low-density lipoprotein cholesterol in 19,589 individuals, MR-MEGAfm and env-MR-MEGAFM (adjusting for urban status) identify five variants with posterior probability > 0.5 within two loci. One variant showed heterogeneity only due to ancestry, while three showed heterogeneity due only to urban status.

Circular RNAs (circRNAs), a unique class of non-coding RNAs, have gained attention due to their stable closed-loop structure and involvement in diverse cellular processes. Their remarkable stability and abundance in biological fluids, particularly when encapsulated within exosomes, make them highly promising candidates for clinical applications. This review offers a detailed insight into the growing field of exosomal circRNAs, starting with the fundamental principles of circRNA biogenesis and the selective mechanisms governing their sorting into exosomes. We explore the complex regulatory network controlled by circRNAs, highlighting their roles as microRNA (miRNA) sponges, interactors with RNA-binding proteins (RBPs), regulators of transcription and RNA stability, and even as templates for translating into novel peptides or proteins. We elucidate how these actions are mediated by exosomal circRNAs when delivered to recipient cells, influencing the behavior of target cells. The review critically evaluates the current and emerging methodologies for the detection, quantification, and functional analysis of exosomal circRNAs. We extensively discuss the clinical implications, highlighting the significant potential of exosomal circRNAs as non-invasive biomarkers for the diagnosis, prognosis, and monitoring of various diseases, with a particular focus on cancer. Furthermore, we explore their therapeutic landscape, including strategies for targeted delivery, their role in modulating drug resistance, and their intricate functional crosstalk with other non-coding RNAs in cancer progression. While recognizing the promising potential of exosomal circRNAs, we also address the existing technical challenges and highlight the future directions needed to leverage their diagnostic and therapeutic applications. Our review highlights the significance of interpreting the circular code of exosomal circRNAs as a vital step toward achieving the objectives of precision medicine.

<h4>Background</h4>Cis-regulatory elements (CREs) control oncogene expression and malignant phenotypes. The high clinicopathological heterogeneity of cancer cannot be explained by gene expression alone, being attributed to CRE heterogeneity. However, characterizing cancer-associated CREs is challenging. To address this issue, we performed a single-cell epigenomic analysis of clinical specimens.<h4>Methods</h4>To map the multicellular ecosystem of breast cancer (BC) and identify candidate CREs (cCREs), we performed a single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq) of 38 prospectively collected BC samples with various clinicopathological characteristics.<h4>Results</h4>First, we performed single-cell chromatin accessibility profiling of a high-quality set of 22,775 cells from BC samples. Cells were annotated using marker gene accessibility and integration with existing single-cell RNA sequencing data. The chromatin accessibility patterns exhibited by cancer cells were consistent with the clinicopathological features of each tumor. We identified 224,585 cCREs across the BC ecosystem. By identifying cluster-specific differentially accessible cCREs (DA-cCREs) and constructing a putative enhancer-promoter network, we mapped the cis-regulatory landscape for cancer cells and the tumor microenvironment. The accessibility of putative enhancers targeting the same gene differed within or between tumors, highlighting intra- and inter-cis-regulatory tumor heterogeneity.<h4>Conclusions</h4>This study provides a valuable resource for future epigenetic research on BC and highlights the diverse regulatory landscape within or among tumor(s), suggesting that cCREs regulate intra- and intertumor heterogeneity.

Neurodegenerative diseases (NDDs), including Parkinson's disease and Alzheimer's disease, are major age-related disorders characterized by progressive neuronal degeneration and a decline in cognitive and motor functions. Managing NDDs poses an increasing healthcare challenge as the global population ages. The onset of NDDs is linked to protein misfolding, oxidative stress, dysfunction of mitochondria and lysosomes, and neuroinflammation. Clinical manifestations of NDDs only appear after substantial neuronal damage has already occurred. This underscores the urgent need for accessible tissue biomarkers to enable early diagnosis, disease monitoring and assessment of therapeutic efficacy. The skin has emerged as a valuable peripheral indicator of neurodegeneration, sharing embryological origin, gene expression profiles, protein alterations and cellular dysfunctions with the brain. Notably, pathological protein deposits, which are hallmarks of NDDs, such as beta-amyloid, tau proteins, and oligomeric alpha-synuclein, have been observed in the skin. Increasing evidence links NDDs with various pathological skin conditions, including melanoma and inflammatory diseases. This review aims to explore the potential of the skin as a window into neurodegenerative processes at an early stage, before clinical signs arise. The main advantages of using skin as a source of NDD biomarkers are its accessibility and the minimally invasive sampling methods such as stratum corneum collection, sebum and volatile compounds analysis, and biopsies. Immunohistochemistry and omics approaches applied to skin samples provide valuable insights into NDD pathophysiology and facilitate biomarker discovery for early diagnosis and disease monitoring. NDDs are multisystemic disorders and new findings in skin research highlight the value of peripheral tissues for investigating central nervous system alterations enabling earlier neuroprotective interventions.

The reactivation of fetal globin genes is the most promising treatment for β-hemoglobinopathies. This implies the reversal of the naturally occurring hemoglobin switching. Here, we show that expression of the orphan nuclear receptor COUP-TFII in adult HUDEP2 erythroid precursor cells activates γ-globin (HbF) at the expense of β-adult globin by specific occupation of the 'adult' δ-β-region within the β-locus. Notably, although COUP-TFII and the main γ-globin repressor BCL11A-XL share a similar DNA binding consensus and a large number of chromatin targets, including the locus control region of the β-locus itself, they bind differentially to the γ and β promoters, eliciting an opposite transcriptional outcome. In addition, we find that COUP-TFII activates Lin28B, a known post-transcriptional repressor of BCL11A-XL. Our work identifies a molecular mechanism that could be leveraged to increase γ-globin levels in patients affected by β-hemoglobinopathies.

γ-Folic acid conjugates are valuable tools for targeting the folate receptor, an important cancer biomarker, but direct conjugation of folic acid is hampered by poor regioselectivity and challenging purification. We report an orthogonal protection strategy that enables controlled regioselective access to γ-folic acid conjugates from readily available building blocks. This approach permits site-selective installation of a range of functional modalities, including an amphiphilic ligand, a fluorescent probe, and a bioorthogonal handle, followed by global deprotection to afford γ-conjugates without the need for reversed phase chromatography.

Small-molecule STAT3 inhibitors face numerous challenges in clinical translation, including poor water solubility, rapid systemic clearance, low bioavailability, poor selectivity, and high cytotoxicity. To address these limitations, we conjugated the potent STAT3 inhibitor-LLL12 to generate six (G6) hydroxyl-terminated (poly(amidoamine)) PAMAM dendrimers using pH-sensitive linkers: sulfonyl carbamate (carbamate), sulfonyl carbamoyl (amide), and hydrazone for intracellular drug delivery. Conjugation greatly enhanced LLL12 solubility (up to 10 mg/mL) and reduced cytotoxicity without altering dendrimer size or surface charge. All three G6-LLL12 conjugates remained stable under neutral pH but exhibited sustained, pH-dependent drug release correlating with <i>in vitro</i> potency and cytotoxicity. Notably, hydrazone-linked conjugate showed an IC₅₀ = 0.42 ± 0.035 μg/mL, comparable to free LLL12 (IC₅₀ = 0.31 ± 0.05 μg/mL) and superior to amide- and carbamate-linked conjugates. In bone marrow-derived immune suppressive myeloid cells, hydrazone-based G6-LLL12 effectively reduciii-derrive, hydrazone-based G6-LLL12 effectively reduced monocytic myeloid-derived suppressor cell expansion and promoted antigen-presenting cell maturation, highlighting a promising pH-responsive delivery system that enhances solubility and safety while retaining potency.

Biotechnology is increasingly relying on high-throughput experiments in which large populations of cells are phenotyped and selected based on performance, e.g., for producing a desired product. Droplet microfluidics enables high-throughput screening, and as such, it lends itself ideally to such experiments, but reliance on a fluorescent readout is limiting. Herein, we report a system integrating droplet electrospray ionization-mass spectrometry (ESI-MS) with a microfluidic voltage-mediated droplet printer to enable online analysis and capture of cell-containing aqueous droplets, ultimately offering the possibility for label-free phenotyping and selection of cells. <i>Escherichia coli</i> (<i>E. coli</i>) cell-containing droplets (20 nL) are split into two volume fractions within a microfluidic splitter chip. One fraction is analyzed using a sheath-flow ESI source with MS, while the sibling fraction is printed onto an agar plate using a custom-built voltage-mediated microfluidic printer. Printed droplets can grow into single microbial colonies that map back to their respective droplet ESI-MS signal with 94-99% accuracy and without carryover. Nearly synchronous and stable system operation is shown for infusion rates in the range of 0.4-1.2 droplets/s, while the achieved droplet spacing and printing precision can enable reliable single-colony retrieval for further analyses or gene sequencing. The system is also shown for phenotyping and selection of an <i>E. coli</i> variant engineered to produce l-lysine among control cells. The method enables screening cell colonies for chemical composition and collecting them for further processing with potential application in synthetic biology and enzyme engineering among others.

Mast cells play complex and context-dependent roles within the tumor microenvironment, yet their molecular characteristics and functional diversity across human cancers remain poorly defined. In this study, single-cell RNA sequencing and spatial transcriptomics data are integrated to comprehensively map the transcriptional and spatial heterogeneity of mast cells across ten cancer types. A distinct proinflammatory mast cell (PMC) population is identified, characterized by strong antigen-presenting features and immune-activating potential. Mechanistic analyses show that retinoic acid (RA) signaling drives the polarization of PMCs through activation of RARα, which promotes CIITA-mediated MHC-II expression, CXCL16 secretion, and T cell recruitment and activation. Across multiple cancer types, tumors with higher PMC abundance are associated with more favorable clinical outcomes. These findings reveal the pivotal role of RA-RARα-CIITA signaling in mast cell reprogramming and suggest that pharmacologic induction of proinflammatory mast cells may represent a promising approach to enhance antitumor immunity.

The systematic characterization of cellular heterogeneity among tissues and cell-type-specific regulation underlying complex phenotypes remains elusive in pigs. Within the Pig Genotype-Tissue Expression (PigGTEx) project, this work presents a single-cell transcriptome atlas of adult pigs encompassing 229 268 high-quality nuclei from 19 tissues, annotated to 67 major cell types. Besides cellular heterogeneity within and across tissues, this work further characterizes prominent tissue-specific features and functions of muscle, epithelial, and immune cells. Through deconvoluting 3921 bulk RNA-seq samples from 17 matching tissues, this work dissects thousands of genetic variants with cell-type interaction effects on gene expression (ieQTL). By colocalizing these ieQTL with variants associated with 268 complex traits, new insights into the cellular mechanisms behind these traits are provided. Moreover, this work highlights that orthologous genes with cell-type-specific regulation in pigs exhibit significant heritability enrichment for some human complex phenotypes. Altogether, this work provides a valuable resource and highlights novel insights in cellular regulation of complex traits for accelerating pig precision breeding and human biomedical research.

The ability of adult epithelial cells to rewire their cell fate programme in response to injury has emerged as a new paradigm in stem cell biology. This plasticity supersedes the concept of strict stem cell hierarchies, granting cells access to a wider repertoire of fate choices. Yet, in order to prevent a disordered cellular response, this process must be finely regulated. Here we investigate the little-known regulatory processes that restrict fate permissibility in adult cells, and keep plasticity in check. Using a 3D regenerative culture system, that enables co-culturing epithelium and stroma of different origins, we demonstrate that oesophageal cells exposed to the ectopic signals of the dermis are capable of switching their identity towards skin. Lineage tracing experiments and histological analysis, however, reveal that the oesophageal-to-skin lineage conversion process is highly inefficient, pointing to the existence of barriers limiting cell fate re-specification. Single-cell RNA sequencing capturing the temporality of this process shows that cells transitioning towards skin identity resist the natural progression towards tissue maturation by remaining in a persistent regenerative state marked by a particularly strong hypoxic signature. Gain and loss of function experiments demonstrate that the HIF1a-SOX9 axis acts as a key modulator of epithelial cell fate plasticity, restricting changes in identity during tissue regeneration. Taken together, our results reveal the existence of lineage conversion barriers that must be resolved for cells to respond to signals instructing alternative fate choices, shedding light on the principles underlying the full regenerative capacity of adult epithelial cells.

Many cellular functions rely on multiprotein complexes and their stoichiometric assembly. Reducing the levels of individual complex components can perturb this process and induce corrective stress responses. In addition to local outcomes, cellular stress in one tissue can induce long-distance responses in other tissues. Here, we used muscle-targeted RNAi to examine the systemic stress responses induced by muscle-specific genetic perturbation of four distinct multiprotein complexes: the sarcomere, mitochondrial respiratory complex I, proteasome, and VCP (valosin-containing protein) complex. Muscle-specific disruption of these four complexes produced largely overlapping transcriptional adaptations in the central nervous system (CNS), and these responses were centered on the upregulation of many proteases and peptidases. Testing in a retinal model of Huntington's disease demonstrated that several stress-induced proteases limit the accumulation of huntingtin-polyQ aggregates during aging, indicating that these proteases protect from pathogenic proteins. We next examined whether the myokine Amyrel is a possible mediator of this stress-initiated muscle-to-CNS signaling because of its previously reported role in inducing protease expression. Consistent with this model, Amyrel expression was transcriptionally induced in muscle by perturbation of each of the four multiprotein complexes. Moreover, experimental upregulation of Amyrel in muscle reduced the amount of pathogenic huntingtin-polyQ aggregates in the retina. Taken together, these findings indicate that Amyrel and protective proteases improve CNS proteostasis following the perturbation of multiprotein complexes in skeletal muscle. Thus, this study provides insight into a muscle-to-CNS signaling axis that conveys information on the stress status of multiprotein complexes.

Obesity is a risk factor for vascular dementia and exhibits sex differences in comorbidities and prevalence. However, its cell-specific effects on the neurovascular unit (NVU) remain unknown. Here we show, using the ob/ob mouse model and single nuclei RNA sequencing, how obesity modulates the hippocampal NVU transcriptome in females. Obesity alters endothelial-specific differentially expressed genes (DEGs) involved in angiogenesis and blood brain barrier permeability. DEGs in common between endothelial cells and other NVU cells are associated with neurotransmission and autophagy. Some gene expression changes from each NVU cell type correlate with behavioral changes. Sex-based analyses comparing to our previously reported male data indicate that the NVU transcriptomic response to obesity is modified by sex in a cell-type specific manner. Endothelial and microglial cells exhibit greater sex-specificity than astrocytes and neurons. The findings from our work may help inform both sex-independent and sex-specific approaches to prevention and treatment for obesity-related dementia.

Repeat expansions (REs) may be Parkinson's disease (PD) risk factors. We screened whole genome sequencing data from the AMP PD Lewy Body Dementia (LBD) and PD cohorts for 37 REs associated with neurological disorders, and identified both interrupted and uninterrupted REs in ATXN2 in 4/2431 PD and 2/2468 LBD cases, but none in controls. These findings support pleiotropy for certain REs in PD.

Tumor endothelial cells (TECs) play a critical role in regulating immune responses within the tumor microenvironment (TME). However, the mechanisms by which TECs modulate immune cell population remain unclear, particularly in non-small cell lung cancer (NSCLC). Here, we investigated how NSCLC cells tweak normal endothelial cells (NECs) into TECs and the subsequent effects on immune regulation. NECs were cocultured with various NSCLC cell lines, using 2D and 3D coculture models to evaluate TEC-mediated effects on immune cells. We show that direct coculture led to significant transcriptomic, proteomic and kinomic alterations in TECs, especially in pro-inflammatory pathways. We identified a downregulation of the co-stimulatory molecule OX40L in TECs compared to NECs, suggesting impaired T-cell proliferation support. While TECs showed a limited effect on CD8⁺ T-cell activation, TECs supported CD4⁺ T-cells polarization into Treg and Th22 subsets. Moreover, TECs also promoted M2-like macrophages polarization, thereby potentially contributing to the TME immunosuppression. State-of-the-art single-cell RNA sequencing of 3D multicellular tumor spheroids (MCTS) revealed formidable heterogeneity in the tumor cells and cancer-associated fibroblast compartments. It also unveiled distinct TEC subpopulations, including an inflammatory subset with an unfolded-protein response signature. This TEC cluster was absent in 2D-cultured NECs but present in freshly isolated and 2D-cultured TECs from NSCLC patients. Importantly, we identified a perivascular M2-like macrophage subset within MCTS that is in close contact with TECs, and is predicted to interact with them through MIF signaling. In conclusion, TECs in NSCLC tumors play a pivotal role in remodeling the TME immune landscape by promoting immune suppression. This study highlights the complex immunoregulatory functions of TECs within different in vitro models that mimic aspects of the TME. Our data may provide new insights into potential therapeutic strategies targeting TECs or regulatory signaling to improve the efficacy of immunotherapy in NSCLC.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is a leading cause of cancer deaths. Given that traditional pathologic features to diagnose LUAD do not fully reflect the biological differences in patients, the search for novel biomarkers is necessary.<h4>Methods</h4>In this study, we obtained immune-related genes (IRGs) from ImmPort and performed cluster analysis on The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) to mine LUAD subtypes with different immune characteristics. Quantitative analysis of IRGs was performed by single-sample gene set enrichment analysis (ssGSEA). Based on the univariate cox and LASSO regression methods, we screened the characteristic genes that significantly affected LUAD and built the model based on the RiskScore coefficients. The relative expressions of characteristic genes in LUAD were determined using qRT-PCR. Transwell and wound healing assays were utilized to verify the practical regulation of these genes on the migration and invasion levels of LUAD. Correlations were established between RiskScore and LUAD drug sensitivity by oncoPredict.<h4>Results</h4>We acquired three LUAD subtypes and demonstrated heterogeneous IRGs scores and clinical features. The molecular subtypes were differentially enriched in bile acid metabolism, fatty acid metabolism, and ECM-receptor interaction. This study identified seven genes (MS4A1, EXO1, CPS1, ZNF750, S100P, NT5E, KCNN4) as a signature affecting prognosis, from the differentially expressed genes (DEGs) among the molecular subtypes, and constructed a RiskScore for the prognosis of LUAD. Cellular experiments verified that 6 of 7 characteristic genes were expression dysregulation in LUAD cell line. Silencing of EXO1 significantly suppressed the migration and invasion of LUAD cell lines. RiskScore and immune checkpoints such as CD276, TNFSF4, and TNFSF9 showed a positive correlation.<h4>Conclusions</h4>This study identified three LUAD subtypes with distinct immune characteristics and constructed a seven-gene prognostic model. This model correlates with immune checkpoint and chemotherapy sensitivity, providing new targets and strategies for clinical diagnosis and treatment.

Venous thromboembolism (VTE) is a major cause of mortality, influenced by genetic and environmental factors. von Willebrand factor (VWF) mediates hemostasis by promoting platelet adhesion, and its plasma levels are associated with thrombotic risk. Although many non-coding variants in ABO are associated with VWF levels, VTE risk, and COVID-19 severity, the mechanisms underlying these associations remain unclear. In this study, we identified the ABO locus as the genomic region with the highest concentration of variants associated with VWF levels. Chromatin conformation analyses in endothelial cells revealed non-coding ABO variants (rs657152, rs9411377, rs660340, and rs505922) associated with VWF levels, VTE risk, and COVID-19 severity, located in spatial proximity to ADAMTS13. ADAMTS13 is a key regulator of VWF activity, and both ADAMTS13 and VWF play crucial roles in coagulation and thrombosis. Chromatin activation (CRISPRa) of the region near the non-coding ABO variant rs657152 increased ADAMTS13 transcription in endothelial cells, suggesting that this variant resides in a regulatory region with the potential to modulate long-range transcriptional control of ADAMTS13. Luciferase assay revealed reduced transcriptional activity driven by the rs505922-C allele in endothelial cells. These findings provide insights into the spatial organization of the ABO locus and its potential role in ADAMTS13 regulation.

Liver fibrosis is a pathological condition marked by excessive extracellular matrix accumulation, potentially leading to cirrhosis. Macrophages play a vital role in regulating inflammatory responses, facilitating tissue repair, and orchestrating extracellular matrix remodeling through their differentiation into proinflammatory and anti-inflammatory phenotypes. Hepatic stellate cell activation and the progression of fibrosis have been linked to ferroptosis, a controlled cell death process triggered by iron and characterized by lipid peroxidation. This review explores the interaction between ferroptosis and macrophage polarization in iron-overload-induced liver fibrosis. It examines how ferroptosis intensifies inflammatory and fibrotic processes through macrophage activity and identifies key macrophage marker proteins involved. Understanding this interplay offers novel therapeutic insights targeting macrophage polarization to mitigate liver fibrosis, particularly in conditions such as hemochromatosis and chronic transfusion-dependent disorders.

Species expand their geographic distribution when environmental conditions are favorable or when mutations arise that allow them to live in previously unfavorable conditions. The European wasp spider, <i>Argiope bruennichi</i>, has expanded its range poleward, and populations at the northern edge show higher tolerance to cold and are genetically differentiated from the core populations. We aimed to investigate the degree and limits of plasticity in a recently cold-adapted Estonian population by exposing overwintering juveniles (spiderlings) to three fixed winter regimes over the course of three months. These regimes differed in absolute and relative day and night temperature: cold (5°C day, -15°C night), moderate (5°C day, -5°C night), and warm (15°C day, -5°C night). We expected a differential response to the winter regimes in survival, lipid content, metabolites, and gene expression patterns. The survival probability of the spiderlings decreased over winter by approximately 20% and their lipid content by 28%, with no significant differences between groups. Spiderlings also did not differ in content of saturated and monounsaturated fatty acids per dry weight. However, in spiderlings exposed to the warm winter regime, short-chain omega-3 PUFAs were less abundant (~57%) and long-chain omega-3 PUFAs more abundant (~66%) compared to the other regimes. The gene expression response was low under the cold regime and much higher under the warm regime, as compared to the moderate regime. The affected pathways suggest a more pronounced stress response under warmer winter temperatures. Taken together, our findings demonstrate that <i>A. bruennichi</i> spiderlings from a northern population can endure very different winter regimes. However, the observed physiological responses to the warmer regime suggest metabolic costs that may reduce spiderling survival probability after emergence from the egg sac. We conclude that, despite remarkable tolerance to different winter regimes, warmer winters have nuanced effects on spiderling physiology beyond survival probability.

This research introduces carboxymethyl kappa-carrageenan-chitosan polyelectrolyte multilayers as a promising and sustainable alternative to heparin, used in surface treatments for blood-contacting medical devices. The polysaccharide-based surface coatings have good cytocompatibility and resist microbial adhesion of both Pseudomonas aeruginosa and Staphylococcus aureus. The blood compatibility of surfaces containing carboxymethyl-kappa-carrageenan was directly compared to similar polyelectrolyte multilayers containing heparin. The carboxymethyl-kappa-carrageenan surfaces inhibit whole-blood clotting like heparin-containing surfaces. Blood clotting is mediated by both protein-surface and cell-surface interactions. The carboxymethyl-kappa-carrageenan surfaces adsorb more fibrinogen and less albumin than the heparin surfaces, and they also exhibit reduced platelet and leukocyte adhesion, compared to heparin surfaces. These results suggest that carboxymethyl-kappa-carrageenan may represent a sustainable alternative to heparin as an anticoagulant surface modification.<h4>Supplementary information</h4>The online version contains supplementary material available at 10.1007/s44340-025-00043-w.

<h4>Abstract</h4>Drug-induced liver injury (DILI) is a diagnostic dilemma, often mimicking other hepatic disorders and posing a significant clinical challenge. While pharmaceuticals are well-documented culprits, herbal and traditional medications, widely perceived as harmless, increasingly contribute to hepatotoxicity, particularly in the eastern parts of the world, where their use is prevalent yet unregulated. We present the case of a 58-year-old female with a 10-day history of jaundice and new-onset abdominal discomfort, with no identifiable risk factors, the only notable detail being her intake of Ayurvedic herbal preparations for joint pains, 3 months before the onset of symptoms. Physical examination revealed icterus, while laboratory work showed hyperbilirubinemia and elevated liver enzymes, which was suggestive of hepatic dysfunction with a cholestatic pattern (R ratio: 1.6). Extensive evaluation for viral, autoimmune, and metabolic causes was negative. Liver biopsy revealed focal hepatocellular necrosis, eosinophilic infiltration, and microvesicular steatosis, which are features of DILI. Applying the Roussel Uclaf Causality Assessment Method (RUCAM), the patient scored 7, indicating a probable link to the herbal medication. Supportive care was initiated, and liver function improved steadily, allowing for a safe discharge within a week. This case highlights the potential hepatotoxicity of certain alternative and traditional herbal medications and drugs, as these preparations may contain undisclosed or poorly regulated ingredients. Hence, thorough history-taking, awareness, and clinical suspicion coupled with structured causality tools like RUCAM can aid in the early diagnosis and timely intervention of DILI.

<h4>Background</h4>Previous studies on acute myocardial infarction (AMI) complicated by atrial fibrillation (AF) have mainly focused on anatomy or underlying disease state, and its prognostic predictors have not been fully explored. Therefore, there is a need for an effective prognosis model for patients with AMI-AF.<h4>Methods</h4>We retrospectively selected 126 patients with acute myocardial infarction complicated with AF hospitalized in Beijing Anzhen Hospital from January 2020 to December 2024 as the case group, and 1719 patients without AF as the control group. The clinical characteristics and laboratory test results of the two groups were compared to determine independent risk factors for AF in patients with acute myocardial infarction. The predictive performance of the model was evaluated by plotting Receiver Operating Characteristic (ROC) for each independent predictor. For the combined model, we used R software to build pattern plots, calibration plots, and Decision Curve Analysis (DCA) based on a multivariate logistic regression model.<h4>Results</h4>Multivariate Logistic regression analysis showed that older age (Odds Ratio (OR) = 1.067, 95% CI: 1.044-1.092), longer hospitalization days (OR = 1.039, 95% CI: 1.013-1.066). The AUCs for age, hospitalization days, history of coronary heart disease, heart rate, International Normalized Ratio (INR), Hemoglobin, and mean platelet volume were 0.721, 0.663, 0.577, 0.614, 0.688, 0.438, and 0.607. The AUC of nomogram model for predicting AF in AMI patients was 0.833 (95% CI: 0.796-0.870, <i>p</i> < 0.001), the sensitivity was 0.817, and the specificity was 0.726. The nomogram model indicated a clinical net benefit when the predicted risk threshold exceeded 0.06.<h4>Conclusions</h4>Multivariable prediction model has good prediction effect. The variables in this nomogram model are easily obtained in clinical practice and can provide reference for individualized prediction of AF in AMI patients.

Cyclin-dependent kinase 5 (Cdk5) is an atypical serine/threonine kinase distinct from classical cell cycle regulators. Its activity is highest in the nervous system and essential for development, but its functions in other tissues, particularly in cancer, are increasingly being elucidated. This review explores the functional duality of Cdk5 by comparing its constructive role in neurodevelopment with its repurposed oncogenic function in cancer. In neurodevelopment, Cdk5 orchestrates nearly every stage of brain construction, including neuronal differentiation, migration, and synaptic plasticity. However, in many cancers, this neurodevelopmental toolkit is repurposed, and aberrantly activated Cdk5 promotes proliferation, metastasis, and therapeutic resistance in diverse solid tumors. Cdk5 also actively shapes the tumor microenvironment by promoting angiogenesis and modulating immunity. Notably, this oncogenic function is not universal, as Cdk5 exhibits its duality even within the context of cancer; it acts as a tumor suppressor in gastric cancer upon nuclear localization. Taken together, these lines of evidence underscore that Cdk5 is a context-dependent kinase whose output is determined by upstream regulation, subcellular localization, and the cellular environment. This review discusses the molecular basis of its dual role and highlights both the potential and complexity of Cdk5 as a therapeutic target in oncology.

Cognitive impairment remains one of the most prevalent and debilitating sequelae of traumatic brain injury (TBI), profoundly compromising long-term quality of life. Nevertheless, effective treatment options are limited, as the complexity of post-TBI pathology often exceeds the protective scope of conventional neuroprotective strategies. Accumulating research has revealed regulated cell death (RCD) as a central driver of neuronal loss and cognitive decline post-TBI. Consequently, targeting RCD pathways has emerged as a promising strategic direction for alleviating post-TBI cognitive impairment. This review provides an analysis of the molecular mechanisms underlying five major RCD forms, including apoptosis, necroptosis, pyroptosis, ferroptosis, and cuproptosis. Furthermore, it critically assesses the therapeutic potential of these pathways while examining their complex interplay in post-TBI cognitive impairment. By systematically synthesizing recent advances in targeted therapeutic strategies, we highlight that targeting RCD pathways paves the way for highly effective and precise therapeutic modalities against post-TBI cognitive impairment, although challenges in multi-target combination therapies and brain delivery warrant further investigation.

Corneal problems, such as delayed and incomplete wound repair, are frequent in diabetes, affecting up to 70% of diabetic patients. In skin, histone deacetylases (HDACs) have been previously found to repress expression of the glycerol channel aquaporin-3 (AQP3), the deficiency of which delays corneal wound healing. We hypothesized that the pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) would improve corneal healing in diabetic mice. Diabetic and normoglycemic C57BL/6J male and female mice were subjected to corneal debridement. Wounds were treated topically with vehicle or SAHA every four hours until they healed. Treatment with SAHA improved wound healing in both normoglycemic and hyperglycemic male mice but, unexpectedly, no changes were detected in female mice. In male mice interleukin-1beta (IL-1β) and tumor necrosis factor (TNF) were significantly increased in diabetic corneas, and SAHA reduced their expression, returning IL-1β and TNF to levels comparable to those in normoglycemic mice regardless of treatment. In normoglycemic male mice, AQP3 levels were not changed in the cornea with SAHA treatment but the expression of AQP3 was increased in the wound's edge relative to the rest of the cornea. In vitro SAHA treatment of human corneal epithelial cells (HCECs) significantly increased protein expression of AQP3, important for corneal wound healing, but had no effect on ROS production. In conclusion, treatment with SAHA improved corneal wound healing, not only in male mice with diabetes and delayed wound healing but also in normoglycemic male mice; therefore, SAHA could potentially be repurposed as a topical treatment clinically to improve corneal wound healing.

Gestational Diabetes Mellitus (GDM) increases the long-term risk for metabolic and cardiovascular diseases in the offspring. However, the underlying mechanisms are not well understood. This study investigates the impact of GDM on fetoplacental vascular function and molecular mechanisms underlying endothelial dysfunction. Clinical data and tissue samples were collected from normoglycemic (NG, n = 33) and GDM (n = 19) pregnancies. Offspring in the GDM group were delivered earlier, had a larger placental size, and had a reduced placental efficiency. Functional analysis using a Mulvany myograph demonstrated a significant impairment of insulin-mediated vasodilation in fetoplacental vessels of GDM patients compared to NG controls. This vascular dysfunction was associated with a reduction in total insulin receptor protein expression. Further investigation revealed an impaired PI3K/AKT/eNOS signaling pathway, as endothelial cells from GDM pregnancies showed a deficient insulin-induced phosphorylation of AKT. These results indicate that maternal GDM induces insulin resistance and endothelial dysfunction in the fetoplacental vasculature through impairment of the AKT/eNOS pathway, providing a key mechanism for its adverse neonatal outcomes and the increased lifelong cardiovascular risk in the offspring.

Osteoporosis remains a substantial healthcare burden in modern times. Current diagnostic methods of osteoporosis detect changes in bone mineral density and microarchitecture, which have already occurred. It is critically important to develop methods of early diagnosis of osteoporosis to be able to plan early interventions in order to stop the disease progression. Genetic screening based on early osteoporosis marker genes appears to be a promising approach for early diagnosis and prevention. However, a significant gap exists in this area of knowledge. Recently, we identified a novel combination of three single nucleotide polymorphisms - <i>FSHR</i> (rs6166) AA, <i>TSHR</i> (rs1991517) CC, and <i>ADRB2</i> (rs1042713) AA, with a high prevalence among osteoporotic patients. Subsequent functional studies using patient-derived mesenchymal stem cell lines revealed impaired osteogenic differentiation capacity. To clarify the role of these polymorphism combinations, this review first examines the physiological aspects of each receptor and the identified single nucleotide polymorphisms at the organismal level. It then analyzes their contribution to the dysregulation of bone remodeling, with a particular focus on osteoblastogenesis. Understanding these mechanisms opens up new opportunities for the development of early osteoporosis diagnosis and stratification of personalized treatments for patients.

The corpus callosum (CC) is the largest interhemispheric commissure in the eutherian brain, enabling inter-hemispheric sensory integration and higher-order cognitive functions. Historically viewed through a neuron- and axon-centric lens, extensive research has established that glial cells (astrocytes, oligodendrocytes, and microglia) are essential regulators of CC ontogenesis. Astrocytic guidepost cells sculpt midline architecture and secrete axonal guidance cues; oligodendrocytes drive callosal axonal maturation and myelination; and microglia regulate their fasciculation and pruning, myelination patterns, and synaptic refinement. In addition to these cell-specific roles, coordinated bidirectional signaling between neurons and glia ensures that axon targeting, maturation, and interhemispheric integration proceed in a precisely orchestrated manner. Disruptions to these glial functions are implicated in congenital and developmental brain pathologies, including malformations and CC agenesis. This review integrates molecular, developmental, and translational insights to provide a comprehensive, mechanistic understanding of glial contributions to CC development and how their dysfunction shapes pathology.

Cardiovascular diseases (CVDs) are a major global health challenge, significantly impacting public health and healthcare systems. Ethnopharmacological remedies and botanical medicines are widely used for the prevention and treatment of CVDs due to their multi-component, multi-target properties. Understanding the mechanisms of these natural products is essential for developing safe and effective therapies. The zebrafish model, an emerging tool in experimental pharmacology, has been increasingly used to evaluate the pharmacological activities of natural products. This review focuses the use of zebrafish models to test the prevention and treatment effects of various CVDs, and to investigate the inhibition of hyperlipidemia, thrombosis, the progress of heart failure, and the promotion of angiogenesis and toxicity. Emphasizing its experimental advantages-including the transparency characteristics of the body, its applicability to the analysis of multiple samples, and its support for real-time monitoring-aims to reveal its potential value in combining traditional cognition with contemporary pharmacological testing capabilities. The literature summary provides strong evidence for the cognitive improvement of the zebrafish system in clarifying the efficacy of natural ingredients in the cardiovascular field.

<h4>Background</h4>Rectal adenocarcinoma has a high incidence and suboptimal therapeutic outcomes. Notably, reliable prognostic biomarkers are currently lacking. Cuproptosis is a newly identified regulated cell death mechanism, and its molecular pathways and implications for rectal adenocarcinoma remain poorly understood. However, compared with colon adenocarcinoma, multi-omics studies of cuproptosis-related genes for rectal adenocarcinoma remain blank.<h4>Methods</h4>This study is the first to construct a cuproptosis-related genes prognostic prediction model for rectal adenocarcinoma by integrating genomics, transcriptomics, and clinical data. Transcriptomic data from the Gene Expression Omnibus and The Cancer Genome Atlas databases were analyzed. Cuproptosis-related genes were curated from prior literature and FerrDb V2. Consensus clustering was employed to identify molecular subtypes based on cuproptosis-related genes expression profiles. Subsequently, functional enrichment and survival analyses were conducted across subtypes. Following univariate Cox regression, LASSO regression, and multivariate analyses, four pivotal cuproptosis-related genes (PPAT, NHP2, INHBB, and MSMP) were selected to construct a prognostic risk model. The associations between risk scores, tumor microenvironment, immune cell infiltration, and drug sensitivity were further investigated.<h4>Results</h4>A total of 30 cuproptosis-related genes were screened, including 16 upregulated and 14 downregulated in rectal adenocarcinoma. These genes were enriched in pathways related to DNA/RNA metabolism, MAPK signaling, and calcium homeostasis. The LASSO regression risk model and nomogram demonstrated robust predictive accuracy for survival outcomes. Notably, cuproptosis-related genes were significantly correlated with immune cell infiltration and checkpoint gene expression, suggesting their dual role in tumor progression and immunomodulation.<h4>Conclusion</h4>The prognostic system based on cuproptosis-related genes constructed in this study not only enhances the accuracy of survival prediction for rectal adenocarcinoma patients, revealing the potential role of cuproptosis in the tumor microenvironment and immune regulation, but also provides a new perspective for risk stratification and the selection of personalized treatment strategies, paving the way for precise oncology approaches in rectal adenocarcinoma.

<h4>Introduction</h4>Breast cancer (BC) prognosis is fundamentally dictated by its pronounced molecular and cellular heterogeneity. While single-cell RNA sequencing (scRNA-seq) has detailed the tumor microenvironment (TME) cellular composition, a critical knowledge gap persists regarding the vascular heterogeneity of tumor endothelial cells (TECs) across different BC subtypes. Current TEC classification remains conventional, obscuring whether subtype-specific TECs possess unique transcriptomic signatures that influence treatment response, particularly to anti-angiogenic therapies. Given the pivotal role of tumor vasculature in proliferation and metastasis, and the existing research bias towards Triple-Negative BC (TNBC), a comprehensive investigation into the complexity of the vascular system is warranted. Furthermore, as lymph node metastasis (LNMT) is a major determinant of cancer mortality, distinguishing the endothelial and immune cell subtypes within LNMT microenvironments from primary tumors is essential for clarifying pathogenesis.<h4>Methods</h4>To address this, we integrated scRNA-seq data from primary BC tumors, ER_LN and healthy tissues, generating a robust dataset of 98,000 cells (~ 12 samples). Employing single-cell transcriptomics, spatial transcriptomics, and immunohistochemistry, we precisely delineated the endothelial cell (EC) heterogeneity within matched tumor and peri-tumoral tissues and scrutinized their intricate interactions with immune populations.<h4>Results</h4>We identified two previously uncharacterized, tumor-enriched endothelial cell subtypes, designated EC4 and EC5, which demonstrate subtype-specific functional adaptations and prognostic significance. EC4 cells, highly prevalent across BC, are principally characterized by antigen presentation, immune cell recruitment, and pro-inflammatory signaling. Conversely, EC5 cells, also enriched in BC, exhibit robust extracellular matrix (ECM) remodeling and potent tumor angiogenesis. We further characterized the functional divergence of EC4 and EC5 within ER tumors relative to HER2 tumors and ER_LN metastases. Our analysis reveals conserved endothelial programming mechanisms across BC subtypes, coexisting with distinct TME-driven transcriptional adaptations. Importantly, interactome analysis highlighted novel and subtype-specific communications between these novel EC subsets and immune cells, particularly CD8+T cells and macrophages. Experimental validation demonstrates that ECs overexpressing APP can mediate the M2 polarization of macrophages, underscoring diverse immunomodulatory roles for EC subsets across different BC contexts.<h4>Conclusions</h4>These findings offer critical and granular insights into the complex interplay between novel EC subtypes and the immune microenvironment in BC progression and metastasis. We establish that ECs are active and heterogeneous modulators of the TME, identifying specific therapeutic vulnerabilities within the tumor vasculature and providing a foundational blueprint for developing future precision immunotherapeutic strategies.

<h4>Background</h4>Intravenous immunoglobulin (IVIG) has been established as the first-line treatment for Kawasaki disease (KD). However, 10%-20% of patients are unresponsive, increasing their risk of coronary artery complications.<h4>Methods</h4>To elucidate the pathogenesis of IVIG non-responsiveness, we performed single-cell transcriptomic profiling on 24 peripheral blood mononuclear cell (PBMC) samples from responsive and non-responsive KD patients before and after IVIG treatment. Finally, the expression of key cytokines was validated using public bulk RNA-seq data and enzyme-linked immunosorbent assay (ELISA).<h4>Results</h4>Non-responders exhibited elevated inflammatory cells, lymphocyte dysfunction, and a stronger inflammatory cytokine response driven by the S100A12-TLR4-MYD88 axis, initiated by Mono_CD14_CD16 cells, which was closely associated with interferon activation. Despite T/NK-cell exhaustion, cytotoxic activity was preserved. All processes appeared to be closely associated with interferon activation. Disrupted Tfh-B-cell coordination was observed alongside plasmablast enrichment. Furthermore, monocytic myeloid-derived suppressor cells (MDSCs) suppressed T-cell function and promoted inflammation. The expression levels of S100A8/A9, S100A12, TNF, TNFSF8/10, and interferon-α were consistent with the transcriptomic data.<h4>Conclusions</h4>Our findings reveal the immune landscape in IVIG non-responsive KD and suggest potential targets for alternative therapies.

Alveolar macrophages (AMs), a type of immune cell, display remarkable plasticity and manifest diverse responses to stimuli by differentiating into distinct subgroups. These phenotypically distinct macrophage subtypes are primarily categorized as either classically activated or inflammatory (M1) macrophages, or alternatively activated or anti-inflammatory (M2) macrophages, the differentiation of which is underpinned by a complex regulatory network. Despite their crucial contribution to the pathobiology of acute lung injury/acute respiratory distress syndrome (ALI/ARDS), the research on AMs is currently limited. This study therefore aims to establish a comprehensive theoretical framework delineating the pathogenic role of AMs in ALI/ARDS, facilitating deeper mechanistic understanding of disease initiation and progression in ALI/ARDS and ultimately identifying novel therapeutic targets.

<h4>Background</h4>Pediatric Acute-onset Neuropsychiatric Syndromes (PANS) and Pediatric Autoimmune Neuropsychiatric Disorders Associated with Streptococcal infections (PANDAS) are characterized by sudden-onset neuropsychiatric symptoms. Growing evidence indicates that gut and oral microbiota may contribute to disease pathogenesis through immune and inflammatory pathways.<h4>Methods</h4>This narrative review analyzed approximately 250 studies published between 2000 and 2024, retrieved from PubMed, Scopus, and Google Scholar. The selected works included clinical, immunological, and microbiome-related studies investigating the role of gut-oral-brain interactions in neuroinflammation or in pediatric PANS/PANDAS.<h4>Findings</h4>Alterations in gut and oral microbial communities appear to modulate neuroinflammation through increased intestinal and blood-brain barrier permeability, immune dysregulation, and altered production of neuroactive metabolites. Specific bacterial families, such as <i>Bacteroidaceae</i>, <i>Rikenellaceae</i>, and <i>Odoribacteriaceae</i>, have been associated with pro-inflammatory states, while oral pathogens may exacerbate systemic inflammation via the gut-oral-brain axis.<h4>Conclusions</h4>The reviewed evidence highlights the potential of microbiome-targeted strategies-including dietary modulation, probiotics, and anti-inflammatory approaches-as promising avenues for future personalized diagnosis and therapy in PANS/PANDAS. However, further controlled studies integrating microbial, immunological, and clinical data are required to confirm causal mechanisms and establish personalized therapeutic protocols.

Conventional methods available for genome editing have proven non-specific, labour-intensive, and time-consuming. In this context, CRISPR/Cas technology represents a significant breakthrough. It is derived from a sophisticated microbial defence system consisting of clustered regularly interspaced short palindromic repeats, or CRISPR, and the RNA-guided DNA endonuclease Cas. Beyond its original role in genome editing, CRISPR continues to play a major role in the field of proteomics, functional genomics, and molecular therapy. Animal models, including mice, <i>Drosophila</i>, zebrafish, <i>etc.</i>, have substantially benefited from CRISPR in uncovering protein function through reverse genetics approaches, including knock-in, knockout, CRISPRi, and indel mutation strategies. On the clinical front, CRISPR gene therapy has also seen successes, including applications in sickle cell disease, hypercholesterolemia, and cancer immunotherapy. However, notable challenges remain, including <i>in vivo</i> packaging and delivery efficiency, toxicity, and genomic off-target effects. Ongoing efforts to overcome these include the development of novel delivery formulations (e.g., nanoparticles, exosomes), artificial intelligence-guided experimental design, and miniaturization of Cas proteins. This review focuses on CRISPR/Cas gene editing mechanisms and explores its state-of-the-art applications in the field of proteomics and theranostics.

The growing interest in utilizing clinical blood biomarkers for non-invasive diagnostics has transformed the approach to early detection and prognosis of respiratory diseases. Biomarker-driven diagnostics offer cost-effective, rapid, and scalable alternatives to traditional imaging and clinical assessments. In this study, we conducted a retrospective analysis of 913 patients from a local respiratory clinic in Hail region, evaluating the diagnostic relevance of 15 blood biomarkers across four respiratory conditions: COVID-19, pneumonia, asthma, and other complications. Through data-driven analysis, statistical correlation assessments, and machine learning classification models (decision tree classifiers), we identified significant biomarker interactions that contributed to disease differentiation. Notably, CRP and HGB demonstrated a strong negative correlation (-55%), supporting the well-established role of systemic inflammation in anemia of chronic disease. Additionally, Ferritin and LDH exhibited a positive correlation (+50%), indicating metabolic stress and cellular injury in severe respiratory illnesses. Other significant correlations included Creatinine and ESR being negatively associated with RBC, while GGT and ALT were positively correlated (+49%). Additionally, bilirubin and HGB were positively correlated (+49%), collectively reflecting systemic inflammatory and metabolic responses associated with respiratory pathology. The machine learning model demonstrated high predictive accuracy, with the following performance metrics: COVID-19: Precision (0.94), Recall (0.96), F1-score (0.95). Pneumonia: Precision (0.97), Recall (0.71), F1-score (0.85). Asthma: Precision (1.00), Recall (0.95), F1-score (0.97). Other Complications: Precision (0.88), Recall (0.90), F1-score (0.90). These findings validate the diagnostic potential of biomarker panels in respiratory disease classification, offering a novel approach to integrating statistical and computational modeling for clinical decision-making. By leveraging biomarker relationships and machine learning algorithms, this study contributes to the development of personalized, non-invasive, and cost-effective diagnostic tools for respiratory diseases, ultimately improving patient outcomes and healthcare efficiency.

Nanomedicine has evolved significantly over the last decades and expanded its applications in pediatric oncology, which represents a special domain with unique patients and distinct requirements. Τhe need for early cancer diagnosis and more effective and targeted therapies aiming to increase the pediatric patients' survival rates and minimize the treatment-related side effects to survivors is profound. Nanoparticles (NPs) come as a beacon of hope to provide sensitive cancer diagnostic tools and assist contrast agents' transport to the malignant tumors. Besides, NPs could be designed to deliver targeted drugs and genes to tumors, minimizing the medicine-related toxicities. Metal and metal oxide NPs could be exploited as sensitizers to enhance chemotherapy and radiotherapy effects. Future research should emphasize pediatric models to gain secure results about NPs' safety and efficiency for pediatric cancer patients. This review presents the recent studies on the use of NPs in pediatric cancer management and highlights their impact on diagnosis, treatment outcomes, and the quality of life of the survivors. The purpose of this study is to investigate the benefits that may arise from the use of NPs in pediatric oncology, address the potential limitations and challenges, and discuss the needs for future research efforts.

Typical 2-Cys peroxiredoxins (2-Cys Prxs, AhpC/Prx1 subfamily) are ubiquitous thiol peroxidases that efficiently reduce H₂O₂ and other hydroperoxides via a reactive peroxidatic Cys (C_P). Under elevated hydroperoxide levels, C_P can be hyperoxidized to sulfinic (C_P-SO₂H) or sulfonic (C_P-SO₃H) acids, leading to enzyme inactivation. Notably, eukaryotic 2-Cys Prxs are orders of magnitude more sensitive to hyperoxidation (sensitive Prxs) by H₂O₂ than their bacterial counterparts (robust Prxs). Sensitivity to hyperoxidation also correlates with the catalytic triad composition: enzymes containing threonine (Thr-Prx) are more prone to hyperoxidation by H₂O₂ than those with serine (Ser-Prx). While hyperoxidation is reversed in eukaryotes by an enzyme (sulfiredoxin), it is generally considered irreversible in bacteria. Here, we compared the hyperoxidation susceptibility of three typical 2-Cys Prxs: human Prx2 (Thr-Prx, sensitive), <i>P. aeruginosa</i> (Thr-Prx, robust) and <i>S. epidermidis</i> (Ser-Prx, robust) to lipid hydroperoxides derived from linoleic acid, containing one or two peroxide moieties per molecule. Employing structural analysis, molecular simulations and kinetic assays, we found that lipid peroxides proved to be potent hyperoxidizing agents for all 2-Cys Prx tested, inactivating the enzymes up to 10,000 times faster than H₂O₂. These results may have implications for understanding bacterial oxidative stress responses and antimicrobial resistance.

Inborn errors of immunity (IEI) are a heterogeneous group of genetic disorders affecting immune function, often presenting with diverse gastroenterological symptoms that pose diagnostic challenges. The latest classification developed by the International Union of Immunological Societies Experts Committee includes 582 IEI disease entities. Unfortunately, 70-80% of cases are still not recognized and do not enter the patient registers. Most cases are also diagnosed too late. This review explores the epidemiology, classification, symptoms, diagnostics, and treatment of gastrointestinal involvement in IEI.

Vertebrate commissural axon (CA) projection and midline crossing require suppression of Slit responsiveness in precrossing axons and its upregulation in postcrossing axons. Differential expression of the Slit receptor Robo1-low in precrossing and high in postcrossing axons-modulates Slit sensitivity and regulates midline crossing. However, the underlying mechanisms controlling this remain unclear. This study demonstrates that the 3' untranslated region (UTR) of <i>Robo1</i> mRNA is regulated by the conserved miR-219a. In the developing spinal cord, gga-miR-219a and cRobo1 exhibit distinct, trajectory-specific expression patterns. miR-219a suppresses cRobo1 protein translation in commissural neurons (CNs) without affecting mRNA levels. Disruption of the miR-219a-<i>cRobo1</i> 3' UTR axis in precrossing CNs triggers premature Slit repulsion <i>in vitro</i> and causes spinal CA defects <i>in vivo</i>. Conditional knockout of mmu-miR-219a in E11.5 mouse embryos elevates mRobo1 protein and causes CA guidance defects. These findings reveal a conserved miR-219a-dependent mechanism that regulates Robo1 expression and Slit-mediated axon guidance.

The gene therapy landscape has evolved substantially in recent years, beginning with the approval of the first adeno-associated virus-based gene therapy, Luxterna, in 2017. Since then, the US FDA has approved nearly 30 new viral gene therapy programs, with notable examples including Zolgensma, Spinraza, Hemgenix, Zynteglo, Lyfgenia, Kymriah, Skysona, and Tecelra. Remarkably, all these products rely on delivery via adeno-associated vectors (AAVs) and lentiviral vectors (LVs). Improvements in viral-mediated gene transfer efficiency and clinical-scale manufacturing, together with immense commercial interest, have greatly propelled the clinical adoption of gene therapy products. In recent years, clustered regularly interspaced short palindromic repeats (CRISPR) and its related Cas proteins (CRISPR-Cas) have made significant advances in gene therapy, offering next-generation approaches for curative gene editing to treat genetic diseases and disorders. In this review, we examine the range of these therapeutics and their viral carriers, focusing primarily on LVs and AAVs. We provide a snapshot of the current status of the field and highlight some of the current challenges in the clinical application of gene therapy, with particular emphasis on viral CRISPR-Cas-based technologies and their future potential.

The effects of non-pathogenic cytosine-adenine-guanine (CAG) repeat sizes on sleep remain unclear, although disrupted sleep has been observed in patients with pathogenic CAG expansions in polyglutamine disease-associated genes (PDAGs), particularly in HTT , ATXN3, and CACNA1A . Here, we assessed the associations between CAG repeat sizes of the three genes and self-reported sleep outcomes in the Netherlands Epidemiology of Obesity study (NEO) and the Netherlands Study of Depression and Anxiety (NESDA). Sleep outcomes included excessive daytime sleepiness (EDS) and Pittsburgh Sleep Quality Index (PSQI) in NEO, insomnia score in NESDA, and sleep duration and chronotype in both. We also stratified by sex, menopausal status in women, and questionnaire-based depression score. We observed 31 associations, of which 26 were specific to women. Larger HTT CAG repeat sizes were associated with lower EDS risk and lower PSQI score in premenopausal women, but higher PSQI score in women with depression. CAG repeats in all three PDAGs were associated with sleep duration, with ATXN3 showing U-shaped effects in all population groups except men. CAG repeat size in CACNA1A was primarily associated with chronotype in women. These findings suggest that non-pathogenic CAG repeats in PDAGs affect sleep differentially by sex and depression status.

The p97 protein is a member of the AAA+ family of ATPases. This protein is encoded by the <i>VCP</i> gene. It is a mechanoenzyme that uses energy from ATP hydrolysis to promote protein unfolding and segregation actively. The unfolded products are subsequently presented to the 26S proteasome for degradation. p97 substrate recognition is mediated by adaptors, which interact with substrates directly or indirectly through ubiquitin modifications, resulting in substrate funnelling into the central pore of the p97 hexamer and unfolding. Here, we engineered synthetic adaptors to target specific substrates to p97, using the extraordinary intracellular binding capabilities of camelid nanobodies fused to the UBX domain of the p97 adaptor protein Fas-associated factor-1 (FAF1). In such a way, we created a p97-directed proteolysis-targeting chimera (PROTAC), representing a novel and unique E3 ubiquitin ligase-independent strategy to promote specific proteolysis. All functional assays were performed in human cell lines to evaluate the system's efficacy and specificity in a physiologically relevant context.

SYTL5 is a member of the Synaptotagmin-Like (SYTL) protein family that differs from the Synaptotagmin family by having a unique N-terminal Synaptotagmin homology domain that directly interacts with the small GTPase RAB27A. Several SYTL protein family members have been implicated in plasma membrane transport and exocytosis, but the specific function of SYTL5 remains unknown. We here show that SYTL5 is a RAB27A effector and that both proteins localise to mitochondria and vesicles containing mitochondrial material. Mitochondrial recruitment of SYTL5 depends on its interaction with functional RAB27A. We demonstrate that SYTL5-RAB27A positive vesicles containing mitochondrial material, autophagy proteins and LAMP1 form during hypoxia and that depletion of SYTL5 and RAB27A reduces mitophagy under hypoxia mimicking conditions, indicating a role for these proteins in mitophagy. Indeed, we find that SYTL5 interacts with proteins involved in vesicle-mediated transport and cellular response to stress and that its depletion compromises mitochondrial respiration and increases glucose uptake. Intriguingly, SYTL5 expression is significantly reduced in tumours of the adrenal gland and correlates positively with survival for patients with adrenocortical carcinoma.

<h4>Objective</h4>To assess the influence of dentin airborne-particle abrasion techniques on the microtensile bond strength, using two adhesive strategies (self-etch and etch&rinse).<h4>Materials and methods</h4>Twenty molars were assigned to two groups (n = 10) according to the bonding system (Clearfil SE Bond or Optibond FL), divided into four parts and submitted to different surface treatments: no pretreatment (control), airborne-particle abrasion with aluminum oxide without irrigation (Al₂O₃), airborne-particle abrasion with aluminum oxide with irrigation (Al₂O₃/Aquasol), and airborne-particle abrasion with bioactive glass 45S5 (BG45S5). Specimens underwent microtensile bond strength (μTBS) testing and scanning electron microscopy (SEM) analysis. Statistical analysis employed a linear mixed regression model, with a 5% significance level.<h4>Results</h4>Statistically significant differences were found for both adhesive strategy (p < 0.001) and pretreatment (p < 0.001). Al₂O₃/Aquasol and Al₂O₃ exhibit statistically significant differences compared to the control (p < 0.001 and p = 0.006, respectively), while BG45S5 did not differ significantly (p = 0.080). SEM images reveal a smear layer with distinct density across groups, and diminished compactation whenever irrigation was used.<h4>Conclusions</h4>Aluminum oxide with simultaneous irrigation resulted in the highest bond strength values, while without irrigation bond strength outcomes depend on the adhesive strategy. Bioactive glass did not interfere with bonding performance.<h4>Clinical significance</h4>Aluminum oxide airborne-particle abrasion with concomitant irrigation presents the preferable surface pretreatment strategy when bonding to sound dentin.

BackgroundCa_V2.3 (R-type) channels mediate calcium signals involved in neuronal excitability, synaptic plasticity, and neurotransmitter release in the hippocampus. However, there is currently no information about their potential changes in Alzheimer's disease (AD).ObjectiveThis study aims to analyze the protein levels, expression, and subcellular localization of Ca_V2.3 channels in the hippocampus of 5xFAD mice.MethodsWe employed histoblot, western blot, qRT-PCR, and immunoelectron microscopy techniques.ResultsUsing the histoblot technique, we observed that the protein content of Ca_V2.3 in female 5xFAD mice was downregulated in the hippocampus and caudate putamen at 5 months, with more pronounced downregulation at 10 months that also affected the septum. The hippocampus was the most affected region, where Ca_V2.3 immunodetection was significantly decreased in a few dendritic layers at 5 months, but this reduction extended to all layers and subregions by 10 months, demonstrating age- and laminar-dependent changes. The pattern in females differed from that seen in males and remained unchanged in APP/PS1 and P301S mice. Immunoelectron microscopy of the hippocampus showed Ca_V2.3 in apical and oblique dendrites of CA1 pyramidal cells, with a significantly higher presence in dendritic spines. Quantification revealed a significant decrease in the density of Ca_V2.3 in oblique dendrites and dendritic spines of CA1 pyramidal cells, whereas apical dendrites and presynaptic compartments were unaffected in 5xFAD mice.ConclusionsOverall, this downregulation in postsynaptic Ca_V2.3 channels could impact neural circuit activity and contribute to the cognitive deficits observed in 5xFAD mice.

CCCTC-binding factor (CTCF) is essential for chromatin organization. CTCF interacts with endogenous RNAs, and deletion of its ZF1 RNA binding region (∆ZF1) disrupts chromatin loops in mouse embryonic stem cells (ESCs). However, the functional significance of CTCF-ZF1 RNA interactions during cell differentiation is unknown. Using an ESC-to-neural progenitor cell (NPC) differentiation model, we show that CTCF-ZF1 is crucial for maintaining cell type-specific chromatin loops. Expression of CTCF-∆ZF1 leads to disrupted loops and dysregulation of genes within these loops, particularly those involved in neuronal development and function. We identified NPC-specific, CTCF-ZF1 interacting RNAs. Truncation of two such coding RNAs, <i>Podxl</i> and <i>Grb10</i>, disrupted chromatin loops in cis, similar to the disruption seen in CTCF-∆ZF1-expressing NPCs. These findings underscore the inherent importance of CTCF-ZF1 RNA interactions in preserving cell-specific genome structure and cellular identity.

Disruption of histone H2A monoubiquitylation (H2AK119ub) causes neurodevelopmental disorders through poorly understood mechanisms. Polycomb complex-deposited H2AK119ub and trimethylation of histone H3 at lysine 27 (H3K27me3) can cooperatively maintain gene repression. However, the extent to which H2AK119ub functions independently of H3K27me3 to repress or potentiate gene expression, and the evolutionary conservation of these roles, remains unclear. Here, we address the interplay among Polycomb-deposited marks and chromatin states in <i>Caenorhabditis elegans</i> embryos. We find that H2AK119ub distribution is distinct from and largely dispensable for H3K27me3 patterns. Unexpectedly, H2AK119ub is enriched at predicted enhancers with developmentally dynamic changes in accessibility, including in neurons. More than two-thirds of well-documented axon guidance genes and neuronal identity transcription factors are associated with H2AK119ub. Many of the genes differentially expressed in H2AK119ub-deficient animals are involved in neuronal differentiation and signaling and feature H2AK119ub-enriched promoters and enhancers, including a subset that is H3K27me3 repressed. We uncover a likely conserved yet underappreciated dual role for H2AK119ub at enhancers and H3K27me3-repressed chromatin, with implications for nervous system development.

Loss-of-function mutations in methyl-CpG binding protein 2 (<i>MECP2</i>) cause Rett syndrome. While we know that MeCP2 binds to methylated cytosines on DNA, the full breadth of the molecular mechanisms by which MeCP2 regulates gene expression remains incompletely understood. Here, using a genetic modifier screen, we identify the super elongation complex, a P-TEFb-containing elongation factor that releases promoter-proximally paused RNA polymerase II, as a genetic interactor of <i>MECP2</i>. MeCP2 physically interacts with SEC subunits and directly binds AFF4, the scaffold of the SEC, via the transcriptional repression domain. Furthermore, MeCP2 facilitates the binding of AFF4 on a subset of genes in the mouse brain regulating synaptic plasticity and concordantly promotes the binding of RNA polymerase II on these genes. Last, while haploinsufficiency of <i>Aff4</i> does not exhibit any behavioral deficits in mice, it exacerbates the impaired contextual learning behavior of <i>Mecp2</i> hypomorphic mice. We propose a previously unknown mechanism by which MePC2 regulates gene expression underlying synaptic plasticity.

Biomolecular condensates, membrane-less assemblies formed by phase separation, are implicated in neurodegenerative disease, but their role in Alzheimer's disease (AD) remains unclear. Here, we report that in the brain of AD patients and animal models, an elevation of poly(C)-binding protein 2 (PCBP2) correlates with biomolecular condensation that involves phase separation. These condensates sequester large numbers of mitochondrial and mRNA-binding proteins, leading to the outside impairment of mitochondrial morphology and function, and BACE1 mRNA decay relative to amyloid deposition. We then identify a small molecule CN-0928 that inhibits the condensates by reducing PCBP2 protein level and mitigates AD pathology and cognitive decline, in which CN-0928 binding to a target protein integrator complex subunit 1 (INTS1) allows to regulate PCBP2 expression. Our findings place PCBP2 condensates as a key player that cooperates the seemingly disparate but important pathways, and show pharmacological modulation of PCBP2 as an effective approach for treating AD.

Understanding how cellular and molecular architecture underpins the large-scale organization of human brain function is a central challenge in neuroscience. By integrating transcriptomic (microarray and single-nucleus RNA-sequencing), molecular imaging, and neuroimaging datasets, we observe spatial correspondences indicating that the distributions of diverse cell types, neurotransmitter systems, and mitochondrial phenotypes align with intrinsic connectivity networks (ICNs). These associations extend beyond local correspondence to reflect network-level structure: inter-ICN similarity networks derived from cellular and molecular profiles recapitulate static and dynamic patterns of functional network connectivity (FNC), mirroring canonical functional domains. Mediation analyses reveal that specific ICNs mediate the relationship between microscale cell-type architecture and domain-specific cognitive processes, while FNCs capture mediating pathways linking cell-type and neurotransmitter similarity networks to cognitive organization. Together, our findings show that the brain's functional architecture systematically aligns with cellular and molecular organization, which may constrain functional network formation and contribute to the neural basis of cognition.

Identification of cell type-specific and temporally dynamic regulatory features of schizophrenia (SCZ) risk genes is essential for advancing mechanistic neurobiological studies. This study systematically dissected the genetic architecture and neurodevelopmental mechanisms of SCZ by integrating genome-wide association study (GWAS) data from European (N = 130,644) and East Asian (N = 30,761) population, alongside multi-omics and single-cell sequencing analyses. Cross-method validation identified seven core genes (e.g., MDK, RERE, ERBB4), with RERE exhibiting a notably high eQTL-SCZ colocalization probability of 92.8-93.9%. Single-cell and epigenetic analyses reveal that the spatiotemporal dynamics of RERE may lead to differences in its mediated SCZ disease risk at the levels of genetic variation and transcriptional regulation. Cleavage Under Targets and Tagmentation (CUT&Tag) confirmed that RERE directly regulates synaptic genes such as PPP4R3B and RPS27L, and its co-expression network was found to be strongly linked to SCZ risk. This study unveils a RERE-mediated epigenetic-neurodevelopmental axis and suggests that GABAergic neurons may be a potential new target for the treatment of SCZ. Future validation using organoid models and exploration of clinical translation are warranted.

Spinocerebellar ataxia type 2 (SCA2) is a polyglutamine disorder, and variants in its disease protein Ataxin-2 act as modifiers in the progression of Amyotrophic Lateral Sclerosis. There are no reliable molecular biomarkers for SCA2. The aim of this study was to define novel molecular biomarker candidates for SCA2. Using cerebellar and cervicothoracic spinal cord RNA/protein from Atxn2-CAG100-KnockIn (KIN) and wildtype mice, a multi-omics study was conducted based on the integration of global transcriptomic, proteomic, and phosphoproteomic data, followed by validation in mice and humans. Venn diagram comparisons across all OMICS datasets indicated that only Serpinb1a-transcript, SERPINB1A-protein and -phosphopeptides were consistently downregulated at terminal stage in 14-month-old KIN mice. Expression studies in cerebellum and spinal cord from 10 weeks (pre-manifest), 6-month-old (early ataxic), and 14-month-old (late ataxic stage) mice confirmed this progressive decrease at mRNA and protein level. SERPINB1 plasma levels were significantly lower in early-stage SCA2 patients, and displayed a significant association with the CAG repeat length at expanded ATXN2 alleles, the age at onset and INAS count. However, these human data from this SCA2 founder population were not robust, so reappraisal in large international studies and at later disease stages of SCA2 is needed. SERPINB1 was identified as novel candidate progression biomarker for SCA2 pathomechanisms.

Genome-wide association studies have identified numerous susceptibility loci in complex diseases, such as chronic immune-mediated inflammatory disorders (IMIDs), yet their impact on pathomechanisms remains poorly understood. Low effect sizes, polygenicity, and predominance within non-coding genomic regions remain major challenges to the functional interpretation of IMID-associated single-nucleotide polymorphisms (SNPs). To address this, we present a novel systems genomics approach which models the cumulative impact of non-coding SNPs on downstream cellular signalling and gene regulatory networks. Applying this to the prototypical chronic IMIDs of Crohn's disease (CD) and ulcerative colitis (UC), both forms of inflammatory bowel disease (IBD), we individually analysed 2,636 patient genomes. Signals from non-coding SNPs were found to propagate towards well-established and novel CD- and UC-associated pathogenic pathways through the signalling and gene regulatory layers. The SNP-propagated gene regulatory networks stratified CD and UC patients into distinct clusters corresponding to cell type-specific gene dysregulation and potential therapeutic response. This approach bridges the gap between genotype and phenotype, laying the foundations for accelerating precision medicine in complex diseases.

<h4>Background</h4>Cytomegalovirus (CMV) is associated with congenital infections that can lead to severe developmental and neurological complications in affected children. Daycare workers, who frequently interact with infants and young children, may be at higher risk of CMV infection. This study aims to investigate the rates of primary infection and reinfection of CMV among female daycare workers in the province of Québec, Canada to better understand the risk of occupational exposure in this population.<h4>Methods</h4>This mixed-method observational study protocol uses quantitative (prospective cohort) and qualitative (semi-structured interviews with daycare workers) components. Female daycare workers (n = 553) are recruited and enrolled in a prospective cohort study, with a non-daycare workers comparison group comprising hospital employees and plasma donors who do not have direct professional contact with children < 36 months of age (low-risk group, n = 1659, 1:3 ratio). Participants are followed for 12 months, with blood for CMV serology, strain-specific antibody profiling with CMV-specific ELISA, CMV quantitative polymerase chain reaction (qPCR) and CMV-Scan assay collected from all participants at the first (V0) and last (V12) study visits. Among daycare workers and hospital employees, saliva samples are collected at these visits and monthly throughout the study for testing via CMV qPCR. Testing enables the identification of primary infections, reinfections, and reactivations, and estimates the association between demographic and occupational factors and CMV infection. Additionally, qualitative data is collected through interviews with daycare workers and daycare managers, focusing on their perceptions of CMV risks and current infection control practices.<h4>Discussion</h4>This study is expected to improve the understanding of CMV epidemiology among a high-risk group (i.e. female daycare workers). By estimating the incidence of CMV infections and patterns of viral shedding, the findings may inform occupational health strategies and public health policies. Based on our findings, guidelines aimed at preventing CMV infections could help protect not only daycare workers but also vulnerable populations, including young children and immunocompromised individuals, with whom they interact.

<h4>Background</h4>The overdose epidemic is presently driven by polydrug use, sparking renewed interest in why people initiate use of certain drugs or drug combinations. Current research privileges the physiological ends of consumption, often ignoring the social and environmental context of use. Framed by social cognitive theory, the purpose of this study was to characterize factors precipitating substance initiation, transition, and combination beyond the immediate effects of the substance(s).<h4>Methods</h4>We conducted 30 semi-structured interviews with people who use drugs across North Carolina, exploring substance use history and risk and protective factors of polydrug use. Participants also completed a visual timeline activity. We used a staged analytic approach, beginning with deductive Structural Coding and ending with inductive Reflexive Thematic Analysis at both the transcript and excerpt levels.<h4>Results</h4>We conceptualized substance transitions as pragmatic responses to environments of constraints and opportunities. Socially, transitions facilitated interpersonal closeness, aligned use with network norms, and responded to ubiquitous drug availability. Transitions also reflected navigation of material constraints, including which substances were locally available, logistically accessible, and financially sustainable.<h4>Conclusions</h4>Beyond the desire for new or enhanced physiological effects, substance transitions serve social and practical functions, like facilitating emotional closeness and responding to market conditions. Interventions to reduce the risks of use should expand viable options by addressing structural barriers and promoting safe, affordable, and accessible supply.

Diabetic cardiomyopathy (DCM) is a significant complication of diabetes mellitus, often leading to heart failure and increased mortality. While its pathogenesis remains incompletely understood, key contributors include chronic hyperglycemia, hyperlipidemia, insulin resistance, myocardial fibrosis, oxidative stress, mitochondrial dysfunction, and aberrant cell death pathways. Emerging evidence highlights ferroptosis, an iron-dependent form of regulated cell death, as a critical player in DCM progression. This review synthesizes current knowledge on the mechanistic links between ferroptosis and DCM, focusing on endothelial dysfunction, myocardial fibrosis, oxidative stress, and mitochondrial damage. We also discuss promising therapeutic strategies targeting ferroptosis to alleviate DCM, including pharmacological inhibitors, natural compounds, and non-coding RNAs, while identifying gaps for future research.

<h4>Background</h4>Emerging evidence indicates that gut microbiota and intestinal injury are crucial in pediatric metabolic dysfunction-associated steatotic liver disease (MASLD), yet the role of key gut microbial metabolites such as tyramine in pediatric MASLD remains largely unknown. In this study, we aimed to explore the role of gut microbial tyramine in intestinal damage and MASLD development in children.<h4>Methods</h4>We investigated the functions and mechanisms of previously isolated Enterococcus faecium B6 (E. faecium B6) and its derived tyramine in a mice model of intestinal injury and MASLD development. An integrative analysis of transcriptomics and proteomics was performed on mouse liver to explore the molecular mechanisms of tyramine in MASLD progression. Targeted metabolomics was performed using fecal samples from a hospital-based population (27 MASLD cases and 27 matched controls) to measure tyramine levels. The association of serum tyramine and MASLD risk was then validated in a school-based population, using serum samples of 294 children in the MASLD group and 235 controls.<h4>Results</h4>E. faecium B6 and its metabolite tyramine significantly disrupted the intestinal barrier and increased intestinal permeability in mice. Tyramine supplementation promoted MASLD-related metabolic phenotype in mice. Multi-omics analysis indicated that the PPAR signaling pathway played an important role in the molecular mechanisms. Tyramine contributed to lipid accumulation mainly by increasing lipid synthesis and lipid uptake but reducing the β-oxidation processes in the mouse liver, as shown by real-time quantitative polymerase chain reaction and western blot. Furthermore, we demonstrated from the hospital-based cohort that tyramine concentration was significantly higher in the MASLD group than in the control group. Consistently, the school-based cohort demonstrated a higher risk of MASLD in the high-tyramine group compared to the low-tyramine group, with adjusted odds ratios (ORs) and 95% confidence intervals (CIs) of 3.65 (95% CI: 2.66-4.32).<h4>Conclusions</h4>These results demonstrated that gut microbial tyramine effectively induced intestinal damage and facilitated MASLD development in mice. Tyramine was positively associated with the risk of MASLD in children. This study offered mechanistic insights into the pathogenesis of MASLD and opened therapeutic opportunities for such metabolic diseases.

<h4>Background</h4>Small cell lung cancer (SCLC) is a highly aggressive malignancy characterized by rapid progression and the frequent emergence of resistance to standard chemotherapeutic agents such as cisplatin (DDP) and etoposide (VP16), resulting in poor clinical outcomes.<h4>Methods and results</h4>To elucidate mechanisms underlying chemoresistance, we conducted a genome-wide CRISPR/Cas9 knockout screen, which identified the histone demethylase KDM6B as a critical mediator of drug resistance in SCLC. Genetic silencing of KDM6B significantly reduced IC₅₀ values of DDP and VP16, particularly in H69-AR cells, and enhanced chemotherapy-induced apoptosis. Consistently, pharmacological inhibition of KDM6B using the dual KDM6A/B inhibitor Gskj1 markedly potentiated the effects of DDP and VP16, while exhibiting minimal cytotoxicity as monotherapy. Overexpression of KDM6B rescued the chemosensitizing effect of Gskj1, thereby excluding confounding contributions from KDM6A. In vivo, the combination of Gskj1 with chemotherapy synergistically suppressed tumor growth without detectable systemic toxicity. To explore the downstream regulatory pathways, we performed transcriptome analysis via RNA-seq followed by KEGG pathway enrichment analysis, which revealed that Gskj1 treatment modulates key oncogenic signaling pathways. Integration of RNA-seq with H3K27me3 ChIP-seq data identified EGR3 as a direct epigenetic target of KDM6B inhibition. STRING analysis further suggested that EGR3 is co-expressed with c-FOS. Functional assays, including qRT-PCR, Western blotting, Co-immunoprecipitation (Co-IP), and dual-luciferase reporter assays, confirmed that EGR3 transcriptionally activates c-FOS, establishing an EGR3/c-FOS regulatory axis downstream of KDM6B. Mechanistically, inhibition of this axis enhanced chemosensitivity by promoting apoptosis, as evidenced by activation of caspase signaling, and by inducing ferroptosis through downregulation of GPX4, upregulation of ACSL4, lipid peroxidation, and modulation of HO-1. Rescue experiments with Z-VAD and ferrostatin-1 further validated that both apoptosis and ferroptosis contribute to the chemosensitizing effects of KDM6B inhibition.<h4>Conclusion</h4>Finally, in vivo experiments using patient-derived xenograft (PDX) models demonstrated that Gskj1 effectively enhances the antitumor efficacy of chemotherapy in SCLC, providing compelling evidence for the clinical potential of targeting KDM6B to overcome chemoresistance.

In this study, we synthesized an ionizable lipidoid, Cdoids, composed of carbon dots (Cdots) conjugated with oleic acid that exhibits superior biocompatibility compared to conventional ionizable lipids while retaining intrinsic fluorescence for real-time tracking and optimal nasal delivery of mRNA cancer vaccines. These properties make Cdoids a promising candidate for lipid nanoparticle (LNP) formulations. Incorporating Cdoids into LNPs enhances mRNA delivery efficiency via intranasal (IN) administration, providing a viable alternative to intramuscular (IM) injection. The unique structural features of Cdoids facilitate the formation of stable nanoparticles that efficiently encapsulate and deliver mRNA leveraging the immunological environment of the nasal mucosa to induce robust systemic and localized immune responses. Notably, Cdoids broaden the range of endocytosis mechanisms beyond conventional LNPs, leading to significantly enhanced mRNA expression within target cells. Comparative analyses demonstrated that Cdoids surpass commercially available ionizable lipids, such as SM-102, in both mRNA delivery efficiency and safety, exhibiting lower cytotoxicity in vitro and improved mRNA expression in vivo following IN administration. Furthermore, when applied as an mRNA cancer vaccine platform, Cdoids-based LNPs elicited strong antigen-specific immune responses, leading to effective tumor growth suppression. This study highlights the potential of Cdoids as an advanced ionizable lipid for LNP-based mRNA therapeutics and introduces an innovative strategy for optimizing intranasal vaccine delivery.

<h4>Background</h4>Patient-derived organoids provide a powerful platform for elucidating mechanisms of drug resistance and tumor evolution in hepatocellular carcinoma (HCC) and identifying novel therapeutic targets.<h4>Methods</h4>Transcriptomic sequencing was used to compare gene expression patterns between organoid-forming and non-forming HCC tissues, as well as between sorafenib-resistant organoids and sorafenib-sensitive counterparts. The TCGA-liver hepatocellular carcinoma (LIHC) cohort was used to screen for the key molecular drivers of HCC evolution from the overlapping differentially expressed genes. The effects of D-amino acid oxidase (DAO) on the growth, migration, invasion, apoptosis, and generation of H₂O₂ were evaluated in HepG2 and SK-Hep-1 cell lines and human HCC organoids. The therapeutic efficacy of DAO against HCC growth and drug resistance was validated with xenograft mouse model and organoids, respectively.<h4>Results</h4>Expression level of DAO was significantly downregulated in HCC tissues that successfully formed organoids compared to those that failed, as well as in sorafenib-resistant organoids versus their parental counterparts. In the TCGA-LIHC cohort, DAO expression was significantly reduced in advanced-stage HCC tissues and was inversely correlated with stemness- and epithelial-mesenchymal transition (EMT)-related molecules. Lower DAO expression was associated with poor overall survival in patients with HCC. In HepG2 cells, DAO knockdown significantly enhanced cell proliferation. Ectopic DAO expression suppressed proliferation, migration, and invasion in HepG2 and SK-Hep-1 cells. D-alanine (D-Ala) supplementation further enhanced the anti-proliferative effect of overexpressed DAO, but did not significantly alter the DAO-mediated suppression of migration or invasion. Ectopic expression of DAO induced apoptosis via the generation of H₂O₂ upon simultaneous supplementation of D-Ala into the culture medium; the addition of catalase, an H₂O₂-degrading enzyme, significantly reversed the D-Ala-induced effects. In BALB/c nude mouse models, HCC cells overexpressing DAO formed significantly smaller tumors than the control cells (P = 0.010), and this tumor-suppressive effect was further enhanced by D-alanine supplementation. Ectopic DAO expression restored sorafenib sensitivity in resistant organoids.<h4>Conclusion</h4>DAO appears to be a novel endogenous stemness repressor. The reduction in DAO is a critical molecular event in the evolution of HCC. Therapeutically, combined DAO and D-amino acid supplementation is a promising strategy for HCC treatment, particularly for reversing sorafenib resistance.

The distribution of adipose depots in different body parts affects pig production value and human health, governed by complex epigenomic mechanisms. Limited studies on pig adipose depots have hindered the genetic improvement of fat-related economic traits and their biomedical applications. To address this issue, we generated epigenomic maps for backfat, belly fat, groin fat, and intermuscular fat (IMF) in Meishan pigs, integrating ChIP-seq, ATAC-seq, RNA-seq, Hi-C, and public whole-genome sequencing data. Our results reveal that belly/backfat share similar chromatin states, while groin fat/IMF exhibit distinct H3K27ac modification, super-enhancer (SE) dynamics, and open chromatin landscapes compared to belly/backfat. The spatially specific expressions of adipogenic transcription factors (TFs), such as lipid synthesis-related TFs PPARA and SOX6, which are highly expressed in back/belly fat, and adipocyte differentiation TF KLF4 was driven by a groin fat specific SE, underlie these chromatin state disparities. These results also suggest enhanced lipid synthesis in belly/backfat and adipocyte differentiation in groin fat. Moreover, candidate functional variants identified in IMF-gained H3K27ac peaks are primarily associated with meat quality traits. Genes linked to pig backfat thickness may also serve as candidate genes for human obesity due to the conserved cis-regulatory elements and gene expression patterns between humans and pigs. Overall, our epigenomic landscape enhances understanding of adipose depot regulation in mammals, facilitating cross-species insights and precision breeding.

Chronic obstructive pulmonary disease (COPD) is a progressive lung disease defined by persistent airflow limitation, chronic inflammation, and ongoing airway remodeling, and poses a substantial global health challenge. Despite its clinical significance, the underlying cellular mechanisms remain poorly defined. Regulated cell death (RCD) incorporates various kinds of cell death that are typically regulated by specific molecular pathways. Numerous new kinds of RCD have been identified outside of the traditional apoptotic pathway, like necroptosis, pyroptosis, ferroptosis, autophagy, cuproptosis, and parthanatos. To date, there is growing evidence indicating that these pathways participate in the regulation of COPD development. However, their specific roles and therapeutic relevance remain poorly understood. In this review, we discuss a summary of the molecular mechanisms by which RCD pathways influence the onset and advancement of COPD. Additionally, the therapeutic benefits of agents that target these cell death pathways in COPD treatment were described. By integrating current insights, this review aims to broaden our knowledge of the pathophysiology of COPD and offer novel approaches to treatment.

<b>Background/Objectives</b>: Cardiovascular disease is the leading cause of mortality in patients with metabolic dysfunction-associated steatotic liver disease (MASLD), which includes simple steatosis (metabolic dysfunction-associated steatotic liver, MASL), metabolic dysfunction-associated steatohepatitis (MASH), and fibrosis. This study aimed to evaluate the association between left ventricular (LV) systolic function, measured by global longitudinal strain (GLS), and liver inflammation and fibrosis in obese patients with MASLD undergoing preoperative evaluation for bariatric surgery. <b>Methods</b>: Intraoperative liver biopsies classified patients into four groups: non-MASLD; MASL; MASH; and MASH with fibrosis. Preoperative transthoracic echocardiography (TTE) was performed, and LV GLS was assessed using automated strain analysis. <b>Results</b>: Ninety-two patients were included: 13 non-MASLD, 34 MASL, 21 MASH, and 24 MASH with fibrosis. Although most patients had normal LV GLS, values were significantly lower in the MASH with fibrosis group compared to the MASL and non-MASLD groups (<i>p</i> = 0.011). In multivariate analysis adjusted for HDL cholesterol and LV mass, LV GLS was associated with inflammation and fibrosis (OR 0.784; 95% CI 0.637-0.965; <i>p</i> = 0.022). <b>Conclusions</b>: LV GLS was significantly lower in patients with MASH and MASH with fibrosis and was associated with hepatic inflammation and fibrosis in obese individuals undergoing bariatric surgery.

Glioblastoma is the most common and lethal type of tumor of the central nervous system, with an average survival of 15 months after first diagnosis. Immune checkpoint inhibitors (ICIs) have been largely investigated for their ability to harness the immune system to combat tumors. However, their efficacy varies a lot depending on tumor type. In glioblastoma, PD-1/PD-L1 immunotherapy has been explored in various studies; however, the unique immunosuppressive environment in the brain and the presence of the blood-brain barrier as well as the large intratumoral heterogeneity have limited its efficacy considerably. In order to improve the clinical efficacy of ICIs, it is important to delve into the different factors affecting the response rate in GBM. Herewith, we summarize the most common causes of resistance to anti-PD-1/PD-L1 immunotherapy as well as possible ways of enhancing its efficacy, particularly through combination with other therapeutic agents in the preclinical and clinical setting. Furthermore, we provide an insight into the most promising methods for modulating the blood-brain barrier, as well as the growing role of molecular imaging and radiogenomics in this field.

<h4>Background</h4>Diabetic nephropathy (DN) is a major microvascular complication of diabetes, and its pathogenesis is closely associated with abnormal epigenetic regulation, particularly the silencing of tumor suppressor genes due to hypermethylation of promoter regions. This study was to investigate the workings of demethylation in diabetic nephropathy by applying bioinformatics methods.<h4>Methods</h4>DN-related datasets (GSE142153 and GSE154881) and demethylation-related genes (D-RGs) were included. Differentially expressed genes (DEGs) (DN vs. normal) were obtained. Candidate genes were obtained from the intersection of DEGs and D-RGs. To identify key genes, the Least absolute shrinkage and selection operator (LASSO) and Boruta algorithm, and expression validation were used for screening. The expression validation was used to identify biomarkers. The receiver operating characteristic (ROC) curve was subsequently utilized to assess the biomarkers' capability to distinguish diseased from normal samples. Subsequently, a predictive nomogram was created to estimate the likelihood of developing DN. In addition, functional enrichment, immune infiltration, subcellular localization, correlation of biomarker expression with renal function, correlation for other diseases, network analysis of molecular interactions and computational drug prediction were carried out. Lastly, Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR) was carried out to confirm the expression levels of biomarkers in blood samples.<h4>Results</h4><i>CXCL2</i> and <i>MLF1</i> were determined to be biomarkers that exhibited notably elevated expression levels in the DN, in contrast to the normal group. Then the nomogram network was built, which had high prediction accuracy. Pathways most significantly enriched by <i>CXCL2</i> and <i>MLF1</i> included cytokine-cytokine receptor interaction and MAPK signaling pathway. Five types of immune cells were identified by immune infiltration analysis. In the RNA binding protein (RBP) -mRNA regulatory network, seven pathways were co-enriched in both biomarkers. In the TF-mRNA regulatory network, TFs shared by both biomarkers include JUN, GATA2, and SRF. Finally drug prediction analysis found a total of 172 target drugs for <i>CXCL2</i> and <i>MLF1</i>. RT-qPCR experiment revealed that both biomarkers showed a notable rise in the DN group relative to the normal group. RT-qPCR results revealed the DN exhibited notably increased expression levels of the two biomarkers (<i>CXCL2</i> and <i>MLF1</i>) compared to the normal group.<h4>Conclusion</h4><i>CXCL2</i> and <i>MLF1</i> were identified as diagnostic biomarkers for DN, offering a new reference for its treatment.

<h4>Background</h4>Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant tumor with poor prognosis. Efferocytosis, an essential process for clearing apoptotic cells, is involved in shaping immunosuppressive microenvironment, facilitating tumor immune evasion. This study aims to evaluate the prognostic value of efferocytosis-related biomarkers in PDAC and elucidate their underlying mechanisms, providing insights for personalized therapy.<h4>Methods</h4>We integrated bulk and single-cell transcriptomic data from public database to construct and validate an efferocytosis-related prognostic model for PDAC. Additionally, we analyzed the specimens from a cohort of 81 PDAC patients alongside cell experiments to elucidate the function of P2RY6. RNA-seq analysis was employed to uncover the effector pathways mediated by P2RY6.<h4>Results</h4>An efferocytosis-based prognostic model, EFFscore, developed based on <i>ADAM9</i>, <i>P2RY6</i>, and <i>CD36</i>, can effectively assess the tumor phenotype of PDAC patients. The EFFscore is strongly associated with tumor evolution, malignant biological characteristics and microenvironmental interactions of PDAC. The essential mediator P2RY6 is significantly upregulated in PDAC tissue and cells, correlating closely with poor prognosis. Functional studies demonstrate that P2RY6 inhibition exhibits tumor-suppressive effects by activating the endoplasmic reticulum stress and enhancing anti-tumor immune responses. The P2RY6 receptor inhibitor, MRS-2578, emerges as a promising therapeutic candidate for PDAC treatment.<h4>Conclusion</h4>The prognostic model EFFscore exhibits remarkable predictive performance, accurately reflecting the malignant potential of PDAC. P2RY6 serves as a key oncogenic factor driver in PDAC, its targeted inhibition significantly suppresses tumor progression, highlighting its dual potential as a diagnostic biomarker and therapeutic target.

<h4>Aim</h4>Physiological bone mineralization and ectopic vascular calcification share similarities in the composition of calcium phosphate minerals. Evidence suggests a connection between the underlying biological mechanisms driving the deposition of bone mineral and cardiovascular calcification. Therefore, understanding the chemistry and composition of bone mineral and vascular calcification may be important for the development of effective treatments and diagnostic tools for cardiovascular diseases, as pharmacological interventions for the treatment of one process might affect the other. The goal of this study was to identify and compare compositional features of calcium phosphates in bone and calcified vascular tissues using phosphorus-31 (³¹P) solid state cross-polarization magic angle spinning (CPMAS) magnetic resonance (MR) spectroscopy, a specialized technique that provides compositional information unattainable through conventional chemical analysis.<h4>Methods</h4>Solid state MR spectra were acquired from biological specimens of human trabecular bone (<i>n</i> = 1), human vascular plaque (<i>n</i> = 1), human calcified aortic valves (<i>n</i> = 5), as well as calcified aortic tissues of apolipoprotein E-deficient mice (<i>n</i> = 1) fed a high cholesterol diet. Synthetic hydroxyapatite [Ca₁₀(OH)₂(PO₄)₆] and synthetic brushite (CaHPO₄·2H₂O) were used to model the solid state ³¹P MR spectra of the phosphate ion PO₄ ^-3 and hydrogen phosphate ion HPO₄ ^-2, respectively. Qualitative spectral features and quantitative metrics derived using Herzfeld-Berger analysis were assessed to characterize mineral composition and maturity.<h4>Results</h4>Solid state ³¹P MR spectra of all human specimens studied suggested a well-ordered crystal structure dominated by unprotonated phosphate (PO₄ ^-3), consistent with mature bone-like mineral. These specimens exhibited long CP time constants (700-900 µs) and modest chemical shift anisotropy. In contrast, the calcified mouse aorta spectrum showed pronounced sidebands, a short CP time constant (∼270 µs), and a more prominent HPO₄ ^-2 component-features indicative of immature, newly deposited mineral.<h4>Conclusion</h4>³¹P solid state MR spectroscopy reveals differences in the phosphate and hydrogen phosphate ion content among the calcified tissues studied. This technique could potentially be an important complement to basic studies of pathological calcification in atherosclerosis and related calcific disorders.

Autophagy is an evolutionarily conserved catabolic process that plays a central role in maintaining cellular homeostasis by degrading and recycling damaged or surplus proteins, organelles, and other cellular macromolecules and components. A growing body of evidence highlights a bidirectional relationship between autophagy and nicotinamide adenine dinucleotide (NAD⁺), a vital metabolic cofactor involved in numerous cellular processes, including energy metabolism, genomic maintenance, stress resistance, and cell survival. Autophagy supports NAD⁺ homeostasis by recycling metabolic precursors, while NAD⁺-dependent enzymes such as sirtuins and PARPs regulate autophagy initiation and lysosomal function. Disruption of this autophagy-NAD⁺ axis has emerged as a common feature in several neurodegenerative diseases, where impaired cellular clearance and metabolic dysfunction contribute to neuronal vulnerability. In this review, we summarize the advances of the molecular links between autophagy and NAD⁺ metabolism, with a particular focus on their roles in mitochondrial quality control, bioenergetic regulation, and cellular resilience. We also discuss the therapeutic potential of targeting the autophagy-NAD⁺ axis to promote neuroprotection in neurodegenerative disease.

<i>Polygonatum kingianum</i>, a medicinally significant herb indigenous to Yunnan, China, exhibits pronounced sensitivity to temperature affecting its bioactive compound accumulation and growth. However, the molecular mechanisms underlying its response to temperature stress remain poorly characterized. To elucidate these mechanisms, we integrated physiological assessments with RNA-seq analysis of <i>P. kingianum</i> under controlled temperatures (10°C, 25°C, 30°C, and 35°C). The results indicate that <i>P. kingianum</i> exhibits optimal root growth at 10-25°C but shows poor adaptation to high temperature. Temperature stress led to inhibited root growth, accompanied by decreased chlorophyll content and leaf yellowing. This stress manifested as increased levels of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in the leaves, along with elevated proline accumulation. In response to oxidative damage, the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as the levels of non-enzymatic antioxidants (ascorbate/ASA and glutathione/GSH), were significantly upregulated. Transcriptome analysis further revealed that these physiological changes are regulated by multiple stress-related pathways, mainly including phenylpropanoid biosynthesis, glycolysis/gluconeogenesis, starch and sucrose metabolism, and plant hormone signaling. Additionally, 19 key genes encoding reactive oxygen species scavenging enzymes were identified and functionally characterized as core regulators of thermotolerance. This study deciphers the physiological and molecular mechanisms of the temperature stress response in <i>P. kingianum</i>.

No abstract available.

<b>Background</b>: Anxiety disorders, depressive disorders, and insomnia often co-occur and impair functioning in psychiatric patients. Virtual reality (VR) is a promising relaxation tool, yet its efficacy relative to classical Schultz autogenic training (AT) remains insufficiently characterized. <b>Methods</b>: Thirty-seven female patients were randomly assigned to four groups: (1) inpatient AT (<i>n</i> = 10), (2) inpatient VR (<i>n</i> = 10), (3) home-based AT (<i>n</i> = 10), and (4) home-based VR (<i>n</i> = 7). Interventions lasted 2 weeks (≥10 sessions). Depressive and anxiety symptoms, sleep quality, and cognitive function were assessed pre- and post-intervention. <b>Results</b>: In the total sample, anxiety and depressive symptoms decreased and sleep quality improved, while cognitive functions showed slight improvement. In subgroup analyses, inpatient AT reduced anxiety and improved sleep, whereas inpatient VR reduced both anxiety and depressive symptoms and improved sleep. In the home-based modality, AT did not significantly affect sleep, anxiety, or depressive symptoms but was associated with modest cognitive gains, while home-based VR improved sleep without significant changes in anxiety or depression. <b>Conclusions</b>: Both methods are straightforward to implement and promote improvement in selected mental health parameters; however, their effect profiles are context-dependent. Interventions delivered in the inpatient setting were more effective than those at home, suggesting a substantial influence of contextual factors (fewer distractions, therapeutic structure, group component). Among the tested conditions, inpatient VR-based relaxation produced the broadest pattern of improvement in anxiety, depression, and sleep. These pilot findings require confirmation in larger, prospectively designed studies.

Idiopathic male infertility (IMI), which accounts for nearly 50% of male infertility cases, remains a major clinical challenge due to the inability of standard semen analysis to reveal underlying molecular defects. Post-translational modifications such as lysine acetylation are increasingly recognized as key regulators of sperm function, affecting antioxidant defense, energy metabolism, and spermatogenesis. In this study, global acetyl-proteomic profiling of spermatozoa from idiopathic infertile patients (IIP) and fertile donors (FD) was performed using immunoprecipitation-based enrichment followed by high-resolution LC-MS/MS. Bioinformatics analyses, including STRING, Cytoscape, and Ingenuity Pathway Analysis (IPA), identified 718 differentially acetylated proteins (DAPs), with significant enrichment in pathways related to redox homeostasis, molecular transport, glycolysis, and mitochondrial metabolism. Hub proteins such as SOD1, PARK7, and PRKACA emerged as key regulators linking oxidative stress with defective motility and impaired sperm physiology. Western blot validation confirmed the downregulation of these hub proteins in IIP, supporting their role in redox imbalance and mitochondrial dysfunction. Our findings highlight dysregulated lysine acetylation as a defining molecular feature of IMI and suggest that targeting acetylation-associated pathways may provide novel diagnostic biomarkers and therapeutic strategies to improve sperm function and male reproductive outcomes.

Ferroptosis is a novel iron-sensitive subtype of regulated cell death (RCD), persisting under extreme lipid peroxidation and iron/redox imbalances. Unlike apoptosis, necroptosis, and pyroptosis, ferroptosis is a signaling-driven process mediated through iron metabolism imbalance, polyunsaturated fatty acid (PUFA) exceeding oxidation, and defects in its protective systems like Xc-/GSH/GPx4. Specifically, this review establishes that iron-driven ferroptosis is a central underlying pathomechanistic factor in a broad range of human diseases. Significantly, whether its modulation is therapeutic, it is entirely conditional on the specific disease context. Thus, its induction can provide a promising antidote for destructive cancer cells when conjoined with immuno-therapies to boost anticancer immunity. Conversely, iron-mediated ferroptosis suppression is a key factor in countering destructive changes in a whole range of degenerative and acute injuries. Current therapeutic approaches include iron chelators, lipid oxidation inhibitors, GPx4 activators, natural and active compounds, and novel drug delivery systems. However, against all odds and despite its intense therapeutic promise, its translation into a practical medicinal strategy faces many difficulties. Thus, a therapeutic agent specifically focused on its modulation is still lacking. The availability of selective biologic markers is a concern. The challenges in the direct pathologic identification of ferroptosis in a complex in vivo systemic scenario remain. Current avenues for its future development are pathogen infections, the discovery of novel regulating factors, and novel approaches to personalized medicine centered on its organ-level in vivo signatures.

Sirtuin 6 (SIRT6), a (Nicotinamide adenine dinucleotide) NAD⁺-dependent deacylase and mono- (adenosine diphosphate) ADP-ribosyltransferase, is increasingly recognized as a pivotal regulator of genomic stability, metabolic reprogramming, and epigenetic remodeling. This review synthesizes current evidence on the dual roles of SIRT6 in cancer, highlighting its context-dependent functions as both a tumor suppressor and promoter across various malignancies. We detail its involvement in DNA damage sensing, repair coordination, glycolytic regulation, and chromatin modification, and discuss how these mechanisms contribute to tumor initiation, progression, and therapy resistance. Emerging therapeutic strategies targeting SIRT6, including small-molecule modulators, genetic interventions, and combination therapies, are critically evaluated. Our analysis underscores the necessity for context-specific therapeutic targeting, and pharmacological modulation of SIRT6 represents a promising avenue for precision oncology.

Ferroptosis is a mechanism of programmed cell death that is characterized by iron-dependent lipid peroxidation, and it is closely associated with organismal metabolism. Modulating ferroptosis may offer therapeutic potential for various diseases. Podocytes are essential intrinsic cells in the renal corpuscle, and podocyte injury contributes to the progression of kidney diseases. A growing number of studies have confirmed that ferroptosis can occur in podocytes, and inhibiting podocyte ferroptosis may provide protective effects in multiple kidney diseases. However, the specific pathogenic molecular mechanisms and treatments for podocyte ferroptosis are still under investigation. In this review, we systematically review the core mechanisms of ferroptosis, including iron metabolism, lipid metabolism, and amino acid metabolism; summarize the molecular mechanisms of podocyte ferroptosis in various types of kidney disease; and discuss the current therapeutic strategies targeting podocyte ferroptosis. By synthesizing this information, we explore the potential of podocyte ferroptosis as a therapeutic target for kidney diseases, aiming to provide insights for both basic research and the clinical treatment of podocyte injury.

In mammals, adipose tissue is involved in energy homeostasis regulation. White adipose tissue (WAT) is the major energy storage site for mammals, whereas brown adipose tissue (BAT) is involved in energy expenditure and thermogenesis. The conversion of triglycerides from WAT into fuel provides calories for the body and aids in reducing WAT accumulation, which is considered an effective strategy for weight loss and improving lipid metabolism. Regulation at the transcriptional level can determine the differentiation and transdifferentiation of WAT and BAT; however, post-transcriptional regulation offers an additional level of coordination to this process. Despite the high number of studies investigating the transcriptional regulatory mechanisms of key thermogenesis-regulated genes, the regulatory roles of these genes at the post-transcriptional level remain poorly understood. Therefore, this review summarizes the post-transcriptional regulatory mechanisms of key thermogenesis-regulated genes, including splicing, m6A modification, RNA degradation, RNA-binding proteins, and ncRNA regulation of pre-mRNAs of these thermogenic genes.

<h4>Background</h4>Metabolic dysfunction-associated steatotic liver disease (MASLD) is a leading cause of liver disease worldwide. We aimed to assess the prevalence and severity of MASLD and fibrosis among asymptomatic individuals with no known history of liver disease in British Columbia (BC), Canada.<h4>Methods</h4>We conducted a cross-sectional, population-based screening study of 2,782 individuals in the Lower Mainland of BC. Baseline demographics were collected, and transient elastography was performed. The prevalence and severity of MASLD and liver fibrosis were calculated.<h4>Results</h4>MASLD affected 53.1% of participants, with 34.0% having severe hepatic steatosis, 6.5% moderate hepatic steatosis, and 12.6% mild hepatic steatosis. Factors associated with a higher MASLD incidence included non-lean BMI (OR 5.50, <i>p</i> <0.001), hypertension (OR 1.29, <i>p</i> = 0.014), diabetes (OR 1.33, <i>p</i> = 0.026), and South Asian ethnicity (OR 1.36, <i>p</i> = 0.014), while female gender was protective (OR 0.81, <i>p</i> = 0.015). Non-lean BMI (OR 5.71, <i>p</i> <0.001), hypertension (OR 1.33, <i>p</i> = 0.002), and diabetes (OR 1.34, <i>p</i> = 0.010) were associated with more severe steatosis. Fibrosis was present in 7.2% of participants, with 4.4% having moderate fibrosis, 1.9% having severe fibrosis, and 0.9% having cirrhosis. Diabetes (OR 1.93, <i>p</i> <0.001) and non-lean BMI (OR 2.37, <i>p</i> <0.001) were associated with a higher prevalence of fibrosis, while East Asian ethnicity was protective (OR 0.50, <i>p</i> <0.001). Non-lean BMI (OR 2.35, <i>p</i> <0.001) and diabetes (OR 1.96, <i>p</i> <0.001) were linked to higher fibrosis severity, while East Asian ethnicity remained protective against severe fibrosis (OR 0.50, <i>p</i> <0.001).<h4>Conclusions</h4>There is a significant burden of liver steatosis and fibrosis in BC, Canada, which highlights the need for comprehensive MASLD screening guidelines.

<h4>Background/objective</h4>Tirzepatide is widely prescribed and generally considered safe. The objective of this report is to describe a patient with tirzepatide-induced liver injury and increase awareness of this rare complication.<h4>Case report</h4>A 60-year-old woman with type 2 diabetes mellitus and obesity presented with epigastric pain 21 days after starting tirzepatide 2.5 mg weekly. She previously received semaglutide and liraglutide (total 7 years) without hepatotoxicity. Isolated right upper quadrant abdominal tenderness was present. Alanine transaminase and aspartate transferase rose from mildly elevated to 1562 U/L and 1466 U/L (normal 0-35 U/L), respectively, in 24 hours, disproportionately versus alkaline phosphatase and bilirubin. Abdominal/pelvic computed tomography scan showed a mildly dilated common bile duct post cholecystectomy. Right upper quadrant ultrasound demonstrated fatty infiltration, confirmed by magnetic resonance cholangiopancreatography. Acute hepatitis panel, acetaminophen level, ceruloplasmin, liver/kidney microsome type 1 Ab, antimitochondrial antibody, α-1-antitrypsin, and alpha-fetoprotein tumor marker were normal. Following drug discontinuation, liver enzymes normalized after 24 days. Home medications were resumed except for tirzepatide, and aminotransferases remained normal at 3 months.<h4>Discussion</h4>Presentations of tirzepatide-induced liver injury include asymptomatic elevation of aminotransferases, nausea, vomiting, and abdominal pain, or, although extremely rare, hepatic failure. Risk factors and pathogenesis are undetermined.<h4>Conclusion</h4>This case documents hepatocellular injury specific to tirzepatide despite previous tolerance of glucagon-like peptide-1 receptor agonist therapy. Clinician awareness is important since initial symptoms may be mild and attributed to common gastrointestinal side effects associated with tirzepatide. Early diagnosis and drug discontinuation resulted in resolution. If unrecognized, although rare, severe hepatic dysfunction with associated complications can result.

The retinal pigment epithelium (RPE) sustains lifelong proteostasis under chronic stress, yet how post-mitotic cells activate autophagy when canonical kinase pathways suppress it remains unresolved. Here we demonstrate that AKT1, conventionally regarded as an autophagy suppressor, drives autophagy through a non-canonical transcriptional mechanism triggered by isoform imbalance. In RPE and age-related macular degeneration (AMD) models, AKT2 hyperactivation destabilizes mTORC2 and engages S6K-mediated IRS-1 inhibition, creating a feedforward autophagic arrest. Compensatory AKT1 activation via mTORC2 phosphorylates telomerase reverse transcriptase (TERT) at Serine 824, driving nuclear translocation that is independent of telomere maintenance. In the nucleus, TERT assembles with FOXO3 and MYC into a tripartite transcriptional complex that occupies the EIF2AK3 promoter, enabling PERK transcriptional activation. This activity converts the unfolded protein response from pro-apoptotic to cytoprotective: PERK-ATF4 signaling drives biogenesis of core autophagy machinery while simultaneously inducing selective ERphagy through the receptors TEX264 and CCPG1, which prevents pathological PERK clustering and preserves tubular ER architecture in diseased RPE. Using C. elegans phosphorylation-deficient mutants, mouse models and AMD patient induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE), we establish that AKT1-mediated TERT phosphorylation is an evolutionarily conserved prerequisite for FOXO/DAF-16 nuclear function and lysosomal homeostasis in post-mitotic cells. Pharmacologic targeting of AKT2 with a first-in-class dual-pocket allosteric inhibitor selectively enhances AKT1 compensation, restoring macroautophagic and ERphagy flux across disease models. These findings reveal a kinase-to-transcription axis that reprograms organelle quality control and identify the AKT1-TERT-PERK-ATF4 pathway as a therapeutic target in proteostasis-driven disease.

<title>Abstract</title> <p> Melanoma is a highly malignant, metastatic skin tumor with complex progression mechanisms. This study reveals that the metabolic enzyme ECI2 is significantly overexpressed in melanoma tissues and is closely associated with poor patient prognosis. Functional experiments confirm that ECI2 significantly promotes the proliferation, migration, and invasive capabilities of melanoma cells both in vitro and in vivo. Mechanistic investigations reveal that ECI2 reshapes arachidonic acid metabolism in tumor cells, leading to the accumulation of the metabolite PGE2. These metabolites activate neutrophils, inducing the formation of neutrophil extracellular trap networks (NETs) via reactive oxygen species (ROS)-dependent pathways. Crucially, this study innovatively demonstrates that NETs are not the final effector. NETs activate the TLR3/COX-2/PGE2 axis in melanoma cells, establishing a metabolic-immune positive feedback loop. This circuitry persistently drives malignant tumor behavior, profoundly reshaping the tumor microenvironment. Our work not only proposes ECI2 as a key oncogene and potential therapeutic target but, more importantly, reveals a novel mechanism whereby tumor cells establish a positive feedback dialogue between metabolic reprogramming and innate immune effector NETs. This provides a novel theoretical framework for understanding melanoma progression and developing combined therapeutic strategies. <bold>Subject terms</bold> : ECI2, arachidonic acid metabolism, neutrophil extracellular traps, melanoma </p>

<h4>ABSTRACT</h4> Cell identity is regulated by chromatin states that encode gene regulatory memory and shape responses to new inputs. To investigate how chromatin context influences inducibility in differentiated cells, we employed engineered synthetic reader-actuators (SRAs), fusion proteins containing the polycomb chromodomain (PCD) that binds H3K27me3. In MCF7 breast cancer cells, we mapped PCD-fusion occupancy by ChIP-seq and used RNA-seq to identify temporally resolved gene activation patterns. ChIP-seq profiling and machine learning models (MLM) demonstrated that PCD-fusion binding was predicted by the presence of H3K27me3, H4K20me1, or H3K36me3 enrichment, suggesting selective accessibility at enhancers and chromatin transition zones. Among genes with PCD-fusion enriched enhancers, the SRA-induced subset was distinguished by promoter features including bivalent histone modifications and enrichment of transcriptional repressors REST and MTA1. Collectively, our results demonstrate that SRA responsiveness depends on a specific chromatin signature beyond H3K27me3 alone. This study demonstrates the power of SRAs to dissect inducible chromatin features in their native genomic context, and suggests that epigenetically repressed regions in differentiated cells can retain regulatory plasticity.

No abstract available.

Psychotic disorders frequently result from repeated ketamine exposure, yet the underlying mechanisms remain elusive. We propose that repeated exposure to ketamine may induce psychotic-like behaviors via DRD1-mediated nuclear signaling pathways. Our investigation focused on phosphorylated DARPP-32 at Thr34, Thr75, and Ser97, alongside transcriptome profiling in both cell and mouse models. We found that DRD1 antagonist mitigated ketamine-induced psychotic-like behaviors and cognitive deficits, whereas DRD1 agonist partially replicated ketamine-like symptoms. In cellular models, ketamine elevated p-Thr34 DARPP-32 levels and facilitated its nuclear accumulation through PKA, while promoting Ser10 H3 phosphorylation by inhibiting PP1 activity. Phosphorylation at Thr75 and Thr97 inhibited p-Thr34 level, with Thr97 enhancing DARPP-32 and PP1 interaction. In vivo, combined approach of RNA-seq and ATAC-seq in the hippocampus indicated that ketamine suppressed neurogenesis. Immunofluorescence showed reduced neonatal neurons and neural stem cells in the dentate gyrus region, while ketamine increased astrocyte numbers. Single-nucleus transcriptome sequencing revealed enhanced neuron-astrocyte interaction post-ketamine treatment. In summary, we identified the DRD1-DARPP-32-Histone H3 pathway as a key mediator of transcriptional abnormalities and impaired hippocampal neurogenesis in ketamine-induced psychotic-like mouse model.

Delay discounting (DD), a person's preference for smaller immediate rewards over larger delayed rewards, is a heritable trait that is associated with psychiatric and physical outcomes, yet the biological mechanisms underlying these links are not known. We performed a GWAS of DD using 134,935 23andMe research participants and identified 11 genome-wide significant loci. We did not replicate our previously reported association with rs6528024 (chrXq13.3, GPM6B; P = 5.30 × 10^-02). The SNP-heritability of DD was 9.85 ± 0.57%. We observed genetic correlations between DD and 73 behavioral, physical, and neuroimaging traits, many of which persisted even after accounting for educational attainment, intelligence, and executive function. Network analysis revealed that the associations between DD and certain traits were explained by both overlapping and trait-specific biological processes. In a hospital-based cohort (N = 66,917), DD polygenic scores were associated with 212 medical conditions. These results demonstrate that DD has a pleiotropic and polygenic common variant architecture, and is genetically associated with numerous outcomes, making it a promising endophenotype for psychiatric and physical health.

Magnetothermal stimulation is key in biomedical applications like tumor ablation, drug delivery, and regenerative therapies. A common method involves injecting magnetic particles that heat under an alternating magnetic field (AMF). However, uncontrolled heating can damage healthy tissues. Maintaining temperatures below 45 °C is critical. Using materials with a Curie temperature (T_c) near this limit offers a self-regulating solution, as magnetization-and thus heating-drops sharply at T_c. This study explores Mn_0.65Fe_1.30P_0.65Si_0.37 (MCM), a magnetocaloric material composed of non-toxic elements and featuring a tunable T_c. It is engineered to exhibit a T_c of 43 °C, close to the safe physiological threshold. MCM particles are encapsulated in a wax matrix to form a composite that responds to AMF exposure. Heat generated by MCM particles triggers the wax phase transition, while the obtained T_c enables the composite to achieve self-limiting thermal regulation under magnetic field exposure. Biocompatibility tests using human umbilical vein endothelial cells (HUVECs) show over 90% cell viability in direct and indirect contact. Stability tests in phosphate buffers at 37 °C confirm controlled degradation over 28 days. These results demonstrate that MCM is a promising, burn-free magnetic material for safe, localized heating, supporting its use in self-regulating, temperature-responsive biomedical systems.

Gastruloids are 3D aggregates of pluripotent stem cells grown in suspension culture that mimic many aspects of gastrulation and early axial elongation. The N2B27 basal medium in which mouse gastruloids are cultured can either be home-made (HM-N2B27) with materials of known origin, or commercially sourced (NDiff227), where the exact formulation is unknown. In this study, we examined whether these formulations resulted in significant differences in gastruloid development. Our results reveal that while both media enable standard gastruloid elongation, HM-N2B27 gastruloids initiate the elongation process earlier, have a higher number of cells and an increased anterior domain. Despite the maintenance of overall gene expression patterns, RNAseq analysis indicated differences in cell fate specification, with HM-N2B27 gastruloids exhibiting higher expression of spinal cord-related genes, while NDiff227 favours mesodermal differentiation. Furthermore, differential gene enrichment analysis suggests that changes in key signalling pathways underlie the differences between HM-N2B27 and NDiff227 gastruloids. These findings highlight the importance of basal media composition for gastruloid development, underscoring the need for careful media selection during in vitro engineering of stem cell-based embryo models.

<h4>Introduction</h4>We diagnosed and managed a patient with pulmonary fibrosis combined with myelodysplastic syndrome (MDS). This paper aims to investigate, through analysis of our clinical case, a review of the existing literature, and relevant genetic analyses, whether the concurrent occurrence of pulmonary fibrosis and MDS represents a clinical coincidence or reflects an underlying shared pathogenetic mechanism.<h4>Case presentation</h4>A 64-year-old male farmer with concurrent MDS and interstitial pulmonary fibrosis underwent whole-exome sequencing, which revealed abnormalities in the MTHFR, PCSK9, and IFIH1 genes.  A literature review demonstrated that these three genes are associated with both MDS and pulmonary fibrosis.  A search of previous literature identified six similar case reports over a 22-year period, suggesting that the concurrent occurrence of these two conditions may be linked to CD68+ cells, myeloperoxidase (MPO)-positive inflammatory cells, VEXAS syndrome, telomere diseases, and other factors.<h4>Conclusion</h4>MTHFR, IFIH1, PCSK9 and CTC1 are involved in key pathways including folate metabolism, immune regulation, inflammatory responses and telomere disorders, which may contribute to the pathogenesis of both MDS and pulmonary fibrosis. The coexistence of these two conditions is likely attributed to complex interactions among multiple gene mutations, environmental triggers, and dysregulated immune processes, rather than a single.

<h4>Background</h4>Ferroptosis is an iron-dependent form of regulated necrosis driven by unchecked lipid peroxidation. Its relevance to cancer biology has become increasingly evident. Our long-standing ferric nitrilotriacetate (Fe-NTA) rat model exemplifies how chronic Fenton chemistry induces DNA damage, genomic instability, and selection of ferroptosis-resistant malignant clones, underscoring the central role of iron in carcinogenesis.<h4>Recent advances</h4>Cancer cells acquire multiple adaptations to maintain iron addiction while escaping ferroptotic pressure, including reinforcement of the xCT-CD44v axis, GPX4 and FSP1 activities, NRF2-mediated antioxidant programs, and mitochondrial remodeling. Ferroptosis can also act as an immunogenic cell death modality by releasing oxidized phospholipids and DAMPs that activate dendritic cells and CD8⁺ T cells, a process termed immunoferroptosis. Low-temperature plasma (LTP) has emerged as a unique modality capable of producing reactive oxygen and nitrogen species to selectively trigger ferroptosis in iron-loaded cancer cells while sparing normal tissues.<h4>Diagnostic and genetic context</h4>HNE-modified proteins currently serve as robust markers of lipid peroxidation in FFPE samples, whereas visualization of catalytic Fe(II)-the executer of ferroptosis-requires frozen tissues or live-cell imaging. Cancer-prone hereditary syndromes such as BRCA1/2 deficiency and Fanconi anemia exhibit ferroptosis resistance, linking defective genome maintenance to impaired ferroptotic signaling.<h4>Outlook</h4>Ferroptosis functions both as an intrinsic tumor-suppressive mechanism and a tractable therapeutic vulnerability. Harnessing ferroptosis offers a promising strategy for targeting iron-addicted cancers.

Proteins interact with lipid membranes to facilitate important cellular processes that underlie health and disease. Transmembrane proteins like ion channels are often composed of bound monomers forming specific contacts within the bilayer. However, molecular mechanisms of channel assembly are scarce. Understanding the role of lipids in this process may help further explain assembly of oligomeric proteins, which are often clinical drug targets. Using the hepatitis C virus p7 hexamer as a representative of proteins with complex transmembrane topology, this work characterizes early lipid-driven dimerization using molecular dynamics simulations. Comparing dimer interactions in aqueous solution versus on a lipid membrane model reveal that protein-lipid interactions critically guide inter-protein residue alignment and binding. Hydrophobic contacts and hydrogen bonding between key residues and phosphatidylcholine/phosphatidylinositol lipids drive essential helix interactions that promote p7 oligomerization, particularly involving the first helix. This study demonstrates that membrane lipids are essential, dynamic contributors to protein binding and aggregation in cellular membranes.

Studies have implicated perturbations in the postnatal development of the gut microbiome as a contributing factor to childhood undernutrition. Compared to a standard ready-to-use supplementary food, a microbiome-directed complementary food (MDCF-2) designed to repair these perturbations produced superior improvements in ponderal and linear growth in clinical trials of Bangladeshi children with moderate acute malnutrition. Here, "reverse translation" experiments are performed where intact fecal microbiomes collected from trial participants before and at the end of treatment are introduced into female gnotobiotic mice just after delivery of their pups. Pups received diets designed to resemble those consumed by children in the trials to recreate "unrepaired" and "repaired" gut ecosystems. Analyses of the abundances of bacterial strains (metagenome-assembled genomes), their expressed genes, and metabolic products, combined with assessments of ponderal growth and intestinal epithelial lineage transcriptomes (single-nucleus RNA-Seq with follow-up immunocytochemistry) disclosed effects of MDCF-2 associated microbiome repair that cannot be determined, in part because "no treatment" control arms cannot be ethically incorporated into these trials. Specifically, microbiome repair in these mice produced significant increases in ponderal growth, changes in microbial gene expression consistent with a less virulent gut ecosystem and alterations in expression of i) components of cell junctions in the enterocytic and goblet cell lineages, ii) pathways for synthesis and secretion of eicosanoid immune effectors in chemosensory tuft cells, and iii) goblet cell pathways involved in glycosylation and secretion of mucin. Experiments of the type described can help formulate and test hypotheses about how microbiome repair affects host biology.

Long-range cerebellar outputs are critical for shaping brain-wide functional architecture, influencing motor, cognitive, and affective domains. Proper cerebellar development and its efferent circuits are essential for brain maturation, and disruptions in these pathways have been implicated in neurodevelopmental disorders, such as autism spectrum disorder and schizophrenia. However, despite the early developmental onset, the spatiotemporal dynamics of cerebellar connectivity remain largely unknown, limiting mechanistic understanding of cerebellar contributions to brain development and disease. Here, we combine viral genetic tracing with high-resolution whole-brain 3D imaging to generate a comprehensive spatiotemporal map of cerebellar output development in mice. This systematic characterization uniquely distinguishes how excitatory and inhibitory cerebellar projections establish, expand, and refine their connectivity across the brain. Both axon types reach their principal brain targets within a narrow perinatal window, coinciding with the earliest formation of presynaptic terminals, and subsequently undergo postnatal expansion followed by region- and cell type-specific refinement. These findings define critical windows in the assembly of cerebellar output circuits, providing a framework to decipher the principles of cerebellar circuit formation and their impact on brain-wide function. Importantly, they also pinpoint periods of heightened vulnerability, when genetic or environmental perturbations are most likely to derail cerebellar-driven circuit maturation. By establishing this developmental blueprint, our study not only advances fundamental knowledge of brain wiring but also lays the groundwork for translational efforts to connect early cerebellar dysfunction with the origins of neurodevelopmental disorders.

Clinical in vitro diagnosis is essential in early diagnosis and prognostic assessment, yet antibody-based assays face limitations in multiplexity and scalability. To address the growing demand for modern molecular diagnostics, a multivalent aptamer corona platform for high-throughput is introduced and multiplexed multi-cancer diagnosis. With clinical serum samples, ovarian cancer-, lung cancer-, and colorectal cancer-based aptamer coronas are obtained through several rounds of alternating positive and negative systematic evolution of ligands by exponential enrichment (SELEX). Leveraging a de novo ProteoFish-SELEX strategy, which integrates nanoparticle-protein corona technology, a multivalent aptamer corona is evolved. Proteomic profiling of protein coronas revealed cancer-associated protein signatures, while aptomic profiling of aptamer coronas identified high-affinity cancer-specific aptamers. With these aptamers, high diagnostic accuracy is demonstrated in multiplexed multi-cancer detection using clinical cohorts. Multivalent aptamer corona's outstanding diagnostic performance, multiplexed detection capability, and sequencing-enabled high-throughput potential position it as a promising versatile tool for novel biomarker discovery and a transformative advancement in precision oncology.

Eosinophils accumulate in different tissues in allergies, cancer, and infectious diseases, adopting distinct properties. Herein, we profiled murine esophageal eosinophils during allergic inflammation using single-cell sequencing, epigenomic mapping, and flow cytometry. Esophageal eosinophils displayed an altered chromatin accessibility profile compared to bone marrow eosinophils, with 761 epi-transcriptionally regulated genes enriched in inflammation, immunoregulation, bacterial sensing, angiogenesis, migration, and apoptosis. The local environment entrains the unique esophageal eosinophil immunophenotype, as suggested by eosinophil-esophageal epithelial co-cultures, esophageal eosinophil transcriptional similarities regardless of the upstream cytokines driving their esophageal localization, and transcription factor gene editing altering esophageal eosinophilia and the associated eosinophil and global esophageal transcriptomes. Finally, the epigenomic and transcriptomic properties of murine esophageal eosinophils are largely conserved in humans. Thus, our data indicate that tissue specialization of esophageal eosinophils is entrained by local environmental cues that induce genome-wide epigenetic reprogramming and regulated by discrete transcription factors and provide a public, epigenetic database of murine tissue eosinophils.

Synapses formed by equivalent pairs of pre- and postsynaptic neurons have similar electrophysiological characteristics, belonging to the same type. However, these are generally confined to microscopic brain regions, precluding their proteomic analysis. This fact has greatly limited our ability to investigate the molecular basis of synaptic physiology. We introduce a procedure to characterise the synaptic proteome of microscopic brain regions and explore the molecular diversity among the synapses forming the trisynaptic circuit in the mouse hippocampus. While we observe a remarkable proteomic diversity among these synapses, we also report that proteins involved in the regulation of the function of glutamate receptors are differentially expressed in all of them. Moreover, neuron-specific gene expression programs would contribute to their regulation. Here, we introduce a combined proteomics and transcriptomics analysis uncovering a previously unrecognized neuron-specific control of synaptic proteome diversity, directed towards the regulation of glutamate receptors and their regulatory proteins.

Medulloblastoma (MB) is the most prevalent malignant brain tumor in children, exhibiting clinical and genomic heterogeneity. Of the four major subgroups, Group 3 tumors (MYC-MB), display high levels of MYC and metastasis rates. Despite treatment with surgery, radiation, and chemotherapy, patients with Group 3 MB are more likely to develop aggressive recurrent tumors with poor survival. To examine resistance mechanisms in this study, we perform single nuclei multiome analysis of matched primary and recurrent tumors. Therapy resistant Medulloblastoma demonstrates an expanded persistent progenitor population. Additionally, distinct chromatin landscapes link to altered transcription and correspond with metabolic reprogramming. In vivo modeling of radiation resistance exhibits similar chromatin-based metabolic reprogramming focused on wild-type isocitrate dehydrogenase (IDH1) activity. IDH1 inhibition reverses resistance-mediated chromatin changes and enables radiation re-sensitization. Ultimately, these findings demonstrate the efficacy of single-cell multiome analysis in elucidating resistance mechanisms and identifying targetable pathways for MYC-driven medulloblastoma.

Genomic therapy has emerged as a transformative strategy for the prevention, diagnosis and treatment of a wide array of diseases, including Alzheimer's disease, amyotrophic lateral sclerosis and other CNS-related diseases. Recent developments in chemical strategies and delivery platforms have enhanced the potential of genomic therapies for brain disorders. In this Review, we summarize such strategies, focusing on advances in delivery platforms such as lipid nanoparticles, polymers and oligonucleotide conjugates to facilitate the brain delivery of DNA-based or RNA-based therapeutics into the CNS. We present an overview of the chemical structures and functional moieties of lipids, polymers and oligonucleotides used in these platforms. Lastly, we provide an outlook on future chemical directions to further improve the delivery of genomic medicines to the brain.

Intestinal stem cells (ISCs) drive the rapid regeneration of the gut epithelium. However, during aging, their regenerative capacity wanes, possibly through senescence and chronic inflammation, albeit little is known about how aging-associated dysfunction arises in the intestine. We previously identified the urokinase plasminogen activator receptor (uPAR) as a senescence-associated protein and developed CAR T cells able to efficiently target it. Harnessing them, here, we identify the accumulation of mostly epithelial uPAR-positive cells in the aging gut and uncover their detrimental impact on ISC function in aging. Thus, both therapeutic and prophylactic treatment with anti-uPAR CAR T cells improved barrier function, regenerative capacity, inflammation, mucosal immune function and microbiome composition in aged mice. Overall, these findings reveal the deleterious role of uPAR-positive cells on intestinal aging in vivo and provide proof of concept for the potential of targeted immune-based cell therapies to enhance tissue regeneration in aging organisms.

<h4>Background</h4>Acute coronary syndrome (ACS) is a major cause of morbidity and mortality worldwide. Identifying biomarkers that predict outcomes is essential for guiding management. This study evaluated whether elevated serum uric acid (SUA) is associated with increased risks of major adverse cardiovascular events (MACE), recurrent myocardial infarction (re-MI), and all-cause mortality (ACM) in patients with ACS.<h4>Methods</h4>This retrospective cohort study enrolled 829 inpatients with ACS admitted to a tertiary referral hospital in Taiwan from 2015 to 2019. Patients were divided into normal (< 7.25 mg/dL, n = 566) and high (≥ 7.25 mg/dL, n = 263) SUA groups based on a receiver operating characteristic-derived cutoff for whole cohort. All patients received standard ACS care, and SUA levels were retrospectively analyzed. The primary outcome was MACE, defined as ACM, re-MI, and target lesion/vessel revascularization (TLR/TVR), assessed up to 60 months. Kaplan-Meier survival analysis, logistic regression, and Cox proportional hazards regression were applied.<h4>Results</h4>The overall incidences of MACE (19.54%), re-MI (2.90%), and ACM (4.46%) were higher in the high SUA group compared with the normal SUA group (MACE: 26.62% vs. 16.25%, p = 0.0005; re-MI: 6.08% vs. 1.41%, p = 0.0002; ACM: 7.22% vs. 3.18%, p = 0.0087). No significant difference was observed in TLR/TVR (overall 11.94%; 11.48% vs. 12.93%, p = 0.5508). Kaplan-Meier analysis at 60 months demonstrated higher event-free rates for MACE, re-MI, and ACM in the normal SUA group (log-rank p = 0.0117, 0.0006, and 0.0261, respectively). Multivariable logistic regression showed that SUA ≥ 7.25 mg/dL was associated with increased odds of MACE (odds ratio = 1.639, 95% confidence interval [CI] = 1.084-2.477, p = 0.0191). Cox regression revealed higher hazards of MACE (hazard ratio [HR] = 1.399, 95% CI = 1.024-1.191, p = 0.0350), re-MI (HR = 3.758, 95% CI = 1.605-8.799, p = 0.0023), and ACM (HR = 1.956, 95% CI = 1.019-3.753, p = 0.0438) in the high SUA group after adjustment for age, uremia, use of drug-eluting stent, and number of diseased vessels.<h4>Conclusions</h4>Elevated SUA is an independent prognostic marker for increased risks of MACE, re-MI, and ACM in patients with ACS. SUA assessment can enhance risk stratification, helping to identify higher-risk patients who may benefit from more intensive secondary prevention strategies and closer follow-up.

<h4>Context</h4>Congenital hypogonadotropic hypogonadism (CHH) is a rare disorder caused by deficient secretion or action of gonadotropin-releasing hormone. While its characteristics are well-documented in adults, data from prepubertal patients remain limited.<h4>Objective</h4>To investigate the clinical, hormonal, and genetic characteristics of CHH in male patients aged < 18 years and assess age-related changes in testicular function.<h4>Design</h4>Retrospective analysis of data from patients with CHH.<h4>Setting</h4>Tertiary pediatric endocrine referral center.<h4>Patients</h4>Overall, 121 male patients with CHH, aged 0-18 years, were included.<h4>Main outcome measures</h4>Hormonal profiles, genetic variants, and testicular function indicators across different age groups.<h4>Results</h4>All patients had micropenis, and 41.3% had cryptorchidism. The > 14-year group had fewer combined cases of both conditions but more isolated micropenis (p = 0.001). Inhibin B, luteinizing hormone, follicle-stimulating hormone, and post-human chorionic gonadotropin testosterone levels were significantly higher in the ≤ 3-year group (p < 0.05). Leydig and Sertoli cell function declined with age. Inhibin B < 33.895 pg/mL and anti-Müllerian hormone (AMH) < 17.545 ng/mL predicted Leydig cell dysfunction with sensitivities of 78.5% and 85.7% and specificities of 82.3% and 73.8%, respectively. Pathogenic variants were identified in 84.6% of cases, with fibroblast growth factor receptor 1, chromodomain helicase DNA-binding protein 7, and prokineticin receptor 2 being the most frequently impacted.<h4>Conclusions</h4>CHH should be suspected in boys with micropenis and cryptorchidism. AMH and inhibin B are key markers for early detection of Leydig cell dysfunction, with genetic testing being essential for diagnosis.

Persistent, unresolved inflammatory processes disrupt the complex homeostatic state and lead to severe disease, ultimately resulting in organ failure, shock, and death. The mediators that monitor and control the programmed mechanisms in tissue homeostasis and repair remain elusive. A recently discovered group of endogenous proteins, termed neuronal guidance proteins (NGPs), has gained attention as important mediators of immunoregulation and tissue homeostasis through their role in the complex interplay between metabolic reprogramming and immunity. In murine peritonitis, the deficiency of the NGP Plexin C1 (PLXC1−/−) led to an increase in neutrophil and Ly6Chi monocyte recruitment to the injury site and a decrease in the phagocytosis rate. This was accompanied by a decrease in the endogenous biosynthesis of specialized proresolving lipid mediators (SPMs) such as Maresin-1 and Protectin DX and finally by a lengthening of the resolution interval. Peritoneal macrophages (MΦs)PLXC1−/− followed specific strategies to adapt their metabolism to catabolic conditions, by reducing fatty acid oxidation and oxidative phosphorylation, increasing aerobic glycolysis and activation of the pentose phosphate pathway, and disrupting the tricarboxylic acid cycle. The decreased MΦsPLXC1−/− citrate levels corresponded with a lipid mediator profile in which PGD2 and PGE2 were greatly decreased in murine PLXC1−/− peritoneal exudates, indicating a lesser effect on lipid mediator class switching and ultimately the formation of SPMs. Analysis of the related signaling networks indicated mTOR and AKT2 phosphorylation pathways are critical determinants of metabolic reprogramming in activated MΦsPLXC1−/−. These findings highlight a novel role for PLXC1 in regulating the crosstalk between immunometabolism and inflammation resolution programs in severe inflammation.

<h4>Background</h4>Non-transfusion-dependent beta-thalassemia (β-NTDT) is characterized by ineffective erythropoiesis, increased intestinal iron absorption, and iron overload. The ferroportin inhibitor, vamifeport, has been shown to improve erythropoiesis via decreases in serum iron and transferrin saturation levels in preclinical models and healthy volunteer studies.<h4>Objective</h4>The objective of this 12-week, double-blind, randomized, placebo-controlled, phase 2a study was to assess the safety and tolerability of vamifeport versus placebo in adults with β-NTDT (primary endpoint). Iron-related pharmacodynamic effects (preliminary efficacy) were also assessed as a secondary endpoint.<h4>Methods</h4>Randomized, adult patients weighing 40-59 kg and 60-100 kg received vamifeport 60 mg and 120 mg (once [QD] or twice [BID] daily), respectively, for 12 weeks. Non-transfusion-dependent thalassemia was defined as transfusion requirements < 5 units of red blood cells during the 24 weeks before randomization.<h4>Results</h4>Twenty-five patients were included (vamifeport QD n = 9, BID n = 12; placebo n = 4); 64% were male and 56% weighed < 60 kg. Baseline serum iron and transferrin saturation levels were similar across treatment groups. All treatment-emergent adverse events were mild/moderate, and rates were similar across groups (vamifeport QD 67%, BID 58%; placebo 75%). There were no deaths or serious treatment-emergent adverse events and no clinically relevant changes in safety parameters. Serum iron and transferrin saturation levels decreased by 2 h after the first vamifeport dose (mean [standard deviation] decreased QD - 12.2 [6.5], BID - 14.5 [12.1] µmol/L and QD - 33.6 [18.9], BID - 37.2 [27.6] %, respectively) and remained below baseline levels at each subsequent visit. There were no clinically meaningful changes in the placebo group.<h4>Conclusion</h4>In this 12-week study, vamifeport had a favorable safety/tolerability profile, with no changes in hemoglobin levels ≥ 1.0 g/dL, and promising pharmacodynamic effects versus placebo in adults with β-NTDT.<h4>Trial registration</h4>ClinicalTrials.gov, NCT04364269. Registered 01 April 2020; Prospectively registered, https://clinicaltrials.gov/study/NCT04364269?term=NCT04364269&rank=1 .

<h4>Background</h4>Helicobacter pylori (H. pylori) infection is a major etiological factor for stomach adenocarcinoma (STAD), yet the key molecular drivers linking infection to tumor progression remain insufficiently defined. This study aimed to identify H. pylori-related hub genes in STAD and validate their functional relevance using integrated bioinformatics and experimental approaches.<h4>Methodology</h4>Differentially expressed genes (DEGs) were identified from two microarray datasets (GSE13911 and GSE54129) comparing H. pylori-positive STAD samples with controls. Common DEGs were used to construct a protein-protein interaction (PPI) network via STRING and Cytoscape, and hub genes were ranked using CytoHubba. Transcriptomic validation was conducted using TCGA-STAD data, followed by analyses of enrichment pathways, promoter methylation, somatic mutations, CNVs, immune subtype associations, and drug sensitivity using GSCA, UALCAN, cBioPortal, and CTRP datasets. miRNA-mRNA regulatory interactions were predicted using miRNet and validated in vitro. Experimental validation included RT-qPCR, Western blotting, CCK-8 proliferation assays, colony formation, and wound-healing assays in MKN45 and AGS cells following siRNA-mediated knockdown of key hub genes. Additionally, AGS cells were infected with live H. pylori to directly assess infection-induced changes in gene expression and malignant phenotypes.<h4>Results</h4>Four hub genes (THBS2, CTNNB1, COL4A1, and E2F3) were identified as commonly upregulated in H. pylori-positive STAD samples and were further validated as highly expressed in STAD tissues and cell lines. Promoter hypomethylation and CNV gains contributed to their overexpression. Pathway analyses linked the hub genes to EMT, cell cycle progression, immune suppression, and oncogenic signaling. miRNA profiling identified hsa-miR-9-3p and hsa-miR-9-5p as common regulators with diagnostic potential. Importantly, H. pylori infection of AGS cells induced strong upregulation of COL4A1 and CTNNB1 and significantly increased proliferation, clonogenicity, and migration, demonstrating a direct infection-driven oncogenic response. Conversely, siRNA-mediated silencing of COL4A1 or CTNNB1 markedly reduced proliferation, colony formation, and wound closure, confirming their functional roles in STAD progression. Immune correlation and drug sensitivity analyses further linked high hub-gene expression to immunosuppressive microenvironments and resistance to multiple therapeutic agents.<h4>Conclusion</h4>This study identifies THBS2, CTNNB1, COL4A1, and E2F3 as key H. pylori-associated oncogenic drivers in STAD. Functional assays demonstrate that H. pylori enhance malignant phenotypes through COL4A1 and CTNNB1, while gene silencing reverses these effects. These findings highlight the hub genes and their regulatory miRNAs as promising diagnostic biomarkers and potential therapeutic targets in H. pylori-related gastric cancer.

Inflammasomes are being increasingly recognized for their critical roles in regulating immune cell infiltration, antitumor immunity, and tumorigenesis in cancer biology. The expression and activation of inflammasomes, which vary according to the tumor type and stage, influence both tumor progression and elimination through pyroptosis. A deeper understanding of the complex functions and regulatory mechanisms of inflammasomes and pyroptosis in diverse aspects of tumor biology is crucial for the development of targeted therapies. In this review, we extensively explore the latest research on various types of inflammasomes, including NOD-like receptor family pyrin domain-containing 3 protein, NOD-like receptor family caspase recruitment domain-containing protein 4, and absent in melanoma 2, and different types of pyroptosis, highlighting their beneficial and deleterious effects on various cancers. We further explore inflammasome-associated immune dynamics, which can drive either tumor suppression or promotion depending on the microenvironmental context. Finally, we discuss emerging strategies to harness inflammasomes for cancer therapy, along with ongoing clinical efforts to develop inflammasome-targeted interventions as adjuncts to conventional antitumor treatments, offering new perspectives for cancer prevention and therapy.

<h4>Background</h4>Several maternal exposures such as acetaminophen during pregnancy have been previously associated with altered placental CpG methylation. Epigenetic gestational age measures biological aging using DNA methylation from gestational tissues such as placenta and cord blood. We hypothesize that placental epigenetic gestational age (eGA) could serve as a biomarker for maternal exposures or pathological processes that influence placental development. To investigate relationships between maternal exposures and placental eGA, we evaluated prenatal medication use (antibiotics, acetaminophen, aspirin, and ibuprofen) in relation to placental epigenetic gestational age acceleration (eGAA) using the Robust Placental Clock (RPC). We also examined associations between these four exposures and the methylation levels of the 558 individual CpGs comprising the RPC. Using data from the Extremely Low Gestational Age Newborns (ELGAN) study (N = 408), we ran linear mixed-effects regression models that accounted for multiple births to the same mother. We further stratified by infant sex assigned at birth to assess effect measure modification.<h4>Results</h4>We observed a positive association between prenatal aspirin use and eGAA at borderline significance (β: + 0.35 weeks gestation; 95% CI: - 0.01, 0.72). When stratified by sex, this finding was significant among females (β: + 0.63 weeks gestation; 95% CI: 0.10, 1.17) but not males (β: - 0.05 weeks gestation; 95% CI: - 0.60, 0.49). Prenatal aspirin use in female pregnancies was further associated with ten differentially methylated RPC CpGs, mapping to genes including Retinoid X Receptor Alpha (RXRA), a nuclear receptor that can function as transcription factor and thereby influence multiple processes critical to placental development.<h4>Conclusion</h4>These findings highlight that prenatal aspirin use is associated with placental eGAA in a sex-dependent manner, and RPC CpG methylation is sensitive to maternal medication exposures. Additional research is required to validate these eGAA observations, and future research should investigate the mechanisms underlying these relationships and their potential long-term clinical implications in the infant.

Advancements in the rapidity, cost efficacy, and sensitivity of next-generation sequencing (NGS) have facilitated molecular risk stratification and precision medicine-based treatment for pediatric leukemia. The benefit of uniform cytomolecular analyses for clinical trial risk assignment is clear. However, the clinical impact of comprehensive NGS for pediatric leukemias at an institutional level is not well described. We report the genomic spectrum of one of the largest cohorts of pediatric and adolescent/young adult (AYA) acute leukemias examined to date (<i>n</i> = 1442) via institutional NGS testing from our large tertiary care center. We evaluated the clinical utility of genomic results for informing prognosis and treatment. We identified high utility of the comprehensive DNA-based mutational panel and RNA-fusion panel, which detected leukemia-associated variants in 99% of specimens. We observed 65% of B-cell acute lymphoblastic leukemia cases and 69% of patients with acute myeloid leukemia harbored a prognostic molecular alteration. In total, 325 of 1134 patients (29%) harbored potentially targetable molecular biomarkers, and 23% of cases with follow-up data received precision medicine-based therapy. Paired diagnostic and relapsed leukemia samples aided in differentiation between treatment-related leukemia versus lineage drift/switch. These findings demonstrate the broad utility of comprehensive molecular sequencing for pediatric/AYA leukemia at an institutional level to improve outcomes.

The female obesity epidemic remains a major global public health challenge, with limited effective treatments available. Daily dietary management serves as an effective strategy for obesity prevention. As a promising functional food, quinoa has been demonstrated to possess anti-obesity properties. However, the mechanisms of quinoa on high-fat diet (HFD)-induced female obesity and its role in regulating adipose tissue remain unclear. In the present study, the effects of different dosages of quinoa on obesity and its underlying mechanisms were investigated in an HFD-induced mouse model of obesity. Six-week-old female C57BL/6J mice were fed an HFD to induce obesity, followed by an 8-week intervention with low dosage of quinoa (LQ), medium dosage of quinoa (MQ), and high dosage of quinoa (HQ), respectively. Then, 16S rRNA gene sequencing was performed to investigate gut microbial community and transcriptome analysis was conducted to characterize differentially expressed genes (DEGs) in gonadal white adipose tissue (gWAT) in mice. The HQ group significantly attenuated body weight in HFD-fed mice. The 16S rRNA gene sequencing analysis revealed that the three intervention groups exhibited distinct bacterial community profiles, and the bacterial taxa significantly enriched in response to quinoa intake were identified. Notably, the HQ group modulated gut microbiota composition, demonstrating significant regulatory effects on the populations of Bacteroidota, Bacteroidia, Bacteroidales, and Muribaculaceae, while concurrently reducing the Firmicutes-to-Bacteroidetes (F/B) ratio. Furthermore, transcriptomic analysis revealed that HQ treatment modulated HFD-induced DEGs associated with pathways of "transmembrane transporter complex," "ion gated channel activity," and "calcium ion binding" in gWAT. These findings suggest that quinoa alleviates HFD-induced obesity by regulating community structure of gut microbiota and the ion channel in gWAT, providing a strong theoretical foundation for the development of quinoa products for the treatment of female obesity.

<i>Fagonia arabica</i> "Dhamasa" is traditionally used in Ayurvedic and Unani medicine across South Asia and the Middle East for its therapeutic properties, notably as a natural blood purifier. It is believed to aid in dissolving blood clots and reducing the risk of brain hemorrhage and cardiovascular events. Given previous reports of saponin content in <i>F. arabica</i>, this study aimed to evaluate its thrombolytic and anticoagulant potential. After confirming its saponin richness through NMR (1D, 2D) and LC-MS, we developed liposomal nanoparticles to enhance bioavailability and target clot dissolution more effectively. Liposomes were characterized using TEM, particle size, PDI, and zeta potential. <i>In vitro</i> thrombolytic and anticoagulant activities were assessed, followed by <i>in vivo</i> testing in adult Wistar rats. The dose-dependent effects of the saponin-rich <i>n</i>-butanol fraction of <i>F. arabica</i> on Antithrombin-III levels were evaluated using an <i>in vitro</i> quantitative immunoturbidimetric assay. Coagulation parameters were evaluated using aPTT and PT assays. A molecular docking study was systematically performed on nine targets using the five structurally elucidated steroidal and terpenoid glycosides to predict and evaluate their binding affinities. These computational findings were designed to complement ongoing <i>in vitro</i> and <i>in vivo</i> investigations. The butanol extract showed significant <i>in vitro</i> thrombolytic and anticoagulant effects at 40 mg mL^-1. The results demonstrate that the saponin-rich fraction modulates Antithrombin-III in a concentration-dependent manner. Liposomal nanoparticles achieved similar efficacy <i>in vivo</i> at just 3 mg kg^-1, compared to 100 mg kg^-1 for the non-nano extract. The liposomal group showed PT of 35 ± 4 s and aPTT >180 s, outperforming the non-nano extract (PT 30 ± 7 s, aPTT >180 s) and controls (PT 12 ± 2 s, aPTT 37 ± 3 s). The docking study provides mechanistic insight into their potential multi-target antithrombotic activities. Besides the butanol extract, liposomal <i>F. arabica</i> enhances antithrombotic efficacy at lower doses, supporting its potential as a natural therapeutic candidate for thrombotic disorders.

A 15-year-old boy presented with pink proliferative plaque over both lips. The child was born to a non-consanguineous couple, at full term, and uneventful antenatal, perinatal history. Examination revealed a cobblestone appearance over tongue with papilliferous projections in both labial mucosa with slate-grey pigmentation. The child had multiple lipomatosis and thyroid swelling. Father had thyroid swelling with multiple epidermoid cysts. A final diagnosis of Cowden syndrome. Genetic analysis could not be performed due to financial constraints. Oral mucosal cobblestone appearance is seen in Darier disease, Cowden's syndrome, lipoid proteinosis, granulomatous cheilitis, mucosal neuroma, Crohn's disease, and mucous membrane plasmacytosis. Diffuse oral mucosal melanosis is described in Peutz-Jeghers syndrome, Laugier-Hunziker-Baran syndrome, Cowden syndrome, and Addison's disease. We report this rare genodermatosis, which presents as oral papillomatosis. Early recognition of Cowden syndrome is crucial, as timely diagnosis enables appropriate cancer surveillance, genetic counseling, and preventive interventions, thereby reducing morbidity and improving long-term outcomes.

A recent explosion in genomic testing has led to the identification of several genetic disorders that mimic CNS-specific autoimmune disorders. Such monogenic disorders, although rare, represent a diagnostic challenge because of their diverse phenotypes and overlapping features. Early recognition of these disorders is crucial not only to prevent overtreatment with immunotherapy but also to ensure that targeted treatments are available for many of these disorders. This review explores some of the monogenic disorders that can masquerade as neuroinflammatory phenotypes. These clinical vignettes are stratified according to neuroanatomical localization along the neuroaxis: supratentorial white matter, gray matter, brainstem, and spinal cord involvement. Through these cases, we discuss how clinical, laboratory, and neuroimaging red flags, such as early onset, relentless progression despite immunotherapy, and lack of CSF markers of inflammation, can guide specific diagnostic workup. In the next section, we highlight the approach to genetic testing in identifying monogenic mimickers. Finally, we discuss a selected list of currently available and emerging therapeutic strategies for some of these disorders. These include JAK inhibitors for Aicardi-Goutières syndrome, anti-TNF therapy for adenosine deaminase 2 deficiency (DADA2), and gene replacement therapy for X-linked adrenoleukodystrophy. By providing a comprehensive and systematic clinical approach, this review aims to equip neurologists with a framework to navigate diagnostic evaluations for such monogenic disorders.

Aging of the gut involves progressive changes in structure, function, and microbial composition, which impact overall health. The vermiform appendix extends from the apex of the cecum; it contains gut-associated lymphoid tissue and serves as a reservoir of gut microbiota. This study investigates histologic and gene expression changes in 20 morphologically normal appendiceal samples obtained from pediatric (<i>n</i> = 5), adult (<i>n</i> = 8), and geriatric (<i>n</i> = 7) patients. Histologic analysis revealed a higher prevalence of lymphoid follicles reduction and the presence of fibrous obliteration of the appendiceal tip in aged samples. RNA sequencing identified 1004 differentially expressed genes (385 upregulated and 619 downregulated; <i>p</i> < 0.05) between the adult and geriatric population. Upregulated pathways were enriched for oxidative stress response, cholesterol metabolism, and mucosal barrier maintenance, including NRF2 targets (NQO1, MGST1), suggesting enhanced antioxidant activity. Downregulated genes were associated with synaptic signaling, ion channel regulation, and neuronal adhesion (e.g., GRIA2, RET, NOS1, NCAM2, CNTN1), reflecting age-related decline in enteric neuronal integrity. Across all age groups, 25 protein-coding genes showed progressive expression shifts with aging, including upregulation of CLDN2, MUC2, and GDF15, and downregulation of NOG and NELL2, indicating barrier loosening, chronic inflammation, and reduced regenerative potential. These findings suggest that aging of the vermiform appendix recapitulates key processes of intestinal aging, including oxidative stress, inflammaging, and neuronal loss, supporting its potential use as a model tissue for studying gut aging mechanisms.

Neurodegenerative and cardiovascular disorders share multifactorial origins, including oxidative stress, (neuro)inflammation, and lipid dysregulation-factors often addressed independently by single-target therapies. In this study, we report a rational multitarget approach through the design and synthesis of novel (benzo)thiazine derivatives that integrate antioxidant, anti-inflammatory, and antihyperlipidemic functionalities within a single molecular framework. The compounds were obtained in good yields via 3-7 step synthetic routes and evaluated through complementary in vitro and in vivo assays. Several derivatives displayed potent inhibition of lipoxygenase (IC₅₀ < 100 μM), significant reduction in carrageenan-induced edema (up to 60%), strong free radical scavenging and lipid peroxidation inhibition, as well as effective iron chelation. In vivo, most derivatives enhanced total antioxidant capacity (by 50-800%) and significantly improved plasma lipid profiles in mouse, while almost all compounds increased the plasma antiatherogenic index by more than 100% with selected compounds exceeding 600%. Notably, several molecules also showed moderate acetylcholinesterase inhibition, suggesting preliminary neuroprotective potential. Altogether, these multifunctional (benzo)thiazine derivatives represent promising lead structures for the development of agents targeting the complex interplay of oxidative, inflammatory, and metabolic pathways underlying neurodegenerative and cardiovascular diseases.

Bacteriocins offer a promising solution to antibiotic resistance, possessing the ability to target a wide range of bacteria with precision. Thus, there is an urgent need for a computational model to predict new bacteriocins and aid in drug development. This work centers on constructing web-based predictive models using the XGBoost machine learning algorithm, based on the physicochemical properties, structural characteristics, and sequence profiles of protein sequences. We employed correlation analyses, cross-validation, and hypergraph-based techniques to select features. Cross-validated feature selection (CVFS) partitions the dataset, selects features within each partition, and identifies common features, ensuring representativeness. On the contrary, hypergraph-based feature evaluation (HFE) focuses on minimizing hypergraph cut conductance, leveraging higher-order data relationships to precisely utilize information regarding feature and sample correlations. The XGBoost models were built using the selected features obtained from these two feature evaluation methods. We also analyzed the feature contributions directly from the best model using SHapley Additive exPlanations (SHAP). Our HFE-based approach achieved 99.11% accuracy and an AUC of 0.9974 on the test data, overall outperforming the CVFS-based feature evaluation method and yielding results comparable to existing approaches. The most influential features are related to solvent accessibility for buried residues, followed by the composition of cysteine. Our web application, accessible at https://shiny.tricities.wsu.edu/bacteriocin-prediction/, offers prediction results, probability scores, and SHAP plots using both cross-validation- and hypergraph-based methods, along with previously implemented approaches for feature selection.

<h4>Introduction</h4>Early prognostication in non-muscle-invasive bladder cancer (NMIBC) is essential for optimizing therapy and follow-up. Epigenetic mechanisms, particularly DNA methylation, have emerged as promising biomarkers for predicting disease outcome.<h4>Materials and methods</h4>A systematic review and meta-analysis were conducted according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines to evaluate the prognostic significance of promoter DNA methylation in NMIBC. Comprehensive searches of PubMed, Web of Science, Embase, MEDLINE, and the Cochrane Library (January 2010-October 2022) identified eligible studies. The Newcastle-Ottawa scale was used for quality assessment, and pooled hazard ratios with 95% confidence intervals were calculated using random-effects models.<h4>Results</h4>Eleven studies with 3,065 NMIBC patients were analyzed. Promoter methylation was significantly associated with poor progression-free survival (pooled hazard ratios (HR) = 2.88; 95% CI = 2.03-4.09; p < 0.0001) and recurrence-free survival (pooled HR = 2.65; 95% CI = 1.93-3.63; p < 0.0001). Although overall survival showed pathway-specific variation (pooled HR = 0.96; 95% CI = 0.36-2.60; p = 0.94), methylation of adhesion and apoptosis-related genes exhibited the strongest associations. Subgroup analyses revealed a greater prognostic impact in Asian cohorts (p < 0.0001), suggesting regional differences in epigenetic susceptibility.<h4>Conclusion</h4>Promoter DNA methylation constitutes a robust prognostic biomarker for recurrence and progression in NMIBC, with stronger effects in Asian populations. Standardization of validated gene panels, assay thresholds, and cross-regional prospective validation will be essential for clinical translation. Integrating methylation-based classifiers into risk stratification models could improve individualized management and long-term outcomes in NMIBC.

<h4>Objective</h4>We aimed to investigate the impact of delayed cord clamping (DCC) on the levels of blood mesenchymal (MSCs), hemopoietic progenitor (HPCs), and immune cells in very preterm neonates.<h4>Methods</h4>We prospectively examined 21 neonates with a median gestational age of 32 weeks (interquartile range, IQR 29-32) who had DCC, and 19 neonates with a median gestational age of 31 weeks (IQR 30-32) who had immediate cord clamping (ICC). We measured the levels of MSCs, HPCs, very small embryonic-like stem cells (VSELs), early endothelial progenitor cells (EPCs), late EPCs, and the immunophenotype in the first, 10th, and 30th day of life, and associated them with late-onset sepsis and bronchopulmonary dysplasia (BPD).<h4>Results</h4>DCC-neonates compared to ICC-neonates had significantly higher values of MSCs (846 vs. 316 cells per million cytometric events, <i>p</i> = 0.003), HPCs (191 vs. 115 cells per million cytometric events, <i>p</i> = 0.034), and lower values of VSELs (21 vs. 37 cells per million cytometric events, <i>p</i> = 0.044) and late EPCs (7 vs. 19 cells per million cytometric events, <i>p</i> = 0.017) at birth. Neonates with late-onset sepsis, in comparison to neonates with no sepsis, had significantly higher values of early (20 vs. 0.3 cells per million cytometric events, <i>p</i> = 0.011) and late EPCs (32 vs. 8 cells per million cytometric events, <i>p</i> = 0.033). In addition, neonates with BPD had significantly higher values of late EPCs compared to neonates without BPD (27 vs. 8 cells per million cytometric events, <i>p</i> = 0.041). DCC, adjusted for gestational age and birth weight, was significantly associated with higher levels of MSCs, HPCs, and lower levels of VSELs and late EPCs.<h4>Conclusions</h4>In very preterm neonates with DCC, MSCs and HPCs are higher, while VSELs and late EPCs are lower in the umbilical cord blood, compared to neonates with ICC. Early and late EPCs were associated with late-onset sepsis and BPD. Further studies are warranted to explore the association of these findings with the long-term clinical outcomes.

<h4>Introduction</h4>Genetic polymorphisms in CYP2D6 and CYP2C19 significantly influence the metabolism, efficacy, and safety of antidepressant medications. Limited data exist on the prevalence of these actionable pharmacogenetic variants in the Central Indian population. This study aimed to determine the frequency of clinically actionable Tier-1 alleles, genotypes, and metabolizer phenotypes and to evaluate their clinical relevance in patients with common mental disorders (CMDs).<h4>Methods</h4>A total of 509 adults diagnosed with depression and anxiety disorders and receiving SSRI & SNRI antidepressant therapy were enrolled from the Department of Psychiatry, AIIMS Bhopal. Genotyping was performed using the KASP qPCR assay, and CYP2D6 copy number variations (CNVs) were determined using the TaqMan qPCR assay.<h4>Results</h4>Among the Central Indian cohort, the most frequent CYP2D6 alleles were *10 (21.6%), *41 (17.3%), and *4 (10.4%), while *3 (5.7%), *6 (1.9%). CNVs, Gene deletions(*5) and Gene duplications(xN) were detected in 4.2% and 4.1% of the cohort. For CYP2C19, the *2 (37.3%), *3 (2.3%), and *17 (16.1%) alleles were observed. Non-normal metabolizer phenotypes were present in 46.2% for CYP2D6 and 74.2% for CYP2C19; CYP2D6 ultra-rapid metabolizers accounted for 5.3%. Overall, 86% of participants had at least one clinically actionable pharmacogenetic phenotype. Overall, 7.5% of patients carried CYP2D6 variants and 20.6% carried CYP2C19 variants, for which CPIC guidelines recommend alternative drug selection or dose modification.<h4>Discussion</h4>This study demonstrates a high prevalence of actionable CYP2D6 and CYP2C19 variants in the Central Indian population, underscoring the need for pharmacogenetic integration in psychiatric prescribing in Indian clinical settings, to enhance treatment efficacy and minimize adverse events.

No abstract available.

Palpitations are a frequent symptom in primary care, with most cases explained by benign etiologies; however, they may represent the initial manifestation of potentially fatal conditions. Psychiatric comorbidities should not be underestimated, as they may coexist with cardiac causes. The diagnostic approach is primarily clinical, requiring a systematic evaluation of history, physical examination, and a 12-lead electrocardiogram (ECG). Ambulatory ECG monitoring is indicated when clinical evaluation suggests arrhythmia or in patients with structural heart disease.

The development of intermuscular bones (IBs) in teleost fish is regulated by osteogenic genes, but the upstream signals that initiate ossification remain elusive. Given the spatiotemporal correlation between IB formation and swimming behavior, mechanical stimuli from muscle activity have been hypothesized as potential regulators. In this study, we investigated the role of the muscle-specific microRNA miR499 in this process. Using CRISPR/Cas9-generated miR499 knockout zebrafish, we demonstrated that the mutant of miR499 induces a fast-twitch muscle phenotype through the upregulation of <i>sox6</i>, accompanied by increased fast-myosin gene expression and fast-twitch fiber proliferation. This shift led to distinct locomotor alterations, characterized by reduced tail-beat frequency and increased stride length. Crucially, these changes resulted in delayed IB ossification onset, slower ossification rates, and simplified morphology. The total ossification area decreased by 27%. Molecular analysis revealed a downregulation of osteogenic genes in IB tissue. Our findings establish miR499 as a key regulator of a muscle-bone functional axis, where miRNA-mediated muscle fiber specification determines mechanical output that subsequently guides IB development. This work provides new insights into the mechanobiological regulation of skeletal development and identifies a promising genetic target for aquaculture breeding programs aimed at attenuated IBs.

Aging and accumulation of α-synuclein (α-syn) oligomers in the brain are indisputably linked to Parkinson's disease (PD). Using an inducible α-syn oligomer mouse model, we demonstrate that the induction of PD-associated α-syn oligomers for the same time span caused PD-associated symptoms only in aged, but not in young mice. Biochemical studies revealed that α-syn oligomer formation precedes motor decline, with age and α-syn expression jointly determining the motor phenotype. Single-nucleus RNA sequencing (snRNA-seq) identified a PD-related transcriptional signature in basal ganglia neurons (BGNs), which overlapped in part with aging-associated changes. Short-term pharmacological inhibition of the small RhoGTPase CDC42 in aged, symptomatic animals improved motor function without reducing oligomer levels. These findings indicate that aging processes strongly influence the susceptibility to PD-like symptoms and that targeting age-related pathways, rather than α-syn oligomer burden alone, may provide effective strategies to improve outcomes in PD.

Radiotherapy often causes severe and irreversible neural damage, including cognitive impairment and depression-like behaviors. Current mitigants are limited, with single-target molecules being ineffective and nanomedicines posing complexity and toxicity risks. Dragon's Blood (DB), a nontoxic, brown-red resin extracted from <i>Dracaena cochinchinensis</i> (Lour.) (S. C. Chen, China), possesses diverse pharmacological properties. Extensive studies demonstrated that the compounds in DB exhibit multiple therapeutic effects, including cardiovascular protection, promotion of blood circulation, and anti-inflammatory effect. Herein, DB's neuronal radiation mitigation effect and mechanism were investigated. In a whole-brain irradiation rat model, DB administration significantly alleviated radiation-induced anhedonia-like behavior, normalized calcium dyshomeostasis, restored mitochondrial membrane potential, mitigated dendritic spine loss, suppressed neuroinflammation (IL-1β and TNF-α), and preserved hippocampal cytoarchitecture. Brain tissue proteomics revealed 23 DB-modulated KEGG pathways, encompassing the glutamatergic/GABAergic synapse, synaptic plasticity, addiction-related pathways, calcium/cAMP signaling, and hormonal regulation. Ensemble analysis integrating proteomics, WGCNA, machine learning, and PPI pinpointed 24 DB radiation mitigation-related proteins. Among these, eight targets (Grin1, Gabra4, Grm2, Grm3, Grm7, Prkcb, Shank3, and Pak7) functioning via ligand-target interactions were dysregulated by radiation and restored by DB. Molecular docking identified three DB ingredients (socotrin-4'-ol, cinnabarone, and 2'-methoxysocotrin-5'-ol) interacted with all eight targets. Plasma proteomics further revealed radiation mitigation-related brain-enriched proteins (Mib1, Gucy1b1, Fkbp1a, Synj1, and Clasp2). PPI between these 5 plasma proteins and 24 brain proteins reveals DB's multitarget radiation mitigation effect on neurotransmission and synaptic regulation, neuroplasticity, and signaling transduction and cellular response. This work nominated DB and its key constituents as promising candidates for mitigating radiotherapy-induced neural injury.

No abstract available.

The degradation of aggregation-prone tau is regulated by the ubiquitin-proteasome system (UPS) and autophagy, which are impaired in Alzheimer's disease (AD) and related dementias (ADRD), causing tau aggregation. Protein ubiquitination, with its linkage specificity determines the fate of proteins, which can be either protein degradative or stabilizing signals. While the linear M1-linked ubiquitination on protein aggregates serves as a signaling hub that recruits various ubiquitin-binding proteins for the coordinated actions of protein aggregate turnover and inflammatory NF-κB activation, the deubiquitinase OTULIN counteracts the M1-linked ubiquitin signaling. However, the exact role of OTULIN in neurons and tau aggregates clearance in AD are unknown. Based on our quantitative bulk RNA sequence analysis of human inducible pluripotent stem cell (iPSC)-derived neurons (iPSNs) from an individual with late-onset sporadic AD (sAD2.1), a downregulation of the ubiquitin ligase activating factors (MAGE-A2/A2B/H1) and OTULIN long non-coding RNA (OTULIN lncRNA) was observed compared to healthy control WTC11 iPSNs. The down-regulated OTULIN lncRNA is concurrently associated with increased levels of OTULIN protein and phosphorylated tau at p-S202/p-T205 (AT8), p-T231 (AT180), and p-S396/p-S404 (PHF-1) in sAD2.1 iPSNs. Inhibiting the deubiquitinase activity of OTULIN with a small molecule, UC495 reduced the phosphorylated tau in iPSNs and SH-SY5Y cells, whereas the CRISPR-Cas9-mediated <i>OTULIN</i> gene knockout in sAD2.1 iPSNs decreased both the total and phosphorylated tau levels. CRISPR-Cas9-mediated <i>OTULIN</i> knockout in SH-SY5Y resulted in a complete loss of tau at both mRNA and protein levels, and increased levels of polyubiquitinated proteins, which are being degraded by the proteasome as confirmed with an inhibitor, Lactacystin. In addition, SH-SY5Y <i>OTULIN</i> ^KO cells showed downregulation of various genes associated with inflammation, autophagy, ubiquitin-proteasome system, and the linear ubiquitin assembly complex (LUBAC) that consequently may prevent development of an autoinflammation in the absence of <i>OTULIN</i> gene in neurons. Together, our results suggest, for the first time, a non-canonical role for OTULIN in regulating the gene expression and RNA metabolism, which may have a significant pathogenic role in exacerbating tau aggregation in neurons. Thus, OTULIN could be a novel potential therapeutic target for AD and ADRD.

Copper is an essential cofactor for neuronal metabolism, enzymatic functions, and neurotransmission. However, copper dyshomeostasis-induced redox activity makes the brain vulnerable to oxidative and proteostatic stress. Cuproptosis, a recently characterized form of programmed cell death, is triggered by copper binding to lipoylated enzymes of the tricarboxylic acid cycle, resulting in proteotoxic stress, mitochondrial dysfunction, and cell death. Given that mitochondria are central to copper handling and the primary site of cuproptosis, we examine mitochondrial pathways and key cuproptosis-related genes. We also assess disease-specific signatures of copper imbalance. In Alzheimer's disease, excess copper binds to amyloid-β, promoting aggregation and neurotoxicity. In Parkinson's disease, copper-bound α-synuclein fosters aggregation, while copper-driven redox cycling elevates reactive oxygen species. Cuproptosis worsens mitochondrial vulnerability in Parkinson's disease and impairs cellular stress responses in Huntington's disease. In amyotrophic lateral sclerosis, superoxide dismutase 1-related defects compromise antioxidant defenses alongside copper-dependent mitochondrial dysfunction. In prion diseases, copper facilitates prion protein misfolding and toxicity. Across these disorders, common features include mitochondrial dysfunction and cuproptosis hallmarks-such as enhanced protein lipoylation, elevated reactive oxygen species, impaired electron transport chain activity, fragile Fe-S clusters, and increased reliance on the tricarboxylic acid cycle-which collectively increase neuronal susceptibility to copper dyshomeostasis. Clarifying and understanding the critical roles of copper metabolism not only elucidates the pathogenesis of neurodegenerative diseases but also offers alternative therapeutic strategies. This review uniquely integrates the mitochondria-centered cuproptosis axis with copper dyshomeostasis across Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and prion diseases, mapping convergent vulnerabilities to mechanism-grounded interventions and outlining testable translational routes.

<h4>ABSTRACT</h4> An expansion of trinucleotide repeats beyond a threshold, at particular loci, is linked to several diseases. The prevalence of these expansion-linked disorders varies across populations, and may be influenced by the prevalence of sub-threshold intermediate alleles, which are prone to expand. The effect of somatic instability of these repeats, adds further complexity to the role of intermediate repeats. We studied the prevalence of intermediate CAG repeat alleles at SCA1, SCA2, SCA3 and HTT in several hundred individuals. These included patients with clinical suspicion of movement disorder (N∼725), those with severe mental illness (N∼ 420), and healthy individuals from the general population (N∼300). Intermediate alleles were defined as those in the range of 36 -38 CAG repeats for ATXN1 , 45 -59 CAG repeats for ATXN3 , 27 - 35 CAG repeats for HTT , and 27-32 CAG repeats for ATXN2 . In those with neuromotor symptoms, we found intermediate alleles in 26 out of 561 (4.6 %) individuals genotyped at ATXN1 , 10 out of 437 (2.2%) individuals at ATXN3 , 16 out of 686 (2.3%) individuals at HTT and 1 out of 484(0.2%) individuals at ATXN2 . In individuals without a diagnosis of movement disorder (patients with SMI and healthy controls), intermediate alleles were detected in 19 out of 725 (2.6%) individuals at ATXN1 and 14 out of 687 (2.0%) individuals at HTT loci. No intermediate alleles were detected at the ATXN2 and ATXN3 loci. The impact of these intermediate alleles on risk of disease in future generations, and prevalence of disease, needs to be evaluated.

Respiratory syncytial virus (RSV) causes severe respiratory infections, with viral replication occurring in cytoplasmic membraneless viral factories formed by liquid-liquid phase separation (LLPS). The interactions between the RSV nucleoprotein N, phosphoprotein P, transcription factor M2-1, and RNA drive these condensates. Here we employed a microfluidic Phase Scan platform, together with biochemical and cellular assays to systematically characterise LLPS involving RSV proteins and RNA. We identified optimal stoichiometric ratios of N oligomers and P tetramers for condensate formation, demonstrated that monomeric N inhibits LLPS, and revealed that M2-1 enhances condensate formation by increasing multivalency. Notably, we discovered that M2-1 preferentially binds 5’ capped RNA, distinguishing it from N, which binds uncapped RNA. These findings elucidate molecular determinants of RSV viral factory assembly and subcompartmentalisation, providing insights into viral replication mechanisms and informing potential antiviral strategies targeting LLPS processes.

Biomolecular condensates are membraneless compartments, crucial for organising and regulating diverse cellular processes. Current approaches to study condensate biology either use simplified recombinant protein systems with limited physiological relevance, or complex live-cell models with restricted experimental control and scalability. Here, we present ExVivo PhaseScan, a droplet microfluidics platform that couples mammalian lysate-based reconstitution with scalable analysis to generate high-resolution phase diagrams of physiological protein condensates. We apply this approach to study two complex multicomponent condensate systems, stress granules and nucleoli, and dissect the physicochemical interactions that influence their stability. We further developed a machine learning pipeline to analyse condensate morphology which we use to reveal how mutations in the ALS-linked protein FUS remodels condensate phase landscapes. We identify liquid-to-solid transitions of mutant FUS within stress granules and nucleoli, and show that these transitions can be reversed by RNA aptamer-based interventions. Together, these findings establish ExVivo PhaseScan as a versatile tool for dissecting the physicochemical and pathological regulation of condensates, with potential to inform therapeutic strategies for diseases driven by aberrant phase transitions.

The order Diptera (true flies) holds promise as a model taxon in evolutionary developmental biology due to the inclusion of the model organism Drosophila melanogaster and the ability to cost-effectively rear many species in laboratories. One of these dipteran species, the scuttle fly Megaselia abdita (Phoridae), has been used in evolutionary developmental biology for 30 years and is an excellent phylogenetic intermediate between fruit flies and mosquitoes, but remains underdeveloped in genomic resources. Here, we present a de novo chromosome-level assembly and annotation of M. abdita and transcriptomes of nine embryonic and four post-embryonic stages. We also compare nine stage-matched embryonic transcriptomes between M. abdita and D. melanogaster. Our analysis of these resources reveals extensive chromosomal synteny with D. melanogaster; 24 orphan genes with embryo-specific expression, including a novel F-box LRR gene in M. abdita; and conserved and diverged features of gene expression dynamics between M. abdita and D. melanogaster. Collectively, our results provide a new resource for studying the diversification of developmental processes in flies.

Avian influenza virus (AIV) remains a persistent threat to global poultry production and public health, owing to its ability to infect a wide range of avian and mammalian species. The global resurgence of H5N1 and the limitations associated with control measures in intensive poultry production have highlighted the need for a paradigm shift in disease management strategies. In this context, the development of AIV resilient poultry lines is gaining momentum as a promising strategy for long-term disease management. In this review, we discuss the ongoing threat of AIV to poultry production and explore genetic approaches to enhance resilience against avian influenza. We primarily focus on intracellular host restriction factors that inhibit viral replication, followed by an overview of virus-targeted strategies. We further discuss the use of genome-wide screening approaches to study host-pathogen interactions to identify high confidence host targets. Finally, we highlight studies that develop transgenic, or genome engineered chickens with the aim to enhance AIV resilience and their potential for transforming AIV control in poultry production.

The intestine is a highly compartmentalized organ, with distinct segments exhibiting both varying susceptibilities and responses to enteric pathogens, although the cellular and molecular bases of these responses remain elusive. Here, we used <i>Salmonella</i> Typhimurium (S. Tm), a prominent enteric pathogen that causes human colitis, to establish a murine model of <i>Salmonella</i> enterocolitis. By integrating bulk RNA-seq, single-cell RNA-seq, and spatial RNA-seq data, we present a comprehensive spatiotemporal single-cell transcriptomic landscape of the colon over a week-long time course of infection. We identified the distal colon as the intestinal segment where most of the host responses were initiated, with distal colonocytes (DCCs) being the most responsive epithelial cells upon the onset of infection. Furthermore, by correlating our findings with human intestinal single-cell transcriptome data, we identified a human colonocyte population that shares many characteristics with murine DCCs. Our study advances the understanding of the cellular and molecular basis of compartmentalized intestinal responses to pathogenic insults and may pave the way for novel preventive and therapeutic strategies to mitigate intestinal damage and combat intestinal infections.

Due to the unique anatomical connection between the gut and liver, metabolites derived from gut microbiota are closely linked to hepatic health and disease. Among these microbial metabolites, amino acid derivatives have emerged as crucial mediators in gut-liver communication, influencing liver pathophysiology through their bioactive properties. Accumulating evidence demonstrates that amino acid metabolites, including amines, indoles, aromatic derivatives, branched-chain fatty acids (BCFAs), and sulphur-containing compounds (SCCs), function as critical signaling molecules within the gut-liver axis. These metabolites modulate immune responses, inflammation, and metabolic homeostasis, thus affecting the progression and severity of liver diseases. Understanding the specific microbial metabolic pathways responsible for producing these metabolites is essential to reveal novel pathological mechanisms and identify promising biomarkers and therapeutic targets. In this review, we summarize recent advances in microbial amino acid metabolism and highlight the diverse roles of amino acid-derived metabolites in liver disease progression. Furthermore, we discuss the translational potential and current challenges associated with targeting microbial amino acid metabolism in clinical settings. A deeper understanding of these metabolic interactions may ultimately facilitate the development of innovative diagnostic tools and effective therapeutic strategies for liver diseases.

Herein, high-contrast colorless-to-colored thermochromic materials are accomplished through novel 3-component mixtures based on catechol/pyrogallol derivatives (color developer - CD), fatty acid phase change materials (PCM), and leuco dyes. Such components ensure: I) always highly colored hot liquid states, promoted by strong stabilization of the dye open form through hydrogen-bonding interactions with the CDs' vicinal -hydroxyl groups, as also demonstrated by density functional theory calculations; II) solid mixtures with fully controllable coloration state, which directly relies on the relative length of the alkyl chain of CD (C_CD) and PCM (C_PCM). As also rationalized by thermal and spectoscopic measurements, colorless solids are obtained when C_CD is larger than C_PCM (ΔC = C_PCM-C_CD<0), due to self-assembly and phase segregation of CD molecules from PCMs, preventing their interaction with the dye. Instead, when ΔC≥0, the CD solubilizes in PCMs, developing the color of the co-dissolved dye. The rule is extendable to several leuco dye families (lactams, lactons, fluorans) and acid PCMs, fostering a tunable hot state color palette and transition temperature. The target transition is also preserved in structured mixtures (solid lipid particles), granting for the preparation of polymer composites and screen-printed thermochromic patterns, of interest for sensors and invisible security inks.

Mutations in the <i>HFE</i> gene, especially non-synonymous single-nucleotide polymorphisms (nsSNPs), are strongly associated with hemochromatosis, an autosomal recessive disorder characterized by intracellular iron overload, a key feature of tumor development. This study examined the structural and functional effects of deleterious nsSNPs in the <i>HFE</i> gene using bioinformatics tools, gene interaction analyses, molecular docking, molecular dynamics (MD) simulations, and assessments of clinical relevance. Functional analyses identified nine deleterious nsSNPs, including C282Y, L183P, and Q283P, which disrupted disulfide bonds, hydrogen bonds, and hydrophobic interactions, destabilizing the protein. Conservation analysis revealed these mutations occur in highly conserved regions, emphasizing their structural and functional importance. Notably, five nsSNPs (R224Q, R224W, I235T, C282Y, Q283P) within the Ig-like C1-type domain were associated with cancer. Gene interaction analyses showed <i>HFE</i>-related genes are linked to immunity and iron balance. Variants in interacting genes, such as <i>HJV, TFR2, TFRC,</i> and <i>B2M</i>, may influence iron disorders, infection risk, and inflammation. Molecular docking showed reduced interface interactions for the C282Y mutant and altered binding to transferrin receptor 1 (TfR1), potentially destabilizing the HFE-TfR1 complex. MD simulations highlighted key differences, with the mutant showing higher RMSD, decreased compactness (Rg), increased flexibility (RMSF), and greater solvent exposure (SASA), confirming destabilization. Furthermore, <i>HFE</i> expression varied across cancers, with elevated levels in twelve tumor types. Higher expression correlated with better survival in breast and gastric cancers but poorer outcomes in lung cancer. These findings highlight how deleterious nsSNPs, especially C282Y, disrupt <i>HFE</i> structure and function, offering insights into disease mechanisms and guiding therapeutic strategies.

Here, we review the use of Saccharomyces cerevisiae as a powerful model organism for studying cellular processes implicated in neurodegenerative disorders, including stress responses, proteostasis impairment, and vesicle trafficking defects. Over the last two decades, baker's yeast models have been developed for complex diseases such as Parkinson's, Alzheimer's, Huntington's, and Amyotrophic lateral sclerosis (ALS). Yeast cells expressing human proteins, such as amyloid-β, α-synuclein, huntingtin, and TDP-43, have become crucial tools for high-throughput drug screening aimed at counteracting disease progression. These yeast models have unveiled key components involved in the metabolism and toxicity of these proteins, enabling the identification of interacting partners and novel factors within each pathway. Importantly, these pathways were subsequently shown to be conserved in mammalian models. Furthermore, drug candidates identified using yeast models have provided significant leads for drug discovery, highlighting their potential for developing treatments for these neurodegenerative diseases.

Liquids are dense repositories of information, challenged only by how well their compositions are defined, preserved, accessed, or measured. The precise spatial patterning of solutes within a bulk liquid is challenging since diffusion disperses local concentrations and thereby attenuates functionality. Herein, a new concept is introduced for writing and preserving information in the liquid state through liquid-in-liquid microdroplet array printing. This technology produces fine resolution, 2D liquid structures, composite of indexed water-in-oil droplet pixels each with a precise composition, a high spatial resolution and a tight inter-pixel pitch. With extreme control over droplet composition and by applying standard and custom encoding schemes, various forms of information are written biochemically such as images, QR codes, text characters and words. As a composite material, reversible phase transitions between dissolved liquid and crystallized solid states control information encryption and decryption. Compared to current liquid printing and chemical encoding paradigms, ours introduces a fundamentally new precedent for deterministically programming information release, exchange or decay without stimuli or physical processing. Further computational principles such as error correction and information storage are demonstrated. These micro-liquid patterns are relevant to any application based on precise liquid handling such as information theory, materials design and biological assays.

Human herpesviruses (HHVs) comprise nine pathogenic members, including herpes simplex virus 1, herpes simplex virus 2, varicella-zoster virus, Epstein-Barr virus, human cytomegalovirus, human herpesvirus 6A/B, human herpesvirus 7, and Kaposi's sarcoma-associated herpesvirus. Clinical manifestations of HHV infection can range from asymptomatic cases to a broad spectrum of neurological complications, spanning from acute conditions such as encephalitis to chronic disorders including Alzheimer's disease and multiple sclerosis. By establishing latency and undergoing repeated reactivation, HHVs maintain lifelong interactions with the human immune system and shape host immune responses, exerting considerable impact on nervous system homeostasis. Individual susceptibility to, and outcomes of, HHV-associated neurological disorders depend on multiple factors, including the infecting HHV strain and host genetics. Recent evidence highlights the pivotal role of the human leukocyte antigen E (HLA-E) pathway-a non-classical major histocompatibility complex class I molecule with immunomodulatory functions-in regulating virus-host interactions. Since some HHVs manipulate HLA-E to evade immune recognition, individual variability in this axis may influence neurological outcomes. In this review, we summarize and discuss current knowledge of the role of HLA-E in herpesvirus-associated neurological diseases.

During brain aging, terminally differentiated neuroglia exhibit metabolic dysfunction and increased oxidative damage, compromising their function. These cellular and molecular alterations impair their ability to maintain myelin sheath integrity, contributing to age-related white matter degradation. Calorie restriction (CR) is a well-established intervention that can slow biological aging and may reduce age-related metabolic alterations, thereby preserving the molecular function of aging glia. Here we present a single nucleus resolution, transcriptomics dataset evaluating the molecular profile of oligodendrocytes and microglia in the brain of aging rhesus monkeys following lifelong, 30% calorie restriction. Oligodendrocytes from CR subjects exhibited increased expression of myelin-related genes and showed enrichment in glycolytic and fatty acid biosynthetic pathways. In CR subjects, a subpopulation of oligodendrocytes upregulated cell adhesion gene, NLGN1 and were in closer proximity to axons. Microglia from CR subjects upregulated amino acid and peptide metabolism pathways and showed a reduced myelin debris signature. Our findings reveal cell-type specific transcriptional reprogramming in response to long term CR and highlight potential protective mechanisms against myelin pathology in the aging primate brain.

The ε4 allele of apolipoprotein E (APOE4) is the strongest genetic risk factor for Alzheimer's disease (AD), increasing AD risk about fourfold in ~ 34 million American and ~ 75 million European females. APOE4 carriers exhibit cerebral metabolic deficits decades before clinical onset. We previously demonstrated that ketogenic diet (KD), a low-carbohydrate, high-fat diet promoting ketone metabolism, confers cognitive benefits in aged C57BL/6 mice, and in the PS1/APP mouse model of early-onset AD. Here, we evaluated the effects of KD in a humanized APOE4 AD mouse model. KD significantly improved composite cognitive performance and spatial working memory, with pronounced effects in females. Synaptic plasticity, measured via long-term potentiation (LTP), was likewise enhanced exclusively in females. Transcriptomic and protein analyses revealed KD-induced activation of CREB pathway, marked by increased phosphorylation of ERK and CREB in female brains. Moreover, KD selectively reduced pro-inflammatory cytokine levels in females. These findings demonstrate sex-specific neuroprotective effects of KD in APOE4 mice and suggest its potential therapeutic role in mitigating AD risk in APOE4-positive women.

<h4>Background</h4>Aging-related genes (ARGs) are prognostic markers in cancers, but their role in lung adenocarcinoma (LUAD) remains unclear. Investigating ARGs in LUAD may provide valuable insights for clinical diagnosis and treatment.<h4>Methods</h4>Gene expression profiles from TCGA and GEO datasets were analyzed to identify ARG-related genes. Univariate Cox regression and LASSO analysis were used to construct a prognostic risk model. Kaplan-Meier survival analysis, ROC curves, and clinicopathological features validated its predictive accuracy. Immune cell infiltration and tumor microenvironment were assessed using CIBERSORT and ESTIMATE. RT-qPCR was used to validate the differential expression of key genes in LUAD and adjacent normal tissues.<h4>Results</h4>Seven prognostic ARGs (RHPN2, BLK, PTPRO, CA4, UBE2C, METTL7A, and H2BC12) were identified. The risk model stratified patients into high- and low-risk groups, with high-risk individuals showing poorer survival, increased immune evasion, and altered immune cell infiltration. These findings were validated in independent datasets. RT-qPCR confirmed elevated RHPN2, BLK, UBE2C, and H2BC12 in tumors, while PTPRO, CA4, and METTL7A were reduced.<h4>Conclusion</h4>A robust ARG-based risk model, leveraging Cox regression and LASSO, effectively predicts survival and immunotherapy responses in LUAD, offering new tools for personalized prognosis and treatment strategies.

<h4>Background</h4>Orthodontic tooth movement (OTM) is a complex biological process triggered by orthodontic forces (OF). This study aims to study the influence of sex and age on the gene expression of the dental pulp (DP) and periodontal ligament (PDL) of human premolars subjected to 7 and 28 days of OF in vivo.<h4>Methodology</h4>Linear mixed and negative-binomial models were used on previously published RNA sequencing (RNA-seq) datasets of DP and PDL tissue subjected to OF for 7 days and 28 days to verify if the effect of OF depends on sex and age. Differentially expressed genes (DEGs) were identified using false discovery rate and functional analysis was performed.<h4>Results</h4>The datasets consisted of 69 DP and 63 PDL samples from 46 and 41 patients respectively, with similar sex and age distribution. RNA-seq showed that sex did not influence the DP's gene expression profile, since only one DEG related to immune response was detected after 28-days of OF. In contrast, sex significantly affected PDL: 505 DEGs were found after 7 days of OF, related to bone homeostasis, osteoclastic activity and immune response. Age impacted both tissues; in DP, 18 DEGs related to Ca²⁺ regulation and DNA damage repair were found at 7 days, and 10 DEGs associated with repair and adaptive capacities emerged at 28 days. In PDL, 181 genes related to bone regeneration were identified at 28 days, with no DEGs noted at 7 days.<h4>Conclusion</h4>Our study demonstrates that under OF, DP's reaction is not sex-based, whereas PDL's is, particularly in the early phase of OTM, with women showing a more pronounced osteoclastic response. Age-related effects in DP tissue primarily influence Ca²⁺ homeostasis and DNA damage repair in early phases, and tissue repair and adaptive responses later. In contrast, age impacts PDL tissue mainly in the later stages of OTM, affecting its regenerative capacity.

In this report, we examine the extensive research landscape of CRISPR with an emphasis on CRISPR therapeutics and showcase our results from an in-depth analysis of the most up-to-date scientific information consisting of more than 53,000 publications encompassing academic journal articles and patents, spanning nearly three decades, extracted from the CAS Content Collection. Our analysis indicates that cancer and infectious diseases are the most explored in the context of CRISPR. Identified gene targets associated with CRISPR-related publications are led by TP53, c-myc, and hemoglobin beta subunit (HBB). Among the many delivery methods, adeno-associated vectors (AAVs) appear to be highly explored. With >140 CRISPR-based therapeutics in the clinical development pipeline and billions of dollars in investment, the field of CRISPR continues to evolve rapidly. We also briefly discuss the ethical implications of CRISPR technology. While some fundamental challenges persist, the future of CRISPR is undoubtedly bright.

Dogs display temperamental and behavioral variation between individuals, just as psychiatric, temperamental, and cognitive traits vary in humans. In both species, these traits are highly heritable, yet causal genes remain incompletely understood. We performed 14 genome-wide association studies (GWAS) for behavioral traits quantified using the Canine Behavioral Assessment and Research Questionnaire (C-BARQ) in ~1,000 golden retrievers, identifying 12 genome-wide significant loci (<i>P</i> < 2.97 × 10^-6) for 8 traits and 9 additional loci exceeding a suggestive threshold (<i>P</i> < 1 × 10^-5). A human phenome-wide association study (PheWAS) showed that most of the 18 canine positional candidate genes identified were associated with one or more of 190 psychiatric, temperamental, or cognitive traits in humans (7/12 genes at genome-wide loci and 5/9 at suggestive loci). For example, a genome-wide significant locus near <i>PTPN1</i> (dog-directed aggression) overlapped with human measures of Intelligence, Educational attainment, and major depressive disorder. The gene <i>ROMO1</i> was within a genome-wide significant locus for trainability in dogs and associated with intelligence, depression, irritability, and sensitivity/hurt feelings in humans. Other genes located at genome-wide significant loci associated with behavioral, psychiatric, temperamental, or cognitive traits in both species included <i>PRDX1(dog-directed fear), VWA8</i> (touch sensitivity)<i>, ITPR2,</i> and <i>ADGRL2/LPHN2</i> (trainability), and <i>ADD2</i> (stranger-directed fear). From suggestive loci we also found cross-species associations for <i>HUNK, and ZC3H12C,</i> (dog-directed fear), <i>SLC35F6</i> and <i>IGSF11</i> (separation-related problems). These results suggest that shared genetic and molecular mechanisms underlie complex behavioral and temperamental states across species and may inform our understanding of emotional states driving undesirable behaviors in dogs.

Myocardial infarction (MI) remains a global health concern. To mitigate subacute and chronic MI pathophysiology, we previously investigated a pro-reparative decellularized extracellular matrix hydrogel. Despite increasing interest in biomaterial scaffolds, single cell and spatially resolved transcriptomics have not been used to probe their therapeutic activity in the heart. Here, we utilize spatial transcriptomics and single nucleus RNA sequencing to delineate the regional and cell-specific bioactivity of extracellular matrix biomaterials. Extracellular matrix hydrogel subacute treatment in female rats induces cardiac resident macrophage preservation, fibroblast activation, and increased lymphatic, vasculature, smooth muscle, and cardiomyocyte development as well as neurogenesis. Chronic treatment in female rats elicits macrophage polarization, neurogenesis, and development of cardiomyocytes, endothelial cells, and fibroblasts. When comparing treatment timepoints, subacute administration has stronger immune modulation, while chronic administration demonstrates higher cardiac development markers. Both subacute and chronic administration are associated with fibroblast activation and vasculature development. Thus, we elucidate undiscovered therapeutic targets of an injectable extracellular matrix hydrogel, further demonstrating the potential of these biomaterials as an MI therapy.

Erectile dysfunction is attributable to numerous biological and psychological issues, and its prevalence increases with age. We conducted genome-wide association studies of erectile dysfunction in AllofUs subjects of European and African ancestry, then meta-analyzed our findings with published datasets [N_European = 913,194 (136,867 cases); N_African = 125,315 (51,599 cases)]. We identified 40 independent variants in Europeans, two in Africans, and 51 cross-ancestry. In all analyses, the strongest effect variants mapped to a non-coding region known to regulate SIM1, previously associated with erectile dysfunction: rs78677597 (Europeans) (p = 5.32 × 10^-139), and rs17185536 (Africans (p = 1.17 × 10^-9) and cross-ancestry (p = 5.3 × 10^-138)). Genetic correlations with psychiatric and health traits were moderate. Positive associations with phenotypes related to sexual drive may reflect ascertainment bias. This study is consistent with indications that erectile dysfunction is a complex trait influenced by multiple factors. Our findings emphasize the need to investigate genetic risk - SIM1 in particular further - to understand the mechanism through which they affect erectile function.

Sarcopenia (SARC), a progressive degenerative disorder associated with aging, is characterized by a gradual loss of muscle mass and strength and imposes major burdens on individuals and society. Current diagnostic and therapeutic methods for SARC remain limited, highlighting the urgent need for novel biomarkers. The present research was designed to investigate the role of lactate metabolism-related differentially expressed genes (LMRDEGs) in SARC, identify key genes and pathways, and develop a diagnostic model for the disease. Differentially expressed genes (DEGs) were identified using data from the Gene Expression Omnibus database. Gene set enrichment analysis was used to determine signaling pathways associated with the DEGs. Pearson's correlation analysis was used to quantify the strength of linear relationships between coding genes and DEGs. Protein-protein interaction networks of DEGs were constructed using the STRING database. Functional annotation of DEGs was conducted using comprehensive enrichment analyses based on Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways to elucidate their biological relevance. Seventeen LMRDEGs were found to be significantly upregulated or downregulated in SARC, underscoring their pivotal roles in disease pathogenesis. GO and KEGG pathway enrichment analyses revealed that these DEGs were primarily involved in metabolic energy regulation and intracellular signal transduction, suggesting their functional importance in SARC development. Immune infiltration analysis suggested substantial variations in immune cell abundance among SARC samples, emphasizing the immune system's potential contribution to disease progression. This study demonstrates the importance of LMRDEGs in SARC and the need for further investigation into their roles as potential therapeutic targets.

This study designs and develops a mobile learning system based on a convolutional neural network to support open teaching strategies. By integrating a temporal convolutional network (TCN), dilated causal convolution (DCC), and reinforcement learning (RL), the study proposes a TCN-DCC-RL model for personalized learning resource recommendations. The model is evaluated using the learning analytics dataset from the UK Open University. Experimental results show that the TCN-DCC-RL model achieves high performance, with an accuracy of 96.49%, F1-score of 90.57%, Mean Average Precision of 92.27%, and Normalized Discounted Cumulative Gain (NDCG@20) of 0.938. These findings demonstrate that the proposed model significantly enhances the personalization and intelligence of learning resource recommendations, offering a novel technical approach for optimizing future intelligent education systems.

Limited knowledge currently exists regarding the shared genetic architecture influencing human cognitive ability-related traits. We utilized Genomic Structural Equation Modeling (Genomic-SEM) along with multiple post-GWAS methods to estimate potentially causal single nucleotide polymorphisms (SNPs) associated with cognitive ability phenotypic variation. Our study identified 3,842 genome-wide significant loci, including 275 novel loci. Applying multiple transcriptome-wide association methods, we analyzed susceptibility gene signal loci highly correlated with cognitive ability GWAS from tissue, cellular, and genomic element perspectives, identifying 13 high-confidence candidate causal genes and related functional element information. Subsequently, we systematically evaluated 80-90% of currently known human common disease data in the IEU database, along with 3,935 brain imaging-derived phenotypes from UK Biobank and cortical morphological features from 51,665 individuals in the ENIGMA database to determine cognitive ability-related susceptibility factors. We applied the BrainXcan pipeline for brain imaging genetic analysis of cognitive abilities. Additionally, we used summary data-based polygenic scoring methods to systematically analyze the genetic contribution of different chromosomes to cognitive abilities and their risk prediction value. Our study, through multivariate GWAS analysis of cognitive ability common genetic factors, contributes to mapping the shared genetic architecture of human cognitive abilities.

<h4>Abstract</h4>Prostate cancer (PCa) presents a significant global health challenge, and cachexia in end-stage PCa further reduces survival time and deteriorates patient quality of life. This necessitates nuanced approaches to prevention, diagnosis, and treatment. This review analyzes the complex role of growth differentiation factor 15 (GDF15) in various aspects of PCa, including metabolism, chemoresistance, metastasis, and clinical implications. Mechanically, the interactions of GDF15 with PCa lipid metabolism and stromal activation are hypothesized to drive PCa progression and promote cachexia. GDF15's role in PCa bone metastasis is mediated through interactions with osteoblasts and osteoclasts. Furthermore, GDF15's role in chemoresistance emphasizes its impact on drug responses and suggests a potential therapeutic target. Clinically, GDF15 has shown potential as a biomarker, offering diagnostic precision and prognostic value, particularly when combined with established markers such as prostate-specific antigen. The review concludes with a discussion on several monoclonal antibodies targeting GDF15 in clinical trials, such as AV-380, NGM120, Visugromab, and AZD8853, highlighting promising strategies for novel therapies and precision medicine.

<h4>Abstract</h4>Ferroptosis is an iron-catalyzed form of regulated cell death mediated by lipid peroxidation. Novel ferroptosis-based therapeutic strategies could selectively target core pathways associated with the proliferation and survival of malignant tumors, not only directly inducing tumor cell death but also overcoming resistance to conventional therapies, paving a new path for precision cancer treatment. Ferroptosis suppresses tumor progression through lethal lipid peroxidation, yet is co-opted by cancers through antioxidant adaptations. This review highlights the dual role of ferroptosis in tumor biology and explores the dynamic interplay between ferroptosis and the tumor microenvironment to modulate both immunogenic cell death and immunosuppressive signaling. We discuss the therapeutic potential of ferroptosis induction through synergistic integration with chemotherapy, targeted therapy, radiotherapy, hyperthermia therapy, and immunotherapy to simultaneously suppress tumor progression, overcome resistance mechanisms, and potentiate antitumor immunity, thereby offering innovative strategies to address treatment-refractory malignancies. Challenges include identifying predictive biomarkers, developing tumor-selective ferroptosis inducers, and modulating ferroptosis-immune interactions within the tumor microenvironment. By addressing these issues, ferroptosis has the potential to overcome treatment-resistant and immune-evasive tumors, reshaping oncology therapeutics.

BACKGROUND: Epithelial compartments play a central role in the progression and metastasis of colorectal cancer (CRC), yet the mechanisms underlying their transcriptional reprogramming across disease stages remain incompletely understood. METHODS: We constructed comprehensive single-cell and spatial transcriptomic atlases encompassing normal colorectal tissue, primary CRC, and liver metastases (LMs), profiling over 200,000 high-quality cells. Specific coexpression modules and transcriptional regulators were identified and spatially mapped. Additionally, the clinical and functional roles of SCAND1 were investigated using western blot, RT-qPCR, and immunohistochemistry in clinical samples, as well as functional assays in CRC cell lines. RESULTS: Distinct coexpression modules were selectively enriched in normal epithelial cells and associated with vesicle transport, ion homeostasis, and barrier function, whereas LM-specific modules were linked to Wnt signalling, ribosome biogenesis, and cell cycle processes. High-CNV epithelial cells exhibited progressive activation of key transcription factors such as CEBPB, E2F, and the AP-1 family, with concurrent suppression of ARGLU1 and MTNR2L8, indicating coordinated remodelling of transcriptional and metabolic programs during metastasis. These changes were spatially validated and correlated with alterations in epithelial composition within the tumour core. Consistently, multiplex immunofluorescence (mIHC) confirmed the expression patterns of CEBPB, ARGLU1, S100A4 and FOSL1, across normal epithelium, CRC, and LM. Notably, SCAND1 was identified as a potential therapeutic target for malignant epithelial plasticity. We demonstrated for the first time that SCAND1 expression is significantly upregulated in CRC patients and is associated with superior diagnostic and prognostic value. Functional experiments revealed that SCAND1 promotes tumour cell proliferation, inhibits apoptosis, and enhances metastatic potential. Furthermore, co-culture of SCAND1-overexpressing tumour cells with T cells showed that SCAND1 overexpression attenuates T cell–mediated cytotoxicity, supporting an additional immune-evasive role. CONCLUSIONS: Our study elucidates the molecular logic of epithelial state transitions during CRC metastasis and underscores the promise of SCAND1 as a novel biomarker and therapeutic target in colorectal cancer.

As a highly fatal and heterogeneous disease, ovarian cancer (OC) lacks sufficient treatment choices. Over the past decade, RNA sequencing (RNA-seq) has profoundly changed research methods in molecular biology, enabling comprehensive analysis of transcriptomes, somatic mutations, and tumour heterogeneity. In OC research, the application of RNA-seq technologies has significantly enhanced scientists' understanding of the biological mechanisms underlying the disease, providing deeper insights into the intrinsic mechanisms of key processes such as tumour initiation, metastasis progression, drug resistance formation, and immune evasion. This breakthrough not only expands our understanding of OC but also holds promise for the discovery of novel diagnostic biomarkers, the development of innovative therapies, and the establishment of precise prognostic assessment systems. This review summarizes RNA-seq technologies and their applications in OC research, while also discussing the challenges and opportunities for their broader implementation in cancer studies.

Single-cell transcriptomics enables precise characterization of cellular heterogeneity, but current pre-trained models relying solely on expression data fail to capture gene associations. We present scKGBERT, a knowledge-enhanced foundation model integrating 41 M single-cell RNA-seq profiles and 8.9 M protein-protein interactions to jointly learn gene and cell representations. scKGBERT employs Gaussian attention to emphasize key genes and improve biomarker identification, achieving superior performance across gene annotation, drug response, and disease prediction tasks. scKGBERT enhances biological interpretability and offers a powerful resource for precision medicine and disease mechanism discovery.

Substantial contamination in the ecosystem (particularly, in waterbodies) due to the disposal of hazardous substances from superfluous industrial byproducts and other human activities is one of the most serious environmental issues. Although water is among the five basic elements required for all the living beings, it is continuously becoming unsafe and unhygienic for the purpose of drinking and household activities. The presence of heavy metal ions and many toxic anions (beyond their permissible concentration) significantly contributes to water pollution. Therefore, it is judicious to detect inorganic ions in order to avoid adverse situations related to the human health and ecological imbalance. Owing to the easy affordability and unique properties of zero-dimensional graphene quantum dots (GQDs), particularly, high/tunable fluorescence, electronic conductivity, electrocatalytic activity, chemiluminescence (CL) and electrochemiluminescence (ECL), GQDs-based/involved platforms are potentially deployed for the detection of inorganic ions with reasonable selectivity and sensitivity. The versatility of these sensing probes includes the possibility of detection through fluorimetric, colorimetric, electrochemical, CL, and ECL techniques. This review aims to comprehensively inspect the deployment of GQDs-based/involved sensors for the recognition of inorganic ions, highlighting different sensing approaches with the development of performance metrics and providing insights into their underlying mechanisms. Furthermore, this collective outlook on GQDs-based/involved sensors for inorganic ions may help to identify shortcomings in the existing knowledge and influence/inspire new research with better prospects.

The Lung Transplant Consortium (LTC), sponsored by the National Heart, Lung, and Blood Institute (NHLBI) is a 20-center collaboration among lung transplant programs in North America conducting both smaller multicenter projects and one consortium-wide prospective observational cohort study called the Prospective Multicenter Research on Donor and Recipient Management Strategies to Improve Lung Transplant Outcomes (PROMISE) Lung Study-for more information about the LTC, visit their website: https://lungtransplantconsortium.org/. The LTC conducted a meeting in April 2025 to strategize how to maximize the benefits of the PROMISE study to advance the field of lung transplant. This paper summarizes the key themes that emerged from the meeting: leveraging PROMISE data elements, including biomarkers, imaging, and PROs; clinical syndrome and consensus definitions; variation in management and management strategies; and future interventional trials leveraging the PROMISE consortium. The PROMISE study will serve as the platform for establishing best practices in lung transplant, inform the validity of newly identified syndromes, and support analysis of patient-reported outcomes, image data, and biosamples. The PROMISE study and LTC create a critically needed research platform and multicenter collaborative structure that may be adapted to conduct future multicenter clinical trials that will advance lung transplant medicine.

Uterine malignancies represent a broad and heterogeneous group of neoplasms, which collectively account for the majority of gynaecological malignancies in developed countries. For a significant proportion of these, mortality rates and clinical outcomes are of major concern, owing to complex molecular profiles and aggressive phenotypes that are largely refractory to conventional therapeutic approaches. Recent advancements in precision medicine and the development of targeted therapies have been transformative for several common cancer types; however, the same benefits are yet to be realised for many uterine cancers. This review comprehensively details the diverse molecular features characterising the various subtypes of uterine malignancies. Furthermore, we have examined the therapeutic approaches actively investigated to target these unique molecular features, identifying pathways and treatments that offer the greatest potential patient benefit. Among increasingly personalised strategies, particular promise is shown by HER2-targeted therapies for HER2-positive malignancies (e.g. trastuzumab deruxtecan). Additionally, targeting <i>TP53</i> wild-type tumours with selinexor, as well as addressing AKT and DNA repair pathways in both uterine carcinomas and sarcomas (i.e. AKT inhibitor and poly(ADP-ribose) polymerase inhibitor combinations), represents key advancements. Furthermore, anti-angiogenic and immune checkpoint inhibitor combinations hold significant promise for future therapeutic strategies.

O-GlcNAcylation, a prevalent reversible post-translational modification, intricately alters non-histone proteins, influencing the organization of gene transcriptional regulation within the accessible chromatin environment. This nucleoplasmic landscape, characterized by histone-free regions, fundamentally enables O-GlcNAc-mediated modulation through dynamic accessibility. However, unraveling the O-GlcNAc-open chromatin interplay that governs sophisticated transcriptional regulatory networks remains constrained by current techniques, which lack the resolution to probe this spatiotemporal crosstalk. Here, we report a general strategy to systematically and chemoselectively profile O-GlcNAc-associated chromatin accessibility on a genome-wide scale (COCA-seq). Through comprehensive validation across low- and high-throughput levels, we demonstrate COCA-seq's dual fidelity in both O-GlcNAc chemoselectivity and open chromatin specificity. We employ it to delve into doxorubicin resistance for breast cancer, scrutinizing pivotal regulatory genes and transcription factors implicated in this complex biological event. By integrating bulk RNA-seq with COCA-seq, we offer a multiomics perspective, shedding light on related biological processes and pathways like drug efflux and stress homeostasis, thereby uncovering potential mechanisms by which O-GlcNAc-associated open chromatin orchestrates tumor drug resistance. COCA-seq emerges as a general and versatile tool across various biological contexts, poised to reveal the landscape of O-GlcNAc-associated open chromatin regions across the genome and decipher the significance of glycosylation behind it.

<h4>Background</h4>Parkinson's disease (PD) is a neurodegenerative disorder marked by dopaminergic neuron loss. Current treatments fail to halt progression and often cause long-term side effects. Traditional Chinese medicine (TCM), recognized for its holistic therapeutic approach, has potential in alleviating symptoms associated with PD. Lingjiao Gouteng decoction (LJGT), a TCM used for tremor symptoms, requires further investigation to clarify its neuroprotective mechanisms and pharmacological basis.<h4>Purpose</h4>This study aims to explore the potential mechanisms underlying the neuroprotective effects of LJGT in the treatment of PD and to identify the key bioactive components contributing to its therapeutic efficacy.<h4>Methods</h4>The chemical components of the LJGT compound were characterized by LC-MS/MS technology. PD-like mouse models were established through 6-hydroxydopamine or adeno-associated virus carrying human SNCA^A53T, followed by LJGT administration. The effects of LJGT on PD-like motor deficits were assessed by rotarod test, pole test, and gait analysis. Dopaminergic neuron loss was evaluated by immunohistochemistry, LC-MS/MS, and Western blotting. To investigate the underlying mechanisms of LJGT in PD treatment, RNA-seq, lipidomics, and oxidized phospholipidomics analysis were conducted. Furthermore, the key bioactive components of LJGT were identified by molecular docking, cellular thermal shift assay, and C11-Bodipy staining.<h4>Results</h4>LJGT significantly ameliorated behavioral impairments and attenuated neuronal damage in PD mouse models. Multi-omics further revealed that the neuroprotective effects of LJGT are mediated through the regulation of phospholipid redox metabolism and inhibition of phospholipid peroxidation. Notably, treatment with LJGT significantly downregulated the expression of ALOX15 in this pathway, while upregulating the expression levels of GPX4 and FTH1. Additionally, the bioactive compounds 3,5-di-caffeoylquinic acid, hesperidin, and isoacteoside were identified as key modulators targeting ALOX15/PEBP1 complex, GPX4/NEDD4 complex, and FTH1, respectively, thereby regulating the phospholipid redox metabolism homeostasis and synergistically contributing to the overall anti-phospholipid peroxidation effect of LJGT.<h4>Conclusion</h4>LJGT exerts neuroprotective effects in PD by modulating the phospholipid redox metabolism, reflecting the holistic therapeutic characteristics of TCM. These findings provide a scientific basis for the clinical application of LJGT and its potential as a multi-component and multi-target therapeutic agent for PD.

<h4>Background</h4>Researches have found that the gut microbiota stimulate inflammation by releasing proinflammatory chemicals, including bile acids (BAs) and L-kynurenine. In the present study, our purpose is to investigate the potential functional roles of gut microbes and their secretions in the comorbidity mechanism between inflammatory bowel disease (IBD) and diabetic retinopathy (DR).<h4>Methods</h4>Specific existing gut microbes among IBD and diabetes mellitus (DM) patients were gained and analyzed to figure out the proinflammatory chemicals secreted by specific existent microbes. The expression data of peripheral blood mononuclear cells (PBMCs) were obtained and underwent differentially expressed genes (DEGs) analysis and weighted gene co-expression network analysis (WGCNA) to find out potential effects of microbes secretions on PBMCs. Single-cell analysis was conducted to elucidate the underlying cytological comorbidity mechanisms.<h4>Results</h4>The results of the study showed that the secretions of abnormal gut microbes in IBD and DM patients, such as BAs and L-Kynurenine, can stimulate abnormal immune response. PBMCs can be activated, mobilized, and disrupt vascular endothelial barriers upon exposure to proinflammatory substances generated by <i>Veillonella</i> and <i>Clostridium</i>, the dysbiotic gut microbiota in IBD, which exacerbate retinal inflammation and worsening DR pathological conditions. Furthermore, genes related to PBMC migration, such as RSU1, FN1, and CD9, are activated in PBMCs after exposure to pro-inflammatory chemicals from abnormal gut microbes, offering a genetic comorbidity mechanism for IBD and DR. We also unraveled that IBD can promote the proliferation and activation of effector memory T (Tem) cells, which also showed elevation in DR patients, thereby providing a cellular basis for the comorbidity between IBD and DR.<h4>Conclusion</h4>The aberrant gut bacteria in IBD patients might secrete proinflammatory substances, such as L-kynurenine and BAs, which may activate mononuclear cells, especially the Tem. The mononuclear cells subsequently migrated into the retina, exacerbating the situation of DR. This finding underscores the potential of comorbidity mechanism for better treatment of DR in the future.

Global warming has intensified the prevalence and severity of heat stress (HS) in poultry, leading to compromised welfare, reduced productivity, and deteriorated meat quality. To address this challenge, this review evaluates astaxanthin (AX), a potent natural carotenoid antioxidant derived from microalgae and yeast, for its application in mitigating HS-induced oxidative damage. AX's unique molecular structure enables efficient scavenging of reactive oxygen species, stabilization of mitochondrial function, and modulation of stress-responsive pathways. Comparative studies highlight AX's superior antioxidant efficacy over conventional vitamins and carotenoids. In poultry, AX supplementation enhances antioxidant enzyme activity, suppresses inflammatory cytokines, and improves immune competence, resulting in better growth performance, feed efficiency, and meat quality traits. This Review synthesizes recent in vivo findings on AX's bioavailability, safety, and integration into commercial poultry diets. Finally, future directions are proposed, emphasizing omics-guided biomarker identification, synergistic antioxidant strategies, and advanced delivery systems to optimize AX application in poultry production.

<h4>Introduction</h4>Workers in the shellfish industry face increased risks of allergy and asthma due to complex bioaerosol exposures in the workplace. This study aimed to assess whether combined exposure to the main components of these aerosols, specifically allergens and microbial agents, can potentiate inflammatory and allergic responses.<h4>Methods</h4>THP-1 monocytes and advanced human alveolar co-cultures model ALIsens® were exposed to shrimp tropomyosin (0.0049, 1.3 and 2.6 µg/mL), and components from Gram-positive bacteria; lipoteichoic acid (0.25-4 µg/mL), and fungi; zymosan (6.25-100 µg/mL), either alone or in combination. The effects on the gene expression and protein secretion of chemokines and cytokines were assessed by RT-qPCR and ELISA or Luminex.<h4>Results</h4>Combined exposure to tropomyosin and lipoteichoic acid resulted in increased <i>CCL20</i>, <i>CCL2, TNF</i> and <i>IL8</i> expression and CCL20 and TNF protein secretion in THP-1 cells, when compared to individual exposure. Similarly, tropomyosin combined with zymosan elicited a response pattern, characterised by increased expression and secretion of chemokines and cytokines in most of the tested combinations. Furthermore, the increased secretion of CCL20 and expression of <i>CCL2</i> following combined exposure to tropomyosin and lipoteichoic acid were confirmed in the alveolar co-culture model, while no effects in combination with zymosan were observed.<h4>Conclusion</h4>These findings suggest that microbial components in shellfish industry bioaerosols may enhance the immunological responses caused by inhaled allergens in an additive manner, highlighting the need to minimise microbial contamination in workplaces where allergen exposure is prevalent.

<b>Background:</b> Venous thromboembolism (VTE) is a multifactorial disorder influenced by both genetic and environmental factors, with substantial variability in susceptibility across populations. Data on VTE-associated genetic variants in Asian populations, including Thais, remain limited. To address this, we developed a 39-single-nucleotide polymorphism (SNP) genotyping panel using the MassARRAY platform and evaluated its association with VTE in a Thai cohort. <b>Methods:</b> A total of 209 individuals, comprising 122 patients with objectively confirmed VTE and 87 age- and sex-matched healthy controls, were genotyped. Allele frequencies were compared, and associations with VTE were assessed. <b>Results:</b> Seven SNPs demonstrated significant associations: five risk alleles (<i>PROC</i> rs146922325, <i>ABO</i> rs8176743, <i>FGG</i> rs2066865, <i>F11</i> rs4253417, and <i>HIVEP1</i> rs169713) and two protective alleles (<i>F5</i> rs4524 and <i>TGFB2</i> rs57615042). To examine cumulative effects, a polygenic risk score (PRS) integrating genetic and clinical factors was constructed. Higher PRS was significantly associated with recurrence, particularly among patients with unprovoked VTE, conferring more than a threefold increase in recurrence risk (HR = 3.53, 95% CI: 1.04-10.2, <i>p</i> = 0.043). These findings provide the first systematic evidence of population-specific genetic risk factors for VTE in Thais and highlight the clinical potential of PRS for recurrence prediction. <b>Conclusions:</b> The MassARRAY-based panel offers a cost-effective, high-throughput strategy for simultaneous SNP detection, supporting scalable genomic studies and personalized risk stratification. Our results contribute to understanding the genetic architecture of VTE and highlight the value of incorporating non-European populations into genetic studies to advance precision medicine.

Influenza A virus (IAV) has a wide range of avian and mammalian hosts, leading to disease outbreaks and increasing the risk of panzootics and pandemics. Subtype H5N1 of clade 2.3.4.4b is causing the current high pathogenicity avian influenza (HPAI) panzootic. Environmental changes are fuelling the spread of HPAI H5N1 in wildlife worldwide, with occasional spillover events from seabirds to cetaceans. Sampling difficulties and limited tests available for diagnosis are a challenge to cetacean virology research. Understanding the risk of HPAI outbreaks in cetaceans requires a comprehensive examination of events of IAV infection. Documented cases relate to IAV subtypes H1N3, H13N2, H13N9, and H5N1 and have been reported in cetaceans sampled in the Pacific, Atlantic, and Arctic Oceans. The number of H5N1 IAV isolated from cetaceans is increasing and affects six host species of the families <i>Delphinidae</i> and <i>Phocoenidae</i> of the suborder <i>Odontoceti</i>. The analysis of 40 molecular markers of viral adaptation to mammals in 21 H5N1 cetacean isolates reveals mutations are present in three viral proteins: hemagglutinin (HA), polymerase basic protein 2 (PB2), and nucleoprotein (NP). Phylogenetic analysis of HA and PB2 sequences isolated from cetaceans and co-occurring cases in seabirds and marine mammals do not support sustained transmission of the virus between cetaceans. IAV H5N1 appears to be reaching cetaceans after spillover from seabirds and other marine mammals. Increasing worldwide surveillance of IAV infection of cetaceans is crucial, as these marine mammals are sentinel species for human pandemic preparedness and key species for marine biodiversity conservation and ecosystem health.

Breast cancer remains a major public health challenge globally, with its etiology influenced by a complex interplay of genetic, hormonal, and environmental factors. While traditional risk factors such as age, reproductive history, and family predisposition are well-recognized, emerging evidence points to heavy metal exposure - particularly iron - as a potential contributor to carcinogenesis. Iron is a vital micronutrient required for cellular metabolism; however, excess iron has been shown to catalyze oxidative stress and deoxyribonucleic acid damage, both of which are implicated in tumor initiation and progression. Occupational and environmental iron exposure is increasingly prevalent due to industrial expansion, especially in metal-processing industries such as welding, mining, and steel manufacturing. Workers in these sectors, as well as individuals living near industrial zones, may experience chronic low-level exposure to iron through inhalation, ingestion, or dermal absorption. When iron accumulates beyond the body's regulatory capacity, it may foster a pro-oxidative and pro-inflammatory milieu conducive to breast tumorigenesis. Furthermore, iron's interaction with estrogen metabolism may have unique implications for hormonally sensitive tissues like the breast.

Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD) are major neurodegenerative disorders that share certain pathological features but differ in their genetic etiology and clinical presentation. Their potential molecular intersections remain incompletely understood. In this research, we conducted a comparative transcriptomic analysis using postmortem brain RNA-seq datasets from AD (GSE53697), PD (GSE68719), and HD (GSE64810) to identify shared and disease-specific transcriptional signatures. Differentially expressed genes (DEGs) were determined and functionally characterized through Gene Ontology (GO) enrichment. Protein-protein interaction (PPI) networks were generated using STRING and visualized in Cytoscape to identify central hub genes, followed by gene-disease and drug-interaction analyses to assess functional and therapeutic relevance. Ten DEGs were found to overlap among the three disorders, exhibiting variable directions of regulation across diseases. Enrichment analysis indicated convergence on immune- and inflammation-related biological processes. Key hub genes, including <i>MMP9</i>, <i>LCN2</i>, <i>CXCL2</i>, <i>CCL2</i>, <i>S100A8</i>, and <i>S100A9</i>, were identified as central nodes within the PPI network. Although the overlap in DEGs was limited, the findings suggest that neuroinflammatory signaling represents a shared molecular theme across AD, PD, and HD, warranting further validation in independent cohorts.

Dynamic mechanical cues are crucial for glial neuromodulation and energy metabolism in neural regeneration, yet the mechanisms underlying mechanotransduction and intracellular organelle responses in glia after neurotrauma remain vague. In this study, we develop mechano-bioactive piezoelectric hydrogel bioelectronics (BaTiO₃-embedded collagen-1 hydrogel) and investigate mechanotransduction in astrocytes and Schwann cells. Ultrasound-driven piezoelectric hydrogel bioelectronics exerts electrical signals from mechanical stimulation and upregulates PIEZO1 channel in astrocytes and PIEZO2 channel in Schwann cells. This mechanoelectrical conversion increases calcium influx to activate ATP synthase subunit and promote MFN/OPA1 mediated mitochondrial fusion. Consequently, it enhances ATP synthesis by forming an efficient energy network as a central bioenergetic hub to promote glia mediated neural repair. Furthermore, this mechano-bioactive piezoelectric hydrogel bioelectronics exhibits therapeutic efficacy for treating central and peripheral nervous injuries in multiple animal models (mice, rats, Beagle dogs, and Rhesus monkeys), demonstrating its wide adaptivity and significant translational potential. The findings elucidate a multilevel mechanobiological energy transduction (mechanical-electrical-bioenergetic conversion) design in neural repair as a promising clinical treatment mode.

Colorectal cancer (CRC) ranks as the third most prevalent malignancy and the second leading cause of cancer-related mortality globally. Despite substantial advancements in CRC therapies over recent decades, drug resistance continues to pose a significant challenge in effective treatment. This comprehensive review investigates the role of long non-coding RNAs (lncRNAs) in the emergence of drug resistance in CRC. Specifically, we analyze the functions of lncRNAs, including HOTAIR and MALAT1, in key processes such as apoptosis, DNA damage response and repair, cancer stem cell regulation, and drug metabolism. By examining the structural and biogenetic characteristics of lncRNAs, this review elucidates their specific contributions to drug resistance. We provide novel insights into the therapeutic potential of targeting lncRNAs to mitigate drug resistance in CRC, thereby offering new perspectives on treatment strategies. Ultimately, this review highlights the clinical implications of lncRNA research, emphasizing their potential as targets for innovative therapeutic interventions in colorectal cancer.