Gene Literature Dashboard

Asthenoteratozoospermia is a common underlying cause of male infertility, with dynein dysfunction playing an important role in the aetiology of the condition. Dysfunction in certain dynein proteins has been implicated in asthenoteratozoospermia, while others exclusively induce asthenozoospermia in the absence of overt morphological abnormalities in the sperm. Dynein axonemal heavy chain 10 (DNAH10), an inner dynein arm heavy chain protein, was identified as being associated with asthenoteratozoospermia in our previous studies. However, the mechanism through which DNAH10 contributes to this condition remains unclear. In this study, we demonstrated that DNAH10 deficiency leads to abnormal morphology of the sperm head and flagella. Additionally, DNAH10 dysfunction leads to impaired manchette function and aberrant localisation of axonemal proteins. Mechanistically, ubiquitin carboxyl-terminal hydrolase L3 (UCHL3) binds to parkin co-regulated gene protein (PACRG) and stabilises it via deubiquitination. In this process, DNAH10 exerts a bridging effect, enhancing the interaction between the UCHL3-PACRG complex to facilitate their involvement in manchette function. Collectively, this study demonstrated the function of DNAH10 in intra-manchette transport, providing important guidance for genetic diagnosis and prognosis in patients with infertility.

Solid tumours present major treatment obstacles because of their immunosuppressive microenvironment and poor response to traditional chimeric antigen receptor (CAR)-based immunotherapies. Recent advances in cellular engineering have introduced CAR-macrophages derived from induced pluripotent stem cells (CAR-iMacs) as a promising approach to get around these obstacles. CAR-iMacs are designed to attack tumours, but their phenotypic plasticity can cause them to transform into M2-like macrophages in the tumour environment (TME), where they may instead suppress immune responses and promote tumour progression and metastasis. Roquin-1 and Regnase-1 are RNA-binding proteins that act as negative regulators of inflammatory genes that contribute to the phenotypic plasticity of macrophages. This perspective highlights a novel approach to augmenting anti-tumour responses of CAR-iMacs by simultaneously knocking out Roquin-1 and Regnase-1 via CRISPR-Cas9 gene editing. This approach drives a shift from an immunosuppressive M2-like state to an M1 state, promoting sustained pro-inflammatory signalling, boosting phagocytic and cytotoxic capabilities within the tumour microenvironment. Addressing a serious constraint in conventional adoptive cell therapies, this dual-targeting platform could provide a potent and scalable immunotherapeutic treatment for solid malignancies.

Human β-glucuronidase (HGUSB), a key lysosomal glycosyl hydrolase for the degradation pathway of glycosaminoglycans (GAGs), plays a crucial role in cell proliferation and inflammation, making it a promising target for novel therapeutic strategies including antibody-drug conjugates (ADCs) with β-glucuronic linkers. In this study, molecular docking and molecular dynamics (MD) simulations were performed to investigate the conformational stability of HGUSB in complex with different ligands, including substrates, inhibitors, and β-glucuronic linkers. Our rationale approach includes the evaluation of commercial substrates and a known inhibitor with different binding stoichiometries to identify the most favorable configuration and the most stable conformation of the enzyme. Based on the binding mechanism of HGUSB to these well-known ligands, the interaction with commercial linkers was evaluated, providing a structural determination of the recognition mechanism between the enzyme and ADCs. MD simulations on HGUSB::Linker complexes revealed that the maleimide-containing hydrophilic β-glucuronide, exhibited the most stable binding making it the best fitting linker among those analyzed in this study. Overall, this study identifies the optimal binding configuration of the HGUSB enzyme for investigating small molecule interactions and, despite the endogenous homotetrameric assembly, justifies the use of a simplified monomeric model for the study of larger macromolecular complexes, like linker analysis, ensuring an efficient and accurate computational approach. These findings lay the groundwork for a rationale optimization of β-glucuronic linker-based ADCs, offering new perspectives for targeted cancer therapies.

Disruptor of Telomeric Silencing 1-Like (DOT1L) has emerged as a critical epigenetic regulator in cancer, primarily because of its role as the sole histone methyltransferase responsible for histone H3 at lysine 79 methylation. This modification affects transcriptional activation, DNA repair, and cell cycle progression. Its dysregulation is associated with both hematological and solid tumors. In Mixed-lineage leukemia (MLL)-rearranged leukemia, DOT1L maintains aberrant gene expression patterns at loci such as HOXA and MEIS1, supporting leukemic stem cell survival and driving oncogenesis. In solid tumors, DOT1L influences diverse processes, including epithelial-mesenchymal transition, angiogenesis, and cell cycle regulation, contributing to tumor growth and metastasis. Therapeutic strategies targeting DOT1L using inhibitors, such as EPZ-5676, have shown promise in preclinical and clinical studies, highlighting their potential as versatile targets for precision oncology. This review summarizes the recent findings on DOT1L's involvement in cancer development and its potential as a therapeutic target.

Based on mRNA Expression Profiles of 57 Patient-Derived Colorectal Cancer Stem-Like Cell (CRC-SC) Lines Compared With Normal Colonic Epithelial Stem-Like Cells (NCE-SCs), we Identified Five CRC Subtypes. The First Subtype of CRC-SCs Showed Markedly Increased Expression of MUC12, PIGR, PLA2G2A, SLC4A4, and ZG16, Which Were Barely Detectable in the Other Subtypes. Importantly, Their Expression Correlated With Favorable Outcomes in Both the Discovery Cohort and Independent Two Test Cohorts From Public Databases. The Remaining Four Subtypes Showed High Expression of DEFA6, BST2, MAGEA6, or IGF2 Compared With NCE-SCs. Although the Expression of Each Gene Individually Influenced Patient Outcomes, Additional Co-Expressed Genes Within Each Subtype Were Also Associated With Prognosis. Furthermore, Integrating the Five Subtype-Specific Signatures Produced a Practical Prognostic Indicator, Designated as the General Colorectal Cancer Signature (GCS), and Provided Individualized Predictive Signatures for Each Patient. The Clinical Significance of GCS Was Further Validated in a Novel Orthotopic Xenograft Mouse Model, Which Recapitulated Patient Outcomes: CRC-SCs With Low GCS Scores Developed Distinct Liver and Lung Metastases, Whereas Those With High Scores Did Not. Apparent Associations Were Observed Between Activating RAS/RAF Mutations and BST2 Expression, and Between the Absence of SMAD4 Mutation and IGF2 Expression, but These Had no Significant Impact on Patient Survival, Suggesting That Driver Gene Mutations May Not Directly Influence GCS. Collectively, Our Findings Provide a Comprehensive Overview of Clinically Relevant Molecular Subtypes of CRC-SCs, Representing the Current Landscape of CRC Molecular Expression Subtypes. They Also Enable Rapid, Low-Cost Outcome Prediction and Suggest Potential Targets for Therapeutics Development.

Copper is an important trace element in the human body and plays an essential role in cells, where it is involved in synthesizing copper‑dependent enzymes, including superoxide dismutase, cytochrome c oxidase, tyrosinase, lysyl oxidase, dopamine‑β‑hydroxylase and other related copper‑containing enzymes. Copper overload or deficiency affects cell activity, leading to the development of neurodegenerative diseases or cancer. Neurodegenerative diseases, including Alzheimer's, Parkinson's and Huntington's disease, as well as cancer, represent significant chronic health burdens. The complexity of their pathophysiological mechanisms, coupled with the limitations of current targeted therapies, complicates the development of effective treatments. This review provides a comprehensive overview of the current understanding of copper's regulatory mechanisms in health and disease, with particular emphasis on its roles in neurodegenerative disorders and cancer. Recent advances in copper‑targeted therapeutic strategies, including copper chelators, ionophores and copper‑based nanoparticles, were highlighted. Furthermore, the clinical potential, key challenges and future prospects of these interventions were assessed. By synthesizing recent preclinical and clinical evidence, this review aims to contribute novel perspectives for improving the treatment of copper‑associated diseases.

Neurodegenerative diseases, including polyglutamine diseases, remain a clinical challenge, partly because of limited animal models that recapitulate human disease. Here, we describe a second-generation transgenic marmoset model of spinocerebellar ataxia 3 (SCA3), a polyglutamine disease, which stably expresses expanded CAG repeats in ataxin 3 (ATXN3). All five offspring of the founder marmoset harbored the transgene with reduced transgene integration sites compared with the founder and without repeat instability or genetic mosaicism, offering improved construct validity. Three of the five marmosets developed progressive motor impairments that segregated into two distinct phenotypes - early onset with rapid progression and late onset with mild progression - accompanied by corresponding patterns in body weight gain and grip strength. Pathological analysis revealed cerebellar Purkinje cell loss, spinal cord neurodegeneration and widespread intranuclear inclusions. The severity of motor phenotypes correlated with transgene expression levels in disease-relevant brain regions, including the cerebellum and spinal cord. By overcoming the translational limitations of rodent systems, our second-generation model offers a powerful platform for investigating disease mechanisms and testing potential therapeutics, advancing the utility of transgenic marmosets as clinically relevant models of neurodegenerative diseases.

Antimicrobial resistance (AMR) is a growing global health concern, especially in the management of bone and joint infections. A bioresorbable scaffold comprising carbonate apatite and 0.4 wt % silver phosphate (Ag₃PO₄) (designated as CASP) was developed in this study to achieve simultaneous osteogenesis, scaffold resorption, and photoactivated antibacterial activity. The CASP scaffold exhibits intrinsic antibacterial activity within 1 h via Ag⁺ ion release, which is further enhanced under visible light irradiation through the generation of reactive oxygen species generation. In vitro studies demonstrate that CASP scaffolds effectively suppress the growth of methicillin-resistant <i>Staphylococcus aureus</i> without compromising osteoblast viability, proliferation, differentiation, and calcification. In vivo implantation in rabbit femoral defects indicates that CASP scaffolds support robust bone regeneration and gradual scaffold resorption comparable to those of scaffolds without Ag₃PO₄ while maintaining long-term antibacterial activity for at least 4 weeks postimplantation. Transient inflammatory responses observed early after implantation are detoxified over time, potentially through the in vivo sulfidation of released silver ions. The findings of this study highlight the potential of CASP scaffolds as promising candidates for nonantibiotic infection control and bone tissue regeneration via visible light-driven photocatalytic therapy, offering an effective strategy to combat AMR in orthopedic applications.

CRISPR-Cas9 technology offers transformative potential in treating Huntington's Disease (HD) by directly addressing its genetic root causes. This manuscript explores the pathophysiological mechanisms of HD, characterized by toxic mutant huntingtin (mHTT) protein resulting from expanded CAG repeats in the HTT gene, and the challenges posed by current therapeutic limitations. We comprehensively review the mechanisms of CRISPR-based therapeutic strategies, including excision of expanded repeats, allele-specific targeting, and epigenome editing, highlighting their efficacy in preclinical studies using animal models and human iPSCs. Delivery methods, such as viral and non-viral vectors, are analysed for their role in optimizing therapeutic outcomes while minimizing off-target effects and immune responses. Ethical and safety considerations, especially regarding precision and long-term impacts, are critically examined alongside emerging strategies to enhance specificity. With ongoing clinical trials and advancements in delivery systems, CRISPR technology represents a paradigm shift in addressing HD and broader neurodegenerative conditions. This review underscores the promise of gene editing in overcoming existing barriers and paving the way for transformative therapeutic approaches.

<h4>Objective</h4>Racial and ethnic disparities in neonatal postoperative outcomes are reported, but differences in cardiac surgery-associated acute kidney injury (CS-AKI) remain unexamined. We assessed racial/ethnic disparities in CS-AKI prevalence and outcomes in neonates undergoing congenital heart surgery.<h4>Design, setting, and patients</h4>Retrospective cohort of neonates across 22 centers (2015-2018) from the Pediatric Cardiac Critical Care Consortium and the Neonatal and Pediatric Heart and Renal Outcomes Network.<h4>Measurements</h4>Race/ethnicity (primary predictor) were categorized into eight groups; CS-AKI was defined using neonatal modified Kidney Disease: Improving Global Outcomes criteria as mild (stage 1), severe (≥ stage 2) and persistent (any stage beyond post-operative day [POD] 3). Outcomes included not achieving net negative fluid balance [FB], respiratory support duration, length of stay [LOS], and mortality. Multivariable regression models examining the association between race/ethnicity and outcomes were also constructed.<h4>Results</h4>Among 2,165 neonates (2% Asian, 18% Hispanic, 1% multi-racial, 0.3% Native American, 14% Non-Hispanic Black (NHB), 59% Non-Hispanic White (NHW), 5% other, and 0.2% Pacific Islander), CS-AKI prevalence (31% mild, 23% severe) did not differ across racial/ethnic groups. NHB neonates had lower birthweight and higher surgical complexity, while NHW neonates had higher private insurance rates. NHB neonates were less likely to not achieve a net negative FB until ≥ POD3 than NHW neonates, but no differences were found in respiratory support duration, hospital LOS, or mortality after multivariate adjustment. Non-white neonates with severe AKI had higher mortality but had similar clinical outcomes.<h4>Conclusions</h4>CS-AKI rates did not vary by race/ethnicity. Differences in fluid balance achievement were noted but no significant disparities were found in mortality, mechanical ventilation duration, or hospital LOS.

Ferroptosis is a novel form of cell death induced by ferrous ions and lipid peroxidation. However, the mechanisms of ferroptosis-related genes (FRGs) in sepsis have not been studied thoroughly. We performed differential analysis using GSE65682, and the differentially expressed genes (DEGs) were intersected with FRGs to obtain the differentially expressed FRGs. We constructed a random forest model to explore characteristic FRGs for sepsis diagnosis using the training set and verified its predictive efficacy using the testing set. There are 43 differentially expressed FRGs and the top five FRGs in the model are MAFG, QSOX1, KLF2, TXN, and PEBP1. Meanwhile, three genes remained after the univariate Cox analysis, survival analysis and nomogram, but only MAFG was validated to be associated with sepsis prognosis. MAFG was selected as the most diagnostically and prognostically significant FRG based on the following evidence: (1) consistently significant overexpression across multiple datasets, (2) the highest MeanDecreaseGini score in the random forest model, (3) the largest hazard ratio in univariate Cox regression analysis, (4) a strong association with patient survival demonstrated by the nomogram, and (5) an AUC of 0.64 (p < 0.05) in GSE185263 in the ROC analysis for sepsis prognosis. Subsequently, TXN was predicted as a ferroptosis-related potential target for the transcription factor MAFG, and their elevation in sepsis was confirmed by RT-qPCR. Ultimately, we discovered that MAFG was mainly localized in monocytes by single-cell RNA-sequencing analysis, which was significantly upregulated in sepsis and non-survivors of sepsis. In this study, we identified MAFG as a potential candidate FRG related to the diagnosis and prognosis of sepsis, although further validation is required, which broadens novel insights into the therapeutic targets of sepsis and the role of ferroptosis in sepsis.

Glioma is one of the most malignant tumors in the brain, with high tumor heterogeneity. Traditional treatment methods, such as surgery, radiotherapy and chemotherapy, face numerous challenges and still pose serious threats to human health. Therefore, it is urgent to explore new treatment directions. The immune checkpoint B7-H3 has attracted much attention due to its significant immunomodulatory function in various cancers. Studies have shown that it is highly expressed in glioma and lowly expressed in normal brain tissue. It not only has immunosuppressive function and induces immune escape in glioma, but also promotes the proliferation, migration and angiogenesis of glioma cells. Moreover, there are many immunotherapy strategies targeting B7-H3, such as monoclonal antibodies, antibody-drug conjugates (ADC) and chimeric antigen receptor T-cell (CAR-T) therapy. Therefore, B7-H3 is regarded as a potential target for glioma immunotherapy. This article systematically reviews the occurrence and development process of B7-H3 in glioma, changes in the immune microenvironment, and related treatment methods. Through analysis of several databases such as TCGA, CTPAC, HPA and GEPIA, this study explores the expression of B7-H3 in glioma, its correlation with prognosis and its correlation with the glioma immune microenvironment, providing a new research target for the diagnosis and treatment of glioma.

Protocols for deriving midbrain dopaminergic (mDA) neurons for Parkinson's disease (PD) modeling and therapy remain incompletely benchmarked against in vivo references. To establish transcriptomic standards, we generated an integrated human fetal whole-brain atlas and a midbrain subatlas. Whole-brain analysis revealed strong region-specific signatures, underscoring the need for global mapping before refined midbrain annotation. We implemented this two-tier strategy, BrainSTEM (Brain Single-cell Two tiEr Mapping), to systematically reassess published single-cell datasets of human midbrain culture models. BrainSTEM confirmed the presence of bona fide midbrain cell types ("on-target"), but also revealed substantial populations aligning with nonmidbrain regions ("off-target"), inflating reported mDA yields across protocols. This unbiased framework enables rigorous evaluation of differentiation outcomes, clarifies current limitations of midbrain-directed models, and provides a foundation for refining protocols toward more faithful in vitro systems for PD research and regenerative applications.

Major depressive disorder (MDD) with atypical features accompanied by psychotic symptoms represents a severe and under-researched subtype of depression and severe mental illness, characterized by significant personal and social impact. This study aims to explore novel biomarkers through a precision medicine approach by combining clinical data, white blood cell (WBC) single-cell RNA sequencing (scRNA-seq), plasma proteomics, and brain organoid models to uncover immunological and neurological alterations in patients with this condition. Patients exhibited elevated stress, anxiety, depression, and increased WBC counts, although the difference in WBC count is not significant after adjusting for age. Plasma proteomic profiling identified an upregulation of proteins implicated in synaptic formation, including Doublecortin-Like Kinase 3 (DCLK3) and Calcyon (CALY), as well as immune-related proteins such as Complement Component 5 (C5). WBC scRNA-seq revealed significant neutrophil and monocyte transcriptomic alterations, suggesting increased inflammation and immune dysregulation. Patient-derived brain organoids display reduced growth and distinct gene expression patterns compared to controls, particularly under dexamethasone-induced stress conditions. Combining WBC scRNA-seq, plasma proteomics, and brain organoid models offers a novel framework for understanding the pathophysiology of psychiatric disorders, which is one of the most complex disorders.

Deep brain stimulation has been used to treat severe, refractory obsessive-compulsive disorder (OCD) with variable outcomes across multiple anatomical targets. To overcome these limitations, we developed an invasive brain mapping paradigm in which electrodes were implanted across the OCD cortico-striato-thalamo-cortical circuit. We then performed extensive stimulation mapping during a multi-day inpatient stay to identify personalized therapeutic targets and characterize their downstream circuit effects. We found two targets within the right ventral capsule (VC) that acutely reduced OCD symptoms. Prolonged VC stimulation suppressed high frequency activity within the structurally and functionally connected orbitofrontal and cingulate cortex, which were identified to be cortical nodes encoding the severity of OCD symptoms. These VC sites were implanted for DBS and combined stimulation of these targets led to a rapid therapeutic response. This case provides the first proof-of-concept that invasive brain mapping can be used to guide a novel personalized, multi-site neuromodulation approach to treat refractory OCD.

Inherited biallelic mutations in the CLN7 gene result in the variant late infantile onset neuronal ceroid lipofuscinosis, a subtype of Batten disease (BD), a severe and fatal childhood neurodegenerative disease. Intriguingly, CLN7 genetic variants have also been associated with retinopathies, amyotrophic lateral sclerosis, and frontotemporal dementia. CLN7 encodes a transmembrane protein localizing to endolysosomal membranes with outward-facing chloride channel activity. Loss of CLN7 function results in cortical neurons accumulating swollen lipofuscin-containing lysosomes, leading to neuroinflammation and neurodegeneration. The molecular mechanisms underlying CLN7 BD neuropathology are not completely understood. We have generated iPSC lines from two CLN7 BD patients and age-matched unaffected controls to interrogate intracellular molecular phenotypes in iPSC-derived neural progenitor cells (iNPC). Taking a multi-omics approach we have identified disease-modified activities in endolysosomal transport in iNPC^BD that lead to lysosomal dysfunction and decreased mitophagy, resulting in the accumulation of metabolically defective mitochondria. We further observe a breakdown in nuclear functions that centre on RNA processing and nuclear export, linking to CLN7 protein interactions at the stress granule. We have identified dual and distinct functions for CLN7, promoting cell survival during the cellular stress response. CLN7 loss of function in BD results in neuronal apoptosis.

Motor neuron hyperexcitability is a broadly observed yet poorly understood feature of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Nuclear depletion and cytoplasmic aggregation of the RNA splicing protein TAR DNA-binding protein 43 (TDP-43) are observed in most ALS and FTD patients. Here we show that TDP-43 dysfunction causes mis-splicing of KCNQ2, which encodes a voltage-gated potassium channel (K_v7.2) that regulates neuronal excitability. Using iPSC-derived neurons and postmortem ALS/FTD brain and spinal cord tissue we find widespread, disease-specific and TDP-43-specific skipping of an exon encoding the KCNQ2 pore domain. The mis-spliced mRNA escapes degradation and is translated into a nonfunctional protein with severely reduced ion conductance that aggregates in the endoplasmic reticulum and causes intrinsic hyperexcitability in ALS neuronal models. This event, which correlates with higher phosphorylated TDP-43 levels and earlier age of disease onset in patients, can be rescued by splice-modulating antisense oligonucleotides that dampen hyperexcitability in induced pluripotent stem cell cortical neurons and spinal motor neurons with TDP-43 depletion. Our work reveals that nuclear TDP-43 maintains the fidelity of KCNQ2 expression and function and provides a mechanistic link between established excitability disruption in ALS/FTD patients and TDP-43 dysfunction.

<h4>Objectives</h4>To prospectively evaluate the diagnostic accuracy of ultrasound-derived fat fraction (UDFF) in quantifying hepatic steatosis, to establish and validate a dual-threshold UDFF classification system, and to investigate its efficacy for risk stratification in body mass index (BMI)-defined subgroups.<h4>Materials and methods</h4>This prospective multicenter study involved 790 suspected metabolic dysfunction-associated steatotic liver disease (MASLD) participants from April 2023 to November 2024 (derivation: n = 553; validation: n = 237). Liver biopsy histopathology (n = 342), MRI proton density fat fraction (MRI-PDFF) (n = 396), or proton magnetic resonance spectroscopy (¹H-MRS) (n = 52) was used as the reference standard. UDFF was compared to noninvasive test Hepatic Steatosis Index (HSI) and Fatty Liver Index (FLI) using area under the curve (AUC). The diagnostic thresholds were optimized to maintain at least 90% sensitivity and specificity in stratifying hepatic steatosis severity. A two-step strategy of UDFF followed by HSI was used to rule in and rule out steatosis at BMI ≥ 23 kg/m² subgroup.<h4>Results</h4>UDFF demonstrated significant correlations with three reference standards (Spearman's ρ = 0.798-0.847). Comparing with HSI and FLI, UDFF showed higher AUC (0.933, 0.948, and 0.914, respectively) for assessing ≥ S1, ≥ S2 and S3. A clinically practical dual-threshold system effectively classified hepatic steatosis severity. A sequential UDFF/HSI strategy achieved a high positive predictive value (PPV = 95.8%) to rule in hepatic steatosis and lowered the proportion of indeterminate cases (from 18.0 to 7.6%) in patients with BMI ≥ 23 kg/m².<h4>Conclusion</h4>UDFF is a highly effective noninvasive tool for quantifying hepatic steatosis. A sequential use of UDFF/HSI could improve hepatic steatosis detection in patients with BMI ≥ 23 kg/m².<h4>Critical relevance statement</h4>The study proposed dual-threshold diagnostic criteria (sensitivity/specificity ≥ 90%) of UDFF for steatosis grading, and established a BMI-stratified risk stratification tool in multi-center cohorts, proving the efficacy of UDFF in noninvasively quantifying liver steatosis.<h4>Key points</h4>Early diagnosis of hepatic steatosis holds critical clinical significance. The study proposed dual-threshold ultrasound-derived fat fraction (UDFF) criteria and BMI-stratified steatosis risk prediction strategy. UDFF provided a non-invasive, accurate diagnostic alternative to liver biopsy and MRI.

<h4>Introduction</h4>Magnesium homeostasis is critical for cellular growth and metabolism, yet its pan-cancer implications remain poorly characterized. This study aims to comprehensively analyze magnesium homeostasis across 33 cancer types, exploring its role in tumorigenesis, immune regulation, and therapeutic potential. Key magnesium homeostasis-related genes (e.g., ANK3, CNNM2) were significantly downregulated in most tumors, correlating with improved prognosis. Magnesium homeostasis scores (MHS) were reduced in cancers and linked to lower tumor mutational burden (TMB), microsatellite instability (MSI), immune dysfunction, and checkpoint gene expression. Single-cell sequencing revealed elevated MHS in CD8 + T cells, suggesting immune modulation roles.<h4>Conclusion</h4>These findings highlight magnesium homeostasis as a regulator of tumor progression and immunity, with MHS serving as a prognostic biomarker. Targeting magnesium pathways may offer novel therapeutic strategies, warranting further clinical validation to advance personalized cancer therapies.

Immunosenescence is the gradual deterioration in the functionality of the immune system that has various clinical manifestations, such as a weakened response to vaccination, higher susceptibility to viral/bacterial/fungal infection, and higher incidence rates regarding the ageing-based disorders. Conventionally used models, like animal or 2D models for unraveling the interactions between ageing and immunity are unable to depict the complexity of bodily environments. However, this major limitation can be addressed via using the organoid technology. For clarification, organoids are tissue-resembling 3D structures that are generated from stem cells; they have the major superiority of preserving the human physiology, multicellular intricateness, and the capability of dynamic interactions between the existing elements. Organoid-based technology has been applied for the study of different organ-specific immunosenescence, such as in the intestine, brain, liver, and skin. Besides, organoids offer the bright innovative future of senolytic CAR T lymphocytes and other regenerative-based therapies. This review narrates the cutting-edge application of organoids and the mechanisms involved in the detailed molecular processes of organ-specific immunosenescence.

<h4>Background</h4>The discovery of RNA 5-methyluridine (m5U) modifications is vital in computational biology due to their essential significance in different biological processes. This study presents a powerful predictor named 5-meth-Uri, which improves the overall accuracy of m5U modification predictions. The proposed method combines a composite of dinucleotide and trinucleotide-based auto-cross covariance with six physicochemical parameters to generate a feature vector. We select essential features using an unsupervised Principal Component Analysis (PCA) technique to enhance the model's efficiency. Finally, an intelligent computation Deep Neural Network (DNN) was utilized to classify RNA 5-methyluridine (m5U).<h4>Results</h4>The model's performance was evaluated on two benchmark datasets, namely mature mRNA and full transcript, using tenfold cross-validation and an independent test. The proposed model achieved an average accuracy of 95.13% and 97.36% on the training set and 95.73% and 96.51% on the independent test sets for the full transcript and mature mRNA datasets, respectively. 5-meth-Uri demonstrated around 7.15% and 3.98% higher accuracy on training data and 5.42% and 3.52% higher accuracy on independent samples for both datasets compared to existing models.<h4>Conclusions</h4>Its high accuracy and reliability make 5-meth-Uri a promising tool for researchers with potential biomedical and pharmaceutical applications.

<h4>Background</h4>Intestinal stem cells (ISCs) sustain epithelial homeostasis through rapid mitochondrial metabolism, however, how they sense nutrient signals to regulate mitochondrial function remains unclear.<h4>Methods</h4>We examined the role of L-glutamate (Glu) in regulating cell mitochondrial biosynthesis using in vivo piglets, ex vivo porcine intestinal organoids (IOs), and in vitro IPEC-J2 cells.<h4>Results</h4>Glu enhanced jejunal development in weaned piglets. Isobaric tags for relative and absolute quantitation (iTRAQ) analysis revealed the significant enrichment of mitochondrial functions and activation of EGFR-MEK-ERK-mTFB2 signaling pathway in the jejunum. In vitro, 5 mM Glu promotes mitochondrial biosynthesis and potentiates the EGFR-MEK-ERK-mTFB2 axis. Whereas inhibition of EGFR with Osimertinib and silencing EGFR abolished these effects in IOs and IPEC-J2 cells. Colocalization and biochemical studies demonstrated interaction between Glu and EGFR in IOs.<h4>Conclusions</h4>Glu promotes mitochondrial biogenesis and ISC expansion by activating the EGFR-MEK-ERK-mTFB2 axis, highlighting a nutrient-sensing mechanism that couples energy availability to ISC function.

<h4>Background</h4>The cycling intestinal stem cells (ISCs) exhibit radiosensitivity, and their death or impaired regenerative capacity following irradiation may result in intestinal barrier dysfunction. The cystathionine-β-synthase (CBS)/H2S axis plays a critical role in regulating cell proliferation, reactive oxygen species scavenging, and the DNA damage response. However, it remains unclear whether the CBS/H2S axis modulates ISC homeostasis and tissue radiosensitivity.<h4>Methods</h4>Intestinal epithelium specific conditional CBS knockout mice were generated by crossing CBS^fl/+ mice with Villin-CreERT2 mice. CAGGCre-ER™ mice were crossed with CBS^fl/fl mice to achieve CBS knockout in multiple tissues and cell types. The Lgr5-Tdtaomato-Flag mice were generated by CRISPR/Cas9 system. The CBS inhibitor AOAA or the H2S donor GYY4137 was used to treat mice or intestinal crypt organoids. Hematoxylin and eosin, immunohistochemistry, immunofluorescence, Western blot, qRT-PCR, et al. were employed to investigate the role of the CBS/H2S axis in ISCs homeostasis and radiation-induced intestinal damage.<h4>Results</h4>Lgr5 + ISCs and progenitor cells expressed higher levels of CBS than differentiated cells. The cecum and colon expressed significant higher CBS levels than the small intestine. Treatment with the H2S donor GYY4137 enhanced the proliferation of intestinal organoids in vitro, while inhibition of CBS by AOAA reduced this effect. Genetic knockout of CBS in the intestinal epithelium or global downregulation of CBS driven by CAGG-CreER™ in vivo did not affect ISC proliferation or differentiation under physiological conditions. Pharmacological regulation of the CBS/H2S axis in vitro failed to protect organoids from radiation-induced damage. Interestingly, administration of AOAA in vivo reduced radiation-induced atrophy of the intestinal mucosa. Furthermore, global downregulation of CBS significantly promoted ISC recovery after irradiation exposure. However, intestinal epithelium-specific CBS knockout did not confer radioprotective effects.<h4>Conclusions</h4>Our findings suggest that the CBS/H2S axis contributes to the regulation of ISC homeostasis and represents a potential target for radiation protection, mediated through the intervention of non-epithelial cells.

We investigate the relationship between glycemic traits-specifically type 2 diabetes mellitus, fasting glucose, fasting insulin, glycated hemoglobin, and 2-hour post-load glucose-and thoracic aortic morphology and diseases. The results indicate an inverse association between elevated glycemic traits and aortic morphology, as well as a reduced risk of thoracic aortic aneurysm. Genetic predictors related to beta-cell proinsulin mechanisms in type 2 diabetes mellitus drive these associations. Key genes such as AGER, GLRX, TCF7L2, and GCK are implicated, highlighting their potential as therapeutic targets for the prevention and treatment of thoracic aortic aneurysm, given their role in glycemic control medication.

Cepharanthine (Cep), a natural alkaloid from Stephania (Menispermaceae), exhibits broad-spectrum anti-cancer activity. In the present study, Cep was found to induce ferroptosis and mitophagy, for which the relationship and upstream targets remain unelucidated. Herein, the role of Cep in the induction of mitophagy was deeply investigated. Cep showed robust anti-lung cancer effects, as confirmed by decreased cell viability, elevated apoptosis, suppressed colony formation and inhibited growth of tumor grafts in Lewis cell-bearing mice. RNA-sequencing analysis revealed that Cep treatment significantly enriched differentially expressed genes (DEGs) in mitophagy and ferroptosis pathways, which were demonstrated in in vitro and in vivo experiments as well. In-depth investigations showed that inhibition of autophagy abolished Cep-mediated ferroptosis, but not vice versa. Moreover, genetic knockdown of BNIP3 dampened the mitophagy and ferroptosis of lung cancer cells induced by Cep. Additional data confirmed that Cep could bind to and thereby inhibit FBXL4, which attenuated the ubiquitination of BNIP3. FBXL4-mediated BNIP3 activation promoted the recruitment of LC3 to mitochondria and autophagic flux in the presence of Cep. Collectively, Our study elucidates a complete mechanistic pathway wherein Cep activates BNIP3-mediated mitophagy by inhibiting FBXL4, ultimately driving ferroptosis and offering a new therapeutic avenue for lung cancer.

<h4>Objective</h4>To identify prenatal risk factors associated with early cord clamping (ECC) in infants born extremely preterm (22-28 weeks) and to use these factors to predict which infants have the highest risk of ECC.<h4>Study design</h4>Stepwise logistic regression and classification and regression tree analysis were performed to identify variables most associated with ECC and to build a parsimonious model to identify infants born preterm who are at the highest risk of ECC.<h4>Results</h4>There were 12 622 infants eligible for analysis (ECC, n = 8465 vs deferred cord clamping [DCC], n = 4157). In the classification and regression tree model, center, lack of antenatal magnesium, cesarean delivery, lower gestational age, and antenatal hemorrhage, in that rank order, were the factors that contributed most to identifying which infants received ECC. Several additional factors were also statistically significant, including time to delivery after admission to the hospital as well as maternal race and ethnicity.<h4>Conclusions</h4>Our results suggest that there are multiple factors among extremely preterm pregnancies, including center, lack of antenatal magnesium, cesarean delivery, lower gestational age, and antenatal hemorrhage, that are associated with infants who are more likely to miss DCC. There are also differences in rates by maternal race and ethnicity in who receives DCC compared with ECC. Identifying these factors now can inform future research and encourage local and larger-scale quality improvement practice changes.

Cancers are characterized by genomic instability events such as aneuploidy, chromosomal arm imbalances, and segmental copy number changes. These genomic features frequently define different subclones within a tumor. Single-cell DNA sequencing (scDNA-seq) identifies these large-scale genomic alterations that define subclonal features. However, scDNA-seq does not provide biological phenotypic information on individual subclones. Single-cell RNA sequencing (scRNA-seq) offers biological information but is less accurate in discovering genomic instability events. We developed a computational framework, scAlign, for integrating scRNA-seq and scDNA-seq from the same specimen and define subclonal cellular phenotypes at the resolution of individual cells. Subclones were defined by aneuploidy and chromosomal arm imbalance among primary and metastatic cancers. Using the cells in the G0/G1 phase, the extensive cellular sampling from both assays characterized the subclonal architecture of these cancers. The scDNA-seq provided a ground truth for copy number-based subclones. From the scRNA-seq data, the epithelial cells in G0/G1 were identified and assigned to specific subclones by the scAlign based on gene dosage. Afterward, we determined the differential gene expression and biological pathway activities of specific clones. Overall, integrative multi-omics analysis of single-cell datasets is more informative than any individual genomic modality, provides deep insights into intratumoral heterogeneity and reveals subclonal biology. scAlign is available at https://github.com/XQBai/scAlign.

<b>Background/Objective:</b> Biochemical recurrence (BCR) after radical prostatectomy (RP) for prostate cancer indicates disease progression. Although type 2 diabetes mellitus (T2D) shows a paradoxical association with prostate cancer risk, the prognostic role of T2D-related genetic variants remains unclear. <b>Methods:</b> We analyzed 113 common T2D susceptibility-related single-nucleotide polymorphisms (SNPs) in 644 Taiwanese men with localized prostate cancer (D'Amico risk classification: 12% low, 34% intermediate, and 54% high) treated with RP. Associations between SNPs and BCR were assessed using Cox regression, adjusting for key clinicopathological factors. Functional annotation was performed using HaploReg and FIVEx, while The Cancer Genome Atlas transcriptomic data were analyzed for C2 calcium-dependent domain-containing 4A (<i>C2CD4A</i>) expression. Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were applied to explore related biological pathways. <b>Results:</b><i>C2CD4A</i> SNP <i>rs4502156</i> was independently associated with a reduced risk of BCR (hazard ratio = 0.80, <i>p</i> = 0.035). The protective <i>C</i> allele correlated with higher <i>C2CD4A</i> expression. Low <i>C2CD4A</i> expression is associated with advanced pathological stages, higher Gleason scores, and disease progression. GSEA revealed negative enrichment of mitotic and chromatid segregation pathways in high-<i>C2CD4A</i>-expressing tumors, with E2F targets being the most suppressed. GSVA confirmed an inverse correlation between <i>C2CD4A</i> expression and E2F pathway activity, with <i>CDKN2C</i> as a co-expressed functional gene. <b>Conclusions:</b> The T2D-related variant <i>rs4502156</i> in <i>C2CD4A</i> independently predicts a lower risk of BCR, potentially via suppression of the E2F pathway, and may serve as a germline biomarker for postoperative risk stratification.

The study of molecular mechanisms underlying late-life depression (LLD) is increasingly important in light of population aging. To date, LLD-related molecular brain changes remain poorly understood. Furthermore, environmental factors such as climate change and geography contribute to LDD risks. One overlooked factor might be deuterium-a stable hydrogen isotope-whose concentration in drinking water can vary geographically (~90-155 ppm) and alter the incidence of mood disorders. Conversely, potential effects of natural variations in deuterium content in drinking water on LLD symptoms and brain gene expression remain unknown. We conducted Illumina gene expression profiling in the hippocampi and prefrontal cortexes of 18-month-old C57BL/6J mice, a model of LLD-like behaviors, compared to 3-month-old controls. Separately, aged mice were allowed to consume deuterium-depleted (DDW, ~90 ppm) or control (~140 ppm) water for 21 days and were studied for LLD-like behaviors and Illumina gene expression of the brain. Naïve old mice displayed ≥2-fold significant changes of 35 genes. Housing on DDW increased their hedonic sensitivity and novelty exploration, reduced helplessness, improved memory, and significantly altered brain expression of <i>Egr1</i>, <i>Per2</i>, <i>Homer1</i>, <i>Gadd45a</i>, and <i>Prdx4</i>, among others. These genes revealed significant alterations in several GO-BP and KEGG pathways implicated in inflammation, cellular stress, synaptic plasticity, emotionality, and regeneration. Additionally, we found that incubation of primary neuronal cultures in DDW-containing buffer ameliorated Ca²⁺ influx and mitochondrial potential in a toxicity model, suggesting the involvement of mitochondrial mechanisms in the effects of decreased deuterium levels. Thus, aging induced profound brain molecular changes that may at least in part contribute to LLD pathophysiology. Reduced deuterium intake exerted modest but significant effects on LLD-related behaviors in aged mice, which can be attributed to, but not limited by ameliorated mitochondrial function and changes in brain gene expression.

Thyroid eye disease (TED) is an autoinflammatory condition characterized by fibrosis in orbital fat and extraocular muscles, primarily driven by TSH receptor antibodies and inflammatory cytokines. While research has predominantly focused on the involvement of fat tissue, the understanding of myopathy in TED remains limited. This study developed a TED mouse model and isolated myoblasts from both control individuals and TED patients for analysis. Single-cell RNA sequencing was used to investigate myofiber type changes in TED and their alterations following treatment with human-derived mesenchymal stem cells. Key regulatory genes involved in myofiber differentiation and fibrosis in myofibroblasts were identified, and their expression balance was validated in myoblasts derived from both normal individuals and TED patients. Our analysis revealed a disease-associated shift in myofiber types and identified Six1 and Eya1 as central regulators of myofiber differentiation and fibrosis suppression. These regulatory effects were validated in primary myoblasts isolated from both control and TED patients. Collectively, our findings uncover a novel role for the Six1/Eya1 axis in modulating muscle remodeling and fibrosis in TED and provide a foundation for the development of targeted therapies for TED-associated myopathy.

<h4>Background</h4>Hyperuricemia (HUA), a metabolic disorder characterized by high serum uric acid (UA) level, presents a growing global health challenge.<h4>Method</h4>In this study, a stable murine model of HUA was established by orally administering adenine (100 mg/kg) and potassium oxonate (600 mg/kg) in C57BL/6J mice, resulting in significant elevation of serum UA and xanthine oxidase (XOD) activity, as well as renal pathological alterations. Given the anti-hyperuricemia potential of <i>Lactiplantibacillus plantarum</i> WLPL04, a strain from a human breast milk was evaluated.<h4>Conclusions</h4>Oral administration of <i>L. plantarum</i> WLPL04 significantly reduced serum UA level and XOD activity in a dose-dependent manner. Moreover, <i>L. plantarum</i> WLPL04 treatment enhanced UA excretion by upregulating <i>ABCG2</i> and downregulating <i>URAT1</i> and <i>GLUT9</i> expression. It ameliorated renal injury and suppressed inflammation via downregulation of the NLRP3 inflammasome pathway. 16S rRNA gene sequencing revealed that <i>L. plantarum</i> WLPL04 restored gut microbial diversity and promoted the enrichment of beneficial genera such as <i>Bacteroides,</i> which was negatively correlated with UA in serum, creatinine, and inflammatory cytokines. Moreover, transcript analysis revealed upregulation of purine salvage genes (<i>hpt</i> and <i>xpt</i>), suggesting enhanced salvage pathway recycling of purine bases and reduced urate production. Those findings suggest that <i>L. plantarum</i> WLPL04 exerted multi-targeted anti-hyperuricemia effects through coordinated regulation of host purine metabolism, urate transport, inflammation, and gut microbiota composition, providing a promising probiotic candidate for HUA management.

<b>Background/Objectives</b>: Fonte Essenziale^®, a mineral water rich in bicarbonate, sulfate, calcium, and magnesium, has shown potential in modulating the gut-liver axis and microbiota in hepatic steatosis. However, its long-term effects on intestinal permeability (IP), systemic inflammation (SI), and oxidative stress-key factors in Metabolic dysfunction-associated steatotic liver disease (MASLD)-remain unexplored. <b>Methods</b>: Eighty-seven MASLD patients were randomized into two groups: group A received Fonte Essenziale^® (400 mL/day, fasting) plus a controlled nutritional regimen for 12 months, followed by a 6-month water washout; group B followed only the controlled nutritional regimen. IP markers, SI (IL-1β, IL-6, TNF-α), oxidative stress (dROMs/BAP), and clinical data (including Controlled Attenuation Parameter-CAP) were assessed at baseline (T0), 12 months (T12), and post-washout (T18). Baseline increased IP (in-IP) was defined by fecal zonulin > 110 ng/mL and serum LBPp > 10 µg/mL; improvement (im-IP) required normalization of both. A ≥30% CAP reduction indicated steatosis improvement. <b>Results</b>: Thirty-eight patients in group A and thirty-nine in group B completed the study. At T12, group A showed significant reductions in fecal zonulin (<i>p</i>: 0.0163) and serum LBPp (<i>p</i> < 0.0001), with increased occludin and claudin 1 (all <i>p</i> < 0.0001). Im-IP prevalence was higher in group A (<i>p</i>: 0.0037). Group A also showed significant reductions in IL-1β, TNF-α, IL-6, LPS, and dROM/BAP (all <i>p</i> < 0.05), especially among those with im-IP. CAP, insulin, and HDL levels improved significantly (all <i>p</i> < 0.0001). Multivariate analysis confirmed water intake (aOR: 2.185, <i>p</i>: 0.001) and im-IP achievement (aHR: 1.267, <i>p</i>: 0.021) as predictors of steatosis improvement. Benefits persisted at T18. <b>Conclusions</b>: Prolonged Fonte Essenziale^® intake improved hepatic steatosis and MASLD outcomes by modulating IP, SI, and oxidative stress. This trial has been registered on clinicaltrials.gov (NCT07211113).

<h4>Introduction</h4>TWIST1, a bHLH transcription factor, regulates mesenchymal specification, differentiation, proliferation and migration during development and in diseases such as cancer. More recently, genome-wide association studies have identified TWIST1 as a causal gene that increases risk for multiple vascular diseases, including atherosclerosis and hypertension. However, its molecular role in the vascular wall remains unclear.<h4>Methods</h4>In this study, we interrogated how TWIST1 dimer composition and discrete TWIST1 domains affect SMC phenotype by expressing forced TWIST1 dimers or TWIST1 variants lacking specific domains, followed by bulk RNA sequencing and proliferation and migration assays in human coronary artery SMCs (HCASMCs).<h4>Results</h4>We found that TWIST1 homodimers had only modest transcriptomic effects but strongly promoted migration and proliferation-effects abolished by deletion of the TWIST1 N-terminus. Heterodimerization of TWIST1 with TCF3-encoded E proteins resulted in larger transcriptomic effects, promoting Rho/ROCK signaling and extracellular matrix production/organization, but had only modest effects on proliferation and no effect on migration. Deletion of the TWIST1 C-terminus resulted in a very large transcriptomic shift with predicted downregulation of angiotensin and Rho/ROCK signaling as well as ECM production/organization pathways, in a manner suggesting a dominant negative effect on TWIST1-E12 function. Comparison with single-cell RNA-seq data from human endarterectomy samples placed the function of these TWIST1 variants in a disease context and showed that deletion of the C-terminal domain prevented a modulated SMC phenotype.<h4>Discussion</h4>These studies demonstrate that TWIST1 influences different aspects of SMC phenotype independently via discrete domains and dimer composition, and link TWIST1 to key signaling pathways that influence SMC phenotype during disease.

Cell membrane-camouflaged nanoparticles (CNPs) have emerged as promising multifunctional platforms for colorectal cancer therapy, integrating drug delivery, immunomodulation, photothermal ablation, and anti-inflammatory effects. This review highlights recent advances in CNP-based strategies, emphasizing their unique capacity to enhance tumor-targeting specificity, potentiate immunotherapeutic efficacy, and overcome the limitations of conventional treatments. We summarize diverse approaches employing immune cell or tumor cell membrane coatings, as well as hybrid systems that combine CNPs with chemotherapy, metabolic modulation, or photothermal therapy. Accumulating evidence demonstrates that CNPs can effectively remodel the tumor immune microenvironment, increase the bioavailability of hydrophobic drugs, and promote synergistic therapeutic outcomes. Despite these encouraging results, clinical translation remains constrained by challenges in biodegradability, biosafety, large-scale manufacturing, and cost. Ongoing clinical trials are evaluating the safety and therapeutic potential of CNP-based nanomedicines. Overall, this review underscores the transformative role of CNPs as a next-generation platform for precision and personalized therapy in colorectal cancer.

Patients with liver cirrhosis experience a significantly reduced health-related quality of life (HRQoL) due to a multifaceted interaction of physiological, psychological, and social factors. Identifying these key determinants is crucial for enhancing care strategies and treatment outcomes. This study systematically explored the factors influencing both negative and positive aspects of HRQoL in patients with liver cirrhosis. A comprehensive literature search was performed across PubMed, Scopus, Embase, and Google Scholar, focusing on original research published since January 1, 2021. Studies involving sarcopenia in cirrhosis and its impact on HRQoL, including various subgroups and influencing factors, were included to ensure a wide-ranging analysis. The review identified multiple significant risk factors associated with poor HRQoL in cirrhotic patients. These included advanced age, female gender, lower socioeconomic status, low body mass index, and the presence of comorbid conditions such as anxiety and depression. Additionally, complications like ascites, hepatic encephalopathy, abnormal endoscopic findings, higher disease severity scores, sarcopenia, disturbed sleep, impaired sexual function, muscle cramps, and elevated levels of bilirubin, prothrombin time, and albumin-bilirubin ratio were linked to diminished HRQoL. On the other hand, interventions such as increased physical activity, liver transplantation, stem cell therapy, mindfulness practices, and the use of probiotics, rifaximin, and lactulose showed positive effects on HRQoL. These insights underscore the importance of adopting a comprehensive management approach for liver cirrhosis that addresses not only physical health but also psychological and social well-being. Future research should aim at developing targeted interventions to reduce the burden of these risk factors and promote overall quality of life in this patient population.

The field of non-coding RNA research is advancing at a breathtaking pace, continually uncovering new layers of regulatory complexity and functional diversity [...].

Teeth are the hardest organs in the human body. As mineralized structures, they possess a unique microstructure composed of orderly arranged piezoelectric materials such as hydroxyapatite crystals and collagen fibers. Teeth exhibit effective piezoelectric coefficients of approximately 1.2-1.6 pC/N. This inherent property enables teeth to function as natural piezoelectric sensors, converting routine mechanical stresses (e.g., chewing and biting forces, typically ranging from 22.4 to 68.3 kg) into localized electrical signals. This characteristic is of great importance in dentistry and materials science, offering new perspectives into a deeper understanding of the physiological functions and pathological mechanisms of teeth. Despite promising advances, challenges regarding the clinical translation, long-term stability, and biosafety of piezoelectric materials in the oral environment remain unresolved. This review highlights the biological functions of the piezoelectric properties of teeth, discusses recent applications and notable advancements of piezoelectric materials in dentistry, and outlines the challenges and research priorities for future clinical applications.

<h4>Background and aim</h4>Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide widely implicated in stress responses, appetite regulation, and emotional behavior. While PACAP has been linked to stress-induced appetite suppression, its role in circadian feeding and stress-related hyperphagia remains poorly defined. This study aimed to clarify the contribution of PACAP to circadian feeding behavior, chronic stress-induced hyperphagia, and emotional regulation in mice.<h4>Materials and methods</h4>Experiments were conducted using PACAP knockout (KO) and wild-type (WT) CD-1 mice. Acute and chronic restraint stress paradigms were applied, and food intake, body weight, and behavioral assays were recorded. Viral-mediated PACAP overexpression in the ventromedial hypothalamus (VMH) was performed using adeno-associated vectors. Emotional regulation was assessed through forced swim and tail suspension tests (TSTs). PAC1 receptor expression was quantified by reverse transcription and quantitative polymerase chain reaction.<h4>Results</h4>PACAP overexpression in the VMH significantly increased nocturnal feeding, demonstrating a circadian-specific effect on appetite regulation. Chronic restraint stress enhanced food intake in both PACAP KO and WT mice, whereas acute stress showed no effect, indicating that chronic stress-induced hyperphagia occurs independently of PACAP signaling. Behaviorally, PACAP-overexpressing mice exhibited reduced immobility in forced swim and TSTs, consistent with enhanced stress resilience and antidepressant-like effects. Importantly, PAC1 receptor expression remained stable throughout the diurnal cycle, suggesting that PACAP's modulatory effects are driven by neuropeptide availability rather than receptor fluctuations.<h4>Conclusion</h4>This study identifies PACAP in the VMH as a key modulator of circadian feeding and emotional behavior, while demonstrating its non-essential role in chronic stress-induced hyperphagia. The findings suggest that PACAP selectively integrates circadian and emotional signals to regulate feeding, independent of compensatory neuropeptide systems that mediate stress hyperphagia. These insights advance the understanding of neuropeptide regulation of energy balance and mood, with implications for stress-related eating disorders and anxiety.

<b>The aim of the study is</b> to analyze the manifestation of selected cellular senescence markers on the models of replicative senescence, stress-induced senescence, and chronological aging of human mesenchymal stem cells and to study the feasibility of predictive models for assessing the age and duration <i>in vitro</i> cultivation based on the transcriptomic data and investigation of cell morphology.<h4>Materials and methods</h4>In the study, the dynamics of expression of individual genes encoding key regulators of cellular aging across various models of cellular senescence, as well as telomere length were investigated by real-time PCR. The analysis of the high- throughput transcriptome sequencing datasets of mesenchymal stem cells from the donors of different ages has been performed. Using regression methods, predictive models based on transcriptomic data were developed to estimate chronological age and the duration of <i>in vitro</i> cultivation. Using microscopy methods and subsequent image analysis by machine-learning algorithms, morphological alterations associated with cellular senescence have been explored and segmentation neural network model has been created for extracting nuclear morphology parameters and classification of the cells based on the duration of cultivation <i>in vitro</i>.<h4>Results</h4><i>CDKN1A</i>, <i>LMNB1</i>, <i>HMGB2</i> genes demonstrated reproducible similar dynamics on the models of replicative or stress-induced senescence and chronological aging of mesenchymal stem cells. The expression profile of the senescence-associated inflammatory phenotype components was variable in different models of cell aging. The analysis of mesenchymal stem cell transcriptomes from the donors of various ages revealed considerable donor-dependent heterogeneity of the cells, which complicates the development of precise transcriptome data-based predictive models. Investigation of the changes in the telomere length has demonstrated its applicability for assessing the dynamics of replicative senescence <i>in vitro</i>. The developed segmentation neural network model allowed for detecting senescence-associated dynamics of nuclear morphology alterations in the process of replicative aging.

<b>The aim of the study</b> was to analyze the composition feasibility of the bioinks based on high-concentration collagen (40 mg/ml) and gelatin granules (6.25 mg/ml) for bioprinting of porous-structured hydrogel constructs using an extrusion 3D bioprinter.<h4>Materials and methods</h4>Bioprinting was performed on a 3D Invivo bioprinter (Rokit, South Korea). We assessed the filament continuity during extrusion, the changes in its thickness after test printing and incubation, as well as the biodegradation of prepared scaffolds. The hydrogel cytocompatibility was studied by the proliferation of adipose-derived stem cells (ADSCs) incorporated into the scaffolds. Flow cytometry was performed to determine the immunophenotype of ADSCs. Cell proliferation in the scaffold structure was studied <i>in vitro</i> during 28 days spectrophotometrically after adding PrestoBlue reagent. The expression of target genes was analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) on day 21 of cultivation. We used the primers for mRNA encoding the synthesis of chondrogenic factors and metabolites (<i>ACAN</i>, <i>SOX9</i>, <i>COL1A1</i>, <i>COL2A1</i>), surface markers (<i>CD29</i>, <i>CD44</i>, <i>CD73</i>, <i>CD90</i>, <i>CD105</i>), as well as hypoxia (<i>HIF1A</i>), proliferation (<i>PCNA</i>), and apoptosis (<i>BCL2</i>, <i>BAX</i>) factors. The morphology of the scaffolds was studied on day 28 of culturing by light microscopy after fixing and staining the histological sections.<h4>Results</h4>The extrusion of the high concentration collagen-based hydrogel composition (40 mg/ml) and gelatin granules (6.25 mg/ml) during printing was stable, there was no filament breakage. When incubated in phosphate-buffered saline, the filament thickness of the hydrogel was statistically significantly higher than the scaffold thickness after printing. The degradation of the scaffolds from the hydrogel and gelatin in the solution of type I collagenase started earlier than the collagen scaffolds. The incubation in phosphate-buffered saline for 14 days resulted in less mass loss when drying the collagen scaffolds with gelatin granules. The cells isolated from human adipose tissue expressed surface markers characteristic of ADSCs. ADSCs proliferation and differentiation in chondrogenic direction were observed in both groups compared. The differences were in the spatial arrangement of the cells. In the collagen scaffolds the most cells were on the surface, while in the scaffolds from collagen and gelatin the cells were distributed throughout the whole volume. The 2^-ΔΔCt quantitative reverse transcription polymerase chain reaction results showed the increased expression of the transcription factor <i>SOX9</i> by the cells in the collagen and gelatin scaffolds, as well as the decreased expression of the anti-apoptotic gene <i>BCL2</i> relative to the collagen scaffolds.<h4>Conclusion</h4>The present study suggested the bioink composition based on high concentration collagen (40 mg/ml) and gelatin granules (6.25 mg/ml) for bioprinting of porous-structured hydrogel constructs. The study showed hydrogel to be appropriate for printing and exhibited the properties of a porous material. The hydrogel provided the uniform distribution of ADSCs in the scaffold volume, contributing to their differentiation in the chondrogenic direction. Thus, the suggested bioink composition appears to be a promising material to be used in tissue engineering.

That males and females age differently has been overlooked while developing aging clocks. Here, we developed a sex-specific metabolic aging clock in 390,941 individuals from the UK Biobank and integrated it with genetic, proteomic and epidemiological data to identify mechanisms accelerating/decelerating metabolic aging in males and females. Our findings reveal dysregulation of cholesterol metabolism, immune system, hemostasis, and cell growth, survival and apoptosis as common mechanisms accelerating metabolic aging in males and females, and upregulation of oxidative stress detoxification, cellular resilience and tissue integrity as common mechanisms decelerating metabolic aging. In females, a further dysregulation of carbohydrate/glucose metabolism, circadian rhythm and hormone metabolism accelerating metabolic aging is observed, while dysregulation of energy metabolism, cancer and longevity pathway is specifically observed in males. Among reproductive factors, late puberty and higher parity manifest as protective factors, decelerating metabolic aging in both sexes, and additionally childbirth at older age decelerating metabolic aging in women. Accelerated metabolic age strongly predicted morbidity and mortality in both sexes, except that the magnitude of association was several folds higher in males, and obesity explained most of the disease associations in females, suggesting that obesity influences metabolic aging and subsequent health outcomes differently in males and females. Consistent with the upregulation of molecular mechanisms involved in cancer in males, accelerated metabolic aging predicted several common cancers in males but not females. Our study provides novel insights into the biological mechanisms underlying aging and disease susceptibility in males and females, underscoring the importance of considering sex differences in healthcare strategies and public health policies.

CXCR7, also known as atypical chemokine receptor 3 (ACKR3), is a naturally-biased, β-arrestin-coupled seven transmembrane receptor (7TMR) that lacks productive coupling with heterotrimeric G-proteins. Despite a critical involvement in cancer metastasis, cardiovascular pathophysiology, and inflammatory disorders, the molecular basis of non-canonical activation and functional divergence of CXCR7 remains elusive. Here, we present a complete landscape of CXCR7 activation using a series of cryo-EM structures, and discover an atypical activation mechanism that is distinct from prototypical GPCRs. CXCR7 is maintained in a basal conformation by a unique tripartite ionic-lock involving TM5-TM6, in contrast to a broadly conserved TM3-TM6 ionic-lock in GPCRs, which is disrupted upon receptor activation. Importantly, activation of CXCR7 results in a constricted pocket and distinct surface topology on the intracellular side compared to prototypical GPCRs. Serendipitously, we capture novel dimeric arrangements of CXCR7 with an inter-protomer stitching by a native phospholipid serving as a molecular glue, and identify previously unanticipated extrahelical allosteric sites on the receptor. Surprisingly, in an intermediate state structure of CXCR7, the second extracellular loop (ECL2) displays a self-blocking conformation, in stark contrast to ECL2-mediated self-activating mechanism reported recently for some orphan GPCRs. Finally, we unequivocally establish CXCR7 as an atypical opioid receptor via a large peptide library screening and structure elucidation in complex with distinct opioid peptides imparting full receptor activation. In summary, our study elucidates an atypical mechanism of CXCR7 activation, and establishes it as an alternative, non-canonical opioid receptor target with potential for novel pain therapeutics.

Differential gene expression establishes the distinct physiology and morphology of cell types in an animal body. Single-cell sequencing and volume EM represent milestones toward the characterization of cell types, yet are difficult to combine for a comprehensive view on the cellular genotype-phenotype link. Here, we map a whole-body single-cell transcriptome into the PlatyBrowser, a multimodal cellular atlas for the marine annelid Platynereis dumerilii , and establish this combination uniquely for an entire animal. We learn that, in the 6-days-old worm, the majority of genes are tightly co-regulated to jointly implement one of eight major cellular morphotypes representing epidermis, gut, vasculature, myofibres, glia, motile cilia, glands, or neurons. Focusing on neurons, we uncover 14 families that by transcription factor identity, axonal projection, or sensory-secretory apparatus resemble conserved neuron types found in vertebrates, insects, or nematodes. We hypothesize that these existed in urbilaterian ancestors and represent the ancient core of nervous system centralization.

<title>Abstract</title> <p>Harmonized phenotyping and diverse population-specific studies are crucial for advancing gene discovery in psychiatric genetics. We conducted a genome-wide association study (GWAS) of DSM-defined lifetime Major Depressive Disorder (MDD) in 64,941 participants (25.7% cases) from the Dutch BIObanks Netherlands Internet Collaboration (BIONIC) consortium. SNP-based heritability was estimated at 13.4%, exceeding recent global meta-analyses, with a high genetic correlation (r = 0.89) to the latest major depression GWAS by the Psychiatric Genetics Consortium (PGC-MD). We identified a novel genome-wide significant locus in PALMD (P = 3.26 × 10⁻⁸), that was confirmed by GWAS-by-subtraction. Polygenic scores (PGSs) based on BIONIC predicted MDD in UKBiobank, and PGSs from PGC-MD predicted into BIONIC, with within-family analyses indicating minimal confounding. Genetic causal inference revealed associations with over 30 phenotypes. Twin concordance for MDD increased with polygenic burden, reinforcing its genetic architecture. This study emphasizes the power of harmonized phenotyping and regional biobanks in uncovering the genetic architecture of MDD, highlighting the value of population-specific studies for improving risk prediction and advancing psychiatric genetics.</p>

<title>Abstract</title> <p> Male infertility is rising globally, yet its causes remain unclear. This study identifies the transcription factor Brn-3b (POU4F2) as essential for spermatogenesis and sperm function. Brn-3b is highly expressed in mature spermatids and infertility in constitutive male Brn-3b knockout (KO) mice is characterised by structural and functional testicular changes such as reduced sperm counts, impaired motility and ultrastructural defects including disrupted acrosomes and defects in the mitochondria and flagella. RNA-seq analyses reveal significant changes in Brn-3b-dependent regulation of genes essential for sperm development, mitochondrial function, and microtubule-based movement. This was confirmed using qRT-PCR with reduced expression of associated genes e.g. <italic>Spata13</italic> , <italic>Dnah6</italic> , Cox7a1 and upregulation of genes linked to inflammation and ECM remodelling (e.g., <italic>Ptges</italic> , <italic>MMP2</italic> ). Human studies showing reduced Brn-3b in infertile men, e.g. with Klinefelter syndrome validated these findings. Exome sequencing identifying potentially deleterious variants in infertile men, suggest Brn-3b as a promising target for understanding and diagnosing male infertility. </p>

Cancer neuroscience, an emerging convergent discipline, offers novel insights into the dynamic interplay between the nervous system and cancer progression. Bidirectional signaling between the nervous system and tumors, particularly within the innervated tumor microenvironment (TME), modulates key cancer hallmarks, including proliferation, immune evasion, angiogenesis, and metastasis. Neural ablation shows heterogeneous outcomes depending on nerve subtype and tumor context, underscoring the importance of nerve-type-specific and context-dependent therapeutic approaches. These mechanistic advances are catalyzing novel therapeutic strategies that target neural-TME interactions through the integration of neuroscience and oncology. Here, we highlight recent progress in cancer neuroscience and propose revised therapeutic frameworks aimed at the neuro-innervated TME. These strategies employ interdisciplinary approaches, such as drug repurposing [β-adrenergic receptor (β-AR) blockers, antipsychotics, antidepressants], and nanotechnology-enabled targeted delivery. Both preclinical and clinical data support the potential of neural-targeted therapies to improve precision, circumvent drug resistance, and enhance clinical outcomes. By bridging neuroscience and oncology, this framework delineates a translational pathway for harnessing neural-tumor crosstalk, presenting a promising avenue for advancing cancer therapeutics and improving patient care.

Parkinson's disease (PD) is a progressive neurodegenerative disorder lacking definitive diagnostic tests. To identify new diagnostic biomarkers, we employed glycoproteomics-mass spectrometry (MS) to investigate dynamic changes in protein <i>N</i>-glycosylation across the serum, urine, and saliva of PD patients. Our comparative analysis of differentially expressed glycoproteins (DEGs) between PD patients and healthy controls (HCs) revealed distinct patterns. Specifically, ATPase phospholipid transporter 11B (ATP11B) was significantly upregulated in the serum of PD patients, while urine and saliva showed an opposite trend. Other key findings included elevated myeloperoxidase (MPO) in urine and clusterin (CLU) in serum. Zinc-α-2-glycoprotein (AZGP1), detected in all three biofluids, displayed increased sialylation and core fucosylation in serum but decreased levels in the saliva and urine of PD patients, along with a distinct bifucosylation pattern in saliva. These glycoprotein expression changes were further validated using enzyme-linked immunoassay (ELISA). Pathway analysis indicated that these DEGs are primarily involved in inflammatory response, complement activation, and synaptic plasticity, suggesting that glycosylation dysregulation may contribute to PD progression by modulating neuroinflammation and protein homeostasis. This study represents the first comprehensive analysis of multibiofluid <i>N</i>-glycosylation in PD. The findings offer potential biomarkers and provide insights into the molecular mechanisms of the disease, which could ultimately inform early diagnosis and the development of targeted therapies.

Accurate binding free energy prediction is vital for drug design, motivating the assessment of new force field models. Here, we evaluate the ABCG2 charge model with nonequilibrium alchemical free-energy simulations. GAFF2/ABCG2 achieves higher hydration free energy accuracy but does not outperform GAFF2/AM1-BCC for protein-ligand binding free energy. Both charge models exhibit comparable accuracy and compound ranking across targets, indicating that property-specific force field optimization does not guarantee improved related-property performance.

Homologous recombination (HR) deficiency upon Breast Cancer Gene 2 (BRCA2) loss arises from defects in the formation of RAD51 nucleoprotein filaments. We demonstrate that loss of the anti-recombinase Fidgetin Like 1 (FIGNL1) retains RAD51 loading at DNA double-stranded breaks (DSBs) in BRCA2-deficient cells, leading to genome stability, HR proficiency, and viability of BRCA2-deficient mouse embryonic stem cells. Mechanistically, we demonstrate that strand invasion and subsequent HR defects upon BRCA2 loss primarily arise from the unrestricted removal of RAD51 from DSB sites by FIGNL1, rather than from defective RAD51 loading. Furthermore, we identify that the MMS22L-TONSL complex interacts with FIGNL1 and is critical for HR in BRCA2/FIGNL1 double-deficient cells. These findings identify a pathway for tightly regulating RAD51 activity to promote efficient HR, offering insights into mechanisms of chemoresistance in BRCA2-deficient tumors.

Spanning two decades, the collaborative ENCODE project aims to identify all the functional elements within human and mouse genomes. To best serve the scientific community, the comprehensive ENCODE data including results from 23,000+ functional genomics experiments, 800+ functional elements characterization experiments and 60,000+ results from integrative computational analyses are available on an open-access data-portal ( https://www.encodeproject.org/ ). The final phase of the project includes data from several novel assays aimed at characterization and validation of genomic elements. In addition to developing and maintaining the data portal, the Data Coordination Center (DCC) implemented and utilised uniform processing pipelines to generate uniformly processed data. Here we report recent updates to the data portal including a redesigned home page, an improved search interface, new custom-designed pages highlighting biologically related datasets and an enhanced cart interface for data visualisation plus user-friendly data download options. A summary of data generated using uniform processing pipelines is also provided.

Hepatic fibrosis, resulting from chronic liver injury, can lead to cirrhosis and liver failure. Curcumin shows anti-fibrotic potential but has low bioavailability. This 16-week double-blind, randomized, placebo-controlled trial evaluated the effects of 80 mg/day nano-curcumin on liver fibrosis, steatosis, liver function test, and anthropometric parameters in 55 adults (30-70 years) with stage ≥ F2 NAFLD-induced fibrosis. Primary outcomes were liver fibrosis and steatosis assessed by FibroScan and FIB-4. Secondary outcomes included changes in liver function tests and anthropometric parameters including body composition. Both groups improved in fibrosis and steatosis, with no significant differences were found between them. The FIB-4 index decreased significantly in the nano-curcumin group (p = 0.022), but between-group differences were not significant (p = 0.135). ALT and AST significantly decreased in the nano-curcumin group (p < 0.001 and p = 0.004), with significant group differences (p < 0.05). GGT reduction was significant between groups after adjustment (p = 0.043), and LDH levels also decreased significantly in the nano-curcumin group (p < 0.001), with a significant between-group difference (p = 0.016). No statistically significant between-group differences were observed in anthropometric parameters. Nano-curcumin improved liver enzymes but showed no significant effect on fibrosis or steatosis compared to placebo. Further research is needed to confirm long-term benefits. Trial registration Iranian Registry of Clinical Trials IRCT20210427051098N2 (Available from: https://irct.behdasht.gov.ir ).

<h4>Background</h4>Cutaneous malignant melanoma is a common cancer in adults but extremely rare in young children, affecting fewer than one child per million each year in Europe. Because of its rarity, most treatments for children are adapted from adult therapies, despite possible biological differences. This study aimed to explore the molecular features of a rare and aggressive melanoma in a 16-month-old patient to understand disease progression and treatment resistance.<h4>Methods</h4>We studied the tumour and metastases of a patient with a melanoma carrying an NRAS mutation, who received chemotherapy and immune checkpoint inhibitor treatment. The patient died 10 months after diagnosis. We used DNA methylation analysis, single-nucleus RNA sequencing, and deep spatial transcriptomic profiling to examine genetic changes, gene activity, and their spatial distribution in both the primary tumour and lymph node metastases.<h4>Results</h4>Here, we show that the tumour displayed high genetic and transcriptomic diversity. We identified increases in MITF and BRAF gene copies as likely key drivers of the aggressive disease, which were not detected at diagnosis. We also found activation of biological pathways, including VEGFA and WNT signalling, and abnormal activity of several genes linked to immune therapy response, with marked variation between tumour regions.<h4>Conclusions</h4>This case demonstrates that paediatric melanoma can harbour complex and spatially variable molecular changes that contribute to rapid disease progression and treatment failure. Our findings support incorporating detailed spatial transcriptional profiling into clinical assessment to better guide therapy in rare paediatric cancers.

Selective autophagy ensures the targeted degradation of damaged or surplus cellular components, including organelles, thereby safeguarding cellular homeostasis. This process relies on selective autophagy receptors (SARs) that link specific cargo to the autophagy machinery. These receptors exist in two distinct forms: soluble SARs that are recruited to the cargo on demand, and transmembrane SARs that are stably embedded in the membranes of organelles they target. While both receptor types converge on the same autophagy core machinery, they differ in how they recognize cargo, are regulated, and recruit this machinery to the site of degradation. In this review, we explore the unique challenges and strategies associated with transmembrane SARs, including how their activity is suppressed under basal conditions and activated in response to stress. We compare their mode of action with that of soluble SARs, highlight key differences in kinase regulation, including the roles of TBK1, ULK1, CK2, and Src, and discuss emerging models of autophagy initiation. We further highlight fundamental principles of organelle-selective autophagy and identify open questions that will guide future research.

Huntington's disease (HD) is a fatal, inherited neurodegenerative disorder marked by the progressive and selective loss of spiny projection neurons (SPNs), resulting in a characteristic triad of motor, cognitive, and psychiatric symptoms. Despite ongoing research, no disease-modifying treatments are available, and existing therapies are limited to symptomatic relief. Stem cell-based approaches represent a promising avenue to restore striatal circuitry by replacing lost neurons and/or delivering trophic support to preserve the remaining neural tissue.In this review, we present a critical analysis of past and current clinical trials exploring cell-based therapies for HD. Early studies using human fetal tissue were hindered by sample heterogeneity and inconsistent outcomes, ultimately limiting their clinical applicability. More recent trials have shifted focus toward mesenchymal stem cells (MSCs), which are valued for their neuroprotective secretome but are not suitable for neuronal replacement. To address these limitations, human pluripotent stem cells (hPSCs) have emerged as a renewable and scalable source for the development of advanced therapy medicinal products (ATMPs). In vitro differentiation protocols mimic key developmental signaling pathways to generate striatal-like neural progenitor cells (NPCs). We review the cellular composition of these hPSC-derived ATMPs and summarize findings from preclinical transplantation studies, including data on graft survival, neuronal maturation, synaptic integration, and functional recovery. In addition, we discuss other emerging strategies such as direct neuronal reprogramming. Finally, we examine the major challenges that remain-such as ensuring graft safety, consistency, and regulatory compliance-and highlight the importance of international collaboration to overcome these barriers and accelerate clinical translation.

<h4>Background</h4>Renal cell carcinoma (RCC) accounts for 90% of all renal neoplasms and is often incidentally detected through unrelated imaging procedures. Differentiating between benign and malignant renal masses remains challenging using imaging alone. Urinary biomarkers may aid in this distinction, yet none are currently implemented in the clinic. Moreover, a comprehensive overview of urinary diagnostic biomarkers for RCC is lacking. Therefore, we aimed to systematically review and summarize existing literature on potential urinary biomarkers with diagnostic properties for RCC.<h4>Methods</h4>PubMed, Scopus and Web of Science were used for the identification of eligible studies evaluating urinary biomarkers in adults with sporadic RCC which reported diagnostic properties compared to controls groups. Standardized data extraction was performed. Risk of bias of was assessed by using a modified STROBE 22-items checklist for observational studies.<h4>Results</h4>In total 136 articles were identified through database search, four via a previous review and 19 through cross-referencing. After screening, 46 articles were included, identifying 105 individual biomarkers: metabolites (n = 40), proteins (n = 29), miRNAs (n = 12), DNA methylation markers (n = 13) and others (n = 11). Additionally, 29 multi-biomarker panels were described. Promising diagnostic markers (AUC ≥ 0.80) included dysregulated energy metabolism markers, proteins AQP1 and PLIN2, and miRNAs; miR-122-5p, miR-15a and miR-30c, however validation is severely lacking.<h4>Conclusions</h4>Various urinary biomarkers for RCC show promising diagnostic potential. The diagnostic ability of multi-biomarker panels often exceeded those of individual markers. However, individual markers and panels require external validation before clinical implementation.<h4>Trial registration</h4>This systematic review was registered on PROSPERO (CRD42023474582), and was designed and written based on the PRISMA guidelines.

<h4>Background</h4>Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML), characterized by a fusion between the PML and RARA genes and by a block in the myeloid maturation at the promyelocytic stage.<h4>Methods</h4>This study investigates the epigenetic landscape of APL by integrating ChIP-seq data on eight histone modifications and RNA-seq in APL as well as non-APL AML. APL showed a distinct chromatin profile that differed from non-APL AML.<h4>Results</h4>We describe APL-specific changes in H3K27ac, H3K9me3, and H3K27me3 with impact on enhancer activity, repression of transposable elements, and Polycomb regulated gene repression. The APL-specific H3K27ac pattern identifies APL-specific enhancer and super-enhancer regions, including a subset of enhancers that are bound by the PML-RARA fusion protein. While chromatin bound specifically by PML-RARA were dominantly active, APL was also characterized by gain of APL-specific heterochromatin states with significant gains of H3K9me3 enriched lamina-associated domains and the transposable elements LINE, LTR, and SINE.<h4>Conclusion</h4>These findings suggest a unique enhancer and heterochromatin profile in APL, with implications for transcription regulation and treatment response. These findings offer novel insights into the pathogenesis of APL.

Canonical splicing variants (±2) contribute significantly to genetic disorders, yet the clinical significance of non-canonical splicing variants (NCSVs) that occur outside of canonical splicing sites remains unknown in male infertility. A comprehensive evaluation of reported studies on hereditary male infertility revealed that the 2,404 pathogenic variants contained 120 canonical splicing variants and 32 NCSVs. Among the remaining 2,252 variants, the splicing variant analytical strategy identified 17 novel NCSVs that disrupt normal mRNA splicing from previously classified missense variants. This expands the contribution of NCSVs by 53.13% (17/32), with NCSVs accounting for 28.99% (49/169) of all the splicing variants. Moreover, thirteen positively validated NCSVs are identified in 12 of 718 idiopathic male infertility patients with negative results by conventional genetic analysis. The first pathogenic variant in the TATA element modulatory factor 1 (TMF1: c.2859+4A>G) results in TMF1 exon 14 skipping and decreased progressive sperm motility and morphological abnormalities in a patient with male infertility. Tmf1 NCSV knock-in mice recapitulated human phenotype, showing significantly decreased sperm count, motility, ultrastructural head defects, and subfertility. This study provides the first comprehensive landscape of NCSVs in male infertility, suggesting that NCSVs may constitute a hidden etiological factor for male infertility.

<h4>Introduction</h4>Evidence-based practices (EBPs) tailored to reduce opioid-related fatalities among minoritized populations may help reduce inequities, but research elucidating the relationship between local-level inequities in social determinants of health (SDH) and selection and implementation of tailored EBPs.<h4>Methods</h4>The HEALing Communities Study was a multisite, wait-listed, community-level cluster-randomized trial with the objective to reduce opioid overdose deaths. Community coalitions selected and partner organizations implemented EBP strategies to increase access to Overdose Education and Naloxone Distribution (OEND) and Medications for Opioid Use Disorder (MOUD). The research team utilized negative binomial and zero-inflated Poisson models to assess whether the relationship between (a) communities' need for tailored OEND and MOUD strategies, as indicated by opioid overdose death rates among minoritized groups; and (b) the selection, implementation, and reach of those strategies was moderated by local-level inequities in SDH, quantified by the Local Social Inequity in Drug Overdoses (LSI-DO) index. The analysis included data from 33 communities across four states.<h4>Results</h4>Eleven of 33 communities implemented tailored OEND and MOUD strategies. Adjusted analysis revealed that LSI-DO scores moderated the relationship between 2021 opioid overdose death rates among minoritized groups and the proportion of community-implemented tailored strategies (p = 0.015). The research team found no moderation for the number of strategies selected or number of individuals reached by EBPs.<h4>Conclusion</h4>Few communities selected EBPs tailored to minoritized populations. For those that did, higher local-level social inequity was associated with decreased implementation of tailored strategies. Continued research on how to address overdose inequities-and local-level social inequity-is vital. Findings highlight the need for policy approaches that pair overdose prevention strategies with investments to reduce local-level social inequities that impede equitable implementation.

Bone marrow-derived mesenchymal stem cell (BMSCs) delivery is a promising therapeutic strategy for bronchopulmonary dysplasia (BPD); however, its efficacy remains suboptimal. Recent studies have reported that extracellular vesicles derived from peroxiredoxin 6 (PRDX6)-downregulated mesenchymal stem cells (MSCs) exhibit diminished anti-inflammatory and anti-oxidant effects through a paracrine mechanism. Nevertheless, whether PRDX6 participates in BPD progression via paracrine signaling remains unclear. Herein, rat BMSCs were first isolated and characterized. The GFP-labeled lentivirus was used to overexpress PRDX6 in BMSCs. PRDX6-overexpressing BMSCs exhibited improved proliferation and migration abilities. Notably, mesencephalic astrocyte-derived neurotrophic factor (MANF) secretion was significantly increased (∼2.5-fold, p < 0.05) in PRDX6-overexpressed BMSCs. Further mechanistic studies revealed that PRDX6 bound to MANF, reducing its endoplasmic reticulum retention by displacing glucose-regulated protein 78, thereby promoting MANF secretion. Silencing MANF expression markedly attenuated the effects of PRDX6 overexpression. PRDX6-modified BMSCs were delivered intratracheally into BPD rat model, which was established by exposing rats in 85 % oxygen for 21 days. PRDX6 overexpression enhanced BMSCs recruitment to injured lung tissues, and significantly reduced apoptosis, inflammation, vascular loss, and pulmonary arterial remodeling. Concurrently, MANF levels in lung tissues, bronchoalveolar lavage fluid, and serum were elevated post-administration. These results suggest that PRDX6-mediated MANF secretion in BMSCs may not only enhance their proliferative and migratory capacities but also directly protect against lung injury. These findings provide a new strategy for amplifying the in vivo efficacy of BMSCs delivery on BPD.

<h4>Background</h4>Tau neurofibrillary pathology is a hallmark of Alzheimer's disease (AD) and can be quantified in vivo using tau-selective positron emission tomography (tau PET). Tau PET signal closely correlates with cognitive decline and disease stage, yet the molecular networks underpinning tau accumulation remain incompletely defined.<h4>Methods</h4>We performed multi-omics integration of proteomics, transcriptomics, and tau PET standardized uptake value ratios (SUVRs), and clinical assessments data from cognitively normal and cognitively impaired individuals. Using Light Gradient Boosting Machine (LightGBM), two-way orthogonal partial least squares, and network-based approaches, we explored key tau-associated proteomic signatures and constructed protein-protein interaction (PPI) modules. Module activities were quantified by gene set variation analysis and related to tau PET and cognition.<h4>Results</h4>Among 60 regions, 15 tau PET imaging biomarkers were selected based on group differences, LightGBM importance, and cognitive relevance. Fifty key tau-associated proteins were identified and organized into four functional modules. PPI modules 1 (metabolic-cytoskeletal) and 3 (adhesion-nutrient sensing) exhibited strong associations with elevated tau PET uptake across selected cortical and limbic regions, as well as with cognitive impairment.<h4>Conclusion</h4>Distinct modules reflected regional tau PET burden and cognitive outcomes in AD, highlighting convergent disruptions in energy metabolism, cytoskeletal stability, and intercellular signaling.<h4>Highlights</h4>Integration of proteomics, transcriptomics, tau positron emission tomography (PET) imaging, and cognition in Alzheimer's disease.Fifteen key tau PET imaging biomarkers were prioritized.Fifty key tau-associated proteins were identified.Four distinct molecular networks contribute to regional tau pathology and cognition.Modules 1 (metabolic-cytoskeletal) and 3 (adhesion-nutrient sensing) strongly associated with tau PET burden and cognitive impairment.

Historically, natural products have been a primary source of new drugs and lead compounds. However, their direct application as therapeutic agents remains limited. Given the multifactorial nature of diabetes, therapeutic agents with multi-targeting properties offer a promising solution for effective management. Herein, we developed a library of seven lipophilic derivatives of caffeic acid (CA) and evaluated their multimodal antidiabetic activities. Among these, the most potent compound, CA14, showed an IC₅₀ of 1.94 μM, representing a 113-fold improvement over acarbose (IC₅₀ 219.70 μM). Further, compared to the parent compound CA (IC₅₀ 12 400 μM), CA14 was 6392 times more potent. Kinetic analysis revealed that CA14 acts as a competitive inhibitor of α-glucosidase. CA14 showed comparable α-amylase inhibition to that of acarbose, while CA showed no inhibition even at 500 μM. Additionally, CA14 exhibited antiglycation activity by inhibiting fructosamine and advanced glycation end products (AGEs). It also showed excellent antioxidant activity (IC₅₀ 13.98 μM), nearly twice as potent as Vitamin C. Notably, acarbose lacked any antioxidant activity. Intrinsic fluorescence quenching and FT-IR analyses further confirmed that CA14 disrupted substrate binding, reducing enzymatic activity. Of note, CA14 did not exhibit any significant cytotoxicity in 3T3-L1 fibroblasts, even at 50 μM. Molecular docking revealed good extent of interactions, while ADMET profiling predicted favorable druglikeness. CA14 was found to be stable under physiological conditions even after 10 days. In summary, CA derivatives demonstrated robust multifaceted antidiabetic potential by inhibiting α-glucosidase, α-amylase, and AGE formation, while also mitigating oxidative stress, which are key contributors to diabetes progression.

Iron (Fe) is an essential micronutrient for metabolic and physiological processes. Its dysregulation is associated with disorders such as Alzheimer's and hemochromatosis. Therefore, the development of cost-effective and selective probes for Fe(iii) detection is of significant clinical importance. In this study, fluorescent carbon dots derived from coconut coir (CCDs) were synthesized <i>via</i> a single-step hydrothermal method. The CCDs exhibited strong blue emission at 450 nm under 350 nm excitation and demonstrated selective fluorescence quenching in the presence of Fe(iii) ions, with a detection limit of 223.2 μM. HR-TEM revealed the particle size of CCDs ranged between 5.64 to 10 nm. XRD confirmed the crystalline nature. FTIR spectra indicated presence of hydroxyl and carboxyl groups contributing to dispersibility and surface passivation. Raman spectroscopy showed distinct D (1354 cm^-1) and G (1582.61 cm^-1) bands, characteristic of low-dimensional carbon nanostructures. Bioimaging and cytocompatibility studies in L929 fibroblast cells confirmed biocompatibility up to 500 μg mL^-1. Collectively, these findings highlight the potential of CCDs as an effective fluorescent probe for Fe(iii) sensing and bioimaging applications in medical diagnostics. The CCDs were cytocompatible at concentrations up to 500 μg mL^-1.

Hepatic stellate cells (HSCs) trans-differentiation into myofibroblasts is central to liver fibrosis. Integrated stress response (ISR) signaling, including metabolic stress, plays a critical role in this process. However, the precise role of ISR signaling in HSCs activation-whether detrimental or protective-remains unclear. Here we identified that the noncanonical cap-binding protein EIF3d-mediated ATF4 expression is significantly upregulated in HSCs from both patients and mouse models of fibrotic livers, with its levels positively correlating with the degree of fibrosis. EIF3d-ATF4 signaling was induced by TGFβ1 in HSCs and was demonstrated to be both necessary and sufficient for promoting HSC survival, proliferation, activation, and extracellular matrix (ECM) production. Furthermore, genetic and pharmacological inhibition of EIF3d-ATF4 effectively prevented TGFβ1-induced HSC activation by suppressing mitochondrial activity and glycolysis. Mechanistically, EIF3d-ATF4 overexpression drove ATF4-dependent S100P transcription, which facilitated metabolic reprogramming and upregulated fibrogenic markers. This EIF3d-ATF4-S100P axis promoted liver fibrosis by activating JNK and NLRP3 signaling in HSCs, thereby inducing HSC activation and conferring resistance to apoptosis. Importantly, mice with HSC-specific ATF4 deletion or treated with our innovative ISR antagonist, ERMT1, were protected from three distinct mouse fibrotic models. These findings underscore the role of the EIF3d-ATF4-S100P signaling axis in liver fibrosis progression and HSC activation, presenting it as a promising therapeutic target for managing liver fibrosis and cirrhosis.

Interferons (IFNs) are essential cytokines in host defense, particularly against viral pathogens. Dysregulation of IFN signaling can result in increased susceptibility to viral infections, systemic inflammation. We report a 6-month-old male who presented with hemophagocytic lymphohistiocytosis (HLH) triggered by cytomegalovirus (CMV) infection. He was admitted to the Pediatric Intensive Care Unit due to fever and respiratory distress. Laboratory findings revealed pancytopenia, elevated inflammatory markers, and laboratory markers suggestive of HLH. Whole-exome sequencing revealed a novel homozygous nonsense variant in ZNFX1 (c.1928G>A, p.Trp643X). During subsequent hospitalization, he developed CMV reactivation, gastrointestinal symptoms, neurologic deterioration, and multiorgan dysfunction. Despite intensive supportive care and immunomodulation with anakinra and corticosteroids, the patient progressed to irreversible organ failure and died from respiratory failure. This case expands the clinical and genetic spectrum of ZNFX1 deficiency and highlights the importance of considering monogenic interferonopathies in children with severe viral infections, HLH, and unexplained systemic inflammation.

Protein aggregates are central to the pathogenesis of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. This comprehensive review explores the mechanisms of protein misfolding and aggregation, their prion-like propagation, and the critical role of oligomeric species in neurotoxicity. It further examines cellular clearance pathways, including the ubiquitin-proteasome system and autophagy, alongside the regulatory functions of molecular chaperones. The review also covers advanced diagnostic imaging and biomarker techniques, as well as emerging therapeutic strategies such as pharmacological agents, gene therapy, and immunotherapy. Controversies regarding the toxicity of aggregates and future directions, including novel degradation technologies and targeted therapeutic approaches, are discussed. By integrating current knowledge, this review aims to provide a broad yet detailed overview of the field, highlighting both established concepts and promising avenues for research and treatment.

Atherosclerosis, the principal pathology underlying cardiovascular diseases, is now recognized as a chronic inflammatory disorder of the arterial wall. This review focuses on the central role of oxylipins, a diverse family of bioactive lipids derived from polyunsaturated fatty acids (PUFAs), in the inflammatory processes driving atherosclerosis. We synthesize evidence that oxylipins produced via cyclooxygenase (COX), lipoxygenase (LOX), cytochrome P450 (CYP), anandamide (AEA) pathways and non-enzymatic transformations of PUFAs are pivotal modulators of vascular function, immune cell recruitment, and plaque stability. The balance between pro-inflammatory mediators and specialized pro-resolving mediators (SPMs) is critical; a shift towards inflammation underlies disease progression. Advances in lipidomics now enable comprehensive oxylipin profiling, revealing distinct signatures with significant diagnostic and prognostic potential for assessing coronary artery disease severity and predicting future cardiovascular events. Therapeutically, while current anti-inflammatory strategies target downstream pathways, this review highlights emerging approaches that modulate the oxylipin system directly. These include promoting SPMs synthesis through omega-3 supplementation, inhibiting pro-inflammatory leukotriene production, and preserving cardioprotective epoxyeicosatrienoic acids (EETs) via soluble epoxide hydrolase (sEH) inhibition. A deeper understanding of these complex oxylipin networks promises to yield novel biomarkers and targeted therapies designed to restore inflammatory homeostasis and combat atherosclerotic cardiovascular disease.

<b>Background:</b> Immunotherapy-based regimens have expanded the treatment landscape for unresectable hepatocellular carcinoma (uHCC); however, real-world data are limited. <b>Methods:</b> This retrospective, observational study used data from electronic medical records from Mayo Clinic sites across the United States. Patients with uHCC who initiated a first-line (1L) systemic therapy between June 2020-October 2022 with ≥2 follow-up visits were included. Treatment patterns, overall survival (OS), and post-index gastrointestinal (GI) bleeding were assessed by GI bleeding risk defined by Child-Pugh Class B or C, pre-index GI bleeding, uncontrolled hypertension, or significant varices and band ligation. <b>Results:</b> Of 186 included patients, 68.8% had GI bleeding risk and 31.2% did not. Atezolizumab plus bevacizumab was the most common 1L systemic therapy in patients with or without GI bleeding risk (72.7% and 29.3%, respectively). Median OS (95% confidence interval) with atezolizumab plus bevacizumab was 12.8 (8.0-19.3) months and not reached in patients with and without GI bleeding risk, respectively. OS rates with atezolizumab plus bevacizumab in patients with or without GI bleeding risk, respectively, were 52.3% and 70.6% at 12 months, 41.6% and 57.8% at 18 months, and 34.6% and 51.3% at 24 months. Post-index GI bleeding with atezolizumab plus bevacizumab occurred in 19.4% and 5.9% of patients with and without GI bleeding risk, respectively. <b>Conclusions:</b> During this study period, atezolizumab + bevacizumab was the most common 1L therapy for patients with uHCC, regardless of GI bleeding risk. OS rates with atezolizumab + bevacizumab were lower in patients with versus without GI bleeding risk. Findings highlight the unmet need for guidance on characteristics-driven treatment decisions.

<i>MYCN</i> amplification defines a highly aggressive subtype of neuroblastoma and is strongly associated with poor clinical outcomes. Due to the intrinsically disordered structure of the N-Myc protein, it remains largely undruggable. This review provides a comprehensive summary of the molecular regulatory network surrounding <i>MYCN</i>, including upstream pathways, key cofactors, and downstream effectors involved in cell cycle control, metabolic reprogramming, and ferroptosis. We further discuss the roles of epigenetic modulators, noncoding RNAs, and positive feedback loops in sustaining <i>MYCN</i>-driven oncogenic programs. Emerging therapeutic strategies such as PROTACs, metabolic inhibitors, immune-based approaches, and RNA-targeting technologies offer promising alternatives to direct <i>MYCN</i> inhibition. This review aims to provide a theoretical foundation for future development of precise and effective therapies targeting <i>MYCN</i>-amplified neuroblastoma.

Aging is a multidimensional process regulated by the interplay of genetic and environmental factors, with epigenetic alterations serving as a central regulatory hub. Aberrant DNA methylation patterns, dysregulation of histone-modifying enzymes (e.g., SIRT1, EZH2), and non-coding RNA-mediated mechanisms collectively remodel gene expression networks, impacting critical pathways such as cellular senescence and mitochondrial homeostasis. This establishes an "environment-epigenome-disease" causal axis, closely associated with pathologies including β-amyloid deposition in Alzheimer's disease, atherosclerosis, immunosenescence, osteoporosis, sarcopenia, and tumorigenesis. Capitalizing on the reversible nature of epigenetic modifications, pharmacological epigenetics has emerged as a cutting-edge field for intervening in aging and age-related diseases. Targeting key epigenetic modifiers such as DNA methyltransferases and histone deacetylases enables the modulation of disease-associated epigenetic states, providing a promising avenue for therapeutic intervention in aging and age-related diseases. This review synthesizes the molecular mechanisms of epigenetic regulation in aging, their role in age-related diseases, and advances in pharmacological epigenetics-from basic research to clinical translation. It further situates key challenges such as target specificity, long-term safety, and tissue-specific delivery within a translational framework, aiming to inform strategies for the diagnosis and intervention of age-related conditions.

As the most common pediatric malignancy, B-cell acute lymphoblastic leukemia (B-ALL) has multiple distinct subtypes characterized by recurrent and sporadic somatic and germline genetic alterations. Identifying B-ALL subtypes can facilitate risk stratification and enable tailored therapeutic design. Existing methods for B-ALL subtyping primarily depend on immunophenotyping, cytogenetic tests, and genomic profiling, which can be costly, complicated, and laborious. To overcome these challenges, RanBALL (an ensemble random projection-based model for identifying B-ALL subtypes) is presented, an accurate and cost-effective model for B-ALL subtype identification. By leveraging random projection (RP) and ensemble learning, RanBALL can preserve patient-to-patient distances after dimension reduction and yield robustly accurate classification performance for B-ALL subtyping. Benchmarking results based on >1700 B-ALL patients demonstrate that RanBALL achieves remarkable performance (accuracy: 0.93, F1-score: 0.93, and Matthews correlation coefficient: 0.93), significantly outperforming state-of-the-art methods like ALLSorts in terms of all performance metrics. In addition, RanBALL performs better than t-SNE in terms of visualizing B-ALL subtype information. We believe RanBALL will facilitate the discovery of B-ALL subtype-specific marker genes and therapeutic targets to have consequential positive impacts on downstream risk stratification and tailored treatment design is believed. To extend its applicability and impacts, a Python-based RanBALL package is available at https://github.com/wan-mlab/RanBALL.

This narrative review underscores the influence of host genetics in actively regulating gut microbiota composition and function, highlighting the distinctive advantages of intestine-specific conditional knockout (cKO) models in gut microbiome research. In contrast to whole-body knockouts or germ-free animals, these precision models, enabled by Cre-loxP technology, eliminate confounding systemic effects to elucidate how localized host genes within intestinal cells regulate the gut microbial ecology. The review identifies three fundamental host-driven regulatory mechanisms through the analysis of specific gene deletions: (1) barrier integrity (e.g., mucus and junction proteins), (2) immune defenses (e.g., antimicrobial peptides and glycan synthesis), and (3) metabolic signaling (e.g., bile acid receptors and glucose transporter). These pathways jointly impose microbial symbiosis, and their disruption leads to dysbiosis characterized by increased abundance of pathobionts (e.g., <i>Escherichia, Proteobacteria</i>), directly connecting host genetics to inflammatory and metabolic disorders. This host-centric viewpoint emphasizes the gut as an active regulator, rather than a passive microenvironment for the microbiota, providing significant insights for creating tailored therapeutics that focus on host pathways to restore microbial balance in disorders such as inflammatory bowel diseases.

Drug resistance remains a significant challenge in achieving successful cancer treatment, often leading to disease recurrence and reduced patient survival. While traditional tissue biopsies provide valuable insights into tumor biology, they are invasive, infrequent, and may fail to capture the full complexity of tumor heterogeneity and dynamic molecular changes. In contrast, liquid biopsy has emerged as a minimally invasive, real-time approach for monitoring tumor evolution through the analysis of circulating biomarkers. Among these biomarkers, circular RNAs (circRNAs) - a distinct class of non-coding RNAs characterized by covalently closed-loop structures - have gained attention due to their remarkable stability, abundance in body fluids, and functional involvement in gene regulation. Increasing evidence supports the role of circRNAs in mediating drug resistance through mechanisms such as inhibition of apoptosis, epithelial-mesenchymal transition, autophagy, and drug efflux, largely via interactions with microRNAs or proteins. Advanced detection methods, including quantitative reverse transcription polymerase chain reaction, droplet digital polymerase chain reaction, and RNA sequencing, combined with computational tools, enable precise profiling of circRNAs in plasma or exosomes. CircRNA-based liquid biopsies offer a dynamic, non-invasive strategy for early detection of therapeutic resistance and may guide personalized treatment decisions. This review highlights the technological advancements, biological relevance, and clinical promise of circRNAs as circulating biomarkers, emphasizing their potential in precision oncology and future collaborative translational applications.

<b>Background:</b> Pediatric acute myeloid leukemia (pAML) is the second most common type of childhood leukemia, behind acute lymphoblastic leukemia. High-throughput technologies have enabled the identification of increasing molecular alterations linked to AML prognosis, revealing genomic heterogeneity among individual patients and providing clinically valuable diagnostic and prognostic information. This study systematically analyzed the correlation between high-frequency mutated genes and prognosis in pAML by performing whole-transcriptome sequencing (WTS) of bone marrow samples from newly diagnosed AML children in Southwest China and mapping their genetic profiles. <b>Methods:</b> pAML patients treated at the Department of Hematology and Oncology, Children's Hospital of Chongqing Medical University, from January 2015 to October 2024, were enrolled, and WTS was performed. The study described the frequency, pathogenicity classification, and risk stratification of mutation genes and fusion genes, and constructed a genetic landscape. For high-frequency pAML mutations, the impact on early induction remission rate (CR) and long-term event-free survival (EFS) was evaluated. <b>Results:</b> A total of 134 pediatric AML patients from Southwest China were included, with a male-to-female ratio of 74:60 and a median diagnosis age of 5.96 years. Based on pathogenicity classification using WTS, fusion genes were categorized into level 1, level 2, and level 3 genes, as well as mutation genes. The study identified five fusion genes of level 1, the most frequent being <i>RUNX1::RUNX1T1</i> (32/134, 23.88%), <i>KMT2A</i> rearrangements (29/134, 21.64%), and <i>CBFB::MYH11</i> (13/134, 9.7%). Sixteen mutation genes of level 1 were detected, seven of which recurred in over 5% of patients, including <i>NRAS</i> (31/134, 23.13%), <i>FLT3</i> (25/134, 18.66%), <i>KIT</i> (24/134, 17.91%), <i>CEBPA</i> (14/134, 10.45%), <i>WT1</i> (13/134, 9.7%), <i>KRAS</i> (11/134, 8.2%), and <i>PTPN11</i> (7/134, 5.22%). Sex-based analysis revealed that <i>PTPN11</i> mutations were significantly more frequent in males (9.45% vs. 0%, <i>p</i> = 0.023), as were <i>KIT</i> mutations (24.32% vs. 10.00%, <i>p</i> = 0.044). Risk-stratified analysis showed that <i>WT1</i> mutations (14.13% vs. 0%, <i>p</i> = 0.031) and <i>FLT3</i>-ITD mutations (13.19% vs. 0%, <i>p</i> = 0.042) were enriched in intermediate- and high-risk groups, whereas <i>CEBPA</i> (25.64% vs. 5.43%, <i>p</i> = 0.012), <i>KIT</i> (35.90% vs. 10.87%, <i>p</i> = 0.003), and <i>KIT</i>-E8 (20.51% vs. 1.10%, <i>p</i> < 0.001) mutations were more prevalent in low-risk groups. Prognostic analysis indicated that <i>PTPN11</i> and <i>KIT</i> mutations did not affect CR or EFS across sexes, nor did <i>WT1</i>, <i>CEBPA</i>, or <i>KIT</i> mutations influence outcomes by risk stratification. However, <i>FLT3</i>-ITD-positive patients had significantly lower CRs (χ² value = 11.965, <i>p</i> = 0.007), although EFS differences were nonsignificant. In contrast, <i>WT1</i> mutations were associated with inferior EFS compared to wild-type (<i>p</i> = 0.036). Furthermore, the univariate and multivariate Cox regression revealed consistent results with the above findings, indicating that <i>WT1</i> mutation was an independent adverse prognostic factor for EFS (HR = 2.400, 95% CI: 1.101-5.233, <i>p</i> = 0.028). The results of univariate and multivariate logistic regression analyses also confirmed that <i>FLT3</i>-ITD mutation was an independent predictor of initial treatment response in our cohort (OR = 10.699, 95% CI: 2.108-54.302, <i>p</i> = 0.004). <b>Conclusions:</b> This study delineated the genetic landscape of pAML in Southwest China and explored the prognostic value of gene fusions and mutations in early and long-term outcomes. These findings provide a foundation for understanding the genetic heterogeneity of pAML and offer evidence for the development of precision medicine approaches.

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that contaminate agricultural commodities, posing risks to food safety, animal productivity, and human health. The gastrointestinal tract is the first and most critical site of exposure, where the intestinal epithelium functions as both a physical and immunological barrier against luminal toxins and pathogens. While extensive research has demonstrated that mycotoxins disrupt epithelial integrity through tight junction impairment, oxidative stress, apoptosis, and inflammation, their effects on the intestinal stem cell (ISC) compartment and epithelial regeneration remain insufficiently understood. This review integrates recent findings from in vivo, cell culture, and advanced 3D intestinal organoid and gut-on-chip models to elucidate how mycotoxins such as deoxynivalenol and zearalenone impair ISC proliferation, alter Wnt/Notch signaling, and compromise mucosal repair. We also discuss dose relevance, species differences, and the modulatory roles of the microbiome and short-chain fatty acids, as well as emerging evidence of additive or synergistic toxicity under co-exposure conditions. By bridging well-established mechanisms of barrier disruption with the emerging concept of ISC-driven regenerative failure, this review identifies a critical knowledge gap in mycotoxin toxicology and highlights the need for integrative models that link epithelial damage to impaired regeneration. Collectively, these insights advance understanding of mycotoxin-induced intestinal dysfunction and provide a foundation for developing nutritional, microbial, and pharmacological strategies to preserve gut integrity and repair.

The treatment of central nervous system (CNS) diseases faces huge challenges, mainly due to the blood-brain barrier (BBB) restricting drug delivery, which leads to many potential treatment methods being unable to effectively reach the target area. In recent years, nasal administration has received extensive attention as a non-invasive drug delivery route because of its anatomical connection with the brain, enabling direct delivery to brain tissue. In particular, the siRNA delivery system based on nanocarriers has shown great promise in the treatment of CNS diseases due to its unique advantages in targeting gene silencing. This article reviews the latest research progress on nasal administration of siRNA nanocarriers, with a focus on the design strategies, administration mechanisms, in vivo and in vitro effects, and safety evaluations of different nanocarriers. The aim is to provide a systematic theoretical basis and future research directions for the application of siRNA nasal administration in the treatment of CNS diseases (see the abstract of the picture).

<b>Introduction</b>: Gene therapy using siRNA is a current area of research in oncology. Although siRNA formulations have not yet been approved for cancer therapy, numerous studies have demonstrated their therapeutic potential for tumor remission. <b>Objective</b>: To provide an overview of the formulations designed and developed to date based on synthetic siRNA for systemic administration to silence cancer genes. <b>Methodology</b>: A thorough search was conducted using the keywords "siRNA", "therapy", and "cancer", with further classification of the resulting works into the various topics addressed in this review. <b>Results</b>: This review encompasses a wide range of aspects, from the design of siRNA using bioinformatics tools to the primary cellular signals and mechanisms targeted for inhibition in cancer therapy. It describes the primary chemical modifications made to siRNA chains to enhance stability, improve bioavailability, and ensure their binding to nanocarrier systems. siRNA formulations ranging from simple conjugates with biomolecules and small molecules to organic, inorganic, and hybrid nanoparticles, which are examined focusing on their advantages and disadvantages. The significance of nanosystems in dual therapy, including siRNA, for developing personalized treatments that achieve better outcomes is emphasized. <b>Conclusions</b>: Personalized cancer therapy appears to be the preferred approach for oncological treatments. To progress, strategies need to be tailored to the patient's genetic profile. siRNA therapies provide a flexible platform for targeting and inhibiting critical oncogenes, enhancing the prospects of genomics-guided, patient-specific therapies.

Since forkhead box P3 (FOXP3) is a hallmark of regulatory T (Treg) cells, the expansion of FOXP3⁺ adult T-cell leukemia/lymphoma (ATL) cells is believed to contribute to immune suppression and the pathogenesis of ATL. However, the mechanisms underlying the expansion of FOXP3⁺ ATL cells remain unclear. OX40, a co-stimulatory molecule, is expressed in ATL cells, and OX40 signaling has been shown to promote the differentiation and proliferation of Treg cells in mouse models. To investigate the mechanisms driving the expansion of FOXP3⁺ ATL cells, we examined the expression of OX40 and its ligand, OX40L. Our findings revealed that OX40 expression was elevated in patients with ATL and with a high frequency of FOXP3⁺ ATL cells. Flow cytometric analysis of peripheral blood mononuclear cells (PBMCs) from patients with acute ATL cultured for 18 h demonstrated that FOXP3^- and FOXP3⁺ cells predominantly expressed OX40L and OX40, respectively. Furthermore, small interfering RNA-mediated FOXP3 knockdown in HTLV-1-infected cell lines increased OX40L expression. These results suggest that interactions between FOXP3^- OX40L⁺ cells and FOXP3⁺ OX40⁺ cells may promote the proliferation of FOXP3⁺ ATL cells.

The advancement of cancer immunotherapy has focused on developing therapies that not only target tumor progression but also enhance immune responses, which could potentially shift the balance within the tumor microenvironment (TME) to promote a more immune-competent environment and improve the effectiveness of antitumor immunotherapies. This study evaluates the internalization of single-stranded RNA origami (ssRNAOG) in coculture models mimicking the TME in vitro and the antitumor efficacy of the combination of ssRNAOG with 5-fluorouracil (5-FU) in pancreatic ductal adenocarcinoma (PDAC) models. The internalization of ssRNAOG triggers the TLR3 signaling pathway, leading to robust innate immune activation. Notably, ssRNAOG induces the overexpression of MHC class I protein on macrophages, which recruits NK cells into the TME. The combination of ssRNAOG and 5-FU significantly suppressed tumor cell colony formation in vitro, demonstrating a synergistic antiproliferative effect. Transcriptomic and proteomic analyses revealed a significant upregulation of inflammatory cytokines and the activation of NF-κB and STAT1, which are indicative of M1-like polarization in macrophages. In vivo administration of both ssRNAOG and 5-FU revealed a marked reduction in tumor burden and an extension of survival in mice bearing xenograft PDAC tumors. Immunohistochemistry revealed a shift in macrophage polarization toward the M1-like phenotype, which is associated with enhanced proinflammatory responses and reduced tumor proliferation. These findings indicate that ssRNAOG, as a potent modulator of the TME that can sensitize resistant tumors to chemotherapy, presents a novel immunotherapeutic strategy for PDAC.

Oligonucleotide therapeutics-including antisense oligonucleotides and duplex RNAs such as small interfering RNAs, small activating RNAs, and microRNAs-hold immense potential for treating both genetic and acquired diseases by modulating gene expression in a target-specific manner. However, effective delivery to extrahepatic tissues, particularly the central nervous system, remains a significant challenge. While <i>N</i>-Acetylgalactosamine conjugation has enabled liver-specific delivery of oligonucleotides leading to several approved siRNA drugs for hepatic indications, there remains a significant unmet need for effective treatment options in the CNS space. We have developed the smart chemistry-aided delivery platform that enables duplex RNA delivery by conjugating to an accessory oligonucleotide, which facilitates protein binding and promotes cellular uptake. Through extensive screening, we identified an optimal SCAD architecture that demonstrates enhanced cell-free protein binding and <i>in vitro</i> activity. In rodent models, local administration of SCAD-siRNA conjugates resulted in broad biodistribution throughout the CNS and sustained mRNA knockdown for over 5 months, with a favorable safety profile. The SCAD platform also exhibited efficient delivery to other extrahepatic tissues, including the eye, lung, and joint. The modular design of SCAD can be easily adapted to any duplex RNA, making it a powerful tool for advancing oligonucleotide therapeutics.

Graphene oxide (GO) is a nanomaterial with excellent physico-chemical properties widely used in a high variety of technological applications. However, conventional protocols for GO preparation rely on wet synthesis, involving extreme chemical conditions. Recently, mechanochemical synthesis has been postulated as a rapid and sustainable dry alternative for the preparation of new materials. In this work, an optimization of the mechanochemical synthesis of GO from graphite is carried out. To characterize the materials, transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-Ray photoelectron microscopy (XPS), elemental analysis, and nitrogen adsorption-desorption isotherms are employed. The GO synthesized via dry protocol (d-GO) and GO prepared via wet synthesis (w-GO) are coupled with magnetic nanoparticles (MNPs) to prepare magnetic graphene oxide sorbents (d-M@GO and w-M@GO). Subsequently, the adsorption properties of the prepared materials toward inorganic and organic pollutants are determined and compared. The results indicated excellent adsorption efficiency for d-M@GO, demonstrating the successful application of the mechanochemical method in magnetic sorbents preparation. To the best of the available knowledge, this is the first work to investigate the applicability of dry mechanochemical GO for the synthesis of a magnetic sorbent (d-M@GO) and its use toward emerging concern and priority pollutants (PFAS and metal ions).

Homoharringtonine (HHT) is widely used in combination regimens for acute myeloid leukemia (AML), yet its direct cellular targets remain undefined, limiting precision application. Here, we identified EWS RNA-binding protein 1 (EWSR1) as the primary target of HHT through chemical proteomics and biophysical validation. HHT bound the RNA recognition motif of EWSR1 with micromolar affinity, inducing an allosteric conformational switch that promoted oligomerization and liquid-liquid phase separation (LLPS). EWSR1 condensates selectively recruited the N⁶-methyladenosine (m⁶A) reader YTHDF2, forming cytoplasmic hubs where HHT disrupted YTHDF2-mRNA interactions. This sequestration attenuated m⁶A-mediated RNA decay, stabilizing key transcripts such as <i>TNFRSF1B</i> and <i>HMOX1</i>, and thereby impairing AML cell proliferation. Integrated transcriptomics and single-cell RNA-seq analyses revealed that <i>EWSR1</i> was markedly upregulated in AML, particularly in hematopoietic progenitor and myeloid subpopulations, and high <i>EWSR1</i> expression correlated with poor prognosis and enhanced HHT sensitivity. In vivo, the anti-leukemic efficacy of HHT was significantly diminished upon EWSR1 knockdown, demonstrating that EWSR1 was required for therapeutic response. Collectively, these findings uncover a phase separation-centric mechanism by which HHT exerts anti-AML activity, establish the EWSR1-YTHDF2-m⁶A <i>axis</i> as a critical regulator of leukemia progression, and position EWSR1 as both a functional target and a predictive biomarker for optimizing HHT-based therapies.

During chronic HBV infection, the massive secretion of HBV envelope proteins (HBsAg) as non-infectious subviral particles (SVPs) remains a significant challenge in achieving a functional cure. Despite this, the HBsAg folding process, essential for HBV and HDV particle morphogenesis, remains poorly understood. DNAJB12 and DNAJB14 are two recently identified co-chaperones implicated in transmembrane protein folding. Utilizing the nucleic acid polymer REP 2139 as a bait, we identified DNAJB12 as a REP 2139 interactor, and its knockdown impedes the morphogenesis and secretion of SVP and HBV virions. Conversely, DNAJB14, which did not interact with REP 2139, selectively impaired the morphogenesis of virions. Additionally, knockdowns of DNAJB12 and DNAJB14 hindered the production of infectious HDV. As DNAJB12 knockdown recapitulated REP 2139 antiviral effects observed in clinical trials, our findings highlight DNAJB12 as the potential primary target of REP 2139 and uncover functional roles for DNAJB12 and DNAJB14 in HBV and HDV life cycles.

Tremor is a common movement disorder associated with several neurodegenerative diseases, yet its mechanisms are not well understood. Using a machine-learning method, Feature Learning-based Leg segmentation and Tracking (FLLIT), we previously characterised gait and tremor signatures in a Drosophila model for spinocerebellar ataxia 3 (SCA3) and found them to be analogous to those in human SCA3. Here, we carried out a functional screen for neuronal populations that underlie tremor and found that dysfunction of a specific population of neurons in the ventral nerve cord (VNC) is necessary and sufficient for tremor. Adult-onset expression of mutant ATXN3 in, or genetic hypo-activation of, these neurons led to tremor, indicating their important role in adult motor control. RNA-sequencing and functional experiments showed that dysfunction of GABAergic neurons, and not that of other neurotransmitter populations tested, causes tremor. Finally, we identified a small subset of ∼30 predominantly GABAergic neurons within the adult VNC that are essential for smooth walking. This study demonstrates that tremor in SCA3 flies arises from GABAergic dysfunction, and that FLLIT can be used to dissect motor control mechanisms.

Poly(ethylene glycol) (PEG) hydrogels crosslinked by orthogonal thiol-norbornene click chemistry have emerged as an ideal platform for tissue engineering and drug delivery applications due to their rapid crosslinking kinetics and excellent biocompatibility. Norbornene-functionalized PEG (PEGNB) is routinely synthesized through the Steglich esterification of 5-norbornene-2-carboxylic acid with hydroxyl-terminated PEG. When crosslinked with thiol-bearing macromers, PEGNB hydrogels undergo slow hydrolytic degradation due to hydrolysis of ester bonds connecting a PEG backbone and a NB moiety. In prior work, we replaced the pungent and nauseous 5-norbornene-2-carboxylic acid with odorless carbic anhydride (CA) for synthesizing PEG-norbornene-carboxylate (PEGNB_CA), a new macromer that could be readily photo-crosslinked into thiol-norbornene hydrogels with faster hydrolytic degradation than the PEGNB counterparts. In this contribution, we employed a modular approach to tune the hydrolytic degradation of PEGNB_CA hydrogels over days to months. We first demonstrated the diverse crosslinking of PEGNB_CA hydrogels using either photopolymerization or enzymatic crosslinking. We characterized the hydrolytic degradation of these hydrogels under different solution pH values and temperatures. <i>Via</i> adjusting crosslinker functionality and the ratio of fast-degrading PEGNB_CA to slow-degrading PEGNB, tunable hydrolytic degradation of PEGNB_CA hydrogels was achieved from under 2 days to over 3 months. Finally, we designed the highly tunable PEGNB_CA hydrogels with varying mesh sizes, degradation rates, and covalent tethering of degradable linkers to afford long-term controlled release of model drugs.

The beet armyworm, <i>Spodoptera exigua</i>, is a significant agricultural pest managed primarily through chemical insecticides, which present environmental concerns and promote resistance. Genetics-based sterile insect technique (gSIT) provides an alternative by inducing male sterility without irradiation. In this study, we cloned the <i>S. exigua</i> boule gene (<i>Sebol</i>), a 741-nucleotide open reading frame encoding a 246-amino-acid protein with an RNA recognition motif. Phylogenetic analysis confirmed its conservation across Lepidoptera and high similarity to orthologs in <i>Spodoptera litura</i> and <i>Helicoverpa armigera</i>. <i>Sebol</i> is predominantly expressed in adult males, particularly in the testis. CRISPR/Cas9-mediated deletion of 190 bp in exon 1 generated a truncated Sebol protein, resulting in complete male sterility. Transcriptome analysis revealed 2069 differentially expressed genes associated with sperm development, with potential interactions involving <i>syf1</i> and <i>ZNFX1</i>. Our findings suggest that <i>Sebol</i> is crucial for spermatogenesis, making it a promising target for gSIT-based pest control.

<h4>Objective</h4>Pyroptosis, apoptosis and necroptosis separately participate in the pathogenesis of ischemic stroke, but the implication of PANoptosis in this disease is still unexplored. This study conducted a comprehensive investigation on PANoptosis in ischemic stroke and glioma.<h4>Methods</h4>Four transcriptomic datasets of ischemic stroke: GSE16561, GSE58294, GSE22255, and GSE37587 were gathered. Consensus clustering analysis was implemented based upon PANoptosis genes. WGCNA was utilized for PANoptosis subtyping-related genes. Multiple well-established machine learning models were also built for feature gene selection. Moreover, survival and receiver operating characteristic (ROC) analysis was performed to evaluated the prognostic value of the feature genes in the glioma.<h4>Results</h4>Fifteen PANoptosis genes were aberrantly expressed in peripheral blood of ischemic stroke than that of controls. Based upon them, ischemic stroke was classified as two PANoptosis molecular subtypes, with heterogeneous molecular mechanisms, and immune cell infiltration. The PANoptosis molecular subtyping-related module genes were determined, and key genes were subsequently via distinct machine learning approaches. Among them, the SVM model had the lowest |residual|, and the AUC of 1 in diagnosing ischemic stroke. The feature genes in the SVM model were regarded as the key genes (comprising TMEM55A, CCPG1, CYP1B1, PJA2, MTPN, SCYL2, BAZ2B, and DPYD). Based upon them, the nomogram was established, which could accurately predict ischemic stroke risk. All of them were in relation to ischemic stroke-related signaling pathways, and immune cell infiltration. Transcription factors were predicted, which transcriptionally modulated them. Additionally, drugs that potentially targeted them were determined. Moreover, the expression and prognostic value of genes in glioma were evaluated in the The Human Protein Atlas (HPA) and The Cancer Genome Atlas database (TCGA). All genes showed a significant prognostic value except PJA2. The diagnostic and prognostic ROC analysis result indicated that BAZ2B can served as potential diagnostic and prognostic marker in glioma.<h4>Conclusion</h4>Altogether, this is the first study to characterize PANoptosis-based molecular subtypes, and diagnostic model for ischemic stroke, providing a promising avenue for patient risk prediction through a non-invasive method. Moreover, the BAZ2B genes could serve as predictive biomarkers in glioma, providing essential implications for patient prognosis.

Autism Spectrum Disorder (ASD) involves a multi-system interaction mechanism among genetics, immunity, and gut microbiota, yet its regulatory network remains undefined. This study conducted a meta-analysis on Genome-Wide Association Study data from four independent ASD cohorts to identify potential genetic loci. By integrating Polygenic Priority Score, brain region, and brain cell eQTL enrichment analyses, and combining summary-data-based Mendelian Randomisation (SMR) analyses of brain cis-eQTL and mQTL, bidirectional Mendelian Randomisation analyses of 473 gut microbiota, and SMR analysis of blood eQTL, SNPs such as rs2735307 and rs989134 with significant multi-dimensional associations were identified. These loci exert cross-tissue regulatory effects by participating in gut microbiota regulation, involving immune pathways such as T cell receptor signal activation and neutrophil extracellular trap formation, as well as cis-regulating neurodevelopmental genes (HMGN1 and H3C9P), or synergistically influencing epigenetic methylation modifications to regulate the expression of BRWD1 and ABT1. The cross-scale evidence chain constructed in this study provides a theoretical foundation for precision medicine research in ASD, holding promise to advance the development of innovative therapeutic strategies.

The study aims to develop a composite coating for magnesium aluminum alloy (MA) to improve its corrosion/infection resistance. MA was coated with carboxymethyl chitosan (CMCS) and nano-hydroxyapatite (HA) through electrophoretic deposition, followed by the deposition of vancomycin-loaded polymer nanoparticles to obtain the ternary composite coating alloy (VAN@PLGA/HA/CMCS-MA). In simulated body fluid (SBF), the average corrosion rate of the coating alloy was 0.27 ± 0.03 mg/cm^-1/day^-1, while maintaining a pH level of approximately 7.2, indicating that the composite coatings effectively mitigated erosion in SBF and ensured a stable physiological pH. In vitro antibacterial experiments showed that VAN@PLGA/HA/CMCS-MA exhibited enhanced antibacterial activity against staphylococcus aureus and enterococcus faecalis compared to single MA owing to sustained antibiotic release. Furthermore, the composite coating promoted alkaline phosphatase activity and induced extracellular Ca²⁺ mineralization, suggesting good bone-promoting ability of the alloy. Finally, the biocompatibility studies confirmed that the composite coating could reduce mild toxicity of the alloy following corrosion, resulting in enhanced cell viability and reduced hemolysis rate. Overall, the ternary composite alloy coating delayed MA degradation and provided long-term effective corrosion/infection resistance.

BackgroundThe HTT protein, mutated in Huntington's disease, is expressed throughout the body, and loss of HTT function as an autophagic scaffold may affect tissues and cellular processes. These processes include lipid metabolism potentially regulated upstream by Apolipoprotein E (APOE) and clearance of APOE itself.ObjectiveTo determine the impact of HTT reduction on autophagy and clearance of APOE in cell culture and in mouse liver <i>in vivo</i>.MethodsWestern blot analysis was performed on liver tissue from tamoxifen-treated mice with and without UBC-Cre expression, required for tamoxifen-induced HTT knockout (KO). siRNA was used to knockdown (KD) HTT in HepG2 immortalized liver cells.ResultsHTT KO in mouse liver reduces levels of LAMP2A, a protein essential for chaperone-mediated autophagy (CMA) which we previously found is required for optimal degradation of APOE and HTT in cultured cells. In turn, APOE levels were increased with HTT KO in mouse liver, while HTT KD in cell culture decreased levels of APOE.ConclusionsIn the context of liver tissue, reduced CMA may contribute to accumulation of APOE and autophagic cargo resulting from a loss of HTT function in autophagy. The extent to which macroautophagy is upregulated to cope with reduced CMA found with HTT KO may be tissue specific, which may relate to the selectivity of tissue pathogenesis observed in Huntington's disease where loss of normal HTT function may be involved. This study may help elucidate the consequences of systemic HTT reduction on autophagy in liver tissue.

Zinc finger NFX1-type containing 1 (ZNFX1) has been established as a critical mediator of the antiviral response in mammals, functioning through dsRNA recognition and priority activation of the retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) signaling pathway. However, the role of its fish ortholog, particularly in relation to aquatic virus interactions, remains elusive. The absence of the RIG-I homolog, a key pattern recognition receptor, in multiple Actinopterygii may compromise their innate antiviral immune responses. Here, ZNFX1 in Epinephelus coioides (EcZNFX1) is identified as an evolutionarily primitive, interferon (IFN)-stimulated dsRNA sensor that compensates for the absence of RIG-I in Actinopterygii. EcZNFX1 is rapidly upregulated by orange-spotted grouper nervous necrosis virus (OGNNV) infection and restricts viral replication in grouper brain-derived (GB) and spleen-derived (GS) cells after binding to viral dsRNA intermediates via its conserved P-loop NTPase domain. Notably, EcZNFX1 exerts a dual immunoregulatory role in modulating virus-induced inflammatory responses in diverse cellular contexts. In GB cells that are highly susceptible to OGNNV and have attenuated regenerative capacity, EcZNFX1 suppresses IFN-I/ISGs production and pyroptosis mediated by viral infection, thereby limiting neurotoxicity by precise tuning of the antiviral response. Conversely, in GS cells with stronger resistance to OGNNV, EcZNFX1 directly interacts with TBK1 to promote its phosphorylation and subsequent nuclear translocation of IRF3, activating a robust IFN-I signaling. Overall, this study elucidates that ZNFX1 is a compensatory receptor for dsRNA sensing in RIG-I-deficient teleost, which exerts context-dependent antiviral effects in cell-type-specific microenvironments, providing mechanistic insights for aquatic virus countermeasures.

RAC1^A159V is a hotspot mutation associated with poor prognosis in several cancers. By gene editing, we generated endogenous homozygous and heterozygous RAC1^A159V mutations, which result in up-regulated RAC1 activity and mammalian target of rapamycin (mTOR) signaling. RAC1^A159V tumors grow faster than RAC1^WT tumors in immune-proficient mice and are resistant to anti-programmed death protein 1 (PD1). Flow cytometry and scRNA-seq analyses reveal that RAC1^A159V cells form "cold" tumors with an immunosuppressive microenvironment and reduced tumor-immune cell interactions. Mechanistically, RAC1^A159V up-regulates glycosphingolipid biosynthesis to activate mTORC1 signaling in tumor cells, which in turn increases glycolysis, impairs key chemokine production, and decreases IFNGR1 expression of the tumor cells. mTORC1 inhibition by rapamycin resensitizes the RAC1^A159V tumors to anti-PD1 treatment by reversing effects of RAC1^A159V mutation. These results demonstrate a mechanism of RAC1^A159V-driven immune evasion and suggest an approach of combining the targeting of RAC1-mTOR signaling with immune checkpoint inhibitor for the treatment of a type of immune-cold tumors.

<h4>Background</h4>Despite the notable success of programmed death ligand 1 (PD-L1)/programmed death 1 (PD-1) immune checkpoint blockade in cancer, resistance remains a substantial challenge. Combining immune checkpoint blockade with direct targeting of effector T cells to tumor cells might improve outcome for a broader spectrum of patients with cancer. Phosphoantigen-responsive Vγ9Vδ2-T cells are potent immune cells that play a pivotal role in tumor immunosurveillance and can coordinate downstream immune activity. Here, we explored whether a Vδ2 bispecific T cell engager (bsTCE) directed against PD-L1 could combine lysis of PD-L1⁺ tumor cells with PD-L1 immune checkpoint blockade.<h4>Methods</h4>PD-L1 specific single domain antibodies (VHHs) were tested for binding to PD-L1 and their ability to interfere with PD-1 binding and function. One PD-L1 VHH was selected for fusion to a Vδ2-T cell receptor specific VHH (PD-L1xVδ2 bsTCE) and tested for its ability to activate Vγ9Vδ2-T cells and lyse melanoma cell lines, as well as patient-derived renal cell carcinoma (RCC) and metastatic melanoma cells. These patient-derived tumor suspensions were also used to explore effects on CD4⁺ and CD8⁺ T cells and myeloid cells. Infiltration of Vγ9Vδ2-T cells and tumor kill was tested in a three-dimensional tumor spheroid melanoma model.<h4>Results</h4>A PD-L1xVδ2 bsTCE was generated and shown to block PD-1 binding resulting in the release of PD-1⁺ cells from PD-L1 mediated inhibition. The PD-L1xVδ2 bsTCE also mediated robust Vγ9Vδ2-T cell activation, efficient lysis of RCC and melanoma cell lines and/or patient-derived tumor cells, and infiltration of Vγ9Vδ2-T cells into a three-dimensional melanoma spheroid model. Of interest, exposure of co-cultures of Vγ9Vδ2-T cells and patient-derived tumor suspensions to the PD-L1xVδ2 bsTCE resulted in upregulation of activation markers on tumor-infiltrated CD4⁺ and CD8⁺ T cells and lysis of PD-L1⁺ myeloid cells with a shift in the myeloid compartment from macrophage-like cells to more mature dendritic cells with costimulatory molecule expression.<h4>Conclusions</h4>A PD-L1xVδ2 bsTCE was generated that acts as PD-1/PD-L1 immune checkpoint inhibitor, enhances Vγ9Vδ2-T cell activation, infiltration and tumor lysis and reshapes the tumor microenvironment towards a more proinflammatory state. By targeting both PD-L1 expressing tumor and myeloid cells, it addresses key challenges of current therapies and thereby offers a promising novel therapeutic strategy.

Lung structures are critical for gas exchange and contribute to the pathogenesis of respiratory diseases, exhibiting notable lobe-specific heterogeneity. To investigate their genetic basis, we apply a deep-learning AI system and Pyradiomics to define lobe-specific lung CT imaging phenotypes, conducting genome-wide analyses in 35,469 participants from the Lung Imaging Genomics Initiative in China. We identify 36 loci associated with voxel intensities and 138 loci linked to three-dimensional shape. Functional annotation reveals significant enrichment of relevant genes in pathways regulating early fetal lung development and loci enriched in fetal lung regulatory elements. Genetic correlations are identified between lung structures and chronic respiratory diseases as well as lung function, with a number of loci showing colocalization. Mendelian randomization analyses suggest a causal role of lung structures in chronic lung diseases and extrapulmonary traits. This study provides new insights into the genetic architecture of lung structures and their links to diverse clinical outcomes.

Regulated pausing of RNA polymerase II (Pol II) is essential for enabling rapid and coordinated transcriptional responses to signalling cues. Pausing also contributes to the formation of nucleosome-free regions with the help of chromatin remodellers. However, if these nucleosome-free regions engage with transcription factors to stimulate the transcription potential of paused promoters is not known. In this study, we demonstrate that ligand-induced estrogen receptor-alpha (ERα) binding is stabilized at Pol II-paused sites. This stabilization results from an increased dwell time of ERα on chromatin, as revealed by single molecule tracking (SMT) experiments. Notably, short chromatin-associated RNAs generated by the paused Pol II contribute to enhancing ERα binding at paused promoters. We also observe that pausing increases H3K27ac levels, which primes paused promoters for robust transcriptional activation upon release. Collectively, these findings suggest that paused Pol II plays a central role in enhancing transcription factor binding through an RNA-dependent mechanism. This, in turn, results in a more vigorous transcriptional response following pausing release, thus contributing to the fine-tuning of ERα-mediated gene regulation.

Exosomes play a crucial role in triple-negative breast cancer (TNBC), influencing various aspects of tumor progression. Given the importance of exosomes in TNBC biology, we proposed a novel exosome-based model that was critically implicated in TNBC. Bulk and single-cell transcriptomics and genetic mutations of TNBC patients were curated for our study. Characteristic exosome genes were selected via LASSO analysis, with subsequent construction of an exosome-based model. The effectiveness in estimating clinical outcomes and treatment responses was then evaluated and validated. MDA-MB-231 and MDA-MB-468 TNBC cells were transiently transfected with FAM129B siRNAs, and cell proliferation and migration were measured via EdU and wound healing assays. The study determined 7 characteristic exosome genes for TNBC: ALCAM, FAM129B, GNB2, KRT6A, PGK1, SERPINE1, and THY1, which were utilized for defining the exosome-based gene signature. It was proven that the signature accurately estimated patient prognosis, and functioned as an independent prognostic predictor. High-risk tumors owned shorter overall survival time, but were suitable for treatment with docetaxel and several small-molecule agents (MK-0752, BRD-K33199242, IC-87114, fumonisin B1, ilomastat, GW-788388, afobazole, and batimastat). High- and low-risk tumors presented the distinct genetic mutation characteristics. The characteristic exosome genes were specifically expressed in the TNBC microenvironment components, indicating their involvement in modulating the microenvironment. High-risk individuals were inferred to better respond to immune checkpoint blockade (CD276, NRP1, TNFRSF4, TNFSF4 or CTLA4). Experimentally, inhibition of FAM129B effectively attenuated proliferative and aggressive phenotypes of TNBC cells. Collectively, our findings proposed the exosome-based gene signature for accurate estimation of clinical outcomes and assisting in individually tailoring therapies in TNBC as well as discovered FAM129B as a potential therapeutic target.

Cholangiocarcinoma (CC) is a highly lethal malignancy that urgently requires reliable prognostic biomarkers. Although MUC1 expression and promoter methylation have been implicated in CC, the clinical significance of promoter methylation pattern composition, beyond average methylation levels, remains unclear. Here, we investigated the relationship between MUC1 promoter methylation heterogeneity, MUC1 mRNA expression, and prognosis in CC. We analyzed bisulfite amplicon sequencing data and mRNA expression of MUC1, DNA methylation-related enzymes (TET1, TET2, TET3, Dnmt1, and Dnmt3a), and tumor microenvironment stress markers in 131 CC tissues. In the neoplastic region, high MUC1 mRNA expression was associated with poor overall survival (HR = 0.131, 95% CI: 0.02 to 0.95, p = 0.042) and correlated with the abundance of completely unmethylated promoter patterns (r = 0.386, p < 0.001). Among the enzymes analyzed, only TET3 expression significantly correlated with the abundance of completely unmethylated patterns in the neoplastic region (Cohen's f² = 0.108, p = 0.009), suggesting a potential region-specific regulatory association. We visualized beta-diversity in methylation pattern composition using t-SNE and classified samples into two groups based on a linear decision boundary in the t-SNE space. This classification stratified prognosis independently of clinical factors (HR = 0.291, 95% CI: 0.06 to 0.94, p = 0.037; multivariate p = 0.021). These findings propose a novel, composition-based epigenetic stratification framework in CC, revealing that MUC1 promoter methylation pattern structure-rather than average methylation level-has prognostic relevance. Our results highlight the potential of pattern-resolved methylation profiling in the development of clinically applicable epigenetic biomarkers.

The histone-fold domain (HFD) is a conserved protein interaction module that requires stabilization through a handshake interaction with an HFD partner. All HFD proteins known to date form obligate dimers to shield the extensive hydrophobic residues along the HFD. Here, we find that the lepidopteran kinetochore protein CENP-T is soluble as a monomer. We attribute this stability to a structural rearrangement, which leads to the repositioning of the HFD helix α3. This brings a conserved two-helical extension closer to the histone fold, where it takes over the position and function of the CENP-T partner CENP-W. This change has no effect on the DNA-binding ability of the lepidopteran CENP-T. Our analysis suggests that the monomeric HFD originated in the last common ancestor of insects, with a possible second independent origin in Acariformes, both of which lack CENP-W. Our study highlights an unexpected structural variation in a protein module as conserved and optimized as the HFD, providing a unique perspective on the evolution of protein structure and the forces driving it.

Heart development relies on topologically orchestrated cellular transitions and interactions, many of which remain poorly characterized in humans. Here, we combined unbiased spatial and single-cell transcriptomics with imaging-based validation across postconceptional weeks 5.5 to 14 to uncover the molecular landscape of human early cardiogenesis. We present a high-resolution transcriptomic map of the developing human heart, revealing the spatial arrangements of 31 coarse-grained and 72 fine-grained cell states organized into distinct functional niches. Our findings illuminate key insights into the formation of the cardiac pacemaker-conduction system, heart valves and atrial septum, and uncover unexpected diversity among cardiac mesenchymal cells. We also trace the emergence of autonomic innervation and provide the first spatial account of chromaffin cells in the fetal heart. Our study, supported by an open-access spatially centric interactive viewer, offers a unique resource to explore the cellular and molecular blueprint of human heart development, offering links to genetic causes of heart disease.

<h4>Background</h4>Metabolic dysfunction-associated steatotic liver disease (MASLD) affects a quarter of the population. Although multiple studies have investigated the mechanisms underlying steatohepatitis and MASLD progression, little attention has been paid to milder cases, particularly those with normal enzyme levels. Our objective was to evaluate the molecular events in patients with MASLD and normal enzyme levels.<h4>Methods</h4>We performed a proteomic analysis on serum samples from 83 healthy controls and 50 patients with MASLD. The major findings were then validated in serum samples from 35 controls and 60 patients with MASLD and normal liver enzyme levels (MASLD-NLE), as well as a male mouse model with metabolic dysfunction-associated liver steatosis and normal liver enzymes (mice-NLE) induced by a high-fat diet (HFD).<h4>Results</h4>Despite having normal enzyme levels, patients with MASLD-NLE exhibited significantly higher levels of ALT, AST, γGGT, hs-CRP, uric acid and creatine compared to controls. Serum proteomics indicated elevated complement-related proteins in patients with MASLD-NLE, evidenced by elevated protein levels of C3 and CFH in both cases using different cutoffs for normal ALT levels. C3 and CFH also showed positive correlations with γGGT, hs-CRP, ALT, and uric acid levels. Additionally, the elevation of C3 and CFH was confirmed in another set of human serum samples, as well as in serum and liver samples of mice-NLE using ELISA.<h4>Conclusions</h4>Our study provides novel evidence that the complement system is already activated in patients with MASLD-NLE, potentially offering new avenues for treating MASLD at an early stage.

BACKGROUND: Elderly patients with type 2 diabetes mellitus (T2DM) often face complex healthcare needs due to comorbidities and polypharmacy, increasing the risk of poor glycemic control and drug-related problems. Comprehensive Clinical Pharmacy Services (CPS) aim to optimize medication management and improve adherence, addressing these challenges effectively. METHODS: This prospective, single-center implementation study was conducted in the outpatient clinic of the Department of Endocrinology – Diabetes of a tertiary care hospital in Vietnam. Elderly patients aged ≥ 60 years with a diagnosis of T2DM for at least one year and an HbA1c level ≥ 8% were included. A trained clinical pharmacist provided a structured CPS intervention, including Medication Therapy Management (MTM) and collaborative goal-setting. The impact of pharmacy services was assessed through the number of Drug-Related Problems (DRPs) identified and resolved in prescriptions and patient medication use behaviors, improvements in HbA1C levels, as well as the satisfaction of both physicians and patients with the services provided by pharmacists. RESULTS: Among 210 patients (mean age 73.5 years; 71% male), the baseline median HbA1c was 8.6% (IQR: 8.2–9.13), with 90.5% experiencing polypharmacy. A total of 231 DRPs were identified, primarily patient-related (66.2%), including unintentional misuse (28.6%) and intentional non-adherence (14.7%). DRPs related to prescribing included inappropriate drug (21.2%) and dose selection (12.6%). Of 78 pharmacist interventions, 93.5% were fully accepted or partially accepted by physicians; 100.0% were deemed clinically significant (potential Adverse Drug Event Score - pADE ≥ 0.1). Following CPS, 22.9% of patients achieved individualized HbA1c targets (p < 0.05), while the proportion of patients with HbA1c ≥ 8.5% declined from 49.2% to 32.2%. Patient and physician satisfaction with CPS was high (95.2% and 100%, respectively), though only 46.3% of patients expressed willingness to pay for services. CONCLUSION: Comprehensive CPS has a significant positive impact on quality use of medicine in elderly outpatients with T2DM. These services should be formally integrated into routine diabetes care to optimize outcomes and reduce healthcare utilization in this population.

<h4>Purpose</h4>Tumor Metabolic Behavior modulates the immunosuppressive microenvironment through multiple pathways, thereby compromising anti-tumor immune responses. To date, there have been limited studies assessing the role of metabolic plasticity or immunometabolism in the tumor microenvironment (TME) during metastasis. Notably, emerging evidence suggests the presence of an immunosuppressive niche in brain metastases. This research aims to delineate distinct metabolic signatures in brain metastatic, investigate the impact of tumor-associated glycolysis on the development of brain metastases in lung adenocarcinoma, and characterize the lactylation regulation in this immunosuppressive microenvironment.<h4>Methods</h4>The GSE131907 and GSE198291 datasets were retrieved for bioinformatic analysis. Combined with the results of proteomic and transcriptomic sequencing conducted on the lung adenocarcinoma brain metastasis model, differentially expressed signaling pathways were systematically identified through KEGG and GO functional annotations. A multimodal approach encompassing immunohistochemical (IHC) staining, immunofluorescence (IF) imaging, enzyme-linked immunosorbent assay (ELISA) quantification, and co-immunoprecipitation (Co-IP) assays was employed to experimentally validate the characteristics of the immunosuppressive microenvironment and the levels of tumor lactate/lactylation. Rescue experiments were performed by adding a lactylation-specific inhibitor (LDHi) or an H3K18la site-specific inhibitor. Finally, immunohistochemical staining was used to verify the expression level of H3K18la in clinical samples.<h4>Results</h4>A total of 86,215 cells were extracted from the GSE131907 dataset, and the metabolic profiles of different cell types were analyzed. The results showed that glycolysis plays a dominant role in tumor cell metabolism. Further analysis revealed that early-stage primary lesions exhibit an inflammatory response signature, while advanced-stage primary lesions and brain metastatic lesions display an immunosuppressive signature. Elevated glycolytic flux showed a significant positive correlation with both the progression of brain metastasis and the immune evasion capacity of brain metastatic lesions. Pathological evaluation of tumor tissues from the LLC-BM (Lewis Lung Cancer Brain Metastasis) model confirmed its immunosuppressive characteristics. Additionally, obvious hypoxia was observed in the tumor tissues, accompanied by intratumoral vascular malformation and dysfunction. Significant lactate accumulation was present in the tumor microenvironment of LLC-BM tumors, and prominent lactylation modifications were detected in the tumor regions. In this model, Rac2 was identified as a potential core mediator of lactylation modification in macrophages, promoting the M2 polarization of macrophages. Meanwhile, CD40, TNFSF13 and CCL22 were identified as key immunoregulatory factors regulated by lactylation signaling. Notably, H3K18la was significantly highly expressed in lung cancer brain metastatic lesion samples.<h4>Conclusions</h4>The glycolytic pathway plays a critical role in the metabolic reprogramming of tumor cells during lung adenocarcinoma brain metastasis. Tumor glycolysis is closely associated with lung cancer progression, brain metastasis, and immune evasion. The Rac2 could be affected by lactylation, and then facilitate the formation of an immunosuppressive tumor microenvironment by induce the M2 polarization of macrophages.

<h4>Background</h4>Iron homeostasis is essential for fundamental biological processes, yet its impact on epigenetic aging and mortality remains poorly understood. This study aimed to investigate associations between dietary iron intake and DNA methylation (DNAm) aging markers and to determine whether these epigenetic signatures mediate iron's effects on mortality outcomes.<h4>Methods</h4>We analyzed data from 2,398 adults aged ≥ 50 years in the National Health and Nutrition Examination Survey (1999-2002) with up to 20 years of mortality follow-up. Dietary iron intake was assessed through 24-h recall interviews. DNA methylation was profiled using the Illumina Infinium MethylationEPIC BeadChip. We employed multiple linear regression to identify iron-responsive DNAm features, Cox proportional hazards models to assess mortality associations, and formal mediation analyses to quantify epigenetic pathways.<h4>Results</h4>Higher dietary iron intake was significantly associated with favorable epigenetic profiles, particularly with reduced levels of mortality-predictive DNAm markers GrimAge2Mort (β = -0.079, FDR = 0.00022), CRPMort (β = -0.072, FDR = 0.0037), and B2MMort (β = -0.057, FDR = 0.042). These iron-responsive DNAm features strongly predicted all-cause mortality (HRs per SD increase: 1.34, 1.21, and 1.08, respectively; all p < 0.05) and cause-specific mortality. Higher iron intake was directly associated with reduced risk of all-cause mortality (HR = 0.93 per SD increase, 95% CI 0.87-0.99), heart disease mortality (HR = 0.87, 95% CI 0.78-0.97), and respiratory disease mortality (HR = 0.72, 95% CI 0.56-0.93). Mediation analyses revealed that GrimAge2Mort mediated 22.7% of iron's protective effect on all-cause mortality and 14.7% on heart disease mortality.<h4>Conclusions</h4>This study establishes dietary iron as a modifiable determinant of epigenetic aging and mortality risk in older adults, with epigenetic recalibration mediating a substantial proportion of iron's protective effects. These findings suggest optimizing iron intake may represent a promising nutritional strategy to promote healthy aging.

<h4>Background</h4>Subcellular RNA localization is crucial for the spatio-temporal control of protein synthesis and underlies key processes during development, homeostasis, and disease. In epithelial cells, RNA can localize asymmetrically along the apico-basal axis. Yet, the localization of most transcripts as well as the diversity of patterns that they adopt remains unexplored.<h4>Results</h4>Here, we use APEX-seq for proximity labeling and MERFISH for spatial transcriptomics to map subcellular transcript localization in intestinal organoids and tissue from adult mice. Many transcripts present localization bias, often localizing in granular structures. We uncover intrinsic and environmental factors that influence the formation of these patterns. Additionally, we identify translation-dependent and -independent localization patterns and pinpoint the role of 3' untranslated regions and RNA-binding proteins.<h4>Conclusions</h4>This subcellular RNA atlas presents a detailed resource for understanding intestinal physiology.

Arrays of pico-to-microliter droplets, organized on a surface, enable chemical and biological workflows at high throughput. Here, a platform employing two-layer droplets is presented to enable flexible manipulation of the droplets' microenvironment for dynamic biological cultivation. Arrays of 6784 agarose droplets (≈2.0 nL per droplet) encapsulating and immobilizing bacterial cells are generated. After that, aqueous droplets (≈3.7 nL) with a defined composition are deposited atop to form a thin liquid layer surrounding the agarose droplets. Chemical exchange between the two layers is extremely fast (equilibrium within 15 s for fluorescein). Moreover, the aqueous layer can be removed, opening the possibility to extract substances from the agarose droplets. Indeed, repeated addition and aspiration of a buffer successfully remove dyes or drugs previously added to the agarose droplets. Therefore, antibiotic drug testing can be performed under both static and transient exposure profiles. The latter reveals that bacterial responses such as bacterial killing and resuscitation are both heterogeneous at the single-cell level. Last, it is exemplified how such droplet manipulation strategy can also be use in long-term experimentation, where medium replenishment, performed at 12-h intervals during a 72-h experiment, enables the cultivation of a slow-growing microorganism in nanoliter droplets.

Cysteine oxidative modifications are critical signaling events regulating cellular functions, but their low abundance and dynamic nature pose technical challenges. We developed the SICyLIA-TMT workflow, which sequentially labels reduced and reversibly oxidized cysteines with light and heavy iodoacetamide (IAA) within the same sample. The inclusion of tandem mass tags (TMTs) enables simultaneous quantification of oxidative modification dynamics and protein levels across multiple conditions using micrograms of material. To improve the detection of low-abundance oxidized cysteines, a dedicated TMT channel serves as a carrier for heavy IAA-labeled peptides (SICyLIA-cTMT), enhancing quantification and enabling precise stoichiometry calculations. We demonstrate the workflow's applicability to cultured cells and full organs under stress. SICyLIA-cTMT achieves unprecedented depth and accuracy in redox proteome analysis while reducing mass spectrometry time. Combining SICyLIA-TMT with latest mass spectrometry technologies further halves the acquisition time without compromising coverage, improving throughput and enabling comprehensive studies of oxidative signaling.

Loss-of-function variants in TBK1, encoding a protein kinase, are strongly associated with familial amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, how haploinsufficiency for TBK1 leads to age-related neurodegeneration remains unresolved. Here, we utilize sets of isogenic induced pluripotent stem cells (iPSCs) with loss of TBK1 or loss of optineurin (OPTN) for quantitative global proteomics and phospho-proteomics in both stem cells and excitatory neurons. We found that TBK1 sustains the abundance and phosphorylation of its interacting adapter proteins, AZI2/NAP1, TANK, and TBKBP1/SINTBAD. Moreover, TBK1 regulates the phosphorylation of endo-lysosomal proteins, such as GABARAPL2, the late-endosome GTPase RAB7A, and selective autophagy cargo receptor proteins-including novel phospho-sites in p62/SQSTM1-in neurons. Finally, we provide a census of the phospho-proteome in nascent human neurons for further studies. Overall, TBK1 serves as a point of convergence in ALS/FTD-linked endo-lysosomal networks that act in a cell-autonomous manner to maintain protein homeostasis in neurons.

Mussels' byssus and their adhesion ability play a crucial role in their attachment and artificial cultivation of mussels. In this study, transcriptomic and proteomic analyses were performed to identify byssogenesis-associated genes in the Mediterranean mussel <i>Mytilus galloprovincialis</i> Lamarck, 1819, seeking to advance our knowledge of the molecular basis of byssal secretion in mussels. Transcriptomic analysis identified 1742 and 1498 differentially expressed genes in the foot tissue of <i>M. galloprovincialis</i> at 9 h and 24 h post-byssal ablation, respectively. Meanwhile, proteomic analysis revealed 1254 and 484 differentially expressed proteins at the same two time points. Integrated analysis identified 121 genes differentially expressed at both transcript and protein levels. Among these genes, 44 were significantly upregulated, and they may constitute high-confidence gene sets associated with mussel byssogenesis. Notably, they included genes encoding tyrosinase-like protein, low affinity immunoglobulin epsilon Fc receptor, and O-methyltransferase MdmC. They were enriched in KEGG pathways, including metabolism of amino acids, lipid metabolism, nucleotide metabolism, and immune system. Quantitative real-time PCR was performed on seven selected genes, confirming that their expression patterns were consistent with those observed in transcriptomic and proteomic sequencing. This study provides novel data and insights for understanding the molecular basis involved in byssus development of <i>M. galloprovincialis</i>.

Gold nanoparticles (AuNPs) are of significant interest due to their unique properties and applications in biomedicine. While hyaluronic acid (HA) has been used to modify pre-formed AuNPs, its thiolated derivative (HA-SH) has been less explored for the direct synthesis and stabilization of AuNPs. This study investigates the use of thiolated hyaluronic acid as a key component in the synthesis of AuNPs. A series of HA-AuNPs (<b>HA-AuNP1-4</b>) were synthesized by reacting HA-SH with HAuCl₄ at different mass ratios. The resulting nanoparticles were characterized using UV-Vis spectroscopy, scanning/transmission electron microscopy (SEM/STEM), X-ray photoelectron spectroscopy (XPS), photon cross-correlation spectroscopy (PCCS), and zeta potential measurements. The chemical transformations of the thiol ligand were studied using NMR spectroscopy. The morphologies and sizes of AuNPs depended on the HA-SH-to-HAuCl₄ ratio, ranging from icosahedral and triangular particles (≥146 nm) to quasi-spherical particles with a bimodal distribution (6-7 nm and 45-60 nm). XPS confirmed the presence of metallic gold (Au⁰) and a Au-S bond, while NMR and XPS revealed the partial oxidation of thiol groups to sulfonic acid. Zeta potential measurements showed that lower HAuCl₄ concentrations resulted in higher negative charge (up to -41.5 mV), enhancing colloidal stability. This work demonstrates a versatile approach to the synthesis of hyaluronic acid-based gold nanomaterials with tunable properties for potential biomedical applications.

Calcium phosphates, including hydroxyapatite, are widely used biomaterials in bone tissue regeneration due to their bioactivity, osteoconductivity, and similarity to the mineral phase of bone. In this study, various apatite calcium phosphate powders were synthesized using three precipitation methods, with controlled pH conditions and reagent ratios, to assess the effect of the synthesis method on their physicochemical and biological properties. Elemental composition (Ca/P ratio), FT-IR spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM) coupled with EDS, and particle size measurements were used to determine the structure, morphology, and stoichiometry of the obtained powders. The results indicated that the synthesis method and pH significantly affect the phase composition of the material, particle size, and Ca/P ratio, which directly influence their solubility and bioactivity. Microbiological tests, NF-κB transcription factor activation, metabolic activity, and cell compatibility of mouse L929 fibroblasts and human hFOB 1.19 osteoblasts showed good biological tolerance of the obtained powders and no cytotoxic effects. The results confirm that a properly selected synthesis method allows for the control of material properties, which is crucial for applications in bone tissue engineering. The materials show potential for use as bioactive components in bone-related biomaterials.

(1) Background: Oral squamous cell carcinoma (OSCC) is the most common type of head and neck malignancy worldwide. Despite the prevalence of modern diagnostic and prognostic techniques, late diagnosis and resistance to treatment still result in a low 5-year survival rate, high recurrence rate, and frequent malignant metastases. Increasing evidence indicates that bacteria of the oral microbiome, such as the Gram-negative anaerobic <i>Porphyromonas gingivalis</i>, may play a crucial role in the initiation and development of OSCC by inducing periodontitis. Indeed, epithelial-to-mesenchymal transition (EMT) and dysregulated immune response have been attributed to the activity of a dysbiotic microbiota. This comprehensive review examines the influence of <i>P. gingivalis</i> on oral carcinogenesis and progression, which has been associated with tumour microenvironment remodelling and the dysregulation of key signalling pathways related to epithelial-to-mesenchymal transition (EMT), cell-cycling, autophagy, and apoptosis. (2) Methods: The article reviews current literature on the possible role of <i>P. gingivalis</i> and induced dysbiosis, periodontitis and a pro-inflammatory environment as key mechanisms driving neoplastic epithelial changes and chemoresistance to anticancer agents in patients with OSCC; the research corpus was acquired from the Pub-Med/Medline/EMBASE/Cochrane Library databases. (3) Results: The identification of virulence factors and key mechanisms used by <i>P. gingivalis</i> to promote the development and progression of OSCC may support traditional diagnostic methods and factors related to treatment response and prevention of OSCC. (4) Conclusions: Emerging evidence suggests a possible association between periodontal bacteria and oral carcinogenesis. <i>P. gingivalis</i> may be an important potential target for future strategies aimed at treating oral cancer.

<b>Background:</b> We aimed to identify key molecular drivers of gastric cancer progression and poor prognosis by integrating multi-omics analyses with experimental validation. <b>Methods:</b> Single-cell RNA-seq data were clustered to delineate major cell types. InferCNV identified tumor epithelial cells, and reclustering revealed a malignant subset with poor prognosis. The overlap between subset markers and The Cancer Genome Atlas Stomach Adenocarcinoma (TCGA-STAD) upregulated differentially expressed genes (DEGs) was modeled with univariate, LASSO-, and multivariate Cox to derive a prognostic signature. Patients were stratified according to signature scores, and group differences in survival and immunologic features were compared. Spatial transcriptomics defined the localization patterns of key signature genes. In vitro functional assays (CCK-8, colony formation, EdU incorporation, flow cytometry, Transwell migration and invasion, and wound healing) confirmed the pivotal role of SRI. <b>Results:</b> Reclustering of tumor epithelial cells yielded seven subsets (C0-C6), with C5 displaying marked malignant features and correlating with poor prognosis in multiple cohorts. Intersecting 208 genes yielded a five-gene signature (ASCL2, REPIN1, CXCL3, TMEM176A, SRI). The signature stratified patients into high- and low-risk groups. The high-risk cohort exhibited significantly poorer survival, distinct immune-infiltration patterns, elevated immune-evasion scores, and a reduced predicted response to immunotherapy. Single-cell and spatial transcriptomics localized TMEM176A to fibroblasts and SRI to the tumor epithelium. Finally, in vitro knockdown of SRI inhibited tumor cell proliferation, migration and invasion. <b>Conclusions:</b> Our multi-omics approach identified a malignant epithelial subset, C5, and a five-gene signature that stratifies gastric cancer prognosis and immune response. Functional assays showed that SRI knockdown impairs tumor cell growth, migration and invasion.

<h4>Introduction</h4>Dementia presents with significant heterogeneity across age groups, particularly in early-onset cognitive decline (EOCD), which poses diagnostic and management challenges. The Cognitive and Neuroimaging for Neurodegenerative Disorders (CogNID) study aims to characterise clinical, cognitive, neuroimaging, and biomarker features across a diverse cohort of individuals with cognitive impairment, with a focus on diagnostic complexity, biomarker utility, and mortality.<h4>Methods</h4>Out of 680 study participants within this prospective cohort enrolled from the real-world clinics within the National Health Service, who consented to take part in the study, we analysed data from 429. Individuals were recruited between December 2018 and November 2024 from the Memory Clinics, including the early-onset dementia service and associated services. Participants underwent structured cognitive assessments, neuroimaging (MRI/CT), and Cerebrospinal fluid (CSF) biomarker evaluation, where available. Diagnoses were made by multidisciplinary consensus. Group comparisons were conducted between early-onset (EOCD, <65 years) and late-onset cognitive decline (LOCD, ≥65 years).<h4>Results</h4>Of the 429 participants, 349 (81.4%) had EOCD and 80 (18.6%) had LOCD. The mean age was 60.05 years, with no significant difference in sex or ethnicity across groups. Depression and anxiety were common (29.6%), as were cardiovascular risk factors. Lumbar punctures were more frequently performed in EOCD (p = 0.03), with 36.4% of tested participants demonstrating biomarker profiles consistent with Alzheimer's disease (A+T+). Functional cognitive disorder (FCD) was more common in EOCD (22.3% vs. 5.0%, p < 0.001). Subgroup analysis revealed significantly lower ACE-III scores and higher pathological CSF findings in Alzheimer's disease versus FCD. Mortality was higher in the LOCD group (11.3% vs. 4.6%, p = 0.03).<h4>Conclusion</h4>The CogNID study highlights the clinical and diagnostic heterogeneity of individuals with cognitive impairment, particularly in younger adults. Incorporating neuroimaging and CSF biomarkers into routine clinical pathways enhances diagnostic precision and reveals distinct phenotypic profiles between EOCD and LOCD. These findings underscore the need for harmonised diagnostic protocols, broader biomarker accessibility, and inclusive recruitment strategies in dementia research and clinical services.

<h4>Background</h4>Severe ischemia-reperfusion (II/R) injury of the intestines is a leading cause of death and disability. According to earlier research, modulating mitochondrial function is the primary mechanism by which II/R injury is ameliorated. In order to further molecular diagnostics and discover possible treatment targets, it is essential to find biomarkers of mitochondria in II/R injury.<h4>Methods</h4>The datasets GSE96733 and GSE37013, along with mitochondrial-related genes (MRGs), were obtained from the Gene Expression Omnibus (GEO) database and MitoCarta3.0, respectively. GSE96733 conducted differential expression gene (DEGs) analysis and weighted gene co-expression network analysis (WGCNA) module screening. In order to find MRGs that were expressed differently, we got their intersection (DEMRGs) and gene enrichment analysis was carried out. The hub genes were screened using machine learning approaches, protein-protein interaction (PPI) network analysis, and Molecular Complex Detection (MCODE). A nomogram was developed for diagnostic evaluation. Furthermore, the relationship between hub gene expression profiles and immune infiltration landscapes was interrogated through immune cell infiltration analysis. The expression patterns of the hub genes were further validated in the II/R injury model through dataset validation and qRT-PCR assays. The procedure concluded in a gene-related hub network, DSigDB prediction of prospective therapeutic compounds, and molecular docking simulations of the drugs' binding affinity with important target proteins.<h4>Results</h4>Hub genes have been found in five different DEMRGs: Pdk4, Yrdc, Bcl2l11, Bcl2a1d and Pmaip1. The nomogram model was beneficial for diagnosis. Dendritic cells (DC) and M2 macrophages are strongly linked to the 5 Hub genes, according to immune cell infiltration research. Afterwards, the regulatory network showed that hub genes and miRNAs had a complicated connection. Additionally, securinine and ABT-737 were anticipated to be possible therapeutic agents for II/R injury. The validation results for the four hub genes (Pdk4, Yrdc, Bcl2l11, and Pmaip1), obtained from both independent datasets and qRT-PCR, were consistent with the initial bioinformatics analysis.<h4>Conclusion</h4>Pdk4, Yrdc, Bcl2l11, and Pmaip1 have been identified as hub genes closely associated with mitochondrial function in eraly II/R injury, thereby providing a theoretical basis for the diagnosis and treatment of eraly II/R injury.

<h4>Background</h4>Breast cancer (BC) is clinically defined as a cold tumor due to its low immunogenicity, which is usually insensitive to immunosuppressive agents. Herein, we investigated the predictive potential of novel immune-related genes (IRGs) in BC, with the objective of more effectively guiding the immunotherapy for patients with BC.<h4>Methods</h4>The least absolute shrinkage and selection operator (LASSO) regression analysis was used to conduct the BC prognostic model based on IRGs, and univariate/multivariate Cox proportional hazards models were employed to establish the BC predictive nomograms. Then, we investigated the expression patterns of these IRGs utilizing The Cancer Genome Atlas (TCGA) database. Moreover, we also performed correlation analyses between IRGs and multiple immune features, including infiltration of immune cells, immune checkpoint members, and immune therapy response.<h4>Results</h4>In our study, six IRGs were finally identified to construct the BC prognostic model, including <i>HMGB3, TNFSF4, CORO2A, SOCS3, TACR1</i>, and <i>FREM1</i>. This model exhibited excellent predictive performance, with area under the curve (AUC) values of 0.676, 0.646, and 0.621 for the 1-, 3-, and 5-year timeframes, respectively. Analysis of expression profiles indicated that <i>HMGB3</i>, <i>CORO2A</i>, and <i>TNFSF4</i> exhibited significant upregulation in BC tissues, displaying strong correlations with diminished overall survival. The three overexpressed genes showed statistically significant correlations with multiple important immune cells, including Tregs, macrophages, and CD4+/CD8+ T cells. Notably, distinct patterns of integrating gene expression and immune infiltration significantly affected the clinical outcomes of BC patients. These upregulated genes demonstrated significant co-expression patterns with key immune checkpoint regulators, suggesting close immunomodulatory interactions in BC. The Tumor Immune Dysfunctional and Exclusion (TIDE) scores were lower in the high-expression groups of <i>HMGB3</i> and <i>CORO2A</i>, whereas the TIDE score was higher in the high-expression group of <i>TNFSF4</i>.<h4>Conclusions</h4>This prognostic model reliably assessed risk for BC patients, providing critical guidance for precision oncology protocols and dynamic surveillance of disease progression.

<h4>Background</h4>Cervical squamous cell carcinoma (CESC) poses significant health risks, contributing to high mortality rates and economic burdens, particularly in developing countries. The p53 signaling pathway plays a crucial role in the initiation, progression, and treatment response of CESC. This study explored the role of <i>SMYD2</i> among genes related to the p53 regulatory pathway in CESC.<h4>Methods</h4>A prognostic model for p53 regulatory pathway-related genes was established by performing least absolute shrinkage and selection operator (LASSO) regression analysis on the training cohort from The Cancer Genome Atlas (TCGA) dataset. By amalgamating the risk scores generated by the model with clinical characteristics, a nomogram was developed to assist in predicting overall survival (OS). In addition, we examined various associations concerning pathway enrichment, treatment modalities, immune cell infiltration, and the prognostic model itself. The functional role of the <i>SMYD2</i> gene was assessed through Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and Gene Set Enrichment Analysis (GSEA). Furthermore, we conducted a comprehensive evaluation of the relationship between <i>SMYD2</i> expression and an array of clinical parameters, including tumor (T), lymph node (N), and metastasis (M) stages, age, and tissue type, utilizing data from the TCGA database. Employing the "rms" R package, we constructed a nomogram model that integrates clinical features with levels of <i>SMYD2</i> expression. Additionally, immunohistochemical assessments were performed on samples from 30 cervical cancer patients to analyze the expression of <i>SMYD2</i>.<h4>Results</h4>We established a prognostic model that encompasses six genes: <i>SMYD2</i>, <i>E4F1</i>, <i>FBXO11</i>, <i>HIPK2</i>, <i>KAT2B</i>, and <i>TP53</i>. The relationships between the alterations in the expression of these genes and patient survival outcomes were validated using Kaplan-Meier survival analysis. Among the six selected prognostic genes, we found that increased <i>SMYD2</i> expression is linked to advanced T and M stages, higher clinical staging, diverse histological types, and poor prognosis. A nomogram was created to integrate <i>SMYD2</i> expression with clinical factors to forecast the OS probabilities of patients at the 1-, 3-, and 5-year marks. Furthermore, immunohistochemical evaluations disclosed that in a cohort of 33 cases, the expression levels of <i>SMYD2</i> in cervical cancer tissues were significantly greater than those observed in adjacent non-cancerous tissues.<h4>Conclusions</h4>This study elucidates specific features associated with the p53 regulatory pathway, particularly <i>SMYD2</i>, and its relationship with the onset and progression of CESC. Furthermore, <i>SMYD2</i> may serve as a prognostic biomarker for individuals diagnosed with CESC, offering new insights for the development of clinical treatment strategies.

<h4>Background</h4>Recent studies have identified a novel cell death mechanism, termed sodium death, triggered by sustained activation of <i>TRPM4</i> ion channels. Prolonged <i>TRPM4</i> channel activity results in excessive sodium ion influx and membrane depolarization, ultimately inducing necrotic cell death. However, the role of sodium death-associated genes (NECSOs) in lung adenocarcinoma (LUAD) remains poorly understood. We used NECSOs to construct a prognostic model of LUAD for prognostic prediction and to explore the features of sodium overload-induced cell death in single cells.<h4>Methods</h4>This study conducted a comprehensive analysis of The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to investigate <i>TRPM4</i>, a NECSO identified in the literature. Through co-expression analysis, 40 NECSOs were identified. Using the AddModuleScore enrichment algorithm, sodium death scores were calculated for distinct cell clusters and categorized into high- and low-score groups. Differentially expressed genes (DEGs) between these groups were subjected to weighted gene co-expression network analysis (WGCNA) to identify gene modules associated with LUAD. Subsequent differential expression and univariate Cox regression analyses were performed on genes within these modules. Consensus clustering analysis was employed to characterize molecular subtypes, while least absolute shrinkage and selection operator (LASSO) regression was used to develop a prognostic model based on NECSOs. Multiple immune infiltration scoring algorithms and cellular communication analyses were applied to evaluate the relationship between sodium overload (NECSO) and the tumor microenvironment (TME), with the aim of identifying potential therapeutic targets for LUAD.<h4>Results</h4>Single-cell and bulk transcriptome analyses identified 126 NECSOs, of which 28 were differentially expressed between tumor and normal tissues. Among these, 14 genes were significantly associated with overall survival (OS). Pseudotime analysis of epithelial cells revealed that risk-associated genes were predominantly upregulated in later stages, whereas protective genes exhibited reduced expression over time. Eight genes were selected to construct a prognostic signature for NECSOs. Patients in the low-NECSOs group demonstrated significantly better prognosis compared to those in the high-NECSOs group. Cellular communication analysis indicated that epithelial cells with elevated NECSOs expression exhibited enhanced activity in the epidermal growth factor (EGF), Midkine (MK), and COMPLEMENT signaling pathways.<h4>Conclusions</h4>The prognostic model developed from NECSOs enables robust survival prediction for LUAD patients and provides insights into the immune landscape of LUAD. Notably, epithelial cells with higher NECSOs scores displayed a greater propensity for transformation into tumor cells, highlighting their potential as therapeutic targets.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is a major subtype of lung cancer with a 5-year survival rate of less than 20%. While immunotherapy has revolutionized cancer treatment, only 10-20% of cases show durable responses to immune checkpoint blockade. Thus, developing accurate methods to predict prognosis and response to immune checkpoint inhibitors (ICIs) is crucial. Programmed cell death (PCD) plays a significant role in maintaining tissue homeostasis and responding to various physiological or pathological conditions. Increasing evidence suggests that PCD is involved in tumor initiation, development, prognosis, and response to immunotherapy. To provide reliable LUAD clinical tools, we developed an immune-related programmed cell death signature (IRPCDS) and validated its ability to predict prognosis and ICI response for precision medicine.<h4>Methods</h4>In this study, we integrated 18 PCD signatures to develop an IRPCDS. We employed 10 machine learning algorithms and 101 algorithm combinations to assess the performance of the IRPCDS. The signature was validated across multiple cohorts to ensure its robustness in predicting clinical outcomes for LUAD patients.<h4>Results</h4>The IRPCDS demonstrated strong performance in predicting the clinical prognosis of LUAD patients, effectively stratifying them into different risk groups for targeted interventions. Notably, the IRPCDS outperformed traditional clinicopathological factors and previously published 52 signatures in predicting overall survival (OS). Patients classified in the low-risk group exhibited high levels of immune infiltration and favorable responses to ICIs, while those in the high-risk group showed a higher overall mutation burden and an increased frequency of mutations in driver genes associated with LUAD. Additionally, we validated the expression of the IRPCDS genes at both the transcriptional and protein levels across multiple datasets and clinical specimens.<h4>Conclusions</h4>The IRPCDS serves as a robust and promising tool for enhancing clinical outcomes and precision medicine for individual LUAD patients. By integrating PCD signatures, this approach provides valuable insights into the prognostic landscape of LUAD, paving the way for more effective immunotherapeutic strategies.

Dendritic cells (DC) are professional antigen presenting cells playing a major role in orchestrating adaptative immune responses. To adapt to various immune challenges, such as different classes of pathogens, specialized subsets of DC have evolved across species. To date, DC are classified as conventional DC (cDC1, cDC2) and plasmacytoid DC (pDC), with the more recent addition of DC3 and transitional DC (tDC) that were discovered in human and mouse thanks to high-dimensional phenotyping and single-cell sequencing technologies. Here, by combining flow cytometry and RNA-seq on the bulk- and single-cell level, we identified the porcine equivalent of tDC in blood as CD14^-CADM1^-CD172a⁺CD4^- cells expressing both Flt3 and CD123 (IL-3RA). This new subset forms a well-defined cluster when mapped onto scRNA-seq data of enriched DC and shares transcriptomic features and abundance with porcine blood cDC2 and pDC. Moreover, we describe putative porcine DC3 as transcriptionally overlapping cells in-between cDC2 and monocytes. With the core functions of tDC and DC3 remaining to be elucidated, our datasets provide a valuable resource for cross-species research on DC heterogeneity in various lymphoid and non-lymphoid tissues.

Hepatocellular carcinoma (HCC) remains one of the leading causes of cancer-related mortality worldwide, characterized by increasing incidence rates and challenging prognoses. This study integrates single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics to unravel the complex molecular and structural landscape of HCC, focusing on the identification of mitochondrial-related genes (MitRGs) and their pivotal role in disease progression. Utilizing scRNA-seq and bulk RNA-seq data, we performed a comprehensive differential expression analysis to highlight MitRGs. A modeling approach using 92 combinations of nine machine learning algorithms was applied, producing a predictive model with good performance. Among the genes analyzed, FLAD1 emerged as significantly upregulated in HCC tissues, correlating with advanced disease stages and poorer patient outcomes, and exhibited exceptional diagnostic accuracy with an AUC of 0.962. Functional enrichment analyses revealed that high FLAD1 expression is involved in crucial biological processes like copper ion detoxification and heme complex assembly. Interaction networks further elucidated the connection between FLAD1 and critical HCC pathways, with its expression levels negatively correlated with key immune effector cells such as CD8+ T cells and DCs. Spatial transcriptomics analysis provided a structural basis for this immune exclusion, demonstrating that an intact tumor capsule can function as a physical barrier that fosters an immune-exempt microenvironment. This analysis also validated FLAD1 upregulation within the spatial context of the tumor. Additionally, DNA methylation analysis indicated a hypomethylation pattern in the FLAD1 promoter region, likely contributing to its overexpression in HCC. Validation of FLAD1 protein levels in an in-house cohort via Western blotting further confirmed these findings. Collectively, our integrative study highlights the utility of MitRGs as potential biomarkers and positions FLAD1 as a dual prognostic and therapeutic target linked to the structural and immune landscape of HCC.

Type 2 diabetes (T2D) is a chronic disease leading to an elevated glucose level in the blood and increased fracture risk. Because T2D individuals tend to have normal to higher areal bone mineral density (aBMD) than healthy individuals, their fracture risk is often underestimated. As an alternative, high-resolution peripheral quantitative computed tomography (HR-pQCT) is an attractive tool to investigate bone morphology in vivo and estimate fracture risk. Based on HR-pQCT scans, bone strength can be estimated using micro finite element ( μ FE) analysis or homogenized finite element (hFE) analysis. While μ FE is computationally expensive, hFE provides an accurate estimation of bone mechanical properties within reasonable efforts. However, the hFE scheme is based on relationships between the local fabric (anisotropy) and elasticity. These relationships have been shown to hold for healthy controls as well as in the case of osteogenesis imperfecta. Nevertheless, whether these relationships are also valid for T2D-diagnosed patients remains unclear. Therefore, the present work aims to compare fabric-elasticity relationships between T2D and non-diabetic controls. The present study collected 56 trabecular bone cores from the femoral head of 28 T2D and 28 control donors. These cadaveric samples were scanned in a micro-CT system at an isotropic 14.8 μ m voxel size. Three cubic regions of interest (ROIs) were selected in each scan. The resolution of these ROIs was coarsened by a factor of 4, mimicking clinical HR-pQCT resolution, and the ROIs were subsequently segmented. Standard morphometric parameters were computed from the segmented ROIs using medtool (v4.8; Dr. Pahr Ingenieurs e.U., Pfaffstätten, Austria). Additionally, their fabric tensor and their apparent stiffness tensors were computed. The ROIs were compared between T2D and control regarding their morphometric and mechanical properties. Finally, ROIs were matched between T2D and control for bone volume fraction ( ρ ) and degree of anisotropy (DA). The matched dataset allowed the comparison of fabric-elasticity relationships between T2D and control samples. No significant difference was observed between T2D and control samples, both regarding their morphology and their mechanical properties. Specifically, fabric-elasticity relationships were shown to hold for both the control and the T2D groups. A comparison of the resulting exponents related to ρ and DA has highlighted different trends but no important difference between T2D and control samples. In conclusion, trabecular bone architecture was similar between T2D and non-T2D donors. Additionally, fabric-elasticity relationships, i.e. morphology-mechanical relationships, are also similar between donors with and without diabetes. Accordingly, HR-pQCT-based hFE analysis could also be used for estimating the bone mechanical properties of T2D patients and for their fracture risk assessment.

<b>Background/Objectives:</b> Hepatocellular carcinoma (HCC) is one of the world's deadliest cancers; however, the mechanisms that contribute to its aggressiveness are poorly understood. In the recent literature, overexpression of the Chromosome 19 MicroRNA Cluster (C19MC) has been associated with an aggressive phenotype and unfavorable prognosis in HCC. However, the molecular consequences of C19MC overexpression in HCC remain poorly understood. <b>Methods</b>: Here, we created a constitutive C19MC-overexpressing HCC model and used two different CRISPR-engineered C19MC-overexpressing HCC models to analyze phenotype and transcriptomic changes. <b>Results</b>: We observed that C19MC overexpression induces cancer stem cell (CSC) phenotypic features in vitro and analyzed transcriptomic changes in genes correlating with stemness, such as NFκB and EMT. <b>Conclusions</b>: C19MC induces changes in HCC that are consistent with stemness and aggression, which provides a better understanding of why C19MC could be a biomarker of poor prognosis.

<title>Abstract</title> <p> <bold>Background</bold> Currently, we lack a clear understanding of the biological mechanisms underlying the different symptom domains of autism, and no approved pharmacological treatments are available. Autism is defined by social communication (SC) difficulties and non-social symptoms, including inflexible, repetitive and restrictive behaviours (RRB) and sensory reactivity. These symptom domains might disperse different biological constructs. Understanding the distinct biological underpinnings of autism symptoms is essential for advancing targeted, precision-based treatments. In our previous work we found correlations between social cognition and serotonin transporter (5-HTT) availability, and here we study other symptom domains in relation to 5-HTT availability in the brain. <bold>Results</bold> Fifteen participants diagnosed with autism and fourteen matched control subjects underwent positron emission tomography (PET) with the radioligand [ ¹¹ C]MADAM which binds to 5-HTT. 5-HTT availability did not correlate to any of our RRB-related measures of behavioural flexibility (cognitive tests (Delis-Kaplan Executive Function System verbal fluency and the Tower) and self-rating scales (Social Social Responsiveness Scale and Autism Spectrum Quotient subscales), and our behavioural flexibility index based on three personality tests). However, in a smaller subsample, we found negative correlations between sensory reactivity (as measured by Sensory Reactivity Test in the Autism Spectrum scale) and 5-HTT binding in most cortical brain regions. <bold>Conclusions</bold> 5-HTT may be less important for RRBs than for SC and sensory symptoms. Further studies investigating the biology of distinct autism symptoms are warranted to aid the development of precision medicine. </p>

<h4>Summary</h4> Previous single-cell profiling studies of the pituitary gland have yielded minimally reproducible insights largely due to their low statistical power and methodological inconsistencies. To address this problem, we generated a uniformly pre-processed Consensus Pituitary Atlas (CPA) using all existing mouse pituitary single-cell datasets (267 biological replicates, >1.1 million high-quality cells). The CPA revealed novel cell typing and lineage markers, including low-expression transcripts that previous analyses could not detect. The scale of the CPA enabled the development of machine learning models to automate and standardize cell type annotation and doublet identification for future studies. Leveraging the curated metadata, we identified sex-biased and age-dependent gene expression patterns at cell type resolution. To identify drivers of cell fates, first we determined consensus cell communication patterns. Secondly, we used RNA-sequencing and chromatin accessibility data to identify transcription factors associated with cell fates across modalities. The epitome platform acts as an interface with the CPA, allowing streamlined user-friendly analyses. <h4>Highlights</h4> Uniform processing of 267 mouse pituitary single-cell datasets (>1.1M cells) The statistical power enabled cell type, sex- and age-specific marker discovery Machine learning models facilitate doublet detection and cell typing in new datasets epitome platform provides programming-free data access and visualizations

Loss-of-function (LOF) Frazzled/DCC mutants disrupt synaptogenesis in the Giant Fiber (GF) System of <i>Drosophila</i> We observed weaker physiology in LOF male and female specimens, characterized by longer latencies and reduced response frequencies between the GFs and the motor neurons. These physiological phenotypes are linked to a loss of gap junctions in the GFs, specifically the loss of the shaking-B(neural+16) isoform of innexin in the presynaptic terminal. We present evidence of Frazzled's role in gap junction regulation by utilizing the UAS-GAL4 system in <i>Drosophila</i> to rescue mutant phenotypes. Expression of various UAS-Frazzled constructs in a Frazzled LOF background was used to dissect the role of different parts of the Frazzled receptor in the assembly of electrical synapses. Expressing Frazzled's intracellular domain in Frazzled LOF mutants rescued axon pathfinding and synaptogenesis. This is supported by the complementary result that Frazzled fails to rescue synaptic function when the transcriptional activation domain is disrupted, as shown by the deletion of the highly conserved intracellular P3 domain or by a construct with a point mutation in the highly conserved P3 domain known to be required for transcriptional activation. A computational model clarifies the role of gap junctions and the function of the GF System. The present work shows how various domains of a guidance molecule regulate synaptogenesis through the regulation of synaptic components.

BRCA1-associated protein-1 (BAP1) is a tumor suppressor protein that regulates DNA transcription through its deubiquitinase activity. Cancer-associated missense mutations within its ubiquitin C-terminal hydrolase (UCH) domain result in structural destabilization and aggregation, contributing to various malignancies such as malignant mesothelioma and uveal melanoma. In this study, we investigated the aggregation mechanisms of highly destabilized BAP1-UCH variants, including N78S, C91W, F81V, and G128R, using Thioflavin T (ThT) binding assays and AmyloFit analysis. Our results reveal that all BAP1-UCH variants follow a secondary nucleation-dominated aggregation model, exhibiting strong concentration dependence and significantly higher aggregation rates, which may be responsible for their impaired nuclear import, leading to increased cytosolic retention. These findings provide critical insights into how specific mutations in BAP1 drive aggregation and compromise its tumor-suppressing functions inside the nucleus, thereby contributing to cancer progression.

AbstractDirect-to-consumer genetic testing has become increasingly popular, offering individuals easy access to genetic data without the involvement of health care professionals. However, as availability grows, clinicians face challenges in interpreting and integrating this information into clinical care. This review provides insights for health professionals about the evolving landscape of direct-to-consumer genetic testing, with a focus on test offerings, limitations, challenges, and ethical concerns. It also highlights issues that can arise with direct-to-consumer genetic testing in the pediatric setting. Clinicians can play a pivotal role in guiding patients and families through the complexities of direct-to-consumer genetic testing, ensuring that decisions are informed and risks are minimized.

The development of smart nanocarriers capable of responding to tumor-specific stimuli represents a promising strategy for improving therapeutic selectivity in oncology. In this work, we present a class of dual-responsive polymeric nanoparticles (NPs) engineered for precision drug delivery in gynecological cancers. Amphiphilic block copolymers of the type P(MAA)-<i>b</i>-P(EG₂MA-<i>co</i>-NIPAM) integrating pH-responsive methacrylic acid (MAA) and thermoresponsive diethylene glycol methyl ether methacrylate (EG₂MA) and <i>N</i>-isopropylacrylamide (NIPAM) units were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization. Fine-tuning of the lower critical solution temperature (LCST) was achieved by modulating the ratio between NIPAM and EG₂MA, yielding copolymers with cloud points within the physiologically relevant range of 30-40 °C. The resulting NPs exhibited sharp and reversible swelling/shrinking behavior in response to pH and temperature stimuli, with sizes below 182 nm and narrow polydispersity indexes. The core-shell architecture was stabilized by a dodecyl-functionalized chain transfer agent, ensuring efficient self-assembly and robust encapsulation of both hydrophilic and hydrophobic drugs. Drug release studies with 5-fluorouracil (5-FU) and the drug-mimetic fluorescein isothiocyanate (FITC) confirmed a marked temperature-triggered release above the LCST and enhanced diffusion in mildly acidic conditions (pH < 6), characteristic of solid tumors. Cellular studies on HeLa and ovarian adenocarcinoma OVCA433 lines revealed rapid internalization, high biocompatibility, and a significant increase in therapeutic efficacy of 5-FU when delivered via NPs, compared to the free drug. These findings highlight the potential of the dual-responsive nanoplatform for targeted and controlled delivery in the treatment of cervical and ovarian cancers.

<b>Background</b>: Huntington's disease (HD) is a progressive neurodegenerative disorder primarily affecting the central nervous system (CNS). However, emerging evidence suggests that peripheral tissues, including skeletal muscle and bone, also undergo pathological changes contributing to disease burden. <b>Objective</b>: To characterize musculoskeletal impairments in the zQ175 knock-in (KI) mouse model of HD, through integrated behavioral, biomechanical, and imaging analyses. <b>Methods</b>: Motor function was assessed using grip strength, rotarod, and open field testing. <i>Ex vivo</i> contractility of the extensor digitorum longus (EDL) and Soleus (Sol) muscles was measured. Muscle fiber cross-sectional area (CSA) was quantified using semi-automated segmentation. Bone microarchitecture was analyzed using high-resolution micro-computed tomography (μCT). <b>Results</b>: Six-month-old homozygous zQ175 mice exhibited significantly reduced muscle strength and impaired contractile properties in both the EDL and Soleus muscles compared to wild-type (WT) controls. µCT analysis revealed decreased trabecular bone volume and alterations in bone structure. <b>Conclusions</b>: These findings provide a comprehensive musculoskeletal phenotyping of zQ175 mice, revealing early-onset muscle atrophy and skeletal fragility. Our study highlights the importance of targeting peripheral manifestations in HD and establishes zQ175 KI mice as a valuable additional model for investigating systemic disease mechanisms.

Parkinson's disease (PD) is a multisystem disorder presenting motor and non-motor symptoms. Motor dysfunction is the most debilitating, caused by the degeneration of dopamine-producing neurons. Increasing evidence indicates that synapse demise occurs years before neuronal death. Yet, the early synaptic dysfunctions in PD remain poorly understood. Leucine-Rich Repeat Kinase 2 (LRRK2), a serine/threonine kinase and GTPase relevant for both familial and sporadic forms of PD, has been increasingly associated with synaptic processes. These include the phosphorylation of key synaptic proteins and interactions with cytoskeletal components. Brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) are fundamental for synapse maturation, maintenance, and plasticity. Recent findings indicate that neurotrophic signaling is impaired in PD. In this review, we critically discuss the significance of identifying and clarifying the early molecular events leading to synaptic dysfunction in PD. We examine how mutant LRRK2 affects these processes and the relationship between LRRK2 and BDNF signaling from both mechanistic and therapeutic perspectives.

Breast cancer is the leading cause of death among women globally. Several genes have been found to be transcriptionally dysregulated in cancer, according to recent studies. TATA-box binding protein (TBP) and its two paralogs, TBPL1 and TBPL2, play roles in human transcription. The TBPL1 gene is implicated in colorectal carcinomas by suppressing the expression of miR-18a. However, its function in breast cancer remains undisclosed. TBPL1 is distantly related to TBP and possesses a 40% similarity with TBP's core domain. In the present study, we explored the potential role of the TBPL1 gene in transcriptome regulation by knocking out the TBPL1 gene through the CRISPR/Cas9 method. Following the knockout of the TBPL1 gene, we examined the gene transcription patterns and compared them to wild-type cell lines. We observed disparate signatures of upregulated and downregulated genes in wild-type and mutated conditions. Healthy breast MCF-12F, and T47D, SKBR3, and MDA-MB-231 breast cancer cell lines were assessed, as these cancer cells exhibit overexpression of the TBPL1 gene. Next-generation sequencing data revealed distinct marker genes regulated by the TBPL1 gene and their potential involvement in cell migration, proliferation, anti-apoptosis, and metastasis. Additionally, we also discovered novel lncRNAs implicated in the transcriptome analysis of the TBPL1 knocked-out gene. Our investigation indicated that this gene might affect varied stages of breast cancer cell lines' cellular properties, such as cell duplication, morphology, and growth. It might also contribute to tumor formation in more aggressive cell lines like MDA-MB-231 in vivo.

Midlife hypertension has been identified as a risk factor for the development of dementia, however, the underlying mechanism of this correlation remains unclear. In this study, we analyzed cerebrospinal fluid (CSF) samples from spontaneously hypertensive rats (SHR) that had been treated with either amlodipine or atenolol for one year and compared these to normotensive Wistar Kyoto rats (WKY). Mass spectrometry-based proteomic analysis of the CSF samples was conducted to identify both the impact of hypertension as well as antihypertensive treatment on CSF proteomics. Both systolic and diastolic blood pressure were increased in hypertensive rats, and both medications lowered blood pressure compared to untreated SHR rats. The analysis of the CSF proteome revealed that hypertension resulted in alterations to processes associated with the development of the central nervous system, inflammation and blood coagulation. The latter included proteins such as YKL-40, KNG1, DAG1, and members of the Serpin family. Amlodipine treatment resulted in changes to proteins involved in gas transport (including CA2, HBB, and HBA1), whereas atenolol treatment led to changes in the complement and coagulation cascade (including CFH, KNG1, APOE, and AHSG). A comparison of the two antihypertensive treatments revealed alterations in pathways associated with cell adhesion, central nervous system development, and vascular development. These findings show that hypertension and long-term treatment with antihypertensive medications elicit distinct effects on the CSF proteome.

The combined use of NGS technologies with bioinformatics tools has significantly advanced research by enabling comprehensive analyses of entire genomes, specific genomic regions of interest, and transcriptomes. Targeted NGS methods, which focus on smaller genome fractions, are widely used to study genetic diseases, epigenetic modifications, microbiomes, and environmental DNA, among other applications. This study aimed to develop a roadmap for detecting and selecting polymorphisms in candidate genes by integrating amplicon NGS-Target techniques with bioinformatics analyses. Sixty-eight genes associated with the hypothalamic-pituitary-gonadal (HPG) axis were selected to develop the amplicon NGS assay, comprising 730 regions that cover a total of 136,274 bp. This method was used to sequence 75 Guzerat cattle, a dual-purpose breed from Brazil, renowned for their high rusticity and adaptability. This Zebu cattle exhibit certain limitations, such as delayed puberty onset, which can reduce reproductive efficiency. Using the GATK protocol a total of 2,600 SNPs and 1,615 indels were detected. A series of consecutive filtering steps (maf, the detection of non-synonymous substitution, phylogenetic amino acid conservation, and biochemical properties) were used, resulting in a subset of 30 candidate SNPs. Then, these polymorphisms were analysed using bioinformatic tools (SIFT, PANTHER, PolyPhen2, and MutPred), identifying 5 SNPs with high effect on the protein. Their structure and stability were estimated using AlphaFold and DDMut. Finally, 3 candidate polymorphisms (IGF1R, LHCGR, TAC3R) with potentially significant effects on the protein remained to be validated through dynamic simulations or in vitro and in vivo experimental assays.

<h4>Background</h4>The lack of understanding of the molecular and cellular characteristics of human retinal progenitor cells (RPCs) has hindered their application in cell therapy for retinal degenerative diseases. This study aims to employ retinal organoids (ROs) derived from a VSX2-enhanced green fluorescent protein (eGFP) reporter human induced pluripotent stem cell (hiPSC) line for positive selection of human RPCs, investigate their features, and facilitate their applications.<h4>Methods</h4>hiPSCs were differentiated into three-dimensional ROs following established protocols. The fidelity of the VSX2-eGFP reporter was confirmed through immunostaining. Fluorescence-activated cell sorting was employed to select VSX2-eGFP-positive (+) cells at distinct developmental stages, followed by bulk RNA sequencing (RNA-seq) analysis to assess their transcriptome profile. Immunostaining and flow cytometry were utilized to validate the identity of VSX2-eGFP+ cells and potential cluster of differentiation (CD) biomarkers for identifying human RPCs.<h4>Results</h4>hiPSCs were successfully differentiated into ROs containing abundant RPCs. The spatiotemporal activity of the VSX2-eGFP reporter recapitulated the dynamic expression of endogenous VSX2 protein. Compared to VSX2-eGFP-negative (-) cells, VSX2-eGFP+ cells mainly exhibited characteristics of RPCs at early stages of retinal development and of bipolar cells at late stages. RNA-seq analysis revealed transcriptional heterogeneity within VSX2-eGFP+ cells across four distinct developmental stages. Moreover, the dynamic expression of 394 known CD biomarkers in VSX2-eGFP+ cells at distinct developmental stages was analyzed herein for the first time. One CD biomarker, TNFRSF1B, which has never been reported to be expressed in RPCs, was found to be highly expressed in RPCs at the early stages and might serve as a candidate CD biomarker for sorting RPCs.<h4>Conclusions</h4>This study provides valuable insights into the molecular and cellular characteristics of human RPCs, especially their expression profiles of CD biomarkers, laying a foundation for research on retinal development and the clinical translation of hiPSC-derived RPCs.

Senecavirus A (SVA) is an emerging virus, classified into the family <i>Picornaviridae</i>. It causes a severe vesicular disease in pigs. SVA 3C protein, a nonstructural protein, in itself is a protease (3C^pro), harboring the typical chymotrypsin-like fold with an S1 subsite. As a protease, 3C^pro plays a dual role in the replicative cycle of SVA. On the one hand, 3C^pro cleaves the viral polyprotein stepwise to generate different proteins. On the other hand, 3C^pro can specifically degrade a variety of host factors. The degradation of host factors may impair one or even more signaling pathways, making SVA not only efficiently propagate in cells but also ingeniously evade the host's immune responses. Herein, we systematically reviewed SVA 3C^pro involved in its structural characteristics, enzymatic properties, and more importantly its functions in cleaving the viral polyprotein and host proteins, aiming to provide a theoretical insight into the development of SVA-related antivirals.

<i>Fusarium oxysporum</i> f. sp. <i>cucumerinum</i>, the causal agent of cucumber fusarium wilt, along with <i>Rhizoctonia solani</i> AG-4 and <i>Pythium ultimum</i>-that causes cucumber damping-off-are soil-borne fungal and Oomycetes pathogens responsible for significant economic losses in agriculture. In this study, the culture filtrate of <i>Streptomyces</i> sp. JCK-6116, isolated from soil, exhibited strong inhibitory activity against the mycelial growth of multiple phytopathogenic fungi in a 96-well microtiter plate assay. In the dual culture assay, JCK-6116 inhibited the growth of 20 species of plant pathogenic fungi and Oomycetes, suggesting a wide antifungal spectrum. Three active compounds-rimocidin A, B, and C-were isolated from JCK-6116 and identified. These rimocidins exhibited antifungal effects against fungi by binding to ergosterol in the fungal membrane. However, none of the compounds exhibited anti-oomycete activity against the tested Oomycetes strains. Among the three compounds, rimocidin A demonstrated the strongest antifungal activity with minimum inhibitory concentration values ranging from 1.25-10 μg/ml. Furthermore, the culture broth of JCK-6116, at 10-fold dilution, effectively suppressed fusarium wilt and the two damping-off diseases in cucumber. Its butanol extract was also effective against the two fungal diseases but showed no activity against <i>P. ultimum</i> damping-off disease. These findings indicate that the culture broth contains metabolites with anti-oomycete activity. This study demonstrates that <i>Streptomyces</i> sp. JCK-6116 has significant potential as a biological control agent for managing soil-borne diseases caused by fungi and Oomycetes.

Extracellular vesicles (EVs) can cross the blood-brain barrier and enter the systemic circulation, potentially acting as peripheral biomarkers of stroke neuropathology. Here, we investigated alterations in EV RNA cargoes extracted from rat brain and plasma before and after stroke induction via middle cerebral artery occlusion and subsequent human neural stem cells (hNSCs) transplantation. EV RNA coexpression profiles were assessed, and digital source tracking was used to determine EV origin. The therapeutic effects of intra-arterial delivery of hNSCs on ischemic rat brains were quantified, focusing on functional recovery, resolution of ischemic lesions, and the microenvironment. Stroke induced distinct EV secretion patterns, with a notable increase in EV secretion from non-neuronal cells. hNSCs transplantation caused minimal immune rejection and transplanted cells survived in the brain for over a week. Stem cell-derived EVs were detected in peripheral blood, indicating prolonged systemic distribution after transplantation. Gene regulatory network analyses identified specific EV miRNAs that play crucial roles in neurogenesis, wound healing, angiogenesis, and blood-brain barrier integrity. An integrated analysis of EV RNAs in brain and plasma samples revealed that stroke increased correlations in RNA expression between brain and plasma and that hNSCs transplantation reversed the effect. Brain- and plasma-derived EVs carry similar molecular information after stroke, suggesting that plasma-derived EV RNAs reflect stroke pathophysiology. Intra-arterial transplantation of hNSCs improved outcomes after stroke in rats, by promoting endogenous neurogenesis and maintaining blood-brain barrier integrity. The identified EV miRNAs provided a new mechanism by which hNSCs transplantation regulates neural regeneration through the miR-204-5p/EFNB3 axis.

Thyroid cancer is the most common endocrine malignancy, with incidence rising steadily worldwide. Although most cases are differentiated thyroid carcinomas with excellent prognosis, a small subset, such as anaplastic thyroid cancer, demonstrates aggressive behavior and poor survival outcomes. Recent decades have witnessed a transformation in thyroid cancer diagnostics and management, driven by improvements in high-resolution ultrasound, fine-needle aspiration biopsy, molecular profiling, and standardized risk stratification systems such as the Thyroid Imaging Reporting and Data System. Despite these advances, overdiagnosis and overtreatment remain key clinical challenges. Accurate risk stratification and management strategies are critical, especially for distinguishing indolent nodules from aggressive subtypes. This review provides a comprehensive overview of thyroid cancer pathogenesis, clinicopathological classification, diagnostic approaches, and evolving therapeutic strategies, ranging from active surveillance to targeted and immunotherapy-based treatments. By integrating molecular diagnostics with conventional parameters, the article underscores how precision medicine can reduce treatment burden, improve outcomes, and guide personalized care. This review offers valuable insight into the biological complexity of thyroid cancer and highlights the need for continued refinement of diagnostic criteria and therapeutic frameworks in clinical practice.

<h4>Background</h4>Huntington's disease (HD) is primarily associated with executive dysfunction, but episodic memory impairment is also present. Traditionally, these memory deficits have been attributed to retrieval difficulties linked to fronto-striatal dysfunction, rather than to disruptions in encoding or consolidation processes. However, the specific nature and diversity of memory impairments in HD remain underexplored.<h4>Objective</h4>To characterize the profile of episodic memory impairment in HD, identify distinct cognitive phenotypes, and examine their clinical, neuroanatomical, and biomarker correlates.<h4>Methods</h4>We assessed episodic memory in HD patients and healthy controls using the Free and Cued Selective Reminding Test (FCSRT), complemented by Item-Specific Deficit Approach (ISDA) indices to quantify encoding, consolidation, and retrieval deficits. Structural MRI was used to identify gray matter volume correlates, and plasma neurofilament light chain (NfL) was measured as a marker of neuroaxonal injury.<h4>Results</h4>Compared to controls, HD patients showed marked impairments in free recall with preserved cued recall, suggesting predominant retrieval deficits. However, nearly one-third of patients exhibited global impairments across all FCSRT components, mainly driven by consolidation deficits consistent with medial temporal lobe dysfunction. This subgroup also showed worse cognitive and functional performance and significant atrophy in the hippocampus, entorhinal cortex, and parahippocampal gyrus.<h4>Conclusion</h4>Episodic memory dysfunction in HD is heterogeneous and includes both retrieval-related and consolidation-driven profiles. These profiles reflect distinct neurodegenerative patterns, emphasizing the importance of cognitive subtyping for improving clinical characterization and biomarker development in HD.

Depression and cardiovascular disease (CVD) frequently coexist, significantly impacting patient prognosis and quality of life. Research indicates that inflammatory responses play a crucial role in the pathogenesis of both conditions. The NLRP3 inflammasome, a key inflammatory signaling platform of the innate immune system, mediates the maturation and release of IL-1β and IL-18 and induces pyroptosis, playing a significant role in both depression and CVD. To explore the mechanisms and therapeutic potential of the NLRP3 inflammasome in the comorbidity of depression and CVD, we systematically reviewed recent literature. Our focus was on its activation pathways, expression changes in animal models and clinical samples, and intervention studies. The results indicate that NLRP3 inflammasome is persistently activated in patients with both depression and CVD, and this activation correlates with disease severity. Furthermore, various pharmaceutical drugs and natural compounds have demonstrated synergistic effects by inhibiting the NLRP3 inflammasome pathway. In conclusion, the NLRP3 inflammasome represents a critical molecular mechanism linking depression and CVD, as well as a potential target for combined therapeutic strategies. This area holds significant research and clinical translational value.

Ischemia/reperfusion injury (IRI) is a common damage due to the restoration of blood flow following an ischemic injury. Its pathogenesis is mainly linked to the production of reactive oxygen species (ROS), which sustain cell damage and promote cell death. The tardigrade damage suppressor protein (Dsup) is a DNA-binding protein that enables tardigrades to tolerate stress conditions, including oxidative stress. We investigated the ability of the Dsup to protect human cells from IRI, using an in vitro model of hypoxia and reoxygenation. We exposed HEK293TT cells transfected with the Dsup to hypoxic injury and analyzed cell viability, oxidative stress, expression of antioxidant proteins using functional assays, and a proteomic approach to dissect the molecular mechanisms modulated by the Dsup. Dsup expression significantly enhanced cell survival following hypoxia-reoxygenation and markedly reduced intracellular ROS levels. Proteomic and Western blot analyses revealed a significant upregulation of antioxidant enzymes in Dsup-expressing cells. Furthermore, the Dsup modulated autophagy and key stress-related pathways, including the MAPK cascade. This study demonstrates that the Dsup protects human cells from IRI by reducing oxidative stress and modulating key cytoprotective pathways. Our results establish the Dsup as a promising candidate for future therapeutic applications against IRI, meriting further exploration in in vivo models.

In recent years, amyloid proteins that perform vital functions in the brain have been characterized. The question of why some amyloids are neurotoxic while others are harmless remains open. Here, we provide a brief overview of pathological and functional brain amyloids and present a comparative analysis of their amino acid sequences based on the percentage of hydrophobic and charged residues, as well as their enrichment in glutamine, asparagine, serine, and glycine. We demonstrate that pathological and functional brain amyloid proteins, along with their amyloidogenic fragments, do not differ in amino acid composition, contrary to previous assumptions. The ability of an amyloid to cause toxicity can instead be explained by the concept of "available targets". Evidence from studies of pathological amyloids demonstrate that their toxicity is determined not only by a loss of function but also by aberrant interactions with specific targets, such as PrP^C or mitochondrial membranes. Binding to these targets triggers pathological cascades that ultimately lead to cell death. In contrast, such targets are inaccessible to functional amyloids, either because of localized translation and protein sequestration within specialized cellular structures, or because their interactions with physiological partners prevent binding to dangerous targets.

<b>Background/Objectives:</b> Gene rearrangements involving oncogenes are major drivers in acute leukemia, influencing disease classification, prognosis, and therapeutic decision-making. Targeted RNA sequencing (RNA-Seq) panels capable of detecting intergenic and intragenic fusions across multiple genes are increasingly used in diagnostic settings. However, comparative evaluation with orthogonal technologies remains limited. <b>Material and Methods</b>: We compared the performance of a 108-gene anchored multiplex PCR (AMP)-based RNA-Seq panel with that of Optical Genome Mapping (OGM) in 467 acute leukemia cases. The cohort included 360 cases of acute myeloid leukemia (AML), 89 B-lymphoblastic leukemia (B-ALL), 12 T-lymphoblastic leukemia (T-ALL), and 6 cases of mixed phenotype acute leukemia (MPAL). <b>Results:</b> Results of both methods were concordant in 175 (74.7%) of 234 detected gene/rearrangement fusions. The concordance rate varied significantly across different leukemia types, ranging from 80.2% in B-ALL to 41.7% in T-ALL (<i>p</i> < 0.001) OGM uniquely detected 37 of 234 (15.8%) clinically relevant rearrangements, whereas RNA-Seq exclusively identified 22 of 234 (9.4%). Enhancer-hijacking lesions, including <i>MECOM</i> and <i>BCL11B</i> rearrangements, <i>CDK6::MNX1</i>, and <i>IGH</i> rearrangements, had a markedly lower concordance (20.6%) compared with all other aberrations (93.1%) (<i>p</i> < 0.001). Conversely, some gene fusions arising from intrachromosomal deletions were interpreted by OGM as simple deletions rather than rearrangements or fusions. <b>Conclusions:</b> Targeted RNA-Seq was effective for detecting chimeric fusion transcripts and showed slightly better performance in identifying fusions resulting from deletions. However, OGM was effective for detecting enhancer-hijacking events that do not generate fusion transcripts. Both methods are complementary for the workup of acute leukemia cases.

<b>Background</b>: Obesity is a multifactorial metabolic disorder influenced not only by excessive caloric intake but also by micronutrient imbalances such as zinc deficiency. Emerging evidence suggests that zinc regulates microRNA (miRNA) biogenesis and expression, linking nutritional status to metabolic regulation. <b>Objective</b>: This review delineates the molecular interplay between zinc and miRNAs in obesity, emphasizing the mechanistic, clinical, and translational relevance of zinc-sensitive miRNAs in adipogenesis, insulin resistance, inflammation, and oxidative stress. <b>Results</b>: Zinc deficiency alters miRNA expression profiles associated with metabolic dysregulation. Key miRNAs-miR-21, miR-34a, miR-122, and miR-144-3p-are consistently modulated by zinc status, influencing inflammation, lipid metabolism, and insulin signaling. Zinc repletion restores several miRNAs (e.g., miR-10b, miR-155, miR-145), suggesting reversibility, while excessive zinc may upregulate miR-144-3p and exacerbate oxidative stress. Circulating and exosomal miRNAs show promise as dynamic biomarkers for zinc intervention efficacy. <b>Methods</b>: A literature search was performed in 4 databases up to August 2025 using keywords related to zinc, miRNAs, and obesity. Eligible studies included both preclinical and human research evaluating zinc status or supplementation and miRNA expression in metabolic contexts. <b>Conclusion</b>: Maintaining optimal zinc levels may normalize miRNA expression and improve insulin sensitivity. The "zinc-miRNA axis" represents a novel frontier for precision nutrition in obesity management.

Nutrient exchanges promote the success of symbioses among reef-building corals, endosymbiotic dinoflagellates (Family: Symbiodiniaceae), and their microbial symbionts. Nutrient dynamics has considerable implications on the metabolism and proliferation of the coral holobiont, with nutrient limitation known to increase the susceptibility of corals to bleaching by disrupting the host-symbiont nutrient exchange. This study examines how two Symbiodiniaceae species, <i>Symbiodinium microadriaticum</i> RT 362 and <i>Cladocopium goreaui</i> RT 152, respond to varying iron (Fe) availability, nitrate (NO₃ ^-), and ammonium (NH₄ ⁺) in batch cultures. Under Fe limitation, phytoplankton growth is reduced when relying on NO₃^- due to the higher Fe requirement for nitrate assimilation enzymes, whereas NH₄ ⁺ uptake is more efficient as it bypasses these Fe-dependent processes. Symbiodiniaceae utilize these nitrogenous compounds to fuel their metabolic processes, with an advantage in using NH₄ ⁺ due to its greater energy efficiency and lower Fe requirement. Due to its role as cofactor of enzymes, Fe is crucial for nitrate reduction and chlorophyll synthesis, NH₄ ⁺ assimilation remains effective even under low Fe conditions. The study reveals that in <i>S. microadriaticum</i> and <i>C. goreaui</i>, chlorophyll production, closely linked to Fe availability, and significantly influences carbohydrate and lipid synthesis, with both species boosting protein and carotenoid production under low Fe conditions. Chlorophyll and the other photosynthetic macromolecule product concentrations continue to increase with NH₄ ⁺ as the N source, even under low Fe conditions. These findings offer critical insights into how these species adapt to varying environmental conditions, improving our understanding of coral resilience.

<h4>Background</h4>Neurogenic bladder (NB) following spinal cord injury (SCI) is a debilitating complication characterized by bladder fibrosis and structural remodeling. Non-coding RNAs (ncRNAs) are emerging regulators of fibrotic processes, yet their roles in SCI-induced NB remain unexplored. This study aimed to delineate the dynamic ncRNA-messenger RNA (mRNA) regulatory landscape in SCI-associated NB using transcriptomic profiling, identifying potential biomarkers and therapeutic targets.<h4>Methods</h4>Twenty female Wistar rats underwent complete T10-T11 spinal cord transection (SCI groups, n=15) or laminectomy-only (NC group, n=5). Bladder tissues were harvested at 1, 2, and 4 weeks post-SCI. Next-generation sequencing (NGS) analyzed mRNA, long non-coding RNA (lncRNA), and microRNA (miRNA) expression. Differentially expressed genes/mRNAs/lncRNAs/miRNAs (DEGs/DEMs/DELs/DEMIs) were identified (|log2FC|>1, P<0.05), and lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks were constructed. Functional enrichment and quantitative real-time polymerase chain reaction (qPCR) validation were performed to confirm key findings.<h4>Results</h4>Histopathology confirmed progressive bladder fibrosis, with collagen deposition increasing from 7.3% (NC) to 34.6% at 4 weeks post-SCI (P<0.001). NGS revealed 3,255/3,449/884 DEMs, 904/870/278 DELs, and 229/77/127 DEMIs in SCI-1/2/3 <i>vs.</i> NC, respectively. Temporal analysis identified 420 shared DEMs and 102 DELs, with lncRNAs enriched on chromosomes 1 and 7. The ceRNA networks implicated transforming growth factor-beta (TGF-β), calcium, and interleukin-17 (IL-17) signaling pathways in fibrosis. Notably, miR-21-5p showed progressive upregulation, while miR-139-5p decreased, correlating with ROCK2-mediated detrusor overactivity (DO). Functional enrichment highlighted inflammatory response and extracellular matrix reorganization as dominant processes. qPCR validated 12 candidate DEGs, including PVT1 and miR-21-5p, aligning with sequencing data.<h4>Conclusions</h4>This first comprehensive transcriptomic atlas of SCI-induced NB uncovers dynamic ncRNA-mRNA networks driving fibrosis through TGF-β and inflammatory pathways. The identified dysregulated ncRNAs and their target genes provide novel biomarkers for early NB diagnosis and potential therapeutic entry points. Our findings establish a foundation for developing ncRNA-targeted strategies to mitigate bladder remodeling post-SCI, addressing a critical unmet need in neuro-urology.

<h4>Background</h4>Cellular metabolic irregularities are intricately associated with the initiation and progression of tumors. Emerging evidence suggests that interactions between intratumoral microbiomes and host mediate this process. However, a comprehensive understanding of the role of metabolism-related intratumoral microbes (MRIMs) in lung adenocarcinoma (LUAD) is still lacking. This study aimed to investigate the characteristics and prognostic significance of MRIMs, as well as elucidate their potential implications in relation to the microenvironment in LUAD.<h4>Methods</h4>Integrated analyses were conducted using accessible datasets of the microbiome, bulk and single-cell transcriptomes. Spearman's coefficient between metabolic activity score and microbial abundance was used to identify MRIMs. An unsupervised clustering approach was utilized to distinguish the MRIMs-featured subtypes in LUAD samples. The Scissor algorithm was executed to select the cell subpopulations featured by MRIMs, and the underlying regulatory network in MRIMs-featured cells was explored. Additionally, a prognostic signature based on the microbial abundance of MRIMs was developed, and comprehensive analyses were subsequently carried out to reveal the correlation between MRIMs and LUAD microenvironment.<h4>Results</h4>Ten microbial species were identified as MRIMs, enabling the classification of LUAD samples into two distinct subtypes that showed significantly associated with clinical features and survival outcomes. The scRNA-seq analysis revealed notable differences in T cells, ciliated cells, mast cells, endothelial cells, and fibroblasts between MRIM+ and MRIM- subpopulations. BCL3, KLF3, and NFKB2 were the regulons in the regulatory network of MRIM-featured cells. Additionally, a microbial prognostic-predictive signature was established comprising <i>Succinimonas</i>, <i>Collimonas</i>, and <i>Marichromatium</i>, which also exhibited potential for indicating immunotherapeutic benefit and predicting drug sensitivity to cisplatin, cytarabine, pyrimethamine, olaparib, bicalutamide and vorinostat in LUAD treatment.<h4>Conclusions</h4>This study identified intratumoral microbes associated with metabolism, revealed distinct subtypes and their roles in LUAD, and established a predictive signature for the prognosis and therapeutic responsiveness of LUAD.

Replication stress (RS) is a major driver of genomic instability and cancer development through impaired DNA replication that can lead to chromosomal instability (CIN). Although RS is mechanistically linked to CIN, its relationship with cellular proliferation is complex. Depending on the context, RS can either promote or suppress cell growth. Existing RS gene expression signatures overlook this complexity, relying on the overexpression of oncogenes such as <i>MYC</i>, which introduces a proliferation bias. To disentangle genuine RS from confounding cell cycle and proliferation transcriptional profiles, we developed and validated a novel gene expression signature that accurately predicts RS independently of oncogene activity. This tumorigenic RS signature (TRSS) captures RS-related transcriptional changes across diverse cellular contexts, enabling a more robust and proliferation-independent measure of RS in both experimental and clinical samples. Applying our signature to patient data, we discovered a link between RS and the non-homologous end-joining (NHEJ) DNA repair pathway. Specifically, we observed that <i>MSH2</i> and <i>MSH6</i> - core components of mismatch repair - are associated with elevated RS and may indicate a shift toward NHEJ-mediated repair under stress conditions. Our study provides a refined approach to quantify RS and sheds light on its broader impact on DNA repair network dynamics.

Chronic rhinosinusitis without nasal polyps (CRSsNP) is a chronic inflammatory disease that lacks a clear pathogenesis/pathophysiology. While large studies focused on elucidating the pathophysiology of CRS with NPs (CRSwNP), this study aimed to use a systemic evaluation approach to identify the redox gene expression profile, its association with oxidative damage in CRSsNP, and the differences between CRSsNP and -wNP. The expression of 84 redox genes was analyzed using real-time PCR array in control and CRSsNP nasal mucosae. Changes in the mRNA and protein levels of these redox differentially expressed genes (DEGs) were verified using a customized real-time PCR array, RT-PCR, and Western blotting in an additional 18 patients. 4-Hydroxynonenal (lipid peroxidation) and 3-nitrotyrosine (protein nitrosylation) expression, representing oxidative stress (OxS) and nitrosative stress (NsS) status, were examined using immunohistochemistry. We found 27 DEGs (24 upregulated and 3 downregulated) in CRSsNP. AKR1C2, GCLM, GPX2, NOS2, and NQO1 were upregulated and LPO was downregulated more than 4-fold. These changes led to a substantial increase in OxS in CRSsNP nasal mucosa. In a comparison of the currently identified 27 DEGs with the 23 previously reported CRSwNP genes, there were 16 unique redox DEGs expressed between CRSsNP and -wNP. A String protein interaction network analysis revealed that CRSsNP possessed "an adaptive antioxidant defense signature", while CRSwNP showed "a pro-inflammatory and -oxidant pathway". Collectively, we systemically performed transcriptomic analysis to profile OxS-related genes in CRSsNP and highlighted the unique redox gene sets and pathway differences between CRSsNP and -wNP.

Alveolar echinococcosis (AE) is a fatal foodborne parasitic disease caused by the larvae of <i>Echinococcus multilocularis</i>. The disease primarily affects the liver. Previous studies have found that Kupffer cells have an immune protective effect, but in the late stages of AE, they are associated with parasite immune escape. The present study analyzed the effects of <i>Echinococcus multilocularis</i> protoscoleces (PSCs) infection on the mitochondrial morphology and function of macrophages, as well as their phagocytic function and apoptosis. Infection with PSCs has been shown to result in the fragmentation of the macrophage mitochondrial network, the impairment of mitochondrial membrane potential, the elevation of mitochondrial reactive oxygen species, and the reduction in mitochondrial DNA copy number. This cascade of events, consequent to the infection, has been demonstrated to promote the apoptosis of macrophages and impair their phagocytic function. Inhibiting mitochondrial fission during PSCs infection has been shown to mitigate mitochondrial dysfunction, suppress macrophage apoptosis, and enhance macrophage phagocytic function. This discovery provides insights into improving macrophage function during the progression of AE.

DNA ligase 1 (LIG1) joins broken strand breaks and seals Okazaki fragments during DNA repair and replication. Huntington's disease (HD)-associated mutation in the <i>LIG1</i> gene, K845N, is associated with delayed symptom onset and predicted to suppress CAG repeat expansion. Yet, how this mutation impacts faithful nick sealing and efficient DNA binding by LIG1 remains unknown. Here, using biochemical analyses, X-ray crystallography, and total internal reflection fluorescence (TIRF) microscopy, we characterized the LIG1 HD-associated K845N variant at biochemical, structural, and single-molecule levels. Our results showed significantly reduced ligation efficiency for nick substrates containing noncanonical mismatches and diminished mutagenic end-joining of damaged DNA, while LIG1 K845N variant exhibits a lack of discrimination against nicks containing 3'-ribonucleotides when compared with the wild-type enzyme. Furthermore, our structures provided an atomic insight into differences in the distances between functional groups of K/N845 and DNA ends, demonstrating similar conformation at the ligase active site. Finally, our single-molecule measurements revealed that the K845N variant binds less frequently to nick, suggesting diminished affinity. Overall, our findings contribute to understanding the mechanism by which LIG1 searches for nick sites on DNA and ensures fidelity to maintain genome stability at the final ligation step in normal versus HD-associated states.

<h4>Objective</h4>Shortened telomere length (TL) in blood cells has been associated with atherosclerotic coronary artery disease (CAD). However, the mechanistic pathways underlying TL attrition in arterial wall tissues of patients with CAD remain unclear.<h4>Methods</h4>TL was measured using quantitative polymerase chain reaction in blood granulocytes and matched arterial wall tissues from 155 Chinese men with CAD. RNA sequencing was performed in 34 patients to profile the arterial wall tissue transcriptome. These patients were dichotomized according to the median TL of either arterial wall tissues or granulocytes, and differential gene expression was examined with respect to TL.<h4>Results</h4>No significant correlation was observed between TLs of arterial wall tissues and granulocytes (R=0.107, <i>p</i>=0.19). We identified 10 significant differentially expressed genes (DEGs) associated with TL attrition in arterial wall tissues (false discovery rate [FDR] <0.045), including upregulation of <i>ADRA2A</i> (FDR=0.043), a gene encoding an adrenergic receptor implicated in vasoconstriction of atherosclerotic coronary arteries. Gene-set enrichment analysis also revealed positive enrichment of the vasoconstriction pathway (FDR=0.012) in association with TL attrition at the arterial wall tissue. However, these 10 DEGs were not associated with TL attrition in granulocytes. Instead, TL attrition in granulocytes was linked to negative enrichment of ATP synthesis pathways, potentially reflecting mitochondrial dysfunction.<h4>Conclusion</h4>Our findings suggest that distinct pathway dysfunctions may underlie TL attrition in atherosclerotic arterial walls versus granulocytes, indicating that TL attrition in these compartments could contribute to CAD pathogenesis through different mechanisms.

Adipose tissue is a dynamic immunometabolic organ whose cellular heterogeneity and functional plasticity are central to systemic energy balance and metabolic regulation. Disruption of immune-adipocyte interactions is closely linked to the development of obesity and related metabolic disorders. In this review, we summarize current advances in understanding of the immune landscape in adipose tissue, with an emphasis on the distinct roles of immune cell subsets. Recent approaches including global and single-cell transcriptomic analysis, spatial profiling, and lineage tracing have expanded our ability to characterize these populations. We further highlight mechanisms through which immune cells influence adipocyte turnover, lipid handling, and thermogenesis, as well as reciprocal signals from adipocytes such as cytokines, lipid mediators, extracellular vesicles, and nutrient exchange. This bidirectional crosstalk governs adipose tissue remodeling and determines the occurrence of metabolic homeostasis or dysfunction. Finally, we provide perspectives into the ways in which these interactions may guide the identification of novel therapeutic targets for obesity and metabolic disease.

Recent advances in sports genomics and epigenetics have significantly enhanced our understanding of the biological underpinnings of sprint, strength, and power performance. While genetic predisposition influences muscle fiber composition, contractile speed, and neuromuscular efficiency, epigenetic mechanisms are increasingly recognized for their role in modulating gene expression in response to training and environmental stimuli. In this narrative review, we synthesize current findings on key genetic variants, including polymorphisms in <i>ACTN3</i> (R577X), <i>ACE</i>, and <i>AMPD1</i>, which have been associated with muscle hypertrophy, anaerobic capacity, and explosive strength. Genome-wide association studies (GWAS) and candidate gene approaches have identified over 150 polymorphisms relevant to power-oriented sports, with <i>ACTN3</i> R577X emerging as a strong predictor of sprinting ability. However, interindividual variability in performance cannot be explained by genetics alone. Epigenetic modifications-such as DNA methylation, histone acetylation, and microRNA (miRNA) activity-regulate skeletal muscle plasticity, recovery, and adaptation. Notably, miRNAs act as key post-transcriptional regulators, influencing pathways involved in oxidative metabolism and neuromuscular remodeling. Furthermore, telomere biology has emerged as a complementary factor, with telomere length serving as a biomarker of biological aging and regenerative capacity, particularly relevant to high-intensity, power-based sports. By integrating genomics, epigenomics, and transcriptomics, this narrative review provides a comprehensive framework for understanding the complex biology of elite athletic performance and highlights future directions for personalized training and talent identification in sport science.

<h4>Rationale</h4>Cocaine use disorder (CUD) is a significant health concern that has devastating impacts on affected individuals and society. There are currently no approved therapies to treat CUD due, in part, to significant gaps in our knowledge about the underlying factors that increase risk. Individual genetic differences contribute to CUD risk. Identifying specific genetic mechanisms that increase risk could reveal novel targets for treatment and prevention.<h4>Objectives</h4>We identified two Collaborative Cross (CC) strains, CC004/TauUncJ (CC004) and CC041/TauUncJ (CC041), that differ significantly for locomotor response and self-administration of cocaine. In the current study, we used mapping crosses generated from both strains to identify genetic loci that are associated with behavioral responses to cocaine.<h4>Results</h4>We bred mice from the low (CC041) and high (CC004) responding strains to C57BL/6NJ mice to produce two F2 populations and identify genetic loci that influence locomotor response to cocaine. We identified three significant loci on chromosomes 7, 11 and 14 in the CC041 F2 mapping cross that collectively explain 14% of the phenotypic variance for locomotor response to cocaine. Bioinformatic analyses identified multiple genes on chromosomes 7 and 11 that are genetically plausible, have functional relevance and are suitable for further exploration.<h4>Conclusions</h4>Genetically defined and phenotypically divergent mouse strains are a useful tool for identifying candidate genes that influence behavioral responses to psychostimulants. Functional and mechanistic analyses of these genes could provide insights into biological processes that increase risk for CUD.

<h4>Background</h4>Colorectal cancer (CRC) is a leading cause of global cancer-related mortality, necessitating the identification of novel therapeutic targets. Integrating genetic and transcriptomic data may reveal key molecular drivers of CRC progression and treatment opportunities.<h4>Methods</h4>We performed a multiomics analysis combining genome-wide association study (GWAS) data (p < 1e-6) and RNA-seq data from the TCGA. Differential expression analysis (Limma) identified 24 consistently dysregulated genes (17 mRNAs, 7 lncRNAs) in CRC. Survival analysis was used to evaluate their prognostic impact on overall survival (OS), relapse-free survival (RFS), and post progression survival (PPS). Drug‒gene interactions were explored via Enrichr, and virtual screening (PubChem) prioritized high-affinity compounds that target PYGL, a metabolic regulator.<h4>Results</h4>Integration of GWAS and RNA-seq revealed that 24 CRC-associated genes, including PYGL, SMAD7, and TCF7L2, are involved in tumor metabolism and Wnt/TCF signaling. Survival analysis revealed that five genes (CDKN2B, BOC, METRNL, etc.) were significantly correlated with OS, RFS, and PPS. Ten small-molecule candidates targeting PYGL exhibited high binding affinity, suggesting their therapeutic potential.<h4>Conclusion</h4>This study identified CRC-linked genes through GWASs and transcriptomics, highlighting their prognostic and druggable relevance. Computational drug repurposing pinpoints PYGL inhibitors as promising candidates, offering a translational framework for CRC therapy development.

Aging is recognized as a significant risk factor for chronic diseases. The decline in intestinal stem cells function is a critical contributor to intestinal aging, resulting in impaired intestinal homeostasis and increased vulnerability to age-related diseases. Medicago sativa L. (alfalfa) saponin are plant-derived bioactive compounds that are shown to have benefits in regulating oxidative stress and gut microbiota. However, the potential of alfalfa saponin (AS) to modulate intestinal aging and enhance intestinal stemness to maintain homeostasis remains insufficiently explored. In this study, the effects of AS on intestinal stemness in naturally aged mice and its underlying mechanisms involving gut microbiota regulation are examined. Antibiotic-mediated depletion of intestinal bacteria and fecal microbiota transplantation are employed to determine the specific role of the gut microbiota in mediating the effects of AS. Comprehensive multi-omics analyses revealed that AS significantly increased the abundance of Lactobacillus intestinalis (L. intestinalis). Notably, L. intestinalis is found to possess bile acids metabolic capabilities, producing ursodeoxycholic acid, which functions as an FXR antagonist to activate the Wnt signaling pathway and enhance intestinal stemness, thereby supporting intestinal homeostasis. These findings are validated in both intestinal organoids and naturally aged mice models. This study provides the first identification of a complete functional axis by which the metabolites of AS and L. intestinalis modulate intestinal stemness to mitigate intestinal aging, offering insights for the development of innovative natural product-based therapeutic strategies to promote healthy aging.

Huntington disease (HD) is a neurodegenerative disease caused by a polyglutamine expansion (polyQ) in the Huntingtin protein (muHTT), which makes it prone to misfolding and aggregation. muHTT aggregates sequester a wide variety of proteins essential for cell homeostasis, including chaperones and transcription factors, and their depletion may contribute to HD pathogenesis. Lysosomes are the main hubs for degradative and signaling activities in cells, and their functionality is crucial for cell homeostasis, especially for neurons. Different forms of cellular stresses, including proteotoxic stresses, can alter lysosome integrity and induce lysosomal membrane permeabilization (LMP). Damaged lysosomes are recognized by galectins, in particular galectin-3 (LGALS3) with activation of the lysosome quality control (LQC) system responsible for repairing, degrading, or replacing leaky lysosomes. The system is transcriptionally regulated by the transcription factors EB and E3 (TFEB and TFE3, respectively). Using HD mouse and cell models, we demonstrated that TFEB and TFE3 are sequestered in muHTT aggregates, and muHTT proteins associates with LMP triggering the translocation of LGALS3 to the lumen of lysosomes, with a close relation between polyQ size and severity of these events. Moreover, we demonstrated that TFEB and TFE3 silencing or overexpression modulate muHTT aggregation. TFEB and TFE3 knockdown worsens muHTT aggregation, while their overexpression reduces muHTT inclusions and concurrently reduces LGALS3 accumulation via lysophagy and lysosome replacement. Our findings suggest that both TFEB and TFE3 are implicated in HD, and their sequestration in muHTT inclusions increase the vulnerability of neurons to lysosome injury, altering LQC and contributing to disease pathogenesis. In physiologial conditions, lysosome membrane permeabilization occurs and activates TFEB and TFE3 triggering a response to induce lysophagy and lysosome biogenesis. In HD, muHTT sequesters TFEB and TFE3 into inclusions and the reduced TFEB/TFE3 bioavailability prevents the activation of lysophagy and leading to the accumulation of damaged lysosomes. Created in BioRender.

Triplet repeat expansion underlies multiple pathologies, including Huntington's disease, often arising in somatic non-dividing tissues such as the brain. Despite identification of genetic modifiers, mechanistic insights remain limited. Using purified human proteins, we show that MutLγ (MLH1-MLH3), stimulated by MutSβ (MSH2-MSH3), incises DNA opposite an extrahelical loop on the 5' side. This activity, with a moderate sequence preference, generates DNA nicks enabling Polδ-mediated displacement synthesis with the loop as a template, leading to expansion. PCNA confines these MutLγ incisions near the loop. FAN1, instead, preferentially targets the looped strand. RFC-PCNA stimulate and direct FAN1 nuclease to the 3' boundary of the loop while restricting its exonuclease activity. No pre-existing nick is required. Following FAN1-RFC-PCNA action, Polδ removes the loop and resynthesizes DNA, causing contraction. FAN1 also directly inhibits MutLγ, preventing its activation by MutSβ. Our study illuminates both repeat expansion and contraction mechanisms and reveals the protective function of FAN1.

Stress responses and neuronal death mediated by the p53 pathway play a central role in the progression of neurodegenerative disease, constituting a common target to extend neuronal function and survival. Interaction of p53 and its signaling network with RNA-binding proteins (RBPs) helps fine-tune its activation and the resulting cell fates. Preclinical therapeutics based on depletion of the RBP STAUFEN-1 (STAU1) protein successfully prevent neurodegeneration, however, the specific mechanisms are not fully understood. STAU1 is pathologically overabundant in multiple neurological disorders and contributes to neurodegeneration by exacerbating autophagy dysfunction, endoplasmic reticulum stress, and RNA-protein condensate accumulation. We previously showed that lowering STAU1 levels mitigates these disease-related features and prevents neuronal death in animal models of amyotrophic lateral sclerosis/frontotemporal dementia (ALS/FTD) and spinocerebellar ataxia type 2 (SCA2). Here, we show by combined transcriptomic and functional analyses that STAU1 reduction results in the inhibition of apoptosis through the p53 pathway. In both proliferating and post-mitotic cell types-human iPSC-derived neurons, mouse cortical neurons, SH-SY5Y cells, and fibroblasts-STAU1 reduction effectively prevented p53-mediated apoptosis and DNA damage induced by Nutlin-3 and etoposide. Further examination in C9orf72-expanded patient-derived fibroblasts and a C9orf72 mouse model of ALS/FTD, which exhibit baseline overabundance of STAU1 and activation of the p53 pathway, confirmed that STAU1 reduction also prevented p53-driven pro-apoptotic signaling. These findings establish STAU1 as a novel modulator of DNA damage and p53-dependent apoptosis, suggesting that targeting STAU1 could be a promising approach to prevent neurodegeneration in ALS/FTD.

Addressing the challenges of energy imbalance and the difficulty in optimizing cluster head selection in clustering protocols for wireless sensor networks (WSNs), this paper proposes a clustering protocol based on an improved zebra optimization algorithm (IZOACP). The method systematically solves the NP-hard problem of cluster head selection by integrating the zebra optimization algorithm (ZOA), Gaussian mutation strategy, and opposition-based learning mechanism, while optimizing the clustering process based on four key metrics: node residual energy, network density, intra-cluster distance, and communication delay. To further enhance data transmission efficiency, a dynamic adaptive inter-cluster routing mechanism is designed, which achieves path dynamic balancing based on node distance, residual energy, and load status. Experimental results demonstrate that, compared to the LEACH, DMaOWOA, and ARSH-FATI-CHS protocols, IZOACP significantly outperforms the comparison schemes in key metrics such as network lifespan (improved by 97.56%), throughput (improved by 93.88%), and transmission delay (reduced by 10.12%). These results validate its superiority in energy consumption control, topology stability, and large-scale monitoring scenarios, providing an efficient and reliable clustering optimization framework for WSN information monitoring systems.

Hair specimens are vital in precision medicine, forensics and environmental monitoring owing to their ability to retain biochemical data over time. Their noninvasive collection and long-term storage suitability make them ideal for diagnostics and investigations, offering historical insights into health and exposure records. In medicine, hair analysis provides a long-term biochemical profile, aiding in monitoring health conditions, nutritional deficiencies, toxin exposure and treatment efficacy. Advances in mass spectrometry, chromatography and spectroscopy have expanded their applications to cancer diagnostics, tuberculosis, HIV, neurological disorders and mental health assessments. In forensic science, the resistance of hair to decomposition and its ability to absorb substances help identify individuals, detect drug use and reconstruct crime scenes. Omics techniques such as genomics, proteomics and metabolomics enhance forensic accuracy by enabling precise substance detection and timeline reconstruction. Despite its potential, challenges such as hair growth variability, contamination and lack of standardized techniques limit the current impact of hair analysis. Addressing these issues could advance its role in diagnostics and forensic investigations. This review explores recent advancements and applications of hair analysis in precision medicine, infectious diseases, mental health, stress assessment and forensic science.

Sodium-glucose cotransporter 2 inhibitors (SGLT2is) are a novel group of oral medications used to treat type 2 diabetes mellitus (T2DM). Nonalcoholic fatty liver disease (NAFLD), a complication of T2DM, is now recognized as one of the most frequent causes of chronic liver disorders. We aimed to investigate the effects of SGLT2is on hepatic function and glucose homeostasis in patients with T2DM and comorbid NAFLD. To our knowledge, this is the first study in the Arab region to compare the effects of SGLT2is and other antidiabetic medications and to evaluate the outcomes after a 6-month follow-up period in this patient cohort. This cohort study involved 100 patients with T2DM. The patients were divided equally into two groups. The exposed group comprised 50 patients receiving any of the SGLT2is (empagliflozin or dapagliflozin); the non-exposed group comprised 50 patients taking any other oral antidiabetic medication (other than glucagon-like peptide-1 receptor agonists or SGLT2is). The outcomes investigated were glycemic control, hepatic function, and liver fibrosis parameters, investigated at baseline and after 6 months. Significant improvements in glycemic control, hepatic function, and fibrosis parameters were observed in the SGLT2i group after 6 months, as evidenced by laboratory and clinical data (p < 0.001). Significant improvements were observed in hemoglobin A1c, Fibrosis-4 index, gamma-glutamyl transferase level, alanine aminotransferase level, and NAFLD fibrosis score (p < 0.05). The findings indicate that 6 months of SGLT2i therapy improved fibrosis and glucose homeostasis in patients with T2DM and NAFLD. Therefore, SGLT2is are effective therapeutic agents in this patient population.

BACKGROUND: Metabolic dysfunction-associated fatty liver disease (MAFLD) affects up to 30% of the global population, yet effective pharmacological treatments remain limited. This systematic review and meta-analysis evaluated the efficacy of GLP-1 receptor agonists and dual GLP-1/glucose-dependent insulinotropic polypeptide (GIP) receptor agonists in managing MAFLD. METHODS: We systematically searched PubMed/MEDLINE, Cochrane CENTRAL, Web of Science, and Scopus, through July 2025. Randomised controlled trials (RCTs) assessing GLP-1 receptor agonists or dual GLP-1/GIP GIP receptor Agonists in managing MAFLD patients were included. Primary outcomes included liver fat content, liver enzymes, and glycemic parameters. Meta-analyses were performed with subgroup analyses by receptor target, treatment duration, control type, and age. In addition, implementing a formal GRADE evaluation framework. RESULTS: Twenty-six trials involving 3,453 participants were included. GLP-1 receptor agonists significantly reduced liver fat content (MD: -3.37%, 95% CI: -4.98 to -1.76, p < 0.001), ALT levels (SMD: -0.47, 95% CI: -0.73 to -0.22, p < 0.001), and AST levels (SMD: -0.29, 95% CI: -0.53 to -0.05, p < 0.05). Significant improvements were observed in HbA1c (SMD: -0.67, 95% CI: -0.99 to -0.34, p < 0.001), fasting glucose (MD: -0.60 mmol/L, 95% CI: -0.92 to -0.27, p < 0.001), HOMA-IR (SMD: -0.34, 95% CI: -0.66 to -0.02, p < 0.05), and total cholesterol (MD: -0.23 mmol/L, 95% CI: -0.30 to -0.15, p < 0.001). Liver stiffness showed no significant improvement (MD: -0.12 kPa, 95% CI: -0.75 to 0.50, p = 0.70). Dual GLP-1/GIP agonists demonstrated superior efficacy compared to mono GLP-1 agonists for reducing liver fat (MD: -7.15, 95% CI: -10.23 to -4.07, p < 0.001 versus MD: -2.44, 95% CI: -4.18 to -0.71, p < 0.01), representing a 2.9-fold greater effect. Long-term treatment (lasting over 48 weeks) demonstrated enhanced benefits across all outcomes. Overall, changes in body weight were not significant (MD: -1.51 kg, 95% CI: -4.07 to 1.06, p = 0.25). CONCLUSIONS: This meta-analysis provides evidence for the effectiveness of GLP-1 receptor agonists in managing MAFLD, with dual GLP-1/GIP agonists demonstrating superior hepatic benefits. Long-term therapy (lasting more than 48 weeks) is necessary for optimal outcomes. These findings support clinical implementation, particularly for patients with concurrent diabetes or obesity, positioning dual agonists as promising advancements in treatment.

<h4>Purpose</h4>To assess the incidence of bronchopulmonary dysplasia in preterm infants born at < 32 weeks of gestation who received delayed cord clamping rather than early cord clamping.<h4>Methods</h4>During this multicenter, prospective, observational cohort study, data of 1283 infants born at < 32 weeks of gestation who received umbilical cord management between January 1, 2020 and December 31, 2021, were collected from 26 tertiary referral neonatal intensive care units in China. The primary outcome was bronchopulmonary dysplasia.<h4>Results</h4>A total of 440 infants (34.3%) received delayed cord clamping. The bronchopulmonary dysplasia rates were 32.5% and 55.6% for infants who received delayed cord clamping and those who received early cord clamping, respectively. After adjusting for potential confounders, the delayed cord clamping group had a lower risk of bronchopulmonary dysplasia (odds ratio [OR], 0.47; 95% confidence interval [CI], 0.35-0.62; P < 0.001). No significant difference in the severity of bronchopulmonary dysplasia was observed between groups (P = 0.09). A subgroup analysis of 227 infants born at < 28 weeks of gestation revealed that 65 (28.6%) received delayed cord clamping, and that 40 (61.5%) of these 65 infants experienced bronchopulmonary dysplasia. After adjusting for potential confounders, the risk of bronchopulmonary dysplasia for the delayed cord clamping group was not decreased (OR, 0.65; 95% CI, 0.30-1.41; P = 0.28).<h4>Conclusion</h4>Delayed cord clamping independently decreased the risk of bronchopulmonary dysplasia in infants born at < 32 weeks of gestation with moderate-to-severe respiratory distress; however, for those born at < 28 weeks of gestation, this benefit was not observed.

BACKGROUND: Understanding the complex molecular mechanisms driving drug resistance in glioblastoma (GBM) is crucial to develop effective therapeutic strategies. While prior studies have identified resistance mechanisms tied to specific drugs or pathways, a multi-modal molecular analysis of resistance across a range of drug classes is lacking. METHODS: We identified highly drug-resistant (n = 5) and drug-sensitive (n = 4) cultures from a cohort of 32 patient-derived glioblastoma stem cell (GSC) cultures screened against a broad panel of ~ 500 anti-cancer drugs. To elucidate the key drivers of drug resistance, we performed integrative profiling of stemness, differentiation capacity, global gene expression, mutation profiles, and DNA methylation patterns between the two groups. RESULTS: Despite heterogeneous gene expression profiles, drug-resistant GSCs showed consistent upregulation of ATP-binding cassette (ABC) drug efflux transporters, stemness, and extracellular matrix (ECM)-related genes. Compared to drug-sensitive GSCs, drug-resistant GSCs exhibited more pronounced stem-like properties and reduced differentiation capacity. Notably, genes linked to axonogenesis displayed significant CpG island hypomethylation in drug-resistant GSCs. CONCLUSIONS: This study suggests a pivotal role for GSC plasticity, stemness maintenance, and ECM-mediated drug evasion in GBM treatment resistance. Our findings highlight the adaptive and dynamic nature of resistance mechanisms in GSCs, emphasizing the need for comprehensive molecular insights to inform targeted therapeutic strategies in GBM.

Soluble tau oligomeric assemblies display neurotoxic properties and may provide a pathogenic link between neurofibrillary tangle evolution and selective neuronal vulnerability in Alzheimer's disease (AD). However, the precise molecular and cellular pathways mediating tau oligomer toxicity are unclear. We combined single-neuron laser capture microdissection with custom microarrays to investigate differences in the molecular signatures of basal forebrain neurons within the nucleus basalis of Meynert (nbM) labeled for p75^NTR, a cholinergic cell marker, or dual-labeled for p75^NTR and TOC1, a tau oligomer marker. Tissue was obtained postmortem from Rush Religious Orders Study participants who died with an antemortem clinical diagnosis of no cognitive impairment (NCI), mild cognitive impairment (MCI), or mild/moderate AD. Using clinical diagnosis as a covariate to isolate tau oligomer-specific mechanisms, we identified 140 differentially expressed genes (DEGs) in p75^NTR + /TOC1 + cholinergic nbM neurons compared to p75^NTR + /TOC1- neurons. STRING interactome and pathway analysis revealed that downregulated genes were associated with pre- and postsynaptic function, with additional enrichment in glutamate and acetylcholine signaling. By contrast, upregulated genes related to cellular stress responses and apoptosis were clustered with a subset of downregulated DEGs regulating mitochondrial metabolism and redox function, indicative of bioenergetic failure. Weighted gene co-expression correlation network analysis of the entire dataset revealed only two significantly correlated modules, which were either negatively correlated with the presence of TOC1 and enriched for synaptic signaling or positively correlated with TOC1 and enriched for cellular responses to hypoxia. These data show with single-neuron resolution that oligomeric tau formation in vulnerable cholinergic nbM neurons, even prior to MCI, is associated with the dysregulation of multiple classes of genes driving cell/mitochondrial stress and synaptic imbalances, which may be amenable for disease-modifying therapeutic approaches.

Cohort studies are essential for elucidating disease progression, guiding research priorities, and identifying gaps in diagnosis and treatment. This manuscript presents the protocol and preliminary findings of the Brazilian Registry on Sjögren’s Disease (BRAS), a national, prospective cohort supported by the Brazilian Society of Rheumatology (SBR). BRAS aims to collect comprehensive data on patients with Sjögren’s Disease (SjD) who meet the 2002 AECG and/or 2016 ACR-EULAR classification criteria, fostering high-quality research initiatives. Data collection is conducted via REDCap and includes demographic, laboratory, and clinical parameters such as disease activity (ESSDAI), damage (SSDDI), comorbidities, cardiovascular risk (Framingham score), labial salivary gland biopsy, salivary gland ultrasound, and therapeutic approaches. Patient-reported outcome measures (PROMs) include ESSPRI, PROFAD, HADS, ESE, IPAQ-SF, and EQ-5D. To date, 1,082 patients have been enrolled (mean age 55.3 ± 13.3 years; 96.6% women). Major findings include xerostomia (93%), xerophthalmia (92%), positive lip biopsy (83.6%), anti-Ro/SSA antibodies (77%), anti-La/SSB (43%), systemic manifestations (71.3%, ESSDAI 4.8 ± 5.8), and organ damage (86.9%, SSDDI 2.3 ± 1.6). Average disease duration was 7 ± 6.3 years with a diagnostic delay of 3.7 years. Treatments included corticosteroids (18%), hydroxychloroquine (46%), immunosuppressants (38%), and rituximab (6.5%). Common comorbidities were hypertension (34.9%), obesity (30.2%), dyslipidemia (29.7%), fibromyalgia (26%), osteoarthritis (24%), hypothyroidism (23.6%), diabetes (12.9%), and cancer (12.7%). Patients reported high symptom burden (ESSPRI 5.4 ± 2.4), anxiety (48%), depression (40%), low quality of life (63.6 ± 22.2), and insufficient physical activity (only 32.7% exercised regularly). Findings highlight the need for a multidisciplinary care program. The findings reveal diagnostic delays, limited healthcare access, high symptom burden, systemic involvement, poor mental health and highlight the need for a multidisciplinary care program. The next phase involves establishing a biorepository for biological samples. Clinical trial number Not applicable.

<h4>Background</h4>The pathophysiology of post-infective fatigue syndromes (PIFS), including Long COVID, is unknown. This systematic review and meta-analysis aimed to investigate if PIFS is associated with persistent immune activation.<h4>Methods</h4>PubMed, EMBASE, and Web of Science were searched for terms related to infection, fatigue, persistent symptoms, and immunological markers.<h4>Population</h4>adults and adolescents; Exposure: documented acute infection; Comparator: those who developed PIFS vs. recovered controls from the same exposure; and Outcomes: immunological biomarkers. Studies which documented acute infection, applied diagnostic criteria for PIFS, and assayed circulating immunologic markers were eligible.<h4>Findings</h4>From 14,985 studies screened, 30 articles were included (n = 5102 participants; 833 PIFS/PIFS-like cases, n = 4269 recovered control participants) with many studies excluded by inadequate quality in eligibility criteria. The meta-analysis (11 studies; n = 413 PIFS cases, analysed with random-effects models) showed PIFS cases had increased: white cell counts at 3-6 months (Cohen's d: 0.41, 95% CI 0.09-0.74); and circulating levels of RANTES and TNFα at 6-12 months (Cohen's d: 0.45 [95% CI 0.16-0.73] and 0.30 [95% CI 0.04-0.57], respectively) compared to controls recovered from the same exposure.<h4>Interpretation</h4>These findings provide cautious support for persistent immune activation in PIFS, but warrant further replication. Future studies should include better documentation of acute infection and PIFS case characterisation.<h4>Funding</h4>ARL is supported by a National Health and Medical Research Council Practitioner Fellowship (Grant 1041897). CXS is supported by a Cancer Institute New South Wales Early Career Fellowship (2021/ECF1310). BZK is supported by the National Institute of Allergy and Infectious Diseases (AI 105781). RAE is supported by the National Institute for Health and Care.

The first gene therapy products for hemophilia A and B have recently been approved by the regulatory authorities. Although this is an important milestone for people with hemophilia, there is still a need to further improve on the efficacy, safety, and stability of expression and to ultimately include pediatric patients before the onset of arthropathy and other complications caused by uncontrolled bleeding. To overcome some of the limitations of conventional gene therapy strategies, gene editing is currently being explored in preclinical studies. Gene editing allows for targeted modifications of the human genome with unprecedented specificity based on zinc finger nuclease, meganuclease, transcription activator-like endonuclease, or clustered regularly interspaced short palindromic repeats (CRISPR) technologies that induce double-strand DNA breaks (DSB). Next-generation gene editing strategies, such as those dependent on CRISPR-derived base or prime editors, allow targeted genetic modification independent of the induction of DSBs, offering a potential safer alternative. Sustained efficacy and production of factor VIII or factor IX can be achieved after gene editing in patient-derived cells or in adult or newborn hemophilia A or B mouse models. These preclinical studies pave the way toward phase I/II clinical trials in patients with severe hemophilia. The potential risk of undesired off-target modifications of the human genome and adverse immune reactions, and the need for efficient delivery of the gene editing components, need to be rigorously addressed before the promise of gene editing for hemophilia can ultimately be fulfilled.

Endoplasmic reticulum (ER) stress plays a pivotal role in tumor progression. As research in tumor biology advances, the relationship between ER stress and tumor initiation, development, and immune regulation has increasingly attracted attention. ER stress activates the unfolded protein response (UPR), thereby affecting key processes in tumor cells, including metabolism, proliferation, invasion, metastasis, and drug resistance. Moreover, it modulates tumor immune responses by regulating the functions of immune cells within the tumor microenvironment. This review consolidates the concept of ER stress as a central signaling hub that dictates cell fate and extensively remodels the tumor ecosystem. From a clinical perspective, this understanding provides a strong rationale for therapeutically targeting the UPR, suggesting that combining ER stress modulators with immunotherapy represents a promising strategy to overcome therapeutic resistance and improve patient outcomes.

<h4>Background</h4>Endometrial cancer (EC) is the most common gynecological cancer. Ferroptosis is a novel type of programmed cell death that is dependent on iron, and mounting evidence suggests that ferroptosis plays an important role in cancer. Long non-coding RNAs (lncRNAs) are known to regulate ferroptosis; however, little is known about the involvement of ferroptosis-related lncRNAs (FerlncRNAs) in EC. This study aimed to determine a FerlncRNA-based prognostic signature associated with the overall survival (OS) and clinicopathological characteristics of patients with EC.<h4>Methods</h4>Tumor transcriptomes and corresponding clinical data from patients with EC were downloaded from The Cancer Genome Atlas (TCGA) database, and the ferroptosis database, FerrDb, was used to identify ferroptosis-related genes (FRGs) (mRNAs). FerlncRNAs in EC were selected based on their correlations with FRGs. Univariate, multivariate, and least absolute shrinkage and selection operator (LASSO) Cox regression analyses were conducted to construct a prognostic model based on the FerlncRNAs signature. The EC patients were grouped into high- and low-risk categories based on the prognostic model risk score. Kaplan-Meier (K-M) survival analysis and time-dependent receiver operating characteristic (ROC) curves were used to evaluate the prognostic value of the risk scores. A predictive nomogram was then established. Gene set enrichment analysis (GSEA) was performed to explore the enriched pathways in the two risk groups. Finally, we compared the proportion of infiltrating immune cells and the expression of potential immune checkpoints between the two groups to understand the tumor immunological microenvironment associated with signature FerlncRNAs.<h4>Results</h4>We constructed a FerlncRNAs model to predict the prognosis of patients with EC. K-M analysis demonstrated that patients in the high-risk group had a worse OS. According to the ROC curves, our prognostic model had a better ability to predict the prognosis of patients with EC than other clinical factors. Moreover, the predictive nomogram suggested that our model could offer an independent prognostic evaluation with high accuracy. GSEA identified several enriched pathways in both groups. Finally, the immune microenvironment, including the infiltrating immune cells and immune checkpoints, showed several differences between the two groups.<h4>Conclusions</h4>This study revealed that a prognostic model based on 10 ferroptosis-related lncRNAs is useful for predicting the prognosis of patients with EC. Our findings provide novel directions for prognostic assessments, immunotherapies, and targeted treatments of EC.

<h4>Background</h4>Cutaneous melanoma (CM) is a highly lethal skin tumor. Some patients respond poorly to existing therapies, and developing new targeted therapies remains challenging.<h4>Methods</h4>We combined the results of eQTLs, pQTLs, and genome-wide association study (GWAS) to identify potential causal effects of two target genes on CM, based on multi-omics Mendelian randomization (MR). Sensitivity analysis, co-localization analysis, and inverse MR analysis were also employed to verify the robustness of this causal relationship. Multi-omics data were then applied to explore the expression patterns of immune infiltration of the target genes and construct nomogram models.<h4>Results</h4>The results showed that the gene prediction levels of EPS15L1 and HGS were associated with an increased risk of CM. Co-localization analysis revealed significant horizontal pleiotropy of the target gene, and reverse MR showed unidirectional causality of the targets. Multi-omics analysis comprehensively demonstrated the expression regulation pattern of the target genes in the CM immune-environment and identified interactions between EPS15L1 (Q9UBC2) and HGS (O14964) and doxorubicin, demonstrating the potential for drug application. The validity of the targets was further verified by molecular biology experiments.<h4>Conclusion</h4>This study provides robust genetic and therapeutic evidence for targeting EPS15L1 and HGS in CM treatment.

Multidrug-resistant (MDR) bacterial infections have emerged as a significant threat to global public health, with antibiotic resistance mechanisms-such as alterations in bacterial membrane permeability-substantially limiting therapeutic options and exacerbating infection persistence. In this study, we developed a novel self-carrier nanodrug formulation by directly assembling antibiotic tobramycin (Tob) and antimicrobial borneol 4-formylbenzoate (BF) into Tob-BF nanodrugs (TBN) <i>via</i> dynamic Schiff base linkages, without the need for additional adjuvants. The resultant TBN exhibits minimal bactericidal activity under physiological conditions but undergoes decomposition and activation specifically within the microenvironment of bacterial infections. Confocal microscopy analysis demonstrates that TBN is preferentially taken up and accumulates intracellularly in multidrug-resistant <i>Staphylococcus aureus</i> (MDRSA) to a greater extent than either Tob or BF alone. In both <i>in vitro</i> and <i>in vivo</i> assays, TBN outperforms Tob, BF, and physically mixed Tob + BF (with equal dose to TBN) in terms of antibacterial efficacy, particularly in promoting the recovery of xenograft infection with MDRSA in mice. Collectively, these findings highlight the potential of our self-carrier nanodrug platform, which integrates two distinct antibacterial agents, as an innovative and effective strategy to overcome tobramycin resistance in MDRSA and eliminate MDRSA infections.

<h4>Background</h4>Medulloblastoma is the most common malignant brain tumor of childhood, accounting for 25% of pediatric central nervous system (CNS) neoplasms. Although the cause of medulloblastoma is unclear, recent findings have contributed to treatment guidelines that emphasize the extent of disease, tumor resection margins, and age of onset. This study synthesizes available evidence to provide further insight into associated genetic markers and associated pathways that may be leveraged for individualized therapy.<h4>Methods</h4>The Search Tag Analyze Resource for NCBI's Gene Expression Omnibus (STARGEO) was utilized to identify 480 medulloblastoma tumor samples and 62 healthy adult and pediatric cerebellum samples. Pathway analysis was conducted using Ingenuity Pathway Analysis (IPA) and restricted to genes with a statistically significant difference (P<0.05) between medulloblastoma and control and an absolute experimental log ratio greater than 0.2.<h4>Results</h4>Overall, 4,142 genes met the inclusion criteria. Genes previously described in the context of medulloblastoma, such as <i>SOX11</i>, <i>TBR1</i>, <i>VSNL1</i>, <i>PVALB</i>, as well as novel gene targets such as <i>LHX2</i>, <i>UBE2C</i> and <i>HEPACAM</i> were among the differentially expressed genes identified. The top canonical pathways associated with medulloblastoma were cell cycle checkpoints, synaptogenesis signaling and pathway, mitotic metaphase and anaphase, glutaminergic receptor signaling pathway, and mitotic prophase. The top upstream regulators were beta-estradiol, <i>TP53</i>, <i>HTT</i>, <i>TBX3</i>, and <i>TGFB1</i>. Some of the diseases and biological functions predicted as activated with medulloblastoma based on the differential genetic expression include motor dysfunction or movement disorder and cell proliferation of tumor cell lines, whereas those predicted as inhibited include coordination and misalignment of chromosomes.<h4>Conclusions</h4>Utilizing STARGEO is an effective method for leveraging genomics metadata to highlight novel and previously described pathways and regulators associated with medulloblastoma. By providing an enhanced understanding of the pathophysiology of medulloblastoma, this study provides a framework for future validation studies-the next step toward identifying target genes and biomarkers for screening, prognostication, and targeted treatment for medulloblastoma.

Organ transplantation is an effective treatment for end-stage organ failure, but rejection remains a major obstacle to transplant success. Neutrophils play a key role in organ transplant rejection, participating not only in early immune responses but also exacerbating graft injury through mechanisms such as the release of neutrophil extracellular traps (NETs). Therefore, in-depth exploration of the immunological role of neutrophils in transplant rejection and their interactions with other immune cells is highly important. This article reviews the latest research progress on the mechanisms of action of neutrophils in transplant rejection and their impact on grafts while also assessing the clinical application prospects of immunosuppressive strategies targeting neutrophils and NETs. By integrating current basic and clinical research findings, this article aims to provide theoretical support and new research directions for the diagnosis and treatment of neutrophil-related rejection, with the goal of improving organ transplant success rates and patient quality of life.

<h4>Purpose</h4>Different hospitals in Sudan detected a rare condition of liver vascular abnormalities characterized by vascular outflow impairment. The study aimed to describe the common radiological features of Budd Chiari syndrome and determine which feature is most frequently employed to characterize this condition during imaging techniques, primarily contrast-enhanced computed tomography (CECT) scans.<h4>Materials and methods</h4>The study was conducted between March 2023 and June 2024 at Kuwaiti Specialized Hospital (KSH) and other diagnostic centers using a liver protocol on a CT machine (Optima 520 GE-CT machine, 16 slices). The study was a retrospective, cross-sectional, and review-based analysis of a rare study type carried out to characterize the state of BCS in Sudan; the CT scan's findings on the liver, HVs, IVC, and abdomen were carefully assessed. The age range of the 61 patients who underwent a successful triphasic CT abdomen for the liver was 2-78 years.<h4>Results</h4>The findings indicate that: the majority of patients 57.4% were male, the most common age groups were 39-52 years old, and the mean age at diagnosis was 45 years. BCS is primarily caused by hepatic veins (HVs) thrombosis, which is observed in 18.03% of cases, and HVs are not seen in 55.73% of cases. Liver parenchymal enhancement appears heterogenous in 27.87%, while heterogeneously enlarged liver was seen in 24.59%, and cirrhotic in 14.75% of BCS patients. In comparison, 59.01 percent of BCS patients arrived without varices. Varices were observed in the splenorenal and gastroesophageal regions in 37.7% of cases. Ascites accounted for the majority of BCS complications 73.77%, with SM vein blockage and squeezed duodenum accounting for 3.27% of each complication. The likelihood of developing ascites increases with age, and it is most common in patients between the ages of 39 and 52 years. Patient age had the greatest effect on the development of ascites.<h4>Conclusion</h4>The common features of BCS as revealed by contrast-enhanced CT of the liver are non-visible HVs, venous occlusion at either level of HVs or IVC, caudate lobe enlargement, heterogenous, normal or enlarged liver, collateral venous varices at the splenorenal and gastroesophageal region, and ascites.

<b>Background</b>: Sarcopenia is a clinical condition linked to various liver diseases, including metabolic dysfunction-associated steatotic liver disease (MASLD). MASLD includes a spectrum from steatosis to steatohepatitis, which may progress to fibrosis, cirrhosis, and hepatocellular carcinoma. The influence of dietary habits and nutrient intake on MASLD and its progression is well-established. However, the association between dietary consumption and sarcopenia in MASLD patients remains underexplored. This study evaluated whether there is an association between sarcopenia and habitual food consumption in MASLD patients. <b>Methods:</b> A cross-sectional study was conducted with outpatients diagnosed with MASLD. Sarcopenia was defined based on the 2019 EWGSOP2 criteria. Dietary intake was assessed using three 24 h recalls per patient, with intrapersonal variance corrected using the Multiple Source Method (MSM) software (Version 1.0.1). Steatosis was diagnosed via upper abdominal ultrasound, and the Fibrosis-4 Index (FIB-4) was used to assess hepatic fibrosis. <b>Results:</b> MASLD patients (<i>n</i> = 76) were evaluated. The mean age was 52.9 (SD, 12.0) years, and 75.0% were female. Two had sarcopenia, and 27.6% (<i>n</i> = 21) had probable sarcopenia (characterized by low muscle strength only). Among probable sarcopenia, F1-F2 were observed in 61.9%, and 23.8% had indeterminate FIB-4 grades. Calcium intake was lower among patients with probable sarcopenia than those no sarcopenia (<i>p</i> = 0.04). <b>Conclusions:</b> In these MASLD patients, only two patients were diagnosed with sarcopenia, and around a third had probable sarcopenia. The majority of MASLD patients with lower calcium, energy, and protein intake, but only lower calcium intake in those with probable sarcopenia.

Periodontitis (PD) is a multifactorial, progressive inflammatory disease affecting the teeth-supporting tissues, characterized by an imbalance of the oral microbiota and the presence of bacterial biofilms leading to host response. Nowadays, reliable biochemical markers for early and objective diagnosis, and for predicting disease progression, are still lacking. Our previous proteomic investigations revealed the significant overexpression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in periodontal pocket tissue, gingival crevicular fluid (GCF), and tooth-surface-collected material (TSCM) from PD patients in comparison to periodontally healthy controls, proposing it as a possible biomarker of PD. This study aimed to evaluate the expression of GAPDH in saliva, a more accessible, non-invasive, and clinically relevant oral sample. The whole saliva was analyzed by a preliminary mass spectrometry-based proteomic approach, identifying significantly increased levels of GAPDH also in salivary samples from periodontal-affected subjects. These data were further validated by enzyme-linked-immunosorbent assay (ELISA). Additionally, protein-protein interaction networks were generated through the Human Protein Atlas database, using different datasets (OpenCell, IntAct, and BioGRID). Bioinformatic analysis provided noteworthy GAPDH-associated networks potentially relevant to periodontal pathology. The scientific significance of this study lies in the detection of salivary GAPDH as a novel strategy to advance periodontal clinical diagnostics from the perspective of a non-invasive screening test. In correlation with other protein markers, salivary GAPDH could constitute a promising set of distinctive and predictive targets to enhance early diagnosis of PD, disease monitoring, and treatment planning in periodontology.

<b>Objectives:</b> This study aimed to assess the dynamics of liver tests (LT) and detect signs of liver fibrosis and steatosis 2.5 years after the first COVID-19 episode in patients without pre-existing liver-related conditions. <b>Methods:</b> The study included 65 adult patients hospitalized with COVID-19 (including 18 with severe or critical illness) in 2020. After 2.5 years, in addition to regular LT, liver health status was assessed by the FIB-4 index, hyaluronic acid, cytokeratin 18 fragment M30 (serum, ELISA), cardiometabolic risk factors, and the multiparametric ultrasound examination. <b>Results:</b> LT abnormalities in the acute COVID-19 period were observed more frequently (<i>p</i> = 0.036) in patients with severe or critical COVID-19 (83%) than in patients with non-severe COVID-19 (55%). LT dynamics in 2.5 years showed an improvement of liver health status in most patients (<i>p</i> = 0.006). Persistent LT abnormalities were associated with LT abnormalities during hospitalization (<i>p</i> = 0.021). After 2.5 years, the presence of cardiometabolic risk factors and signs of liver fibrosis were associated with the severity of the first COVID-19 episode. However, regression analyses did not support disease severity as a predictor for LT abnormalities and liver stiffness. The latter was predicted by cardiovascular diseases in the anamnesis. <b>Conclusions:</b> In most patients, LT normalized despite potential risk factors. Simultaneously, in some patients, signs of liver fibrosis after COVID-19 might be stimulated by COVID-19-related metabolic dysfunction and the presence of cardiovascular diseases.

SARS-CoV-2 infection remains a global health concern, with its impact on host immune responses not fully understood. In a case-control study, we examined how COVID-19 affects DNA methylation patterns in the upper respiratory airway of hospitalized individuals. DNA methylation arrays were performed on nasopharyngeal samples at inclusion/hospitalization and 6 weeks post-inclusion. We found a distinct DNA methylation pattern in COVID-19 patients compared to healthy controls, identifying 510,099 differentially methylated CpGs. Within the transcription start sites (TSSs) and gene body, COVID-19 patients displayed a higher number of genes/CpGs with elevated methylation levels. Enrichment analysis of TSS-methylated genes revealed effects of SARS-CoV-2 on genes associated with type I interferons, anti-viral and inflammatory responses, and immune functions. Some CpG methylations were transient, and normalized at group level by 6 weeks post-inclusion. Several IFN-regulated genes, including OAS1, OAS3, IFIT3, and MX1, were identified. Among the top regulators were IL17A and ERK1/2, both involved in inflammatory processes. Networks nodes included IGF1 and EGF, associated with processes including tissue repair and activation of immune responses. Overall, our data suggests that COVID-19 can impact the upper airway by modifying gene methylation patterns. This could have implications for conditioning of the airways, how individuals respond to future airway infections, and therapeutic interventions.

Ferroptosis, an iron-dependent form of regulated cell death characterized by lipid peroxidation, has emerged as a pivotal vulnerability in oral squamous cell carcinoma (OSCC). This review provides an overview of ferroptosis mechanisms and their implications for OSCC pathobiology and therapy. OSCC cells exhibit heightened reliance on anti-ferroptotic defenses such as GPX4, SLC7A11, FSP1, and Nrf2, and disrupting these pathways suppresses tumor growth and restores sensitivity to chemotherapy, radiotherapy, and immunotherapy. Genetic and epigenetic regulators, including p53, PER1, circ_0000140, and STARD4-AS1, critically modulate ferroptotic sensitivity, while metabolic enzymes such as ACSL4, LPCAT3, and TPI1 link ferroptosis to cellular plasticity and resistance. Preclinical studies highlight the promise of small-molecule inhibitors, repurposed agents (e.g., sorafenib, artesunate, trifluoperazine), natural compounds (e.g., piperlongumine, Evodia lepta, quercetin), and nanomedicine platforms for targeted ferroptosis induction. We further address ferroptosis within the tumor microenvironment, highlighting its immunogenic and context-dependent dual roles, and summarize genomic and transcriptomic evidence linking ferroptosis-related genes to patient prognosis. Beyond cancer, ferroptosis also contributes to non-malignant oral diseases, including pulpitis, periodontitis, and infection-associated inflammation, where inhibitors may protect tissues. Despite these advances, clinical translation is constrained by the lack of safe ferroptosis inducers and validated biomarkers. Future research should focus on developing pharmacologically viable GPX4 inhibitors, refining biomarker-driven patient stratification, and designing multimodal regimens that combine ferroptosis induction with standard therapies while preserving immune and tissue integrity. Ferroptosis therefore represents both a mechanistic framework and a translational opportunity to reshape oral oncology and broader oral disease management.

<h4>Background</h4>Colorectal cancer (CRC) represents a huge global health challenge characterized by significant morbidity and mortality. The urgent need to identify biomarkers through integrative validation research to enhance diagnostic accuracy and prognostic stratification has prompted the exploration of immune and prognostic genes. This study aimed to systematically identify differentially expressed genes (DEGs) associated with both immunity and prognosis in CRC, validate their clinical significance, and construct a reliable prognostic model.<h4>Methods</h4>This research sought to identify DEGs associated with immunity and prognosis in CRC. We examined clinical and RNA sequencing data from 698 CRC patients obtained from The Cancer Genome Atlas (TCGA). Utilizing the Xiantao Academic Platform, we conducted differential expression analysis and identified hub genes associated with immunity and prognosis through Least Absolute Shrinkage and Selection Operator (LASSO) and Cox regression analyses, alongside five machine learning algorithms to construct a prognostic model. The hub genes were validated using the Gene Expression Omnibus (GEO) database, molecular docking, molecular dynamics simulation, single-cell and spatial transcription analyses.<h4>Results</h4>LASSO and Cox regression analyses, along with five machine learning algorithms, were employed to identify significant genes linked to immunity and prognosis, yielding three hub genes: <i>ULBP2</i>, <i>INHBB</i>, and <i>STC2</i>. Validation of these genes in the GEO dataset GSE21815 demonstrated significant diagnostic performance, with area under the curve (AUC) values of 0.908, 0.742, and 0.934, respectively. A prognostic model integrating clinical factors and hub genes was developed, demonstrating high predictive accuracy for 1-, 3-, and 5-year survival rates. Further analysis revealed significant enrichment in the TGF-β signaling pathway and natural killer cell-mediated cytotoxicity, as evidenced by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The single-sample Gene Set Enrichment Analysis (ssGSEA)-based immune infiltration analysis revealed immune infiltration differences between groups with high and low immune phenotype scores. Molecular docking and dynamics simulations revealed valproic acid, cyclosporine, and genistein as potential therapeutic compounds with strong binding affinities to the hub genes. Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics provided insights into hub gene expression patterns and interactions within the tumor microenvironment.<h4>Conclusions</h4>This comprehensive study highlights the potential of <i>ULBP2</i>, <i>INHBB</i>, and <i>STC2</i> as promising biomarkers for CRC, emphasizing their roles in regulating tumor progression and immune responses. Future studies should focus on targeted therapeutic strategies that utilize these biomarkers to enhance treatment efficacy and patient prognosis.

<h4>Background</h4>CCT4 is a key molecular chaperone and its dysfunction may impair actin or microtubule assembly, potentially driving hepatocellular carcinoma (HCC) progression. However, its roles in multiple cell types and tumor microenvironment infiltration in HCC remain unclear. This study aimed to investigate the biological functions of <i>CCT4</i> in HCC by integrating single-cell and bulk transcriptomic data.<h4>Methods</h4><i>CCT4</i> expression in HCC was analyzed using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Functional analyses were conducted via Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). Furthermore, Cox regression analysis and nomogram construction were employed to assess patient prognosis. Immune infiltration was evaluated with CIBERSORT, and drug sensitivity was tested using the CellMiner database. Hepatocyte features were analyzed using single-cell RNA-seq tools, including Cellular Trajectory Reconstruction Analysis using gene Counts and Expression (CytoTRACE) for stemness and CellChat for cell-cell communication.<h4>Results</h4><i>CCT4</i> showed significant overexpression in HCC tissues and was associated with clinical features such as age and tumor node metastasis (TNM) stage. Functional analyses linked co-expression genes of <i>CCT4</i> to ribosomal biogenesis and protein folding. Cox regression and nomograms confirmed <i>CCT4</i> as an independent prognostic factor for HCC. Correlations with nine immune cell types were observed through immune infiltration analysis. Drug sensitivity analysis revealed that HCC patients with high expression of <i>CCT4</i> were resistant to sorafenib and other drugs. Single-cell analysis revealed that elevated <i>CCT4</i> expression was enriched in pro-metastatic hepatocyte clusters, associated with increased hepatocyte proportions and poor differentiation, with weakened T/natural killer (NK) cell interactions in tumors.<h4>Conclusions</h4><i>CCT4</i> is overexpressed in HCC, linked to poor prognosis, and may drive tumor progression via pro-metastatic hepatocyte dedifferentiation and immune shifts.

With an ageing global population, the triad of type 2 diabetes (T2D), obesity and dementia poses a growing public health challenge. India harbours a notable proportion of younger patients with T2D with a non-obese or lean phenotype. However, data remain scarce on the impact of adiposity on cognition in T2D, particularly when assessed using a comprehensive, culturally and linguistically adaptable cognitive battery in a cohort free from major confounding factors. The aim of this study is to examine the relationship between various anthropometric indices and cognitive performance in patients with T2D. This cross-sectional observational study was conducted at the diabetic clinic of a tertiary hospital from 2022 to 2024. Eligible participants were patients with T2D aged 20-60 years with at least primary education. Exclusion criteria included non-T2D diagnosis, inability to communicate in Bengali and conditions known to impair cognition. A total of 125 patients with T2D were recruited. Demographics, diabetes-related variables and anthropometric measurements were recorded. Cognitive function was assessed using the Bengali version of the Addenbrooke's Cognitive Examination (ACE-III). Statistical analysis was performed using Jeffreys' Amazing Statistics Program (v.0.19). ACE-III total scores showed significant positive correlations with height, weight and neck circumference (NC) (p<0.001 for each). Attention was positively associated with height, weight, NC, neck-height ratio (NHR) and negatively with weight-adjusted waist index (WWI) (p<0.05 for all). Memory correlated positively with height and weight (p<0.05 for both). Language was positively related to height, weight and NC (p<0.05 for all) and negatively to WWI (p=0.011). Visuospatial ability positively correlated with height, weight, waist circumference (WC), hip circumference (HC), NC and NHR (p<0.05 for all). Lean patients with T2D had significantly lower visuospatial scores (p=0.040), lower ACE-III total scores (p=0.049) and a greater prevalence of cognitive impairment (p=0.032). In multiple linear regression, height (p=0.014) and HC (p=0.024) were independent predictors of ACE-III total score. This is the first Indian study to evaluate the association between anthropometric measures and cognition in T2D. Cognitive impairment and dementia were more prevalent in lean than in obese patients with T2D. Future studies incorporating imaging-based body composition analysis are warranted to identify modifiable anthropometric risk factors for cognitive decline in T2D.

To mitigate the adverse effects of doxorubicin (DOX) during the treatment of liver cancer, this study aimed to develop a novel DOX-encapsulated active-targeted nanodelivery system. The auxiliary materials hydroxyapatite (HAP), polysuccinimide (PSI), galactose-modified polyethylene glycol (Gal-PEG), PSI covalently linked with three different mole ratios of polyethylene glycol (PEG-PSIs), and galactose covalently linked with PEG-PSIs (Gal-PSIs) were synthesized and structurally characterized. The Box-Behnken and Three-Level Factorial Design response surface methodologies were employed to optimize the formulations and synthesis protocols for DOX@HAP/PSI (DOX@DC, where DC denotes the drug carrier), DOX@PEG-DCs, and DOX@Gal-DCs. The in vitro drug release and in vivo tissue distribution of each formulation were examined. Furthermore, in vitro studies were conducted to examine the effects of these formulations on the proliferation, apoptosis, and migration of Huh-7 liver cancer cells. The formulations and synthesis protocols for different DOX-based preparations were optimized. All nanoparticles gradually released DOX at pH levels >5, with the release rate increasing with the pH value. Among the tested formulations, DOX@Gal-DC20 (mole ratio of Gal-PEG to PSI = 1:20) showed the best hepatic targeting in mice in vivo. Furthermore, in vitro pharmacodynamic experiments indicated that Gal-DC20 had low cytotoxicity, could be taken up by cancer cells, and could significantly inhibit the proliferation of Huh-7 cells. Hemolysis experiments confirmed that none of the prepared formulations induced hemolysis. The novel nanodelivery system established in the study (DOX@Gal-DC) is simple to prepare and shows significant hepatic targeting.

The vascular system plays a crucial role in maintaining homeostasis, ensuring the supply of oxygen and nutrients to tissues, while facilitating the removal of metabolic waste. Additionally, it contributes to immune defense, temperature regulation, and the transport of hormones and signaling molecules. Vascular anomaly (VA) arises due to developmental abnormalities or functional defects in the vessels. This review describes venous malformations (VM), a rare disorder predominantly caused by somatic mutations. Advances in recent research have substantially improved our understanding of the molecular mechanisms underlying these malformations, largely through the identification of their genetic origins and the study of animal models and endothelial cells derived from patients. Most of the somatic mutations associated with venous malformations affect genes within oncogenic growth factor signaling pathways, making it possible to repurpose certain cancer therapies to treat these VAs. This article summarizes the key molecular findings and explores emerging therapeutic strategies aimed at novel targets.

Sepsis-associated acute kidney injury (SA-AKI) is a prevalent and life-threatening complication in critically ill children, contributing to high mortality rates (up to 30%) and long-term renal dysfunction in pediatric intensive care units. This review synthesizes recent advances in the signalling pathways underlying SA-AKI, emphasizing pediatric-specific mechanisms, biomarkers, and therapeutic targets. This review covers inflammatory cascades via TLR/NF-κB leading to cytokine storms (IL-6, TNF-α); apoptosis and necrosis involving mitochondrial Bcl-2 dysregulation and OLFM4; and emerging processes like pyroptosis (NF-κB-mediated), metabolic reprogramming (choline deficiency and Nrf2-mitophagy), and novel routes such as cGAS-STING and TGF-β signalling. Biomarkers like urinary OLFM4, DKK3, NGAL, and serum suPAR, alanine, and Penkid enable early diagnosis and risk stratification, with models like PERSEVERE-II enhancing prognostic accuracy. Therapeutic strategies include fluid optimization, renal replacement therapies (CRRT, SLED-f), and pathway-targeted interventions such as choline supplementation, oXiris for cytokine removal, Humanin for immunomodulation, and investigational cGAS-STING inhibitors. Despite progress, challenges persist in translating animal models to pediatric trials and addressing heterogeneity. Integrating multi-omics and precision medicine holds promise for improving outcomes, underscoring the need for multicenter studies in children.

This paper presents the design of a Delay-Locked Loop (DLL) with a simple architecture and a wide input clock duty cycle range. The design is tailored to meet the increasing data rate and stringent clock requirements of modern semiconductor chips, with particular applicability to dynamic random-access memory (DRAM) systems. The structure features two Bang-Bang Phase Detectors (BBPDs) to adjust the rising and falling edges of the divided clock. Implemented using a 65 nm CMOS process, the design was verified through simulation. At a working frequency of 3.2 GHz, the input clock duty cycle range spans from 18% to 72%, with a maximum output clock duty cycle error of just 0.6%, a peak-to-peak jitter of 15.73 ps, and a power consumption of 12.7 mW.

At birth, major circulatory and pulmonary adaptations are required for a successful foetal-to-neonatal transition. In newborns with transposition of the great arteries (TGA), this transition is often impaired, leading to severe hypoxemia. This may also result in persistent pulmonary hypertension of the newborn (PPHN), worsening hypoxemia, and increasing the risk of urgent invasive interventions after birth. Physiological-based cord clamping (PBCC), delaying cord clamping until after lung aeration and ventilation are established, promotes a more stable circulatory transition and has shown benefits in both preterm and term neonates. PBCC may also provide advantages in infants with congenital heart disease. In TGA, combining PBCC with early supplemental oxygen may reduce the incidence and severity of PPHN, decrease related complications, and minimize the need for invasive interventions. To evaluate the feasibility, safety, and clinical outcomes of the stabilisation approach, we have initiated a stepwise quality improvement initiative comprising 2 sequential studies. Study phase 1, a single-centre study at Leiden University Medical Centre will assess feasibility, protocol adherence, and safety. Study phase 2, an observational cohort study across multiple European centres will evaluate clinical outcomes, focusing on incidence and severity of PPHN and the need for urgent interventions. All TGA newborns will be stabilised with an intact umbilical cord while receiving 2 L/min nasal high flow (fraction of inspired oxygen 1.0) as supplemental oxygen. Cord clamping will occur once the infant is considered stable, defined as a heart rate >100 bpm and preductal SpO₂ >75% with supplemental oxygen. This study will inform guidelines for delivery room management and early preoperative care in TGA newborns.

<h4>Background</h4>Immunometabolism plays a vital role in the immunopathogenesis of people living with human immunodeficiency virus type 1 (HIV-1) (PLWH). However, the precise relationship between metabolic profiles and T-cell dysfunction in this population remains unclear. This study aimed to investigate the metabolic reprogramming and underlying mechanisms contributing to T-cell dysfunction in PLWH, highlighting potential pathogenic mechanisms during chronic HIV-1 infection.<h4>Methods</h4>This study re-analyzed single-cell RNA sequencing data from the Genome Sequence Archive of the Beijing Institute of Genomics Data Center, Chinese Academy of Sciences. The dataset comprised samples from healthy donors (HD), HIV-1-infected treatment-naive patients (TN), and patients undergoing antiviral therapy. Various analytical approaches-including functional analysis, transcription factor analysis, network analysis, and enrichment analysis-were performed to assess T-cell functional and metabolic characteristics, as well as to identify potential targets within metabolic-epigenetic or non-epigenetic regulatory axes involved in T-cell dysfunction.<h4>Results</h4>By analyzing the transcriptional profiles, a total of 58,752 CD4⁺ T cells and 68,907 CD8⁺ T cells were identified and annotated. Among these, the naive subset CD8-CCR7 was significantly reduced in TN patients compared to HD (<i>P</i> < 0.05), whereas CD4-CCR7 showed a decreasing trend. Conversely, the effector subset CD8⁺ activated effector/memory T cells (CD8-EMRA) were significantly increased in TN patients (<i>P</i> < 0.05), while cytolytic CD4⁺ T cells (CD4-CTL) displayed an increasing trend. ART did not effectively reverse these alterations. Additionally, naive subsets and CD8-EMRA cells were associated with disease progression. Further analysis revealed that naive subsets exhibited hyper-activation and increased differentiation, whereas effector subsets showed excessive activation and a strong interferon (IFN) response in PLWH compared to HD (<i>P</i> < 0.05). Intriguingly, we observed substantial metabolic alterations linked to immune dysfunction within the four T-cell subsets. Specifically, elevated levels of the methyltransferases absent, small, or homeotic-like 1 (<i>ASH1L</i>) and SET domain containing 1B (<i>SETD1B</i>) may have promoted the differentiation and exhaustion of the CD4-CCR7 subset via the ASH1L/SETD1B-H3K4me3-FOXP1 axis. These enzymes were also associated with the exhaustion of CD8-CCR7 cells in TN patients through ASH1L/SETD1B-H3K4me3 axis. Additionally, isocitrate dehydrogenase 2 (IDH2)-mediated production of α-ketoglutarate (α-KG) may have contributed to the dysfunction of CD8-CCR7 cells by activating Janus kinase (JAK)-signal transducer and activator of transcription (STAT)3-dependent interferon signaling during HIV-1 infection. Conversely, increased activity of SET domain containing 2 methyltransferase was closely linked to hyperactivation, a strong type I interferon response, and cellular senescence in CD4-CTL cells from TN patients. Furthermore, heightened expression of solute carrier family 7 member 5 correlated with exhaustion of effector subsets in TN individuals. The IDH2-STAT1 axis may have also played a crucial role in driving the over-activation and exhaustion of CD8-EMRA cells through interferon signaling pathways.<h4>Conclusion</h4>These findings indicate that amino acid- and IDH2-related metabolism may contribute to the dysfunction of both naive and effector subsets by metabolic-epigenetic or non-epigenetic regulatory axes in PLWH.

<title>Abstract</title> <p> Background: Telomerase reverse transcriptase ( <italic>TERT</italic> ) promoter mutation is among the most frequent genetic alterations in hepatocellular carcinoma (HCC), closely associated with tumor initiation, progression, and early recurrence; however, its prognostic role remains unclear. This study aimed to investigate the prevalence of <italic>TERT</italic> promoter mutations in HCC and evaluate their association with early recurrence, with emphasis on clinicopathological subgroup differences.. <h4>Materials and Methods:</h4> We retrospectively analyzed 108 patients with histologically confirmed HCC who underwent curative hepatectomy. <italic>TERT</italic> promoter mutations were identified via Sanger sequencing. The risk of early recurrence, defined as tumor relapse within 24 months after surgery, was assessed using Cox proportional hazards models, including subgroup analyses. <h4>Results:</h4> <italic>TERT</italic> promoter mutations were present in 45.4% of patients. While not predictive of early recurrence in the overall cohort (HR = 1.39; 95% CI: 0.76–2.56; p = 0.282), mutation status was significantly associated with reduced RFS in specific subgroups: patients <60 years (HR = 3.45; 95% CI: 1.22–9.82; p = 0.020), males (HR = 2.03; 95% CI: 1.02–4.02; p = 0.043), and those with high mitotic activity (HR = 3.60; 95% CI: 1.01–12.80; p = 0.047).. <h4>Conclusion:</h4> Although <italic>TERT</italic> promoter mutations did not predict early recurrence in the overall cohort, they were significantly associated with poorer recurrence-free survival in defined clinical and pathological subgroups. These findings suggest the context-dependent prognostic value of TERT promoter mutations and may support their potential role in individualized risk stratification following curative resection in HCC. </p>

Spalt-like (SALL) proteins are C2H2 zinc-finger transcription factors important for embryogenesis, with mutations in SALL1 and SALL4 causing rare congenital disorders Townes-Brocks and Okihiro syndromes, respectively. While SALL proteins are known to associate with one another, the biological significance of the resulting complexes is unknown. Here we define a conserved glutamine-rich region that mediates SALL1/4 homo- and heterotetramerisation and find that complex formation is indispensable for DNA binding. Modelling a patient mutation that abolishes SALL4 multimerisation led to gene misregulation and, in mice, embryonic lethality, therefore phenocopying a complete Sall4 knockout. Furthermore, a common disease-causing SALL1 truncation, which retains multimerisation but lacks DNA-binding domains, sequesters SALL4 into heterotetramers that are defective in DNA binding, thereby providing a mechanistic explanation for the dominant-negative effects of many Townes-Brocks mutations. Together, our findings establish tetramerisation as a prerequisite for SALL function, linking complex formation to developmental gene regulation and human disease.

Avian influenza viruses (AIVs) are a global public health risk; human infection is typically associated with high mortality. While the relationship between several mammalian adaptive mutations and host factors have been described, it is unknown whether additional uncharacterised mutations lead to adaptation. Here, we combine phylogenetic analysis and complementary experimental methods to quantify the impact of novel mutations that emerge at the avian-mammal interface. We constructed phylogenetic trees of mammalian and avian influenza sequences for the polymerase (PA, PB1, PB2) and nucleoprotein (NP) segments and identified potential avian to mammal spillover events. We found >6500 mutations across the polymerase and NP, including known signatures of mammalian adaptation such as PB2 E627K and D701N which occurred independently in mammals 143 and 56 times respectively. We selected 95 mutations which were mostly undescribed and emerged independently multiple times in a range of species and subtypes. Using a minigenome assay in an avian H5N1 backbone to measure the effect of these mutations in human cells we identified PA P28S, NP I425V and G485R as novel mutations leading to polymerase adaptation. In addition, to determine the mechanism of adaptive mutations, we measured polymerase activity in cells lacking a key host factor, ANP32, and cells overexpressing host restriction factors MxA and BTN3A3. Our combined approach revealed novel mammalian adaptive mutations and demonstrated the benefit of combining phylogenetic and molecular approaches in validating novel adaptive mutations.

Complete degeneracy of aliphatic ¹³C chemical shifts of the same type of amino-acid residues precludes unambiguous NMR assignments of asparagine (Asn) and glutamine (Gln) carboxamide NH₂ groups in intrinsically disordered proteins with conventional techniques. Here, a pair of triple-resonance NMR experiments is described that correlate the ¹H/¹⁵N chemical shifts of Asn and Gln carboxamide NH₂ groups with the backbone amide ¹⁵N chemical shifts of the same and the next residue. These experiments exploit a by far superior dispersion of backbone amide ¹⁵N chemical shifts, and permit unambiguous assignments of all Asn/Gln side-chain NH₂ groups in the intrinsically disordered protein α-synuclein and the majority of Gln NH₂ groups in the disordered huntington exon-1 protein with a 7-residue glutamine repeat, htt^ex1Q₇.

Surgical site infections remain a critical concern in dental and orthopedic procedures. To combat surgical site infections, gentamicin-loaded carbonate apatite (GM-CAp) granules, a novel antibacterial biomimetic bone substitute is developed. GM-CAp granules rapidly release the loaded gentamicin, leading to swift suppression of bacteria and proactive prevention of infection. In vitro studies validate the bactericidal effects of GM-CAp granules against a spectrum of odontogenic bacteria. In vivo investigations confirm the effectiveness of GM-CAp in suppressing Staphylococcus aureus infection, mitigating inflammation, especially neutrophil recruitment, and stimulating bone regeneration. Transcriptomic analysis reveals that early-stage neutrophil dynamics may influence the inflammatory milieu and subsequent bone healing in the infected bone defect following CAp or GM-CAp implantation. Overall, GM-CAp granules represent a potential strategy for combating surgical site infections and increasing the success rate of bone healing in dental and orthopedic surgeries. Additionally, it is proposed that the status of early postoperative neutrophil recruitment may serve as a potential parameter for evaluating the antimicrobial efficacy of biomaterials.

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by CAG repeat expansion in the HTT gene. Existing toxin-induced and genetic models provide important insights, but none fully replicate the progressive pathology of HD. An AAV9-mediated striatal mouse model expressing mutant HTT with 82 CAG repeats was established to reproduce hallmark neuropathological changes and behavioral deficits. Male C57BL/6 mice received bilateral intrastriatal injections of AAV9-HTT-82Q or control AAV9-GFP. Behavioral performance was assessed by rotarod, balance beam, open field, and Y-maze tests. Neuropathology was examined with HE/Nissl staining, TUNEL assay, and immunofluorescence for mHTT, DARPP-32, GFAP, and Iba1. AAV9-82Q mice exhibited progressive motor coordination deficits on the rotarod from Week 4 and impaired beam traversal from Week 18. Open field testing revealed persistent hyperactivity from Week 8, while anxiety-like and cognitive measures showed only mild, non-significant trends. Histological analysis demonstrated extensive mHTT aggregation in the striatum, accompanied by neuronal pyknosis, vacuolization, and significant loss of Nissl-positive neurons. TUNEL staining confirmed increased apoptosis. Immunofluorescence further revealed selective reduction of DARPP-32+ medium spiny neurons, along with marked astrogliosis and microgliosis, indicating robust neurodegeneration and inflammatory responses. The AAV9-82Q model induces adult-onset, progressive HD-like pathology with early motor impairments, neuronal loss, and glial activation. It complements existing models and provides a reproducible platform for mechanistic studies and preclinical therapeutic evaluation.

Tumor-derived osteochondral defects (OCDs) pose significant clinical challenges, necessitating integrated strategies for local tumor ablation while preserving the regenerative niche and enabling coordinated cartilage and subchondral bone repair. Herein, we introduce a spatiotemporal microneedle (MN) platform that synergistically combines protoporphyrin IX (PpIX)-mediated sonodynamic therapy (SDT) for precise tumor eradication with chondral organoid (CO)-driven regeneration, leveraging differential mechanical cues for lineage-specific guidance. COs were engineered from bone marrow stem cells (BMSCs) via agarose microwells, yielding uniform aggregates that matured into hyaline-like phenotypes over 30 days and demonstrated superior resilience to oxidative stress compared to monolayer BMSCs, as evidenced by reduced ROS accumulation, preserved proliferation, and modulated inflammatory/antioxidant pathways. MN was fabricated from gelatin methacryloyl (GelMA) in a bilayer architecture: stiff, discrete 10 %GelMA tips to facilitate penetration and promote osteogenesis, and a softer, monolithic 5 %GelMA base to enhance chondrogenesis. PpIX and COs were co-encapsulated for targeted delivery. <i>In vitro</i>, MN(+PpIX) under ultrasound induced robust ROS generation, apoptosis, and proliferation inhibition in giant cell tumor of bone (GCTB) cells via NF-κB modulation. <i>In vivo</i>, in a subcutaneous GCTB xenograft model, MN-mediated SDT significantly reduced tumor volume and enhanced apoptosis. In a rabbit knee OCD model, MN(+PpIX/CO) achieved superior integrated osteochondral repair, with improved bone volume fraction, trabecular thickness, Safranin O/H&E histology, and higher O'Driscoll/ICRS scores compared to controls. Biosafety was confirmed by normal major-organ histology and serum chemistry at 12 weeks. This platform innovatively sequences SDT and regeneration, using stiffness gradients to guide CO fate, offering a minimally invasive solution for tumor-associated OCDs.

This review presents recent advancements in cold atmospheric pressure (AP) plasma (CAP) processes for the synthesis and surface treatment of polymer films and metal nanoparticles (NPs) in biomedical applications. We discuss the properties and applications of atmospheric pressure plasma (APP) processes, including dielectric barrier discharge (DBD) and plasma jet methods, highlighting their effectiveness in controlling surface characteristics such as wettability and functionalization.

<h4>ABSTRACT</h4> Huntington’s disease (HD) is a fatal neurodegenerative disorder caused by a CAG repeat expansion in the Huntingtin ( HTT ) gene. Although transcriptomic and proteomic changes have been characterized in patient-derived neurons, the contribution of post-translational modifications (PTMs), such as phosphorylation, remains poorly understood. Here, we present the first phosphoproteomic analysis by mass spectrometry (P-MS) of human induced neurons (iNs) directly reprogrammed from HD patient fibroblasts. We identified 177 phosphopeptides with significantly altered abundance in HD-iNs, mapping to phosphoproteins associated with key signaling pathways known to be affected in HD, such as splicing and autophagy. By integrating P-MS data with previously published proteomic and transcriptomic data from the same donors, we identified distinct subsets of ON-OFF phosphopeptides that exhibited a complete loss of phosphorylation in either HD- or Ctrl-iNs, without corresponding changes at the RNA or protein level. An exception was MXRA8, previously described in glial cells as a mediator of blood-brain barrier integrity and astrocyte-mediated neuroinflammation. This protein showed increased protein abundance despite the absence of phosphorylation in HD- iNs, suggesting a compensatory mechanism - a pattern also observed in human post mortem cortical HD tissue. Additionally, MXRA8 showed altered protein-protein interactions with lysosomal and metabolic regulators in HD-iNs, highlighting its potential role in autophagy impairment as well as in neurovascular dysfunction. These findings uncover a distinct layer of post-translational dysregulation in HD, suggesting that phospho-switch proteins such as MXRA8 may be candidate effectors of pathology and thus site-specific phosphorylation loss may contribute to impaired signaling and proteostasis in human HD neurons.

Carbon-based nanomaterials (CBNs), including graphene oxide (GO), carbon nanospheres (CNSs), carbon nitrides (CNs), carbon nanotubes (CNTs), carbon dots (CDs), nanoporous carbon, and nanocomposites, possess exceptional thermal, mechanical, electrical, and optical properties with highly versatile surface chemistries. Their tunable size, shape, and surface functionalities facilitate strong π-π interactions and semiconductor-like behavior, enabling efficient light absorption and intimate biomolecular interfacing. These attributes have positioned CBNs as leading candidates in biomedical engineering, inspiring biomimetic designs that integrate organic and inorganic functions within unified architectures. This review highlights recent advances in the use of CBNs for next-generation biosensing and theranostics. Approaches such as physicochemical engineering, deoxyribonucleic acid (DNA) origami templating, peptide- or enzyme-assisted assembly, polysaccharide anchoring, and lipid modification have enhanced their biocompatibility, selectivity, and catalytic activity. Innovations in fluorescence switching, aptamer-CNT photophysics, and spacer-controlled energy transfer enable ultra-sensitive detection of metal ions, metabolites, neurotransmitters, pathogens, drugs, and cancer biomarkers. Furthermore, integrated CBN-based platforms, including field-effect transistors and laser-scribed graphene electrodes, demonstrate capabilities for single-virus or single-cell diagnostics and responsive therapeutic intervention. Finally, translational challenges related to scalable synthesis, biosafety, regulatory harmonization, and public acceptance are discussed, and interdisciplinary strategies combining flexible electronics, organ-on-chip models, and AI-guided design to advance clinical translation of CBN-enabled precision diagnostics and personalized theranostics are proposed.

In HFE-hemochromatosis, transferrin saturation (TSAT) indirectly reflects plasma non-transferrin-bound iron (NTBI), which drives iron overload and tissue damage. TSAT above ~75% suggests harmful NTBI levels. Despite its clinical relevance, NTBI remains understudied due to the limitations of current assays. High TSAT levels persist during maintenance therapy, yet some guidelines omit TSAT monitoring, though it correlates with symptoms like fatigue and joint pain. Monitoring TSAT may improve outcomes. Randomized trials comparing ferritin-only versus ferritin-plus-TSAT-guided treatment are needed. Consideration should be given to approaches that minimize plasma NTBI in managing and monitoring patients with hemochromatosis. Until reliable NTBI testing has been established, TSAT should be included in the management of HFE-hemochromatosis.

Centromeres have been the focus of extensive research for almost a century, so it may come as a surprise that a consistent definition and nomenclature for these structures remains elusive. In recent times, centromeric chromatin is most frequently defined by the presence of nucleosomes containing the H3 variant CENP-A and is typically synonymous with the site of the inner-kinetochore. However, crucial mammalian centromere proteins including CENP-B and INCENP have well defined distributions that show very little overlap with CENP-A. Additional protein localisations spanning the primary constriction or forming a band below CENP-A chromatin have been reported. Together, these observations suggest a complex and multi-layered chromatin organisation that is not well served by the canonical dichotomy of 'centromeric' and 'pericentromeric' chromatin. Strikingly, this is not a new observation but was made soon after the discovery of CENP proteins, including in a 1991 publication titled 'When is the centromere not a kinetochore?'. Here we revisit this question, which has become more pertinent following technical innovations in long-read sequencing and super-resolution microscopy. We present a model of centromere organisation for monocentromeres that incorporates additional complexity. We then use this model to reconceptualise diverse centromere forms in other eukaryotes including regional centromeres, holocentromeres and centromeres that lack key proteins including CENP-A. In this way, we hope to move towards a unified understanding of centromeric chromatin.

Ferroptosis, an iron-dependent form of regulated cell death, is increasingly leveraged in nanomedicine to sensitise tumours and overcome drug resistance. Driven by the Fenton reaction, ferroptosis results in lipid peroxidation through elevated intracellular iron levels and excessive production of reactive oxygen species (ROS). In this review, we outline the molecular markers of ferroptosis and define the criteria necessary to attribute ferroptosis induction to nanoparticles (NPs). We emphasise the importance of distinguishing targeted ferroptosis from non-specific ROS-mediated nanotoxicity and other types of programmed cell death. This distinction requires the use of lipophilic radical-trapping antioxidants (eg, ferrostatin-1, liproxstatin-1), iron chelators, and evidence implicating glutathione peroxidase 4 (GPX4) or the system Xc^- antiporter. Morphology is considered supportive but non-diagnostic, requiring converging evidence from both biochemical and genetic sources. We then compare various nanosystems designed to induce ferroptosis, such as iron-based nanoparticles, lipid nanocarriers, light-triggered nanoparticles, and magnetically induced nanocarriers, highlighting mechanistic patterns, efficacy determinants, and common pitfalls that often occur during biological investigations. Finally, we discuss translational challenges, including tumour microenvironment heterogeneity, NP protein corona dynamics, clearance and off-target effects. We aim to provide a framework that links NP design to ferroptotic mechanisms and clinically relevant outcomes, offering clear criteria and priorities for future research.

Pulmonary arterial hypertension (PAH) is a fatal disease with no cure. Until recently, most specific therapies for PAH had aimed at enhancing cyclic nucleotide (cAMP and cGMP) pathways, taking advantage of the vasorelaxant and antiproliferative properties of these key intracellular messengers. This process can be achieved by inhibiting phosphodiesterases (PDEs), which are intracellular enzymes responsible for cyclic nucleotide degradation. To date, only inhibitors of PDE type 5 (PDE5) have been approved for the treatment of PAH. Because the PDE superfamily comprises 11 families that encompass many variants, substantial experimental investigation has been conducted to assess the relevance of inhibiting other PDE families, aiming to offer therapeutic alternatives. This review synthesizes the main research work conducted on in vivo or ex vivo models, as well as on biological resources from patients. It helps provide evidence for the expression of PDE isoforms in the lung vasculature, as well as the efficacy and limitations of various pharmacological compounds tested for inhibiting pathological processes ongoing in the disease. Perspectives and suggestions for future research orientation are proposed.

Disulfidptosis is a novel Nicotinamide Adenine Dinucleotide Phosphate (NADPH) deficiency-driven cell death pathway characterized by cystine overload and aberrant disulfide bond formation in actin cytoskeletal proteins, distinct from apoptosis, ferroptosis, and other programmed cell death modalities. In gynecological tumors (ovarian, cervical, and endometrial cancers), this process is orchestrated by dysregulated SLC7A11 expression, impaired thioredoxin system function, and Rac-WRC-Arp2/3-mediated actin network collapse. Bioinformatic analyses of The Cancer Genome Atlas (TCGA)/Gene Expression Omnibus (GEO) datasets have revealed that disulfidptosis-related genes (e.g., SLC7A11, GYS1, NCKAP1) and lncRNAs (e.g., PRDX6-AS1, EMSLR) correlate with patient prognosis, chemoresistance, and tumor immune microenvironment (TME) remodeling. Therapeutic strategies to induce disulfidptosis include glucose deprivation to limit NADPH supply, inhibition of NADPH-generating enzymes (e.g., G6PD inhibition), and nanodelivery systems (e.g., FeOOH@Fe-Ap@Au) that synchronize disulfidptosis with ferroptosis. Preliminary evidence proposes that disulfidptosis inducers may synergize with immune checkpoint inhibitors (ICIs) through TME modulation, though experimental validation remains ongoing. Beyond malignancies, disulfidptosis-related pathways have been implicated in endometriosis, where disulfidptosis-related genes (DRGs; e.g., PDLIM1, ACTB) regulate ectopic lesion progression via immune-metabolic crosstalk. This review comprehensively summarizes the molecular mechanisms, disease associations, and translational potential of disulfidptosis in gynecological disorders, proposing targeted therapeutic paradigms and future research directions.

The oxidative modification of proteins induced by hydrogen peroxide (H₂O₂) results in the formation of disulfide bond between two cysteines and affects protein conformation and biological function. Transcription factor FOXM1 participates in the development and progression of cancers and its levels are upregulated by the oxidative stress of H₂O₂-treated condition. In this study, we found that Peroxiredoxin-1 (PRDX1), one of the most H₂O₂-reactive antioxidant enzymes, interacted with FOXM1 and led to its oxidation under H₂O₂ stimulation through generating an intermolecular disulfide bond with FOXM1 C539, which was subsequently transferred to form an intramolecular disulfide bond between C167 and C175 in the oxidized FOXM1. The PRDX1-mediated oxidative modification enhanced the protein stability and transcriptional activity of FOXM1, which stimulated the transcription of FOXM1 target gene X-ray cross-complementing protein 1 (XRCC1) and improved the repair of H₂O₂-induced DNA damage in cancer cells. The disruption of PRDX1-mediated FOXM1 oxidation impaired the colony formation ability of cancer cells <i>in vitro</i> and the growth and DNA damage repair ability of cancer cells <i>in vivo</i>. The analysis of The Cancer Genome Atlas (TCGA) breast cancer patient data confirmed that PRDX1 and FOXM1 together facilitated clinical cancer progression. Overall, we established an H₂O₂-PRDX1-FOXM1 oxidation pathway that likely contribute to the development and progression of cancers.

Brain arteriovenous malformations (bAVMs) consist of a tangled nidus of abnormal dilated vessels characterized by direct connections between arteries and veins that lack an intervening capillary bed, creating a high-to-low flow pressure system that is predisposed to spontaneous hemorrhage with significant associated neurologic morbidity and mortality. Treatment options for bAVMs include the following: surgical resection, intravascular embolization to obliterate blood flow through the AVM, and radiosurgery. Understanding the molecular mechanisms of bAVM formation and factors that predispose it to hemorrhage can lead to novel treatments that can improve the prognosis for patients. This review summarizes emerging insights into the complex and dynamic molecular mechanisms of bAVMs. Dysregulation in key VEGF, TGF-β/BMP9/10-ENG-ALK1-SMAD4, Notch, and MAPK/ERK signaling pathways drive abnormal angiogenesis in both syndromic and sporadic forms, with KRAS/BRAF/MAPK21 mutations specifically linked to the latter. Advances in bAVM-induced animal models have corroborated many of the genetic profiles found in humans, and they continue to provide novel insights into bAVM mechanisms. Collectively, these mechanistic findings are guiding translational advances, with targeted therapies and liquid biopsy approaches emerging as avenues for precision treatment and improved patient outcomes.

Endoscopic injection of a cyanoacrylate-lipiodol mixture is widely regarded as the first-line treatment for gastrointestinal bleeding secondary to ruptured gastric varices. Although generally effective, this technique carries the risk of rare but potentially life-threatening complications, including pulmonary glue embolism and interstitial pneumonitis. We present the case of a 65-year-old woman with cirrhosis who presented with hematemesis and melena and underwent endoscopic cyanoacrylate-lipiodol injection for bleeding gastroesophageal varices. Within 24 hours, she developed a persistent fever and bilateral crackles. Chest X-ray, CT imaging, and bronchoscopy findings were consistent with glue-induced pneumonitis. Corticosteroid therapy resulted in rapid clinical and radiological improvement. This case highlights the importance of recognising pulmonary complications following cyanoacrylate injection for gastric varices and underscores the need for prompt diagnosis and appropriate management to mitigate morbidity.

Phosphatase and tensin homolog (PTEN) is a critical inhibitor of the PI3K/AKT signaling pathway, yet its direct upstream regulators remain poorly defined. In this study, we investigated the role of peroxiredoxin 4 (PRDX4) in alveolar macrophages (AMs) activation and pulmonary fibrosis. Analyses of lung tissues from silicosis patients by transcriptomic and histological analyses revealed that PRDX4 is selectively upregulated in AMs and positively correlated with profibrotic and inflammatory gene expression. Consistent results were observed in silicosis model mice, where PRDX4 expression co-localized with the macrophage marker F4/80 and correlated with fibrotic indicators. Functional studies demonstrated that macrophage-specific silencing of PRDX4 using adeno-associated virus improved lung function and reduced inflammatory infiltration and fibrosis. PRDX4 upregulation aberrantly activated AMs and promoted epithelial-mesenchymal transition and fibroblast-myofibroblast transition. Mechanistically, PRDX4 enhanced AKT/NF-κB signaling with minimal effects on PI3K. Biochemical interaction assays further demonstrated that oligomeric PRDX4 disrupted PTEN homodimer formation, with mutational analyses identifying Cys124 and Cys245 as essential residues. Notably, Conoidin A alleviated crystalline silica-induced fibrosis in mice, with its therapeutic effect likely mediated by disrupting PRDX4 oligomerization. These findings identify PRDX4 as a novel upstream regulator of PTEN, establish a mechanistic PRDX4-PTEN axis in macrophage activation, and highlight PRDX4 as a promising therapeutic target for idiopathic pulmonary fibrosis and silicosis-associated fibrosis.

The norbornadiene/quadricyclane (NBD/QC) photoswitch is a promising candidate for molecular solar energy storage. Isomerization to the energy-storing, metastable QC occurs upon UV light irradiation, while the back-switching can be triggered in various ways. However, inducing complete, on-demand energy release across multiple switching cycles remains a significant challenge in molecular solar thermal (MOST) research. We demonstrate the implementation of perylene diimide (PDI) as covalently connected, photoactive redox-catalyst to enable NBD regeneration upon 475 nm irradiation. We investigated the NBD/QC interconversion of several imide- and ortho-connected NBD-PDI hybrids upon irradiation at 310 nm and 475 nm, revealing that the connection position on PDI only minimally affects the interconversion efficiency. Concentration-dependent studies identified an alternative intermolecular back-isomerization mechanism in addition to the previously known intramolecular pathway, for which adequate flexibility of the linker proved crucial for effective back-isomerization to NBD. This study advances the exclusively photoinduced isomerization of the NBD/QC system within novel NBD-PDI dyads, establishing a framework for the targeted manipulation of NBD hybrids with potential application in energy storage. The presented switches operate independently, without the need for further additives to facilitate interconversion, which renders them promising candidates for molecular solar energy storage research.

<h4>Objective</h4>Conditions presenting with both epilepsy and movement disorders (EPIMDs) range from relatively benign cases to severe developmental encephalopathies. However, the full clinical and genetic spectrum still needs to be better defined. The aim of this study is to describe the presentation of EPIMDs in pediatric patients with known genetic etiologies, correlating these features with age at onset and underlying pathological mechanisms to identify patterns that could improve patient management.<h4>Methods</h4>We retrospectively analyzed the clinical records of pediatric patients who underwent genetic testing for EPIMDs at our institution between 2009 and 2022. Genetic testing included single-gene Sanger sequencing, multigene next-generation sequencing panels targeting epilepsy and/or MDs, and array comparative genome hybridization. Demographic, clinical, and electroencephalography (EEG) data were collected. Statistical analyses were conducted using multivariate and cluster analyses.<h4>Results</h4>A total of 97 subjects were included. The mean age ± SD at epilepsy onset was 3.6 ± 4.4 years, whereas the mean age at MD onset was 5.2 ± 4.9 years. Based on the mechanism of genetic dysfunction, we identified six groups: transportopathies (n = 18), synaptopathies (n = 11), channelopathies (n = 18), metabolic disorders (n = 15), intracellular trafficking defects (n = 8), and unknown/complex function disorders (n = 27). Cluster analysis identified three distinct groups: (1) early-onset epilepsy and paroxysmal MDs with normal intelligence quotient (IQ) and mild intellectual disability, associated with transportopathies (n = 38); (2) early-onset epilepsy with dystonia and myoclonus, low IQ, and developmental and epileptic encephalopathy, associated with channelopathies (n = 31); (3) epilepsy onset between 1 and 5 years of age with developmental delay, parkinsonism, and low IQ, associated with metabolic etiology (n = 26).<h4>Significance</h4>Our study demonstrates that, despite the phenotypic and genetic pleiotropy observed in EPIMDs, a precise characterization of semiological and cognitive features remains essential to support variant interpretation and to refine the diagnostic process and patient management in these rare conditions.

<h4>Background</h4>In patients with metabolic dysfunction-associated steatotic liver disease (MASLD), non-Gaussian diffusion-weighted imaging (DWI) has been proposed for the diagnosis of liver fibrosis, but its measurement is partially confounded by steatosis. We therefore asked whether a fat-corrected approach could improve fibrosis assessment.<h4>Purpose</h4>To evaluate the diagnostic performance of non-Gaussian diffusion coefficients for the assessment of fibrosis in MASLD patients with a method accounting for intravoxel fat.<h4>Study type</h4>Prospective single-center cross-sectional study.<h4>Population</h4>A total of 289 participants with Type 2 diabetes, hepatic steatosis, and elevated aminotransferases were enrolled from October 2018 to June 2021. Among them, 222 participants (mean age 59 ± 10 years; 149 men) underwent liver biopsy and MRI and were included in the final analysis.<h4>Field strength/sequence</h4>3 T, DWI using spin-echo echo-planar imaging, MR elastography (MRE) using gradient echo sequence and fat fraction imaging using a multiple gradient echoes sequence.<h4>Assessment</h4>Diffusion coefficients were estimated using two non-Gaussian models: a shifted apparent diffusion coefficient (sADC) and a non-linear least squares fit (ngADC), both computed without and with intravoxel fat correction (corr) using fat fraction on PDFF. Fibrosis was staged histologically. Quantitative parameters were compared across fibrosis stages. Diagnostic performance for F0 versus ≥ F1 was evaluated and compared to liver stiffness on MRE.<h4>Statistical tests</h4>Group comparisons used Kruskal-Wallis tests (α = 0.05), and diagnostic performance was assessed via receiver operating characteristic (ROC) curve analysis with 95% confidence intervals, with p < 0.05 considered statistically significant.<h4>Results</h4>ngADC_corr was significantly different between fibrosis stages (Kruskal-Wallis p < 0.05). ROC curve analysis indicated comparable performance in discriminating fibrosis stages F0 versus F1-F4 for ngADC_corr and stiffness (AUC = 0.66, 95% CI: [0.59, 0.7], p < 0.05 and 0.68 [0.62, 0.74], p < 0.05, respectively).<h4>Data conclusion</h4>Fit-based non-Gaussian DWI with fat correction could potentially be used with similar diagnostic accuracy as MRE for detecting fibrosis in patients with MASLD.<h4>Evidence level</h4>3.<h4>Technical efficacy</h4>Stage 2.

Thyroid cancer remains one of the most prevalent cancers worldwide, with lymph node metastasis (LNM) playing a critical role in determining patient prognosis and treatment. MicroRNAs (miRNAs) are essential post-transcriptional regulators of gene expression and have been implicated in cancer progression, particularly in epithelial-mesenchymal transition (EMT). This study aimed to identify miRNAs involved in the lymph node metastatic process in thyroid cancer and to evaluate their potential as therapeutic targets. RNA-seq datasets from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) were used to identify dysregulated miRNAs in LNM from papillary and medullary thyroid carcinoma (PTC and MTC). The candidate miRNAs were transfected in thyroid carcinoma cell lines, TPC-1 and BCPAP, followed by quantification of target genes and analysis of cell migration and invasion assays in vitro. Three miRNAs miR-199a-5p, miR-199a-3p, and miR-148a-5p were identified as downregulated in thyroid cancer with LNM. Transfection of TPC-1 and BCPAP cells with mimetics reduced cell migration and invasion. Additionally, these miRNAs modulated the expression of EMT-related genes, particularly in BCPAP cells. Our results indicate that miR-199a-3p, miR-199a-5p, and miR-148a-5p are involved in key metastatic processes, suggesting their relevance as potential biomarkers and/or therapeutic targets for thyroid tumors. Further in vivo validation and clinical studies are required to explore their translational applications.

Dietary selenium (Se) deficiency elevates the risk of cardiometabolic disorders driven by excessive caloric dietary intake. A high-fat diet leads to redox imbalance and elevates lipid deposits in the heart regardless of sex. Selenium maintains redox homeostasis by regulating the levels of antioxidant selenoproteins, such as glutathione peroxidases 1 and 4 (GPX1,4) and thioredoxin reductases 1 and 2. Se-dependent tissues such as the heart have a recycling mechanism to maintain selenoprotein synthesis, especially when Se is limiting, carried out by the enzyme selenocysteine lyase (SCLY). Despite being highly reliant on Se to function properly, it remains unclear if SCLY is required to modulate Se and selenoprotein synthesis in the heart, particularly in obesity. In this study, we analyzed cardiomyocyte (CM) function and molecular modifications in the hearts of male and female whole body <i>Scly</i> knockout (<i>Scly</i> KO) mice fed a Se-deficient, obesogenic diet. We found sex-specific differences demonstrating that the loss of SCLY activity is more detrimental to the hearts of female mice on an obesogenic diet than in male mice, due to reduced glutathione (GSH) peroxidase 1 and 4 expression and decreased circulating Se levels. In addition, overall GPX activity and total GSH levels in <i>Scly</i> KO female hearts resulted in significantly reduced CM contractility. In summary, the loss of <i>Scly</i>, coupled with Se deficiency and a high-fat diet reduces the ability of the GSH system to mitigate oxidative stress in female hearts, increasing redox imbalances that favor the establishment of cardiometabolic disorders.<b>NEW & NOTEWORTHY</b> Global loss of selenocysteine lyase controls the glutathione system in the heart, impacting glutathione peroxidase protein levels and activity, and glutathione levels in a mouse model of obesity. Loss of selenocysteine lyase and selenomethionine deficiency impacts cardiomyocyte function and selenoproteins in a sexually dimorphic nature, with females being affected more than males when mice are on a high-fat diet.

This study aimed to establish the correlation between activated partial thromboplastin time (aPTT) in seconds and ratio, first derivative peak (Max1), second derivative peak (Max2), second derivative minimum peak (Min2), and delta-OD of clot waveform analysis in patients diagnosed with cirrhosis, sepsis, and a control group. The study involved a series of cases using the Bayesian ANOVA model with Bayes factor (BF) estimated using the Markov Chain Monte Carlo sampling method with 2 chains. Each chain consisted of 10 000 repetitions and started 5 000 times to predict the variables aPTT (s), aPTT (ratio), Max1, Max2, Min2, and delta-OD respectively across the 3 study groups. The model used was variable ~ stage - 1. This analysis illustrates the adjustment of the posterior proportion for multiple testing by setting the prior probability of the initial hypothesis to 0.5 for all comparisons. There is insufficient evidence that aPTT (s), aPTT (ratio), and Min2 differ between the study groups (0 < BF < 3). There is compelling evidence for the Max1 value in both the sepsis group and the control group (BF10 = 44.042). Additionally, between the sepsis group and the cirrhosis group, there is substantial evidence for the Max1 value (BF10 = 5.842). Moreover, there is convincing evidence for the Max2 value in the sepsis group compared to the control group (BF10 = 38.782). There is almost confirmatory evidence that the delta-OD value in the sepsis group is higher than that in the control group (BF10 = 1251.781). Finally, there is convincing evidence that the sepsis group has a higher delta-OD value than the cirrhosis group (BF10 = 35.555). The Max1, Max2, and delta-OD indices effectively reflect coagulation disorders in patients with cirrhosis and sepsis, even when the aPTT test results are within normal limits.

β-Acyloxyl alkenyl amides (AAAs) are vinylogous esters with high activity toward acylation reactions. However, only one synthetic procedure consisting of five independent operation steps involving highly sensitive amino alkynone intermediate is available for their synthesis to date, depressing rigidly the application of AAAs in peptide synthesis and expanded acylation reactions. Herein, by combining consecutive trifold defluorination, acyl migration, and multicomponent reactions, a considerably simpler one-step protocol has been established for AAA synthesis with the α-trifluoromethyl ketones, amines, and carboxylic acids/amino acids as starting materials using Cs₂CO₃ as the only promoter. The resulting AAAs have been verified with excellent performance in the synthesis of esters, amides, and peptides. In all investigated examples of peptide synthesis, no racemization or epimerization has been observed, documenting the high application potential of AAAs as a complementary option in peptide synthesis and other related acylation reactions.

Understanding how genetic and epigenetic factors influence predisposition, susceptibility, protection, severity, and recovery potential in sports-related muscle, bone, and brain injuries is gaining increasing attention, as these injuries represent a major concern for athletes' health and performance. This narrative review explores the role of specific genetic variants, such as those in <i>ACE, ACTN3, AMPD1, VDR,</i> and <i>APOE</i>, in predisposing athletes to injury. In parallel, the emerging role of microRNAs, including miR-21, miR-133a, miR-208a, and miR-214 in regulating gene expression related to muscle regeneration, inflammation, and neuroprotection, is highlighted. These small noncoding RNAs are influenced by environmental stimuli such as sports activity and training, and may serve as promising biomarkers for injury risk and recovery. This review is based on a narrative approach, involving targeted literature searches in databases such as PubMed, Scopus, and Web of Science, with no restrictions on language or publication date. Articles were selected based on relevance using predefined keywords and were categorized by injury type (muscular, bone, and concussion-related). However, several limitations remain, including population heterogeneity, small sample sizes, and the lack of functional validation of identified variants, which currently limits their clinical application. Ethical considerations surrounding the use of genetic data in athletes also need to be addressed. In the future, better integration of multiomic data could support the development of personalized and ethically sound strategies for injury prevention and athlete care.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and play critical roles in various cellular processes. Increasing evidence suggests that miRNAs are involved in the development and progression of atherosclerosis, which is the leading cause of myocardial infarction and stroke. These molecules influence key pathological mechanisms, including lipid metabolism, endothelial dysfunction, vascular inflammation, and plaque stability. This review summarizes the role of miRNAs in atherosclerosis-induced cardiac and cerebral infarction and explores their potential as therapeutic targets. We discuss emerging miRNA-based interventions, such as miRNA mimics and inhibitors, which offer promising strategies for disease prevention and treatment. Understanding the regulatory functions of miRNAs in cardiovascular and cerebrovascular events may provide new insights for developing innovative therapies aimed at reducing the burden of atherosclerosis-related diseases.

We investigated the relationship between incidence of gastrointestinal tract cancers, metabolic dysfunction-associated steatotic liver disease (MASLD), and alcohol-related steatotic liver disease in diabetic population. A nationwide cohort of 2,616,828 individuals with diabetes under Korean National Health Insurance Service from 2015 to 2016 was divided into four subgroups: no steatosis (group 1), MASLD alone (group 2), MASLD with heavy alcohol intake (group 3), and alcoholic liver disease (group 4). We used fatty liver index to assess the probability of hepatic steatosis using cutoff scores of 30 and 60. We analyzed incidences of esophageal, stomach, colorectal, biliary, and pancreatic cancers until 2022. Compared with group 1 (reference), group 2 showed increased hazard ratios for stomach, colorectal, and biliary cancers, with a decreased hazard ratio for esophageal cancer (adjusted hazard ratio [95% confidence interval]: 1.10 [1.06-1.13], 1.13 [1.10-1.16], 1.10 [1.05-1.16], 0.88 [0.79-0.97], respectively). Probability of hepatic steatosis was positively correlated with all gastrointestinal tract cancers except esophageal cancer in non-drinkers, but only with stomach, colorectal, and biliary cancers in mild drinkers (p_trend < 0.001). In conclusion, MASLD increases gastrointestinal tract cancer risk, except esophageal cancer, in diabetic population. For non or mild drinkers, probability of hepatic steatosis serves as a predictor of gastrointestinal tract cancer risk.

This study investigated the impact of intermittent fasting (IF) on older rats by administering plasma from 12-month-old male Sprague Dawley rats subjected to 35 days of IF, with 18-hour fasting intervals followed by 6-hour feeding periods. Over this period, 0.5 ml of plasma was transferred bi-daily to 24-month-old male rats, totaling 15 infusions. Infrared spectroscopy-based qualitative and quantitative analyses revealed significant changes in lipid, protein, and nucleic acid profiles, varying by tissue type and fasting regimen. Histological examination showed structural enhancements in the ileum and colon, along with reduced inflammatory markers TNF-α and COX-2, particularly with IF plasma. Additionally, plasma transfer markedly improved gut microbiota composition, increasing alpha diversity and adjusting the Firmicutes to Bacteroidetes ratio. High-Performance Liquid Chromatography results indicated elevated levels of key cecal short-chain fatty acids, suggesting enhanced gut health. These findings highlight the potential of plasma from intermittently fasting rats in modulating biomolecular profiles, intestinal tissue structure, gut microbiota, and SCFA production, offering integrated insights into intestinal health improvements.

This study aimed to characterize transcriptomic alterations and the effects of demographic and behavioral risk factors on disease-associated gene expression patterns in late-stage keratoconus (KC). Corneal stromal tissues from 31 KC patients and four normal donors underwent bulk RNA sequencing. Differentially expressed genes (DEGs) and weighted gene co-expression network analysis (WGCNA) were used to identify key molecular changes. Subgroup analyses based on sex, allergy status, eye rubbing intensity (ERI), body mass index (BMI), and childhood socioeconomic status (SES) were conducted using gene set enrichment analysis (GSEA) and WGCNA. Key genes from WGCNA modules were validated by quantitative real-time PCR (qRT-PCR) and three public transcriptomic datasets. Enrichment analyses based on the 4469 identified DEGs (Padj < 0.05) revealed suppression of extracellular matrix (ECM) organization, axon guidance, synaptic signaling, and immune activity (Padj < 0.05). Subgroup GSEA demonstrated that high ERI correlated with ECM pathways, elevated BMI correlated with mitochondrial dysfunction, and low childhood SES correlated with upregulated mitochondrial metabolism and immune activation (Padj < 0.05). Chromatin regulation and antioxidant activity were enriched in females (Padj < 0.05). No transcriptomic associations were detected for allergy status. WGCNA revealed ERI-related modules involving Wnt and PI3K-Akt signaling and SES-related modules implicating cholinergic synapse and axon guidance pathways (Padj < 0.05). Validation using qRT-PCR and public datasets confirmed the downregulation of all 15 selected genes in KC (Padj < 0.05). qRT-PCR also confirmed differential expression of five subgroup-associated genes across KC subgroups (P < 0.05). These findings emphasize the neural-immune-stromal axis dysregulation in KC and elucidate the contribution of common risk factors to disease progression.

INTRODUCTION: Cirrhosis is a major cause of morbidity and mortality worldwide [1]. While serum ferritin is recognized as a marker of liver injury and inflammation, its prognostic value for cirrhosis-related complications is not well established. We investigated whether serum ferritin levels could predict complications in cirrhotic patients. MATERIALS AND METHODS: In this retrospective observational cohort study, which included both descriptive and analytical components, serum ferritin levels were measured in adult cirrhotic patients admitted to the gastroenterology department of a University Hospital Center between March 2017 and March 2024. The normal reference range was defined as 22–275 ng/mL for men and 4.63–204 ng/mL for women. Patients were monitored for the occurrence of complications over a one-year period. Statistical analyses were performed to assess the association between serum ferritin levels and complications, and to determine whether elevated ferritin was an independent predictor of adverse outcomes. RESULTS: The study included 200 cirrhotic patients (111 females and 89 males) with a mean age of 58,06 ± 15.21 years (range: 19–95). The most common etiologies were metabolic-associated steatohepatitis (MASH) (49.7%) and viral hepatitis (19%). Serum ferritin levels were significantly higher in decompensated patients compared to compensated patients (173 ± 295.22 ng/mL vs. 121.32 ± 251.98 ng/mL). During one-year follow-up, 68.5% of patients with elevated ferritin levels developed complications, compared to 27.7% of those with normal ferritin levels. Serum ferritin levels were significantly higher in Child-Pugh B patients (424.9 ng/mL) compared to Child-Pugh A (74.8 ng/mL, p = 0.002). Receiver operating characteristic (ROC) curve analysis demonstrated that serum ferritin has moderate discriminative power in predicting the occurrence of complications in cirrhotic patients (AUC = 0.670). Elevated ferritin (≥ 45 ng/mL) was associated with a significantly increased risk of complications (p < 0.001). Multivariate analysis revealed the following predictors of hepatic decompensation: elevated serum ferritin (aOR 5.76, 95% CI 2.08–15.90, p = 0.001), hyponatremia (aOR 0.39, 95% CI 0.18–0.85, p = 0.017), and vitamin D deficiency (aOR 6.52, 95% CI 1.86–22.92, p = 0.003). These findings highlight the complex and multifactorial nature of hepatic decompensation and support the inclusion of serum ferritin in a comprehensive prognostic assessment. CONCLUSION: Elevated serum ferritin levels are independently associated with an increased risk of complications in cirrhotic patients. Given its wide availability and low cost, serum ferritin may serve as a simple prognostic biomarker to guide risk assessment and optimize clinical management in this population.

PURPOSE: To evaluate the occurrence of uremic pruritus among peritoneal dialysis (PD) patients and explore its correlation with serum iron levels. METHODS: A cross-sectional study was carried out on 189 PD patients at a single center in China. The severity of pruritus was assessed through the validated 5-D Itch Scale questionnaire. The relationship between clinical factors and pruritus was examined using multivariate logistic regression, restricted cubic spline analysis, piecewise regression, and cluster analysis of iron metabolism patterns. RESULTS: Moderate/severe pruritus affected 27.5% of participants, mainly on lower limbs (52.9%) and back (46.6%). Serum iron was significantly higher in the moderate/severe pruritus group (18.43 ± 8.50 vs. 15.84 ± 6.09 µmol/L, p = 0.029). After adjustments, each standard deviation increase in serum iron corresponded to higher odds of moderate/severe pruritus (odds ratio (OR) = 1.45, 95% confidence interval (CI):1.03–2.08, p = 0.037). Analysis revealed a threshold association(p = 0.043) with an inflection point at 13.7µmol/L, above which each unit increase in serum iron raised pruritus odds by 10% (OR = 1.10, 95%CI:1.03–1.18, p = 0.006). Cluster analysis identified that patients with high transferrin saturation (TSAT), moderate ferritin, and high serum iron had the highest pruritus prevalence (52.8%). CONCLUSION: In PD patients, we identified a threshold effect of serum iron on uremic pruritus. Iron levels above 13.7 µmol/L independently worsened pruritus, especially in patients with high TSAT, moderate ferritin, and high serum iron, where prevalence reached 52.8%. These findings highlight the importance of assessing iron status and personalized management.