Gene Literature Dashboard

Stigma is defined as the perception of an undesirable attribute that leads to discrimination against individuals and groups. Stigmatisation is often triggered due to visible physical or cognitive differences. Although the literature consistently highlights the (fear of) stigmatisation as a significant concern among individuals living with hereditary conditions, no studies in Portugal have specifically provided evidence on this issue. This study aims to address this gap by examining the experiences and impact of stigma on individuals and families affected by hereditary diseases in Portugal. After receiving ethics approval, a total of 216 participants, including affected individuals, asymptomatic carriers and family members from families with a range of hereditary conditions, were recruited through patient support associations. Participants completed an online questionnaire via Limesurvey. Data were analysed through Exploratory Factor Analysis (EFA), median comparison tests, and thematic analysis. Of the participants, 78.7% were women, 55.6% had a university degree, and 20.4% were aged between 42 and 47 years. Findings indicate that stigma impacts individuals across various domains, including social interactions, institutional settings, the workplace, and healthcare. EFA identified a bi-factorial model of stigma, comprising Stigma Experiences and Perceived Support subscales, and the overall scale demonstrated high internal consistency (α = .879). Women and younger participants reported higher levels of stigma. Religiosity and humor emerged as key coping strategies. This study is the first in Portugal to assess stigma among individuals living with hereditary conditions. Our findings contributed to validating a measurement instrument, identified sociodemographic variations, and examined the psychosocial dimensions of stigma among affected patients. These findings highlight the need for comprehensive strategies to address and mitigate stigma, improve support systems, and enhance the well-being and healthcare experiences of individuals and families impacted by hereditary diseases.

Cancer-associated fibroblasts (CAFs) play a crucial role in the progression of pancreatic ductal adenocarcinoma (PDAC). Here, integrated single-cell RNA sequencing analysis is utilized to comprehensively map CAFs in the human PDAC tumor microenvironment (TME). Normal fibroblasts (NFs) and nine distinct CAF subtypes are identified including newly identified CAF subtypes, CDCP1⁺FTL⁺ CAFs, transitional CAFs (tCAFs), interferon simulated genes (ISG)⁺ myofibroblastic CAFs (myCAFs), and proliferative CAFs (pCAFs). CDCP1⁺FTL⁺ CAFs, pCAFs, and ISG⁺ myCAFs are associated with unfavorable clinical outcomes. CDCP1⁺FTL⁺ CAFs exhibit enhanced glycolysis and iron metabolism, resisting ferroptosis. The antigen-presenting CAFs (apCAFs) show high heterogeneity, consisting of multiple subtypes expressing distinct immune cell signatures. The CAF subtypes display differentiation plasticity, transitioning from early normal-like CAFs (nCAFs) to inflammatory CAFs (iCAFs) and myCAFs, ultimately leading to more invasive pCAFs. AP-1 family members FOS and JUN regulate the malignant phenotype conversion of NFs to nCAFs, while transforming growth factor-β (TGFβ) and interferon-γ (IFNγ) signals trigger the interconversion between classic myCAFs and iCAFs, respectively. A close interaction between CAFs and myeloid cells (especially neutrophils) is further observed in PDAC-TME, mainly mediated by CXCR4-CXCL12 chemotaxis. This work depicts a detailed CAF map and its dynamic interconvertible shift, providing important insights for combined targeted CAFs therapy.

<h4>Background</h4>Hepatocellular carcinoma (HCC), the most common form of liver cancer, presents a challenging malignancy with scarce treatment options. Liver progenitor cells (LPCs) play a pivotal role in both liver regeneration and the progression of liver cancer, yet the specific functions of LPCs from different origins in liver cancer remain to be fully elucidated.<h4>Methods</h4>We explored the liver progenitor-like cells derived from human hepatocytes (HepLPCs) on the proliferation of HCC both in vitro and in vivo. The mitochondrial function was assessed through electron microscopy and functional experiments. Transcriptomic sequencing and western blot unveiled the fundamental mechanisms at play, whereas metabolomic sequencing pinpointed crucial effector molecules involved in the paracrine secretion of HepLPCs.<h4>Results</h4>By employing a co-culture system of HepLPCs and HCC cells, we found that HepLPCs markedly inhibited HCC growth by prompting mitochondrial dysfunction, which further led to the co-inhibition of the Notch1 and JAK1/STAT3 signaling pathways through paracrine actions involving S-adenosylmethionine (SAM) and Nicotinic acid (NA).<h4>Conclusions</h4>This study has uncovered that HepLPCs have a suppressive influence on the proliferation of HCC cells. This is achieved through the impairment of mitochondrial function and the inhibition of key signaling pathways, namely, Notch1 and JAK1/STAT3, which are critical drivers of cancer progression. The secretion of the metabolites SAM and NA by HepLPCs appears to be instrumental in mediating these effects. These findings provide a solid foundation for identifying new therapeutic targets and clarifying the mechanisms through which HepLPCs can be harnessed to effectively treat HCC.

Cerebellar information processing is critically shaped by several types of inhibitory interneurons forming various intra-cerebellar feed-forward and feed-back loops. Evidence gathered over the past decades has focused interest on a non-uniform set of cortical inhibitory interneurons distinct from "classical" Golgi, basket or stellate cells, summarily referred to as PLIs (for Purkinje cell layer interneurons). Similarly, cerebellar nuclear inhibitory interneurons have gained increasing attention. Our understanding of the functions of these cells is still fragmentary. For humans, we lack functional data, and even any dependable morphological classification for these cells. Here, I used publicly available single cell based gene expression data to compare inhibitory interneurons from the cerebellar cortex and inhibitory nuclear neurons of humans and mice. Integration of nuclear and cortical cells revealed transcriptomic similarities between subsets of these cells and suggest known characteristics of cortical cell types may be helpful to devise strategies for the further characterization of nuclear inhibitory interneurons. Comparison of human and murine PLIs indicate that these strongly differ by the expression of genes used to characterize these cells in mice. This limits their utility to identify and classify human PLIs, and leaves the question open as to the number and characteristics of non-Golgi inhibitory interneurons resident in the cerebellar granule cell and Purkinje cell layers in humans.

<h4>Purpose</h4>Colorectal cancer (CRC) is the third most common cancer globally, necessitating novel biomarkers for early diagnosis and treatment. This study proposes an efficient pipeline leveraging an integrated bioinformatics and machine learning framework to enhance the identification of diagnostic and prognostic biomarkers for CRC.<h4>Methods</h4>A selection of methylated differentially expressed genes (MeDEGs) and features (genes) was made using both statistical and Machine learning (ML) approaches from publically available datasets. These genes were subjected to STRING network construction and hub genes estimation, separately. Also, essential miRNAs (micro-RNAs) and TFs (Transcription factors) as regulatory elements were revealed and findings were validated through scRNA-seq analysis, promoter methylation, gene expression levels correlated with pathological stage, and interaction with tumor-infiltrating immune cells.<h4>Results</h4>Through an integrated analysis pipeline, we identified 27 hub genes, among which CTNNB1, GSK3B, IL-1β, MYC, PXDN, TP53, EGFR, SRC, COL1A1, and TGBF1 showed better diagnostic behaviour. Machine learning approach includes the development of K-Nearest Neighbors (KNN), Artificial Neural Networks (ANN), and Random Forest (RF) models using TCGA datasets, achieving an accuracy range between 99 and 100%. The Area Under the Curve (AUC) value for each model is 1.00, signifying good classification performance. The high expression of some diagnostic genes was associated with poor prognosis, concluding IL-1β as both a prognostic and diagnostic biomarker. Additionally, the NF-κB and microRNAs (miR-548d-3p, miR-548-ac) and TFs (NFκB and STAT5A) play a major role in the comprehensive regulatory network for CRC. Furthermore, hub genes such as IL-1β, TGFB1, and COL1A1 were significantly correlated with immune infiltrates, suggesting their potential role in CRC progression.<h4>Conclusion</h4>Overall, the elevated expression of IL-1β coupled with abnormal DNA methylation, and its consequent effect on the PI3K/Akt signaling pathway are relevant prognostic and therapeutic marker in CRC. Additional molecular candidates reveal insights into the epigenetic regulatory targets of CRC and their association with immune cell infiltration.

This study investigated whether octacalcium phosphate (OCP) enhances bone regeneration through its synergistic effect with stromal-derived factor-1 (SDF-1). Recombinant SDF-1 (0.5-5.0 μg) was combined with OCP granules through lyophilization. OCP/SDF-1 granules were implanted into a rat femoral standardized defect for 2 and 4 weeks and subjected to histomorphometry, C-X-C motif chemokine receptor 4 (CXCR4) and osteocalcin immunohistomorphometry, and tartrate-resistant acid phosphatase (TRAP) staining. Calcium-deficient hydroxyapatite (CDHA) was used as a control for in vitro analyses. Mesenchymal stem cell (MSC) migration was estimated using a Transwell system with OCP/SDF-1. SDF-1 release from OCP/SDF-1 into the supernatant was determined without cells. SDF-1 adsorption in 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer onto OCP, the chemical structure of OCP immersed in the medium using Fourier transform infrared spectroscopy, and the degree of supersaturation of the medium were determined. Bone regeneration and OCP degradation were enhanced the most by 1.0 μg of OCP/SDF-1 at 2 weeks after implantation by CT analysis and increasing CXCR4-positive, osteocalcin-positive, and TRAP-positive cells accumulation around the OCP. MSC migration increased until 48 h in the following order: SDF-1 only, CDHA/SDF-1, and OCP/SDF-1, with the greatest effect with 1.0 μg of SDF-1 than from OCP. CDHA promoted a greater release than OCP at 48 h. The physicochemical analyses indicated that SDF-1 interacted with OCP through Freundlich-type adsorption and that the adsorption controlled SDF-1 release from OCP during the hydrolysis into CDHA. Therefore, leveraging its molecular affinity for the OCP surface, OCP/SDF-1 facilitates MSC migration and enhances bone formation by ensuring the controlled, sustained release of SDF-1 from OCP.

The goal of this study was to analyze copy number variations (CNVs) in spontaneous abortions with a euploid karyotype, irrespective of the method used for CNV detection. This systematic review was performed in accordance with the PRISMA guidelines. Articles published between 2006 and 2023 were selected through the PubMed database. Studies were included if they involved CNV analysis in spontaneous abortions using any CNV detection method. The pathogenic significance of CNVs was interpreted based on the American College of Medical Genetics and Genomics (ACMG) guidelines. Nineteen publications met the inclusion criteria. A total of 1425 CNVs were identified in 550 samples from 3953 euploid spontaneous abortions, representing 14% of the cases. Among the detected CNVs, 9% were classified as pathogenic, and 7.5% were likely pathogenic. The most frequently observed pathogenic CNVs included 22q11.2 deletion/duplication, 16p13.11 deletion, 15q11.2 deletion/duplication, 1p36.33 duplication, and 17p13.3 duplication. The genomic regions with the highest frequency of CNVs, regardless of their pathogenic effect, were 8q24.3, 16p13.3, 21q22.3, Xp22.33, Xp22.31, and Xq28. No clear associations were found between specific CNVs and pregnancy loss. However, deletions in the 22q11.2 region emerged as the most likely candidates contributing to lethality during the early stages of embryonic development.

<h4>Aim</h4>To review the aetiopathogenesis of infantile epileptic spasms syndrome (IESS) and mechanisms of action of adrenocorticotrophin hormone (ACTH)/corticosteroids established in humans.<h4>Method</h4>MEDLINE, PubMed, and Embase were systematically searched from inception to December 2023 to identify studies related to IESS aetiology and treatment response. Mechanistic themes were identified and through consensus meetings refined and grouped into five overarching hypotheses.<h4>Results</h4>Five hypotheses were generated from 17 mechanistic themes: (1) gene and epigenetic regulation altering expression of 'vulnerability' genes; (2) stress and hypothalamic-pituitary-adrenal axis activation; (3) neuroinflammation and altered immune function; (4) altered neuronal transmission and pathways; and (5) dysfunction of metabolic pathways.<h4>Interpretation</h4>The evidence that ACTH/corticosteroids alter these processes remains limited. It is plausible that these processes interact with one another, rather than existing independently, and affect maturational and regulatory processes in the central nervous system, consistent with proposals that IESS is a neurodevelopmental disorder. Understanding how ACTH/corticosteroids work in IESS may facilitate disease-modifying treatments and improve neurodevelopmental outcomes.

Soft-shelled turtles, or Trionychia, are an enigmatic and fascinating group due to their specific morphological features and ecological adaptations. Based on mitochondrial DNA (mtDNA) and/or nuclear markers, previous studies showed the incongruent phylogenetic topologies within Trionychia (e.g., the Palea and its closely related species). In order to resolve the equivocal relationships and obtain some "genome-level" common evolutionary characters of soft-shelled turtles, in this study, we assembled and annotated the complete mitochondrial genomes of Palea steindachneri and Pelodiscus axenaria, both naturally distributed in Asia. The sizes of the two mitochondrial genomes were 16,811 bp and 17,143 bp, respectively. Typical vertebrate animal mtDNA features were observed, such as the usual gene components and arrangements (37 genes with a non-coding control region) and the A + T biased nucleotide compositions on the light strand (61.5% and 62.7%, respectively). All conserved blocks common to the vertebrates control region except for the extended terminal associated sequences (ETAS2) were found in the two soft-shelled turtles. The ω ratio averaged over all sites of each protein-coding gene (PCG) was below 1, which indicated purifying selection at the gene-wide level. However, a positive selection site at the 350-codon position in the cytb gene was detected, as estimated by Bayes empirical Bayes (BEB) analysis. Compared with the gene subsets, the mitogenomes provided the most robust phylogenetic resolution. The monophyly of the clades Amydona, Gigantaesuarochelys, and Apalonia was well supported. Topology discrepancies were observed among different datasets (e.g., the positions of Lissemys and Palea), reflecting the heterogeneous phylogenetic signals in the soft-shelled turtle mitogenomes. Precise date estimation based on Bayesian relaxed clock analyses indicated that the crown group age of extant Trionychia was approximately 115.84 Ma (95% HPD: 91.33-142.18 Ma). Paleoclimate changes, especially the Eocene - Oligocene transition, could be responsible for the speciation in these groups. Our results reiterated the necessity and effectiveness of incorporating entire mitochondrial genomes to delineate phylogenetic relationships in chelonian phylogeny studies.

<h4>Background</h4>The parkin-interacting substrate (PARIS, also known as ZNF746) is a transcriptional repressor, whose accumulation and phosphorylation play central pathological roles in Parkinson's disease (PD). PARIS-induced transcriptional repression of PGC-1α or MDM4 contributes to mitochondrial dysfunction and p53-dependent neuron loss in PD. Despite the important role of PARIS in PD pathogenesis, unbiased transcriptomic profiles influenced by PARIS accumulation in dopaminergic neurons remain unexplored.<h4>Methods</h4>We engineered Tet-Off conditional transgenic mice expressing PARIS in dopaminergic neurons, driven by DAT-PF-tTA driver mice. The conditional PARIS transgenic mice were characterized by PD-associated pathologies, including progressive dopamine cell loss, neuroinflammation, PGC-1α repression, and mitochondrial proteome alteration. Motor impairment was assessed using pole and rotarod tests. L-DOPA and c-Abl inhibitors were administered to PARIS transgenic mice to evaluate their therapeutic efficacy. The transcriptomic profiles and gene ontology clusters were analyzed by bulk and single-nucleus RNA-seq for the ventral midbrains from PARIS transgenic and age-matched controls.<h4>Results</h4>Conditional dopaminergic PARIS expression in mice led to the robust and selective dopaminergic neuron degeneration, neuroinflammation, and striatal dopamine deficits, resulting in L-DOPA-responsive motor impairments. Consistent with the results of previous reports, PARIS suppressed dopaminergic PGC-1α expression, disturbed mitochondrial marker protein expression, and reduced COXIV-labeled mitochondria in dopamine neurons. Pharmacological inhibition of c-Abl activity in PARIS transgenic mice largely prevents PD-associated pathological features. Unbiased transcriptomic analysis revealed PARIS-regulated differentially expressed genes (DEGs), both collectively and in a cell-type-specific manner, along with enriched biological pathways linked to PD pathogenesis. Single-cell resolution transcriptomic analysis confirmed repression of PGC-1α and several mitochondria-related target genes in dopaminergic cells. Additionally, we identified distinct glial cell subpopulations and DEGs associated with PD pathogenesis.<h4>Conclusions</h4>Conditional PARIS transgenic mice recapitulate robust and dopaminergic neuron-selective pathological features of PD, allowing the preclinical evaluation of antisymptomatic and disease-modifying therapeutic strategies within a couple of months. Based on this new PD mouse model, we provide unbiased bulk and single-nucleus transcriptomic profiles that are regulated by PARIS and potentially contribute to PD pathogenesis. A PD mouse model with flexible pathology induction capacity and a whole transcriptome could serve as a useful resource for translational PD research.

<h4>Background</h4>Necrotizing enterocolitis (NEC) is a critical gastrointestinal disease in preterm infants, for which no specific treatment is established. We previously demonstrated that thrombin-preconditioned mesenchymal stromal cell-derived extracellular vesicles (thMSC-EVs) enhance protection against other neonatal tissue injuries. Therefore, this study aimed to evaluate the therapeutic potential of thMSC-EVs in modified in vitro, in vivo, and organoid models of NEC.<h4>Methods</h4>In vitro, the effects of thMSC-EVs and naïveMSC-EVs were compared in hyperosmotic, ischemic, and hypothermic (HIT)-stressed IEC-6 cells and LPS-treated peritoneal macrophages. In vivo, NEC was induced in P4 mouse pups by three cycles of formula feeding, oral LPS administration, hypoxia, and hypothermia, followed by overnight dam care. 2 × 10⁹ thMSC-EVs were intraperitoneally administered daily for three days, and the therapeutic effects were assessed macroscopically, histologically, and biochemically. NEC mouse-derived organoids were established to evaluate the thMSC-EVs' effect in mature enterocytes. LC-MS/MS was performed to analyze the EV proteomics.<h4>Results</h4>In vitro, compared with naïveMSC-EVs, thMSC-EVs significantly improved cellular viability in HIT-induced IEC-6 cells and reduced pro-inflammatory (IL-1α, IL-1β, TNF-α) but increased anti-inflammatory (TGF-b) cytokine levels in LPS-treated peritoneal macrophages. In vivo, thMSC-EVs significantly attenuated clinical symptoms, reduced intestinal damage, and retained intestinal stem cell markers, showing more significant localization in NEC-induced intestines than in healthy intestines. In NEC mouse-derived organoids, thMSC-EVs significantly increased OLFM4 and claudin-4 expression and reduced stress-related markers such as sucrase-isomaltase, defensin, and chromogranin A. Proteomic analysis revealed that thMSC-EVs were greater enriched in anti-apoptotic, anti-inflammatory, cell adhesion, and Wnt signaling pathways than naïveMSC-EVs.<h4>Conclusion</h4>thMSC-EVs improved cellular viability, reduced apoptosis, attenuated inflammation, and upregulated key intestinal stem cell markers, collectively suggesting their tissue-protective effects and highlighting their potential as a treatment for NEC.

<h4>Background</h4>Alcohol use disorder (AUD) is a prevalent neuropsychiatric disorder that is a major global health concern, affecting millions of people worldwide. Previous studies of AUD used underpowered single-cell analysis or bulk homogenates of postmortem brain tissue, which obscure gene expression changes in specific cell types. Therefore, we sought to conduct the largest-to-date single-nucleus RNA sequencing (snRNA-seq) postmortem brain study in AUD to elucidate transcriptomic pathology with cell type-specific resolution.<h4>Methods</h4>Here, we performed snRNA-seq and high-dimensional network analysis of 73 postmortem samples from individuals with AUD (n = 36, n_nuclei = 248,873) and neurotypical control individuals (n = 37, n_nuclei = 210,573) in the dorsolateral prefrontal cortex from both male and female donors. Additionally, we performed analysis for cell type-specific enrichment of aggregate genetic risk for AUD as well as integration of the AUD proteome for secondary validation.<h4>Results</h4>We identified 32 distinct cell clusters and found widespread cell type-specific transcriptomic changes across the cortex in AUD, particularly affecting glial populations. We found the greatest dysregulation in novel microglial and astrocytic subtypes that accounted for the majority of differential gene expression and coexpression modules linked to AUD. Differential gene expression was secondarily validated by integration of a publicly available AUD proteome. Finally, analysis for aggregate genetic risk for AUD identified subtypes of glia as potential key players not only affected by but also causally linked to the progression of AUD.<h4>Conclusions</h4>These results highlight the importance of cell type-specific molecular changes in AUD and offer opportunities to identify novel targets for treatment on the single-nucleus level.

<h4>Background & aims</h4>SLC11A2 (DMT1) and SLC40A1 (ferroportin) are essential for dietary iron absorption, but their role in manganese transport is debated. SLC30A10 deficiency causes severe manganese excess due to loss of gastrointestinal manganese excretion. Patients are treated with chelators but also respond to oral iron, suggesting that iron can outcompete manganese for absorption in this disease. Here, we determine if divalent metal transport 1 (Dmt1) and ferroportin can transport manganese using Slc30a10-deficient mice as a model.<h4>Methods</h4>Manganese absorption and levels and other disease parameters were assessed in Slc30a10^-/- mice with and without intestinal Dmt1 and ferroportin deficiency using gastric gavage, surgical bile collections, multiple metal assays, and other techniques. The contribution of intestinal Slc30a10 deficiency to ferroportin-dependent manganese absorption was explored by determining if intestinal Slc30a10 deficiency increases manganese absorption in a mouse model of hereditary hemochromatosis, a disease of iron excess due to ferroportin upregulation.<h4>Results</h4>Manganese absorption was increased in Slc30a10-deficient mice despite manganese excess. Intestinal Dmt1 and ferroportin deficiency attenuated manganese absorption and excess in Slc30a10-deficient mice. Intestinal Slc30a10 deficiency increased manganese absorption and levels in the hemochromatosis mouse model.<h4>Conclusions</h4>Aberrant absorption contributes prominently to SLC30A10 deficiency, a disease previously attributed to impaired excretion, and is dependent upon intestinal Dmt1 and ferroportin and exacerbated by loss of intestinal Slc30a10. This work expands our understanding of overlaps between manganese and iron transport and the mechanisms by which the body regulates absorption of 2 nutrients that can share transport pathways. We propose that a reconsideration of the role of Dmt1 and ferroportin in manganese homeostasis is warranted.

Alzheimer's disease (AD) is the most common form of dementia and is characterized by progressive memory loss and cognitive decline, affecting behavior, speech, and motor abilities. The neuropathology of AD includes the formation of extracellular amyloid-β plaques and intracellular neurofibrillary tangles of phosphorylated tau, along with neuronal loss. Although neuronal loss is an AD hallmark, cell-cell communication between neuronal and non-neuronal cell populations maintains neuronal health and brain homeostasis. To study changes in cell-cell communication during disease progression, we performed snRNA-sequencing of the hippocampus from female 3xTg-AD and wild-type littermates at 6 and 12 months. We inferred differential cell-cell communication between 3xTg-AD and wild-type mice across time points and between senders (astrocytes, microglia, oligodendrocytes, and OPCs) and receivers (excitatory and inhibitory neurons) of interest. We also assessed the downstream effects of altered glia-neuron communication using pseudobulk differential gene expression, functional enrichment, and gene regulatory analyses. We found that glia-neuron communication is increasingly dysregulated in 12-month 3xTg-AD mice. We also identified 23 AD-associated ligand-receptor pairs that are upregulated in the 12-month-old 3xTg-AD hippocampus. Our results suggest increased AD association of interactions originating from microglia. Signaling mediators were not significantly differentially expressed but showed altered gene regulation and transcription factor activity. Our findings indicate that altered glia-neuron communication is increasingly dysregulated and affects the gene regulatory mechanisms in neurons of 12-month-old 3xTg-AD mice.

<h4>Background</h4>Lung adenocarcinoma (LUAD) is a highly heterogeneous cancer type with a poor prognosis. Accurate subtype identification can help guide its treatment. The traditional subtype identification methods using a single-omics approach make it difficult to comprehensively characterize the molecular features of LUAD. Identification of subtypes through multi-omics association strategies can effectively supplement the shortcomings of single-omics information.<h4>Methods</h4>In this study, we used the Generative Adversarial Network (GAN) to mine transcriptomic, proteomic, and epigenomic information and generate an integrated data set. The newly integrated data were then used to identify LUAD immune subtypes. In the improved GAN (MOGAN) method, we not only integrated multiple omics datasets but also included the interactions between proteins and genes and between methylation and genes. Thus, we achieved effective complementarity of multi-omics information.<h4>Results</h4>Two subtypes, MOGANTPM_S1 and MOGANTPM_S2, were identified using immune cell infiltration analysis and the integrated multi-omics data. MOGANTPM_S1 patients displayed higher immune cell infiltration, better prognosis, and sensitivity to immune checkpoint inhibitors (ICIs), while MOGANTPM_S2 had lower immune cell infiltration, poorer prognosis, and were insensitive to ICIs. Therefore, immunotherapy was more suitable for MOGANTPM_S1 patients in clinical practice. In addition, this study developed a LUAD subtype diagnostic model using the transcriptomic and proteomic features of five genes, which can be used to guide clinical subtype diagnosis.<h4>Conclusions</h4>In summary, the MOGAN method was applied to integrate three omics data types and successfully identify two LUAD immune subtypes with significant survival differences. This classification method may be useful for LUAD treatment decisions.

Diabetic nephropathy (DN) is one of the leading causes of end-stage renal disease. The most popular biomarkers in current research on DN are microRNAs. There are studies showing that while the expression of SOX6 increases, hsa-miR-342-3p expression decreases in kidney tissues with DN. The current study evaluated hsa-miR-342-3p expression by Real Time PCR and SOX6 expression by immunohistochemistry in a cohort of 110 DN biopsies, as well as their relationship with various clinical and histomorphological parameters. An inverse relationship between expression of hsa-miR-342-3p and SOX6 was demonstrated. SOX6 genetic expression was correlated with serum creatinine and tubular basement membrane thickening. Immunohistochemically, SOX6 staining was observed in mesangial cells and podocytes in 21 patients, with tubular staining in 45, and interstitial staining in 27 patients. Tubular staining was associated with proteinuria, interstitial fibrosis and inflammation; interstitial staining was associated with creatinine; and staining in the glomerular compartment was associated with advanced DN class. Our study is the first in the literature in which SOX6 was applied immunohistochemically in human kidney tissue, and its relation with DN classes was examined. We demonstrate its correlation with laboratory and histomorphological parameters, and provide a rational basis for future studies on larger patient groups that may result in the development of new biomarkers to predict the progression of DN and enhance its treatment.

The prevalence of papillary renal cell carcinomas (PRCCs) is estimated to be between 10% and 15%. At present, there is no effective therapeutic approach available for patients with advanced PRCCs. The molecular biomarkers associated with PRCC diagnoses have been rarely studied compared to renal clear cell carcinomas; therefore, the necessity for the identification of novel molecular biomarkers to aid in the early identification of this disease. Bioinformatics and artificial intelligence technologies have become increasingly important in the search for diagnostic biomarkers for early cancer detection. In this study, three genes-BCL11A, NTN5, and OGN-were identified as diagnostic biomarkers using the Cancer Genome Atlas (TCGA) database and deep learning techniques. To identify the differential expression genes (DEGs), ribonucleic acid (RNA) expression profiles of PRCC patients were analyzed using a machine learning approach. A number of molecular pathways and coexpressions of DEGs have been analyzed and a correlation between DEGs and clinical data has been determined. Diagnostic markers were then determined via machine learning analysis. The 10 genes selected with the highest variable importance value (more than 0.9) were further investigated, with six upregulated (BCL11A, NTN5, SEL1L3, SKA3, TAPBP, SEMA6A) and four downregulated (OGN, ADCY4, SMOC2, CCL23). A combined receiver operating characteristic (ROC) curve analysis revealed that the BCL11A-NTN5-OGN genes, which have specificity and sensitivity values of 0.968 and 0.901, respectively, can be used as a diagnostic biomarker for PRCC. In general, the genes introduced in this study may be used as diagnostic biomarkers for the early diagnosis of PRCC, thus providing the possibility of early treatment and preventing the progression of the disease.

Due to the minimal survival benefits of existing therapies for pediatric diffuse midline glioma (DMG) patients, new therapeutic modalities are being investigated. Immunotherapies such as CAR-T cells and oncolytic viruses (OVs) are part of these efforts, as evidenced by the increasing number of clinical trials. αβ T cells engineered with a high-affinity γ9δ2 T-cell receptor (TEGs) are immune cells designed to target metabolic changes in malignant or virally infected cells via BTN2A1 and BTN3A. Because the expression of BTN2A1 and BTN3A can be altered in tumor and infected cells, combining TEGs and OVs could potentially enhance the anti-tumor response. We investigated this hypothesis in the following study. We demonstrate that TEGs can indeed target DMG, which expresses BTN2A1 and BTN3A at varying levels, and that OVs can further enhance the expression of BTN3A-but not BTN2A1-in DMG. Functionally, TEGs killed DMG cell cultures, and this killing was further increased after OV infection of the DMGs with either adenovirus Δ24-RGD or reovirus R124 under suboptimal conditions. However, this additive effect was lost when γ9δ2 TCR-ligand interaction was boosted by pamidronate. This study demonstrates the additive effect of combining OVs and Vγ9Vδ2 TCR-engineered immune cells under suboptimal conditions and supports a combination strategy to enhance the efficacy of both therapeutic modalities.

Diabetes mellitus (DM) is a chronic metabolic disorder and one of the most significant global health burdens worldwide. Key pathophysiological mechanisms underlying its onset and associated complications include hyperglycemia-related stresses, such as oxidative stress and endoplasmic reticulum stress (ER stress). Long non-coding RNAs (lncRNAs), defined as RNA transcripts longer than 200 nucleotides and lacking protein-coding capacity, play crucial roles in various biological processes and have emerged as crucial regulators in the pathogenesis of diabetes. This review provides a comprehensive overview of lncRNA biogenesis and its functional roles, emphasizing recent findings that link stress-related lncRNAs to diabetic pathology and complications. Also, we discuss how lncRNAs influence diabetes and its complications by modulating pathways involved in cell death, proliferation, inflammation, and fibrosis, which contribute to pancreatic β cell dysfunction, insulin resistance, diabetic nephropathy, and retinopathy. By analyzing current research, we aim to enhance understanding of lncRNA involvement in diabetes while identifying potential therapeutic targets and guiding future research directions to elucidate the complex mechanisms underlying this pervasive condition.

Functional iron-deficiency anemia (FIDA) is a side effect of many cancer treatments, occurring when chemotherapy drugs damage bone marrow cells, which are responsible for producing red blood cells, due to the myelosuppressive effects of chemotherapy, or to the cancer itself. This study was performed to compare the effects of darbepoetin alfa alone, or in combination with ferric derisomaltose in cancer patients with FIDA, and to elucidate the mechanism underlying the effects in F36E cells. F36E cells treated with darbepoetin alfa showed increased cell viability. AML and GC cells treated with darbepoetin alfa, ferric derisomaltose, or ferric derisomaltose plus darbepoetin alfa showed no induction of apoptosis. The effects of these drugs on the anticancer efficacy of PTX chemotherapy were examined by analyzing cell viability and induction of apoptosis. Darbepoetin alfa, ferric derisomaltose, and ferric derisomaltose plus darbepoetin alfa showed no significant inhibitory effects on the apoptosis-inducing activity of PTX in GC cell lines. Patients with chemotherapy-induced FIDA in Group I receiving ferric derisomaltose plus darbepoetin alfa showed higher hemoglobin levels, transferrin saturation, and ferritin levels compared to those in Group II, treated with darbepoetin alfa alone. In cancer patients with FIDA, the prognosis of anemia treatment was better in the ferric derisomaltose plus darbepoetin alfa combination group than in the group receiving darbepoetin alfa monotherapy.

As the blood-brain barrier (BBB) prevents molecules from accessing the central nervous system (CNS), the traditional systemic delivery of chemical drugs limits the development of neurological drugs. However, in recent years, innovative therapeutic strategies have tried to bypass the restriction of traditional drug delivery methods. In vivo gene therapy refers to emerging biopharma vectors that carry the specific genes and target and infect specific tissues; these infected cells and tissues then undergo fundamental changes at the genetic level and produce therapeutic proteins or substances, thus providing therapeutic benefits. Clinical and preclinical trials mainly utilize adeno-associated viruses (AAVs), lentiviruses (LVs), and other viruses as gene vectors for disease investigation. Although LVs have a higher gene-carrying capacity, the vector of choice for many neurological diseases is the AAV vector due to its safety and long-term transgene expression in neurons. Here, we review the basic biology of AAVs and summarize some key issues in recombinant AAV (rAAV) engineering in gene therapy research; then, we summarize recent clinical trials using rAAV treatment for neurological diseases and provide translational perspectives and future challenges on target selection.

Cancer, ranking just below cardiovascular diseases, is a leading cause of mortality worldwide. The key to enhancing survival rates among cancer patients lies in the early detection, removal, and treatment of tumors. However, the broad-spectrum nature of current treatments, including chemotherapy and radiation therapy, results in significant collateral damage to healthy cells and tissues. In this context, hyperbranched polymers present a promising avenue for more targeted therapy. These polymers can be loaded with chemotherapeutic drugs and modified with specific ligands to selectively target cancer cells via glucose transporters, which are overexpressed in many cancer types. To enhance the delivery of drugs to cancer cells, we have engineered an N-acetyl glucosamine conjugated version of this polymer. The characterization of these nanocarriers was evaluated using various techniques, including ¹H NMR, dynamic light scattering, and FTIR spectroscopy. Additionally, confocal microscopy was utilized to compare the accumulation of doxorubicin in cancer cells using both the N-acetyl glucosamine-conjugated and unmodified versions of H40 Boltorn™. Our observations indicated a superior accumulation of doxorubicin in cells treated with the modified H40 polymer. Further evaluation of the drug-loaded nanocarriers was conducted on MDA-MB-231 and 4T1 breast cancer cell lines, focusing on their cytotoxic effects. This suggests that the targeted delivery of anticancer drugs using the modified H40 Boltorn™ nanocarriers significantly enhances the ability to kill breast cancer cells, offering a more efficient and selective approach to chemotherapy that minimizes impact on healthy tissues and cells.

<h4>Introduction</h4>This study investigates the subclasses of metabolic syndrome (Mets) and their relationship with non-alcoholic fatty liver (NAFLD) and the probable predictive role of serum vitamin D and CRP levels.<h4>Methods</h4>This community-based, cross-sectional study was performed on adults in the framework of the Amol cohort prospective study (AmolCPS). Mets was defined as Adult Treatment Panel III criteria (ATP III) and ultrasound was used to diagnose NAFLD. Anthropometric and blood pressure measurements were conducted, and biochemical measurements were assessed after fasting. Data analysis included Latent class analysis, two-tailed χ2 statistics, one-way analysis of variance, and logistic regression using Mplus (version 7.4) and spss (version 26) softwares.<h4>Results</h4>The study involved 2308 participants, with a mean age of 43.17 ± 12.30 years. Mets prevalence was 25.64%, with three identified classes: Mets with Hypertension (HTN), Mets without HTN (Non-HTN), and Low Risk. Mets with HTN had a high probability of at least four components, particularly high SBP. Non-HTN had at least three high probable components, especially high TG and low HDL but not high SBP and DBP. The low-risk class had a low probability of all components except low HDL in women. Serum vitamin D and CRP levels did not significantly predict Mets classes in men, while CRP level significantly predicted the HTN class in women (OR:1.03, CI:1.004-1.067). Both HTN, and Non-HTN Mets classes significantly increased the odds of NAFLD compared to the low risk class, especially in women (HTN class OR: 4.20 vs 2.94; non-HTN class OR: 5.60 vs 3.12 in women and men respectively).<h4>Conclusion</h4>The latent class analysis in northern Iran identified three Mets classes: HTN, Non-HTN, and low-risk, with hypertension playing a crucial role in determining these classes. These classes were stronger predictors of NAFLD in women. Serum CRP and vitamin D levels did not emerge as significant predictors of the classes, except for serum CRP in the HTN class among women.

<h4>Background</h4>A comprehensive review of 45-year trends in intrahepatic cholangiocarcinoma (ICC) and extrahepatic cholangiocarcinoma (ECC) in the United States has not been published. Given their rising incidence, our study aimed to analyze trends in incidence and survival, comparing ICC and ECC.<h4>Methods</h4>We extracted a 45-year dataset (1975-2020) from the Surveillance, Epidemiology, and End Results database. Age-adjusted incidence rates were calculated using SEERStat®. Annual Percent Change (APC) was estimated via weighted least squares. Relative survival (1- and 5-year) was calculated using the Ederer II method and compared across sexes and races.<h4>Results</h4>A significant rise in ICC and ECC incidence was observed in both sexes (APC 3.71 for ICC vs. 6.16 for ECC; P<0.001). In females, ECC incidence increased more than ICC (APC 5.96 vs. 4.09, P<0.05), whereas males showed a fluctuating ECC trend and a steady ICC rise. Survival rates significantly improved across all races and sexes (P<0.05). ICC survival rose from 17.45% to 41.41% (1-year) and 2.83% to 10.99% (5-year), while ECC increased from 30.33% to 41.12% (1-year) and 5.96% to 10.44% (5-year). Among white and other-race females, ECC showed less improvement than ICC. Black individuals lacked statistically significant data.<h4>Conclusions</h4>Our study highlights disparities in ICC and ECC incidence, with higher rates in males, but better survival for ECC in males and ICC in females. The underrepresentation of Black individuals warrants further study to explore contributing factors such as risk, access to care, and treatment.

TAR DNA-binding protein 43 kDa (TDP-43) proteinopathies are a group of neurodegenerative diseases (NDs) characterized by the abnormal accumulation of the TDP-43 protein in neurons and glial cells. These proteinopathies are associated with several NDs, including amyotrophic lateral sclerosis, frontotemporal lobar degeneration, and some forms of Alzheimer's disease. Yeast models have proven valuable in ND research due to their simplicity, genetic tractability, and the conservation of many cellular processes shared with higher eukaryotes. For several decades, <i>Saccharomyces cerevisiae</i> has been used as a model organism to study the behavior and toxicity of TDP-43, facilitating the identification of genes and pathways that either exacerbate or mitigate its toxic effects. This review will discuss evidence showing that yeast models of TDP-43 exhibit defects in proteostasis, mitochondrial function, autophagy, and RNA metabolism, which are key features of TDP-43-related NDs. Additionally, we will explore how modulating proteins involved in these processes reduce TDP-43 toxicity, aiding in restoring normal TDP-43 function or preventing its pathological aggregation. These findings highlight potential therapeutic targets for the treatment of TDP-43-related diseases.

Methamphetamine (METH) use is frequent among people with HIV (PWH) and appears to increase the risk of neuronal injury and neurocognitive impairment (NCI). This study explored in vivo the effects of a 12 week (long-term), low-dose METH regimen in a transgenic animal model of neuroHIV with inducible expression of HIV-1 transactivator of transcription (Tat). Seven months after transient Tat induction and five months after METH exposure ended, we detected behavioral changes in the Barnes maze (BM) spatial memory task in the Tat and METH groups but not the combined Tat + METH group. The novel object recognition (NOR) task revealed that Tat extinguished discrimination in female animals with and without METH, although METH alone slightly improved NOR. In contrast, in males, Tat, METH, and Tat + METH all compromised NOR. Neuropathological examination detected sex-dependent and brain region-specific changes of pre-synaptic terminals, neurites, and activation of astrocytes and microglia. RNA-sequencing and quantitative reverse transcription polymerase chain reaction indicated that METH and Tat significantly altered gene expression, including factors linked to Alzheimer's disease-like NCI. In summary, chronic low-dose METH exerts long-term effects on behavioral function, neuropathology, and mRNA expression, and modulates the effects of Tat, suggesting sex-dependent and -independent mechanisms may converge in HIV brain injury and NCI.

Stroke, particularly ischemic stroke, is a leading cause of global mortality and disability. It is caused by blood flow obstruction and reduced oxygen delivery to brain tissue. Conventional treatments, such as tissue plasminogen activator (tPA) and mechanical thrombectomy (MT), have limited efficacy in repairing neural damage and carry risks of adverse effects. As a result, stem cell therapies, including mesenchymal stem cells (MSCs), have emerged as promising approaches for enhancing neural recovery and offering neuroprotection in ischemic stroke management. MSCs offer multifaceted benefits, such as reducing inflammation, protecting neurons, and promoting angiogenesis and neurogenesis. Recent evidence highlights the importance of MSC secretomes-extracellular vesicles (EVs) and exosomes rich in neuroprotective factors, such as microRNAs, proteins, and cytokines. These bioactive molecules demonstrated considerable efficacy in preclinical models by reducing neuroinflammation, preserving neurovascular integrity, and promoting cellular repair in ischemic environments. Preclinical <i>in vitro</i> and <i>in vivo</i> studies demonstrate the potential of the MSC secretomes to restore brain function after ischemic stroke. This is achieved by enhancing neuronal survival through mechanisms such as angiogenesis or vascular recovery, neuroprotection including modulation of immune or inflammatory responses, apoptosis, and autophagy, and promoting post-stroke neurogenesis. This review explores the translational challenges and future potential of integrating conventional ischemic stroke therapies with stem cell-based or cell-free approaches. The present study synthesizes current insights into the role of MSC-derived secretomes from both <i>in vitro</i> and <i>in vivo</i> studies.

<h4>Background</h4>The elevated health burden of thromboembolic events necessitates development of blood-based risk monitoring tools.<h4>Objectives</h4>We explored the potential of mass spectrometry-based plasma proteomics to provide insights into underlying plasma protein signatures associated with treatment and occurrence of thromboembolic events.<h4>Methods</h4>Utilizing a high-throughput, data-independent acquisition, discovery-based proteomics workflow, we analyzed 434 plasma proteomes from different groups of individuals with elevated risk of thromboembolic events, including individuals I) on vitamin K antagonists (VKAs; <i>n</i> = 130), II) with a prior venous thromboembolism (<i>n</i> = 10), III) with acute cerebral venous sinus thrombosis (<i>n</i> = 10, and IV) with SARS-CoV-2 infection (<i>n</i> = 67). Plasma protein levels measured with mass spectrometry were correlated with international normalized ratio and conventional clinical laboratory measurements. Plasma profile differences between different groups were assessed using principal component analysis, moderated <i>t</i>-test, and clustering analysis.<h4>Results</h4>Plasma protein levels were in agreement with conventional clinical laboratory parameters, including albumin and fibrinogen. Levels of vitamin K-dependent proteins inversely correlated with international normalized ratio. In the individual studies, we found decreased levels of vitamin K-dependent coagulation proteins in patients on VKAs, alterations in inflammatory signatures among CVST patients and a distinctive signature indicative of SARS-CoV-2 infection. However, no protein signature associated with a thromboembolic event could be identified neither in individual nor combined studies.<h4>Conclusion</h4>Although VKA treatment-specific and disease-specific signatures were captured, our study highlights that the challenges of discovering biomarkers in patients at risk of thromboembolic events lie in the heterogeneity of individual plasma profiles in relation to treatment and etiology.

Hepatitis E virus (HEV) is an emerging zoonotic pathogen and it is associated with chronic hepatitis E virus infection (CHE) in immuno-suppressed patients due to failure of viral clearance. A network of the CHE-associated-differentially expressed genes (DEGs) is known. Hence, a comprehensive assessment of the known protein-protein interaction (PPI) network is of interest. After network clustering, the hub gene-associated microRNAs (miRNAs) were explored and subsequently, these identified miRNAs (miR-129-2-3p, miR-130a-3p, miR-138-5p, miR-212-3p, miR-221-3p, miR-27b-3p and miR-29c-3p) were undertaken for enrichment analysis. It should be noted that these miRNAs are significantly associated with Hepatitis E virus infection for further validation and analysis.

Pulmonary arteriovenous malformations (PAVMs) is a rare condition involving an abnormal connection of the pulmonary vasculature between the artery and vein, bypassing the capillary structure and causing a right-to-left shunt (RLS) of blood flow. This case report describes a 17-year-old female patient presenting with recurrent nose and tongue bleeding. Cyanosis and clubbing of the fingers were observed, along with visible telangiectasis on the skin and mucosal surfaces. Hereditary hemorrhagic telangiectasis (HHT) was diagnosed based on the Curaçao criteria. The RLS manifestations suggested a high probability of PAVM, confirmed by an agitated saline test showing a positive bubble appearance after four beats from the pulmonary vein to the left ventricle. An increased intraventricular wall diameter (19 mm) with a granular sparkling appearance indicated potential cardiac amyloidosis (CA). Speckle tracking echocardiography (STE) revealed a 'cherry-like' appearance in a 'bull's eye' pattern. This case illustrates a diagnostic approach for PAVM in an adolescent HHT patient with suspected CA in a rural setting using limited resources.

<h4>Background</h4>Non-small cell lung cancer (NSCLC) is the leading cause of cancer morbidity and mortality worldwide. The prognosis of patients has been significantly improved by chemotherapy, but acquired drug resistance remains a major obstacle to NSCLC treatment. Circular RNAs (circRNAs), which act as miRNA or protein sponges, are critically associated with the development and chemotherapy resistance of NSCLC.<h4>Methods</h4>CircRNA sequencing was performed to analyze the differential expression of circRNAs between A549 and A549-GR cells. Chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) technologies were used to detect the expression of hsa_circ_0125356, miR-582-5p,and FGF9 in NSCLC tissues and para-carcinoma tissues. Fluorescence in situ hybridization (FISH), dual-luciferase reporter assays and RNA immunoprecipitation (RIP) were conducted to evaluate the expression and regulation of hsa_circ_0125356, miR-582-5p, and FGF9. Furthermore, the regulation of hsa_circ_0125356/miR-582-5p/FGF9 on gemcitabine sensitivity was confirmed by TUNEL, Transwell, EdU, CCK8 and immunohistochemistry.<h4>Results</h4>We identified a novel hsa_circ_0125356 as a therapeutic target against gemcitabine resistance. Hsa_circ_0125356 was significantly elevated in clinical samples of patients with NSCLC. Moreover, hsa_circ_0125356 overexpression promoted gemcitabine resistance to NSCLC by upregulating FGF9 via sponging miR-582-5p in vivo and in vitro. Notably, WNT canonical (ERK/GSK3β/β-catenin) and non-canonical (Daam1/RhoA/ROCK2) signaling pathways were activated due to hsa_circ_0125356 acting as an endogenous miR-582-5p sponge to regulate the expression of FGF9, and thereby enhancing gemcitabine resistance via promoting DNA damage repair and inhibition of apoptosis. The results were further confirmed by two small molecule antagonists, WAY 316606 and XAV-939,which could inhibit the activation of WNT signaling pathway induced by hsa_circ_0125356.<h4>Conclusion</h4>We first demonstrated that hsa_circ_0125356 was significantly upregulated and served as a biomarker for gemcitabine resistance in NSCLC by sponging miR-582-5p/FGF9 axis to regulate the WNT canonical and non-canonical signaling pathways, which provided a new direction for identification of therapeutic targets for the treatment of gemcitabine resistance of NSCLC.

<h4>Aims</h4>A large number of surgical operations are available to treat osteochondral defects of the knee. However, the knee joint arthroplasty materials cannot completely mimic the articular cartilage and subchondral bone, which may bring some obvious side effects. Thus, this study proposed a biocompatible osteochondral repair material prepared from a double-layer scaffold of collagen and nanohydroxyapatite (CHA), consisting of collagen hydrogel as the upper layer of the scaffold, and the composite of CHA as the lower layer of the scaffold.<h4>Methods</h4>The CHA scaffold was prepared, and properties including morphology, internal structure, and mechanical strength of the CHA scaffold were measured by scanning electron microscopy (SEM) and a MTS electronic universal testing machine. Then, biocompatibility and repair capability of the CHA scaffold were further evaluated using a rabbit knee cartilage defect model.<h4>Results</h4>The CHA scaffold was well suited for the repair of articular cartilage and subchondral bone; the in vitro results showed that the CHA scaffold had good cytocompatibility. In vivo experiments demonstrated that the material had high biocompatibility and effectively induced cartilage and subchondral bone regeneration.<h4>Conclusion</h4>The CHA scaffold has a high potential for commercialization and could be used as an effective knee repair material in clinical applications.

The physiological basis of cognitive decline remains largely uncharacterized. We identified a protein panel signature, in living humans, that correlates to improvement in neurocognition over a period of 5 years. Our signature is composed of complement proteins, coagulation cascade, and extracellular matrix regulators. In our cohort, SERPINF1 is associated with greater maximal oxygen uptake after an aerobic exercise intervention. Sleep quality is also a key factor in relation to inter-alpha-trypsin inhibitor heavy chain H2, which was associated with greater sleep efficiency. Additionally, we validate that the coagulation profile of decliners' plasma contains procoagulant agonists, leading to greater platelet activation. ANN NEUROL 2025;97:1007-1018.

No abstract available.

No abstract available.

Skin cutaneous melanoma (SKCM) is one of the most lethal cancers translating into 75% of skin cancer-related deaths. Despite the advances in SKCM management and treatment strategies, the overall survival of patients remains unsatisfactory due to the metastatic properties of SKCM as well as the absence of effective prognostic biomarkers. Recent studies have shown that overload copper renders accumulation of mitochondrial proteins and fuels a form of cell death at odds with known death mechanisms and is hinged on mitochondrial respiration, the so-called cuproptosis. However, the exact role of cuproptosis in SKCM development and progression is unknown, and painting a clear picture of its functions in SKCM is fraught with challenges. A more systematic investigation is justified. In this study, we were posed to dissect the clout and latent regulatory mechanisms of cuproptosis-related genes (CRGs) in reining in SKCM progression. Also, we identified three CRGs that stood out were used to construct a prognostic model, which could be employed to predict the prognosis of patients with SKCM. Finally, through pan-cancer analysis, we found that the four cuproptosis key genes play a role in multiple tumors, suggesting that cuproptosis may impact tumor progression at the pan-cancer level. Taken together, these findings may not only contribute to the development of treatment strategies but also provide clues for treatment decision-making.

Newly produced platelets acquire a low activation state, but whether the megakaryocyte plays a role in this outcome has not been fully uncovered. Mesenchymal stem cells (MSCs) were previously shown to promote platelet production and lower platelet activation. We found that healthy megakaryocytes transfer mitochondria to MSCs, which is mediated by connexin 43 (Cx43) gap junctions on MSCs and leads to platelets at a low energetic state with increased LYN activation, characteristic of resting platelets with increased LYN activation, characteristic of resting platelets. On the contrary, MSCs have a limited ability to transfer mitochondria to megakaryocytes. Sickle cell disease (SCD) is characterized by hemolytic anemia and results in heightened platelet activation, contributing to numerous disease complications. Platelets in SCD mice and human samples had a heightened energetic state with increased glycolysis. MSC exposure to heme in SCD led to decreased Cx43 expression and a reduced ability to uptake mitochondria from megakaryocytes. This prevented LYN activation in platelets and contributed to increased platelet activation at steady state. Altogether, our findings demonstrate an effect of hemolysis in the microenvironment leading to increased platelet activation in SCD. These findings have the potential to inspire new therapeutic targets to relieve thrombosis-related complications of SCD and other hemolytic conditions.

Depletion or inhibition of core stress granule proteins, G3BP1 in mammals and TIAR-2 in <i>Caenorhabditis elegans</i>, increases the growth of spontaneously regenerating axons. Inhibition of G3BP1 by expression of its acidic or "B-domain" accelerates axon regeneration after nerve injury, bringing a potential therapeutic strategy for peripheral nerve repair. Here, we asked whether G3BP1 inhibition is a viable strategy to promote regeneration in injured mammalian central nervous system (CNS) where axons do not regenerate spontaneously. G3BP1 B-domain expression was found to promote axon regeneration in the transected spinal cord provided with a permissive peripheral nerve graft (PNG) as well as in crushed optic nerve. Moreover, a cell-permeable peptide (CPP) to a subregion of B-domain (rodent G3BP1 amino acids 190 to 208) accelerated axon regeneration after peripheral nerve injury and promoted regrowth of reticulospinal axons into the distal transected spinal cord through a bridging PNG. G3BP1 CPP promoted axon growth from rodent and human neurons cultured on permissive substrates, and this function required alternating Glu/Asp-Pro repeats that impart a unique predicted tertiary structure. The G3BP1 CPP disassembles axonal G3BP1, G3BP2, and FMRP, but not FXR1, granules and selectively increases axonal protein synthesis in cortical neurons. These studies identify G3BP1 granules as a key regulator of axon growth in CNS neurons and demonstrate that disassembly of these granules promotes retinal axon regeneration in injured optic nerve and reticulospinal axon elongation into permissive environments after CNS injury. This work highlights G3BP1 granule disassembly as a potential therapeutic strategy for enhancing axon growth and neural repair.

Mesoscale building blocks are instrumental in bridging multilevel hierarchical mineralization, endowing macroscale entities with remarkable functionality and mechanical properties. However, the mechanism orchestrating the homogeneous morphology of mesoscale mineralized motifs in collagen-based hard tissues remains unknown. Here, utilizing avian tendons as a mineralization model, we reveal a robust correlation between the mesoscale mineralized spherules and the presence of phosvitin. By designing a phosvitin-stabilized biomineral cluster medium, we replicate the well-defined mesoscale spherical structure within collagen matrix in vitro and ex vivo. In-depth studies reveal that phosvitin undergoes a conformational transition in the presence of biominerals at physiological concentrations, and self-assembles into mineral-dense amyloid-like aggregates. The spatial binding of these mineral-dense aggregates to collagen serves as a template for guiding the formation of mineralized spherules on the mesoscale. On the nanoscale, this binding facilitates mineral precursor release and diffusion into the fibrils for intrafibrillar mineralization. This discovery underscores the pivotal role of phosvitin-biomineral aggregates in templating hierarchical mineralization from the mesoscale to the nanoscale. This study not only elucidates the intricate mechanism underlying the collagen-based mineralization hierarchy but also promotes a cutting-edge advance in highly biomimetic material design and regenerative medicine.

Head and neck squamous cell carcinoma (HNSC) is a common malignant tumor with high incidence and mortality rates. ETS variant transcription factor 4 (ETV4), an important transcription factor, plays a key role in various cancers. However, the role of ETV4 in HNSC remains unclear. This study aimed to explore the potential prognostic value and oncogenic effects of ETV4 in HNSC. We analyzed ETV4 expression in HNSC patients' data from the TCGA database, alongside clinical pathological characteristics. The STRING and GEPIA databases were utilized to explore ETV4's interaction proteins and expression related genes. Gene Set Enrichment Analysis (GSEA) was performed on the stratified TCGA-HNSC cohort based on ETV4 expression levels. The correlation between ETV4 expression and immune cells, immune checkpoints, immune regulatory genes was further analyzed using R packages and TISIDB database. Finally, knockdown ETV4 in nasopharyngeal carcinoma cells (NPCs) using siRNA and evaluate cell proliferation, migration, and invasion using CCK-8, wound healing, clone formation, and Transwell assays. ETV4 was significantly overexpressed in HNSC and closely related with clinical pathological characteristics and prognosis. GSEA enrichment analysis showed significant enrichment of ETV4 in multiple immune suppression pathways. Further immune-related analysis indicated that ETV4 negatively correlated with most immune cells, immune checkpoints, tumor-infiltrating lymphocyte type characteristic molecules, immunoinhibitors, immune activators and MHC molecules. Knocking down ETV4 significantly inhibited the proliferation, migration and invasion of NPCs. ETV4 may serve as a prognostic biomarker and immunotherapy target in HNSC. High expression of ETV4 may have a negative regulatory effect on the immune level, matrix components and immune regulatory molecules.

<h4>Objectives</h4>Nerve growth factor (NGF) is a key mediator in osteoarthritis pain signaling. Clinical studies revealed that anti-NGF antibodies are often accompanied by progressively worsening cartilage degeneration, although they exhibit significant analgesic effects. However, the relationship between NGF expression and cartilage destruction remains unclear. Our study aimed to investigate the effects of NGF on chondrocytes and to elucidate the underlying mechanisms involved.<h4>Methods</h4>The ATDC5 cells were induced to differentiate into chondrocytes and stimulated with NGF at different concentrations (0.5-10 ng/mL). The cell counting kit-8 assay (CCK-8) was used to measure the effects of NGF on chondrocyte proliferation. Chondrocytes were subsequently stimulated with varying doses of NGF to identify the expression levels of the extracellular matrix. Chondrocytes were pretreated with GNF5837 (a tropomyosin receptor kinase A inhibitor) or LY294002 (a phosphoinositide 3-kinase inhibitor) before exposure to 5 ng/mL NGF to analyze associated signaling pathways. Western blotting and immunofluorescence staining were employed to analyze expression of related proteins.<h4>Results</h4>Alcian blue, toluidine blue staining, and type II collagen immunofluorescence staining demonstrated that ATDC5 cells differentiated into functional chondrocytes after 14 days of chondrogenic induction. The CCK-8 assay confirmed that cell proliferation was unaffected. NGF (0.5-5 ng/mL) was found to enhance chondrocyte matrix synthesis in a dose-dependent fashion, particularly in the expression of aggrecan, type II collagen, Sox9, and through the activation of the PI3K/AKT signaling pathway. The highest promoting effects were exhibited at 5 ng/mL of NGF. Further analysis indicated that GNF5837 (TRKA inhibitor) or LY294002 (PI3K inhibitor) could reverse the protective effects of NGF on chondrocyte matrix synthesis.<h4>Conclusion</h4>Our study identified a potentially beneficial role of NGF at concentrations of 0.5-5 ng/mL in chondrocytes, enhancing extracellular matrix synthesis, with significant involvement of the PI3K/AKT signaling pathway in this process.

<h4>Background</h4>Bladder cancer (BLCA) represents one of the most prevalent urological malignancies worldwide. Bridging integrator 1 (BIN1), a well-characterized tumor suppressor that interacts with and inhibits oncogenic Myc transcription factors, has demonstrated crucial roles in various cancer types. However, its specific functions and underlying molecular mechanisms in BLCA development and progression remain poorly understood. This study aims to elucidate the role of BIN1 in regulating BLCA cell proliferation, metastasis, and cancer stem cell properties.<h4>Methods</h4>Using urinary proteomics analysis, we identified BIN1 as a significantly dysregulated protein in BLCA. The clinical significance of BIN1 was further validated through comprehensive analyses of public databases. BIN1 expression levels defined distinct molecular and immunological subtypes of BLCA. Through proteomic profiling of BIN1-overexpressing UMUC3 cells and corresponding controls, we identified ALDH1 as a key downstream effector in the BIN1-regulated ALDH1/NOTCH signaling axis. We employed multiple experimental approaches, including Western blot analysis, quantitative RT-PCR, immunofluorescence staining, wound healing assays, transwell migration assays, colony formation assays, tumor sphere formation assays, flow cytometry, CCK8 proliferation assays, and cell transfection experiments.<h4>Results</h4>We observed significant downregulation of BIN1 in both BLCA tissues and cell lines compared to normal adjacent tissues and SV-HUC-1 cells, respectively. BIN1 overexpression inhibited cancer cell proliferation by promoting apoptosis and suppressed epithelial-mesenchymal transition (EMT), thereby reducing local invasion and distant metastasis. Additionally, BIN1 regulated cancer stem cell properties through modulation of ALDH1 expression, with NOTCH2 acting as a crucial downstream mediator of ALDH1 signaling.<h4>Conclusion</h4>Our findings demonstrate that BIN1 functions as a tumor suppressor in BLCA and suggest its potential utility as both a diagnostic biomarker and therapeutic target for BLCA treatment.

Human α1,4-galactosyltransferase (A4galt) is a glycosyltransferase existing in humans as two isoforms, widespread A4galt (named A4G) and its rare variant with p.Q211E substitution (A4Gmut). Both isoforms produce Gb3 (Galα1→4Galβ1→4Glc-Cer) on glycosphingolipids and P1 glycotope (Galα1→4Galβ1→4GlcNAc-R) on glycoproteins, which serve as receptors for Shiga toxin types 1 and 2 (Stx1 and Stx2). Stx1 is bound by Gb3 and P1 glycotope, while Stx2 is recognized solely by Gb3. To elucidate the role of these receptors, CHO-Lec2 cells expressing human A4G and A4Gmut were modified by disrupting the hamster B4GALT5 gene using CRISPR/Cas9 technology. The B4GALT5 gene encodes β1,4-galactosyltransferase 5 (B4galt5), synthesizing lactosylceramide, the key substrate for Gb3 synthesis. Consequently, B4GALT5-deficient CHO-Lec2-expressing A4G and A4Gmut cells lacked Gb3 glycosphingolipid but retained the ability to synthesize glycoprotein-based P1 glycotope. Both B4GALT5-deficient CHO-Lec2 cells expressing A4G and A4Gmut demonstrated no binding of Stx1B and Stx2B. The cytotoxicity assay showed that B4GALT5-deficient CHO-Lec2 cells expressing A4G were completely resistant to Stx1 holotoxin while A4Gmut-expressing cells revealed reduced sensitivity to Stx2. The glycoengineered CHO-Lec2 cells obtained in this study provide a valuable model for studying receptors for Stxs, enabling a detailed assessment of their roles in toxin binding and cytotoxicity.

<h4>Objective</h4>Supplementing animal feed with Allium mongolicum Regel powder (AMRP) additives can promote muscle production and improve meat quality. Here, we explored the effects of dietary AMRP supplementation on the performance, meat quality, and muscle transcriptome profile of Angus calves.<h4>Methods</h4>Twelve healthy female black Angus calves (average body weight = 280.4±15.74 kg, average age = 14±0.6 months) of the same genetic background were randomly assigned to two feed groups: control (CON; basal diet without any supplementation) and AMRP (basal diet supplemented with 20 g of AMRP per calf per day).<h4>Results</h4>In general, compared with the control group, dietary AMRP supplementation significantly increased the longissimus thoracis muscle area (p = 0.027) and pH24h (p = 0.027) but significantly reduced Warner-Bratzler shear force (p = 0.009) and cooking loss (p<0.001). Moreover, 1,284 differentially expressed genes (DEGs) were identified in AMRP-supplemented Angus calves. Pathway analysis revealed that the DEGs were involved in multiple pathways related to muscle development and fat deposition, such as the focal adhesion and MAPK pathways.<h4>Conclusion</h4>Dietary supplementation of AMRP improved muscle growth and development in Angus beef cattle. It also significantly modulated meat quality, possibly altering signaling pathways by influencing key gene expression. Our results provide novel insights into the development of the meat industry and indicate the mechanism through which AMRP regulates muscle development and improves meat quality at the molecular level.

Neurodegenerative diseases encompass a number of very heterogeneous disorders, primarily characterized by neuronal loss and a concomitant decline in neurological function. Examples of this type of clinical condition are Alzheimer's Disease, Parkinson's Disease, Huntington's Disease and Amyotrophic Lateral Sclerosis. Age has been identified as a major risk in the etiology of these disorders, which explains their increased incidence in developed countries. Unfortunately, despite continued and intensive efforts, no cure has yet been found for any of these diseases; reliable markers that allow for an early diagnosis of the disease and the identification of key molecular events leading to disease onset and progression are lacking. Altered adult neurogenesis appears to precede the appearance of severe symptoms. Given the scarcity of human samples and the considerable differences with model species, increasingly complex human stem-cell-based models are being developed. These are shedding light on the molecular alterations that contribute to disease development, facilitating the identification of new clinical targets and providing a screening platform for the testing of candidate drugs. Moreover, the secretome and other promising features of these cell types are being explored, to use them as replacement cells of high plasticity or as co-adjuvant therapy in combinatorial treatments.

Autism spectrum disorder (ASD) is a complex multifactorial neurodevelopmental disorder. Despite extensive research involving genome-wide association studies, copy number variant (CNV) testing, and genome sequencing, the comprehensive genetic landscape remains incomplete. In this context, we developed a systems biology approach to prioritize genes associated with ASD and uncover potential new candidates. A Protein-Protein Interaction (PPI) network was generated from genes associated to ASD in a public database. Leveraging gene topological properties, particularly betweenness centrality, we prioritized genes and unveiled potential novel candidates (e.g., <i>CDC5L</i>, <i>RYBP</i>, and <i>MEOX2</i>). To test this approach, a list of genes within CNVs of unknown significance, identified through array comparative genomic hybridization analysis in 135 ASD patients, was mapped onto the PPI network. A prioritized gene list was obtained through ranking by betweenness centrality score. Intriguingly, by over-representation analysis, significant enrichments emerged in pathways not strictly linked to ASD, including ubiquitin-mediated proteolysis and cannabinoid receptor signaling, suggesting their potential perturbation in ASD. Our systems biology approach provides a promising strategy for identifying ASD risk genes, especially in large and noisy datasets, and contributes to a deeper understanding of the disorder's complex genetic basis.

Malaria remains a major global health problem that has been exacerbated by the impact of the COVID-19 pandemic on health systems. To combat this, the World Health Organization (WHO) has set a target of driving forward research into innovative treatment methods such as new drugs and vaccines. Quinones, particularly 1,4-naphthoquinones, have been identified as promising candidates for the development of antiprotozoal drugs. Herein, we report several methods for the preparation of 2-benzyl-1,4-naphthoquinones. In particular, the silver-catalyzed Kochi-Anderson radical decarboxylation is well suited for the preparation of these compounds. The antiprotozoal activity of all synthesized compounds was evaluated against <i>Plasmodium falciparum</i> NF54 and <i>Trypanosoma brucei rhodesiense</i> STIB900. Cytotoxicity towards L6 cells was also determined, and the respective selectivity indices (SI) were calculated. The synthesized compounds exhibited good antiplasmodial activity against the <i>P. falciparum</i> (NF54) strain, particularly (2-fluoro-5-trifluoromethylbenzyl)-menadione <b>2e</b>, which showed strong efficacy and high selectivity (IC₅₀ = 0.006 µM, SI = 7495). In addition, these compounds also displayed favorable physicochemical properties, suggesting that the benzylnaphthoquinone scaffold may be a viable option for new antiplasmodial drugs.

Anxiety disorders and depression often co-occur and lack broadly available treatments. Gardenia extract significantly associated with treatment of anxiety-like depression. Based on the dose effect hypothesis and previous studies, it is speculated that crocin I, the main component of gardenia, is significantly related to the treatment of anxiety-like depression. The present study aimed to verify the reversal effect of crocin I on chronic corticosterone-induced anxiety-like depression, and to further explore its metabolic process <i>in vivo</i>. Ultimately, a new method for rapid and sensitive detection of trace substances was established. In this study, the rat model of anxiety-like depression was induced by chronic corticosterone. The effects of crocin I were explored by combining behavioral, pathological sections and ELASA data. It is the first time that crocin I can reverse the morphological changes of hippocampus induced by corticosterone in rats. In terms of behavior, crocin Ⅰ can significantly improve the anxiety-like depressive behavior exhibited by model rats in water maze and sugar water preference experiments. It can also repair neuronal cell damage in the Dentate gyrus, CA1, and CA3 areas of the hippocampus. It also regulates the expression levels of monoamine neurotransmitters in the rat brain, thereby exerting an anti-anxiety-like depression effect. Pharmacokinetic analysis was performed to determine the metabolic process <i>in vivo</i>. Further integrating Surface-Enhanced Raman Scattering (SERS) technology, a highly sensitive and rapid detection method for trace substances had been established. It was first discovered that crocin I can reverse the changes in rat hippocampal morphology caused by corticosterone. It was determined that crocin Ⅰ can reverse the anxiety-like depression induced by chronic corticosterone and exert its therapeutic effect by regulating the levels of neurotransmitters in the brain. <i>In vivo</i> pharmacokinetic experiments revealed that crocin Ⅰ could not pass through the intestinal barrier into the blood, but its metabolite crocetin could pass through the intestinal barrier into the blood. Finally, by synthesizing silver nanoparticles, a detection method for trace amounts of the metabolite crocetin in blood samples was established for the first time.The calculated enhancement factor is 4.49 × 10¹¹. The method was stable and reproducible over a week. This series of studies revealed the great potential of crocin I in treating comorbid anxiety and depression. It shortens the distance from theoretical research to clinical application.

<h4>Introduction</h4>CAR T cell therapy provided transformative outcomes for patients with B-cell lymphoma; however, a large fraction of patients remains at risk for relapse, underlying the need to uncover mechanisms of resistance and predictive biomarkers. Herein, we leveraged the ZUMA-7 phase III randomized trial of relapsed/refractory large B-cell lymphoma (LBCL) patients treated with axicabtagene ciloleucel (axi-cel; CD19-targeting CAR T cells) to discover tumor gene expression signatures (GES) associated with outcome.<h4>Methods</h4>With tumor transcriptomics from 134 axi-cel patients, we employed multivariate penalized Cox models analyzing event-free survival (EFS), progression-free survival (PFS), and duration of response (DOR).<h4>Results and discussion</h4>We identified two novel GES, a six-gene/transcript signature (6-GES; CD19, CD45RA, CCL22, KLRK1, SOX11, SIGLEC5) correlated with improved outcome after axi-cel (HR: 0.27, 95% CI: 0.16-0.44 for EFS), representing lymphomas with abundant target antigen (CD19) expression, adhesion molecules, and relatively low immune infiltration mostly composed of cytotoxic lymphocytes (T and NK cells) and DCs, and secondly, a 17-gene/transcript signature (17-GES; CD45RO, BCL2, IL-18R1, TNFSF4 [OX40L], KLRB1 [CD161], KIR3DL2, ITGB8, DUSP5, GPC4, PSMB5, RPS6KB1, SERPINA9, NBN,GLUD1, ESR1, ARID1A, and SLC16A1) correlated with disease progression after axi-cel (HR: 6.12, 95% CI: 3.57-10.50 for EFS), consistent with high immune inflammation and escape mechanisms, such as the upregulation of genes involved in repair of damaged DNA or chromatin remodeling, inhibition of apoptosis, and a metabolically restrictive environment. These signatures did not correlate with outcome in the standard-of-care arm of ZUMA-7 (chemotherapy, followed by transplant) or frontline therapy, supporting their predictive rather than prognostic value. The findings were technically reproduced in a subset of ZUMA-7 samples profiled by RNA-seq (axi-cel, n=124; SOC, n=125). The 6-GES was reduced, whereas the 17-GES was elevated at progression post axi-cel, consistent with the notion that these signatures represent features relevant for response and resistance to CAR T-cell therapy.<h4>Conclusion</h4>Our transcriptomic analysis identified gene expression signatures potentially predictive of outcome with CD19-directed CAR T-cell therapy, and these findings are informative for risk stratification and development of next-generation products.

For more than 15 years, the International Livestock Research Institute (ILRI) has been striving to understand and address One Health challenges at the intersection of livestock, humans, and the environment. We present an overview of ILRI One Health projects implemented with partners across Asia and Africa, reflecting on key learnings and future directions for One Health research and food systems transformation. Drawing on a review of peer-reviewed and grey literature, we analyzed processes and outcomes of ILRI-led and supported initiatives using a realist evaluation framework (context, mechanisms, outcomes), and present insights within select One Health topic areas such as zoonoses, food safety, antimicrobial resistance. Our findings emphasize the need for stronger cross-sectoral collaboration, greater engagement with policymakers to translate research findings into actionable strategies, and the development of adaptable and context-specific interventions.

<h4>Objectives</h4>The identification of methods to improve the endometrial receptivity (ER) is increasingly of interest. The effect of the electromagnetic field associated with Quantum Molecular Resonance (QMR) on ER was investigated here.<h4>Methods</h4>Ishikawa cells were used to evaluate the effects of QMR both on the expression of a group of genes involved in ER, i.e., <i>HOXA10</i>, <i>HOXA11</i>, <i>LIF</i>, <i>ITGB3</i>, and <i>ITGAV</i>, and on cell toxicity. Endometrial samples were obtained from six patients during routine diagnostic procedures, four of which were subsequently used to assess the transcriptional response to QMR through microarray.<h4>Results</h4>Compared to unexposed controls, a single exposure of Ishikawa cells to QMR for 20 min was associated with a significant and power-dependent up-regulation of all the selected ER-related genes up to 8 power units (PU). Repeated exposure to QMR, up to three consecutive days, showed a significant up-regulation of all the selected genes at power values of 4 PU, from day two onwards. Negligible cytotoxicity was observed. Gene set enrichment analysis, on microarray data of endometrial biopsies stimulated for three consecutive days at 4 PU, showed a significant enrichment of specific gene sets, related to the proteasome system, the cell adhesion, the glucocorticoid receptor, and cell cycle pathways.<h4>Conclusions</h4>Our results suggest a possible favorable impact of QMR on ER.

Kratom, an opioid substitute derived from the leaves of <i>Mitragyna speciosa</i>, is commonly used for its opioid-like effects in East Asia and is gaining popularity in the United States. A rare side effect of chronic kratom use, previously noted in the literature, is skin hyperpigmentation. We present a unique case of diffuse, photodistributed hyperpigmentation in a patient with over a decade of kratom use. This case is accompanied by an evaluation of the adverse effect, focusing on its clinical presentation, histopathological findings, and potential mechanism. Additionally, we provide a brief literature review on the topic.

<h4>State the purpose</h4>Obtaining high-quality samples to diagnose streptococcal pharyngitis in pediatric patients is challenging due to discomfort associated with traditional pharyngeal swabs. This may cause reluctance to go to the clinic, inaccurate diagnosis, or inappropriate treatment for children with sore throats. Here, we determined the efficacy of CandyCollect, a lollipop-inspired open-microfluidic pathogen collection device, to capture Group A <i>Streptococcus</i> (GAS) and compare user preference for CandyCollect, conventional pharyngeal swabs, or mouth swabs in children with pharyngitis and their caregivers.<h4>Results</h4>All child participants (30/30) were positive for GAS by qPCR on both the mouth swab and CandyCollect. Caregivers ranked CandyCollect as a good sampling method overall (27/30), and all caregivers (30/30) would recommend CandyCollect for children 5 years and older. Twenty-three of 30 children "really like" the taste and 24/30 would prefer to use CandyCollect if a future test were needed. All caregivers (30/30) and most children (28/30) would be willing to use CandyCollect at home.<h4>Conclusion</h4>All participants tested positive for GAS on all three collection methods (pharyngeal swab, mouth swab, and CandyCollect). While both caregivers and children like CandyCollect, some caregivers would prefer a shorter collection time. Future work includes additional studies with larger cohorts presenting with pharyngitis of unknown etiology and shortening collection time while maintaining the attractive form of the device.

Diabetes involves multi-organ complications that seriously threaten human life and health, and has become a major public health problem of global concern. Unfortunately, clinical management strategies for diabetic complications are still in their "infancy", restricted by a limited understanding of their complex pathological mechanism. As is well established, lipid metabolism disorder is the characteristic pathological factors of diabetes, but the detailed molecular mechanisms driving the progression of multi-organ complications remain obscure. Protein S-acylation (often referred to as S-palmitoylation) is a reversible lipid modification that reversibly binds fatty acids to protein-specific cysteine (Cys) residues through palmitoyl acyl transferases (PATs, also known as DHHCs) and deacylation enzymes, which is involved in the pathological progression of a variety of complex diseases such as cancer, neurological disorders and metabolic syndrome. Notably, recent studies have shown that protein S-acylation drives the progression of diabetes and its multiple complications, and targeted intervention in the protein S-acylation process significantly alleviates the progression of diabetes and its complications, suggesting that protein S-acylation may be a common pathological link and intervention target of diabetes complications. Therefore, this review systematically comprehends the contribution of protein S-acylation to the progression of diabetes and its complications, summarizes the influence of the diabetic environment on S-acylation related enzymes, as well as providing an in-depth analysis of current drugs, measures, and challenges in targeting S-acylation. Finally, the accessibility of targeting protein S-acylation to prevent diabetes and its complications and the focus of future in-depth studies are envisioned, with a view to providing comprehensive and in-depth references and rationale for future novel strategies targeting protein S-acylation to prevent and treat diabetes and its multi-organ complications.

No abstract available.

<h4>Background</h4>RNA methylation is a potential target for cancer therapy, while anoikis, a form of programmed cell death, is linked to cancer metastasis. However, the prognostic and immune significance of RNA methylation- and anoikis-related genes in colorectal cancer (CRC) remains unknown.<h4>Methods</h4>Transcriptomic and clinicopathological data for CRC were obtained from TCGA and the GEO databases. A novel signature was constructed based on RNA methylation- and anoikis-related genes using univariate and multivariate Cox regression as well as LASSO Cox regression methods. CRC patients were stratified into low- and high-risk groups based on this signature. Differences in prognosis, immune infiltration, and drug sensitivity between two groups were analyzed. Finally, immunohistochemistry, western blot, and RT-qPCR were employed to validate the expression of the key gene SERPINE1 in CRC tissues and cells, as well as the effect of FTO on its expression.<h4>Results</h4>We identified 79 differentially expressed RNA methylation-associated anoikis-related genes (RMRARGs) in both cancerous and normal tissues. A signature composed of 9 key genes (BID, FASN, PLK1, CDKN3, MYC, EPHA2, SERPINE1, CD36, PDK4) was established. Kaplan-Meier analysis revealed a poorer prognosis in the high-risk group. Compared to the other three published models, this signature demonstrated superior predictive performance based on the ROC curve analysis. Functional analyses highlighted differences in drug sensitivities and signaling pathways between risk groups. Furthermore, immune analysis results showed that risk score was associated with some immune cells and immune checkpoints. Immunohistochemistry showed high SERPINE1 expression in CRC tissues, with FTO expression positively correlated with SERPINE1. Furthermore, RT-qPCR and western blot indicated FTO knockdown markedly downregulated SERPINE1 levels.<h4>Conclusion</h4>Our findings underscore the prognostic value of this signature in CRC patients and its utility in assessing immune status. Additionally, the m6A demethylase FTO regulates the expression of the anoikis-related gene SERPINE1.

How mixed cryoglobulinemia (MC) affects cancer risk in chronic hepatitis C patients with sustained virologic response (SVR) remains unclear. In a 12-year prospective study, post-SVR MC was assessed every 3‒6 months. Among the 891 SVR patients, 265 (29.7%) had baseline (24 weeks after completing anti-HCV therapy) MC, and the 12-year cancer cumulative incidence was 19.7%. Among the 73 patients who developed cancer, 37 (50.7%) had hepatocellular carcinoma (HCC), with the following associated baseline variables: for cancer, male sex, age and alanine aminotransferase (ALT) levels; for HCC, male sex, age, and cirrhosis; and for non-HCC cancer, rheumatoid factor levels. Among patients with post-SVR HCC, the mean time to HCC was longer in those with than in those without baseline MC (1545.4 ± 276.5 vs. 856.9 ± 115.2 days, <i>p</i> = 0.014). Patients with baseline MC had decreased circulating interleukin-10 (IL-10)-positive B cell (CD19+IL-10+cells/CD19+cells) (31.24 ± 16.14 vs. 40.08 ± 15.42%, <i>p</i> = 0.031), regulatory B cell (Breg) (CD19+CD24hi CD27+cells/CD19+cells) (10.45 ± 7.10 vs. 15.76 ± 9.14%, <i>p</i> = 0.035), IL-10-positive Breg (CD19+CD24hiCD27+IL-10+cells/CD19+cells) (5.06 ± 4.68 vs. 8.83 ± 5.46%, <i>p</i> = 0.015) and HCC-infiltrating Breg (18.6 ± 10 vs. 33.51 ± 6.8%, <i>p</i> = 0.022) ratios but comparable circulating and HCC-infiltrating regulatory T cell ratios relative to patients without baseline MC. In conclusion, old male SVR patients with elevated ALT levels or cirrhosis require intensive monitoring for cancer development, especially HCC. Tailored HCC follow-up is needed for SVR patients according to their baseline MC, which might downregulate Bregs to decelerate HCC development for almost 2 years.

Glioblastoma (GBM) is the most aggressive and lethal type of glioma, characterized by aberrant expression of noncoding RNAs including circular RNAs (circRNAs). CircRNAs may impact cellular processes by interacting with other molecules-like RNA-binding proteins (RBPs). The diagnostic value of circRNA and circRNA/RBP complexes is still largely unknown. To explore circRNA and RBP transcript expression in GBM, we performed and further analyzed RNA-seq data from GBM patients' primary and recurrent tumor samples. We identified circRNAs differentially expressed in primary tumors, the circRNA progression markers in recurrent GBM samples, and the expression profile of RBP genes. Furthermore, we demonstrated the clinical potential of circRNAs and RBPs in GBM and proposed them as stratification markers in de novo assembled tumor subtypes. Additionally, we experimentally validated the subcellular localization of select circRNAs and their interactions with FUS. Subsequently, we showed that circARID1A may play a role in promoting GBM cell proliferation. Overall, we described circRNA-RBP interactions that could play a regulatory role in gliomagenesis and GBM progression and provided a list of molecular players in GBM for further extensive studies.

<h4>Abstract</h4>In recent years, the roles of mitochondrial RNA and its associated human diseases have been reported to increase significantly. Treatments based on mtRNA metabolic processes and nuclear gene mutations are thus discussed. The mitochondrial oxidative phosphorylation process is affected by mtRNA metabolism, including mtRNA production, maturation, stabilization, and degradation, which leads to a variety of inherited human mitochondrial diseases. Moreover, mitochondrial diseases are caused by mitochondrial messenger RNA, mitochondrial transfer RNA, and mitochondrial ribosomal RNA gene mutations. This review presents the molecular mechanisms of human mtRNA metabolism and pathological mutations in mtRNA metabolism-related nuclear-encoded/nonencoded genes and mitochondrial DNA mutations to highlight the importance of mitochondrial RNA-related diseases and treatments.

The ligand Netrin mediates axon guidance through a combination of haptotaxis over short distances and chemotaxis over longer distances.

This study investigates the effects of different treatment sequences involving demineralization, brushing, and remineralization on the calcium and phosphorus (Ca/P) content, surface roughness, and microhardness of dentin specimens. Bovine dentin samples were subjected to the following five treatment conditions: control, demineralization, demineralization followed by remineralization, demineralization followed by brushing then remineralization, and demineralization followed by remineralization then brushing. X-ray fluorescence spectroscopy was then utilized to assess the elementary composition changes, while scanning electron microscopy provided microstructural analyses. Surface roughness and microhardness were also quantified to assess the physical changes in dentin. The control group retained significantly higher Ca/P content compared with all treated groups, indicating that demineralization, regardless of subsequent treatment, leads to a substantial loss of hydroxyapatite. Among the treated groups, those that underwent remineralization immediately after demineralization manifested higher Ca/P retention compared with those that included brushing before remineralization. Additionally, microhardness measurements indicated that post-demineralization brushing negatively affected dentin's microhardness. The sequence of demineralization, brushing, and remineralization treatments significantly affects Ca/P retention in dentin, along with its surface roughness and microhardness. Pre-remineralization brushing diminished mineral recovery, whereas exposure to mineral-rich beverage immediately after demineralization resulted in greater mineral deposition.

Horse domestication revolutionizes human civilization by transforming transportation, agriculture, and warfare patterns. Despite extensive studies on modern domestic horse origins, the intricate demographic history and genetic signatures underlying pony size remain unexplored. Here, a high-quality genome assembly of the Chinese Debao pony is presented, and 452 qualified individuals from 64 horse breeds worldwide are extensively analyzed. The authors' results reveal the conservation of ancient components in East Asian horses and close relationships between Asian horses and Western pony lineages. Genetic analyses suggest an Asian paternal origin for European pony breeds. These pony-sized horses share close genetic affinities, potentially attributed to their early expansion and adaptation to local environments. In addition, promising cis-regulatory elements influencing horse withers height by regulating genes such as RFLNA and FOXO1 are identified. Overall, this study provides insightful perspectives on the dispersal history and genetic determinants underlying body size in ponies, offering broader implications for horse population management and improvement.

Mammalian genomes are folded through the distinct actions of structural maintenance of chromosome (SMC) complexes, which include the chromatin loop-extruding cohesin (extrusive cohesin), the sister chromatid cohesive cohesin and the mitotic chromosome-associated condensins^1-3. Although these complexes function at different stages of the cell cycle, they exist together on chromatin during the G2-to-M phase transition, when the genome structure undergoes substantial reorganization^1,2. Yet, how the different SMC complexes affect each other and how their interactions orchestrate the dynamic folding of the three-dimensional genome remain unclear. Here we engineered all possible cohesin and condensin configurations on mitotic chromosomes to delineate the concerted, mutually influential action of SMC complexes. We show that condensin disrupts the binding of extrusive cohesin at CCCTC-binding factor (CTCF) sites, thereby promoting the disassembly of interphase topologically associating domains (TADs) and loops during mitotic progression. Conversely, extrusive cohesin impedes condensin-mediated mitotic chromosome spiralization. Condensin reduces peaks of cohesive cohesin, whereas cohesive cohesin antagonizes condensin-mediated longitudinal shortening of mitotic chromosomes. The presence of both extrusive and cohesive cohesin synergizes these effects and inhibits mitotic chromosome condensation. Extrusive cohesin positions cohesive cohesin at CTCF-binding sites. However, cohesive cohesin by itself cannot be arrested by CTCF molecules and is insufficient to establish TADs or loops. Moreover, it lacks loop-extrusion capacity, which indicates that cohesive cohesin has nonoverlapping functions with extrusive cohesin. Finally, cohesive cohesin restricts chromatin loop expansion mediated by extrusive cohesin. Collectively, our data describe a three-way interaction among major SMC complexes that dynamically modulates chromatin architecture during cell cycle progression.

Bone regeneration is one of the most effective methods for treating bone defects. In this work, tricarboxylic cellulose/sodium alginate loaded with hydroxyapatite (HA) and/or graphene oxide (GO) was coagulated by calcium ions to create beads as scaffolds. In the first, cellulose was oxidized to water-soluble tricarboxylic cellulose (TCC) by 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), periodate, and chlorite oxidation. HA was extracted from eggshells via microwave treatment, and GO was synthesized using the Hummer method. The structural behavior of the formed beads was meticulously investigated through various characterization techniques such as Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM). The SEM images confirmed the formation of particles of micrometric size without any specific morphology. Incorporating GO or HA does not affect the morphologies of the materials on the micrometric scale. The cytocompatibility of different bead preparations was studied on murine mesenchymal stem cells. Moreover, the swellability in water and biodegradability by cellulase enzyme of prepared beads were studied. The results show that the prepared beads may be promising for bone tissue engineering.

Protein-protein interactions (PPIs) are ubiquitous in biology, yet a comprehensive structural characterization of the PPIs underlying cellular processes is lacking. AlphaFold-Multimer (AF-M) has the potential to fill this knowledge gap, but standard AF-M confidence metrics do not reliably separate relevant PPIs from an abundance of false positive predictions. To address this limitation, we used machine learning on curated datasets to train a structure prediction and omics-informed classifier (SPOC) that effectively separates true and false AF-M predictions of PPIs, including in proteome-wide screens. We applied SPOC to an all-by-all matrix of nearly 300 human genome maintenance proteins, generating ∼40,000 predictions that can be viewed at predictomes.org, where users can also score their own predictions with SPOC. High-confidence PPIs discovered using our approach enable hypothesis generation in genome maintenance. Our results provide a framework for interpreting large-scale AF-M screens and help lay the foundation for a proteome-wide structural interactome.

<h4>Purpose</h4>The complexity and heterogeneity of the tumor immune microenvironment (TIME) are linked to the development and poor prognosis of hepatocellular carcinoma (HCC). However, the cell type within the TIME that is most closely associated with HCC development remains unclear. Herein, we aimed to identify cell clusters that significantly contribute to HCC development and their underlying mechanisms.<h4>Method and results</h4>Using single-cell RNA sequencing (scRNA-seq), we analyzed changes in the TIME of normal and tumor tissues, identifying plasma cells as the key cluster in HCC development. Based on plasma cell-related genes (PCRGs), we constructed and validated an eight-gene prognostic model (ST6GALNAC4, SEC61A1, SSR3, RPN2, PRDX4, TRAM1, SPCS2, CD79A) using internal and external datasets and a nomogram. Functional enrichment, miRNA network construction, and transcriptional regulation analyses were performed to explore underlying mechanisms. TIDE scores and the GDSC database were used to predict immunotherapy and chemotherapy sensitivity in different risk groups. Finally, SSR3's biological function was validated in vitro in HCC cell lines.<h4>Conclusion</h4>Plasma cells are key clusters in HCC development. A prognostic model based on the PCRGs can accurately predict the prognosis of patients with HCC and guide clinical treatment.

<h4>Background</h4>Metabolic dysfunction-associated fatty liver disease (MAFLD) is a global concern. The gut microbiota and hepatic fatty acid beta-oxidation have been shown to be important factors in the development of MAFLD. Independently, exercise and berberine can significantly ameliorate symptoms of MAFLD, although the specific mechanism is not clear; moreover, it is not known whether the combination of exercise and berberine produces a better therapeutic effect.<h4>Methods</h4>Over an experimental period of 17 weeks, the effects of exercise, berberine, and a combined (exercise/berberine) intervention on the composition of the gut microbiota and the expression of hepatic fatty acid beta-oxidation related proteins were examined. The mice were divided into five treatment groups: CON (control group, n = 10), HFD (high-fat diet, n = 10), HFE (high-fat diet + exercise, n = 10), HFB (high-fat diet + berberine, n = 10), and HBE (high-fat-diet + exercise + berberine, n = 10). The dose of BBR administered for oral gavage was 300 mg/kg, once per day, for 8 weeks. Mice were subjected to treadmill exercise, 5 days per week for 8 weeks, and the intensity was increased gradually.<h4>Results</h4>Serological and histopathological results showed that exercise, berberine and a combined (exercise/berberine) intervention effectively improved liver lipid accumulation caused by a high-fat diet. Analysis of 16S rRNA showed that the three interventions restored the species and number of gut microbiota in MAFLD mice. The functional prediction of gut microbiota revealed significant differences in beta-oxidation-related units among groups. Simultaneously, exercise and berberine intervention regulated the expression of hepatic fatty acid beta-oxidation-related proteins ACOX1, HMGCS2, and CPT-1α, with the combined intervention having a more significant effect than each intervention alone.<h4>Conclusion</h4>Our findings indicate that exercise and berberine ameliorate MAFLD by regulating the gut microbiota and hepatic fatty acid beta-oxidation, suggesting that their combination may be a potential therapy for MALFD.

<b>Background:</b> Graft versus host disease (GVHD) and the graft versus tumor (GVT) effect after allogeneic hematopoietic cell transplantation (allo-HCT) result from complex interactions between the donor immune system and the recipient environment. High-temporal longitudinal monitoring might be necessary to identify triggering events of GVHD and GVT and to intercept these events before their occurrence. But it would require an overall considerable amount of blood by venipuncture, which is unfeasible in such a fragile population. <b>Methods:</b> In this study, we implemented a targeted multiplex microfluidics q-PCR-based transcriptional fingerprint assay (TFA) on 50 µL of blood collected by a simple fingerstick to evaluate post-allo-HCT systemic immune perturbations associated with the development of GVHD. Fluctuations of a panel of 264 genes were measured in 31 allo-HCT patients by frequent (weekly or biweekly) analysis of 50 µL serial blood samples. Cross-sectional and longitudinal analyses correlated with detailed clinical annotations were performed. <b>Results:</b> Signatures of neutrophil activation and interferon (IFN) characterized the onset of acute GVHD, while an ongoing cytotoxic response was modulated in chronic mild GVHD and protein-synthesis and B-cell-related signatures characterized late acute/overlap GVHD. An unexpected erythroid signature distinguished patients with acute and mild chronic GVHD. <b>Conclusions:</b> Our micro-invasive approach unveiled the molecular heterogeneity of GVHD and identified hierarchically important biological processes conducive to different forms of GVHD. These findings increase our understanding of GVHD and reveal potentially targetable alterations. This approach might be implemented clinically to intercept GVHD before its occurrence and to modulate therapeutic interventions accordingly.

The somatic cell count (SCC) is widely used to assess milk quality and diagnose intramammary infections. Several whey proteins have been shown to correlate significantly with SCC and are considered potential indicators of udder health. However, the relationship between milk whey proteins and SCC has not been fully elucidated. In this study, milk samples were grouped into five categories based on SCC levels. Subsequently, whey proteins were identified using a label-free proteomics approach, and the differential abundance of proteins was validated through a selected reaction monitoring (SRM) method. The levels of various proteins, including azurocidin 1 and kininogen-2, exhibited an increase, whereas topoisomerase I, tropomyosin-1, and desmin showed a significant decrease depending on the SCCs. Principal component analysis unveiled that these proteins contributed to the developmental alterations in milk proteins. A majority of these differentially abundant proteins were associated with response to stimulus, localization, and defense response. Our results provide fundamental information on the SCC that can be utilized for evaluating milk quality and serve as potential indicators for detecting intramammary infections.

Selenium (Se) is an essential micronutrient, recognized for its role in cellular redox systems and its therapeutic potential in cancer treatment. Organic selenium compounds, particularly selenocystine (SeCys), have demonstrated anticancer efficacy due to the ability to induce apoptosis and enhance the effects of chemotherapy agents. Recent studies have shown that SeCys exhibits selective toxicity against cancer cells while sparing normal cells. Unfortunately, its clinical application is limited by stability and solubility concerns. A possible solution to overcome these hurdles comes from recent advances in functionalized nanomaterials. In this study, we investigate the possible incorporation of SeCys with hydroxyapatite nanoparticles (HASeCys) via various methods (adsorption, co-precipitation, and co-precipitation through thermal decomplexation), resulting in the formation of nanocomposites with elemental selenium. The highest elemental selenium yield was achieved with a thermal decomplexing co-precipitation, highlighting the influence of synthesis parameters on Se allotrope formation. Finally, as a preliminary investigation, the HASeCys samples were tested on a panel of cancer cell lines, showing an interesting activity when the hydroxyapatite nanocrystals were functionalized with both crystalline gray and amorphous red selenium.

Bone infection remains a challenging condition to fully eradicate due to its intricate nature. Traditional treatment strategies, involving long-term and high-dose systemic antibiotic administration, often encounter difficulties in achieving therapeutic drug concentrations locally and may lead to antibiotic resistance. Bone cement, serving as a local drug delivery matrix, has emerged as an effective anti-infective approach validated in clinical settings. Calcium phosphate cements (CPCs) have garnered widespread attention and application in the local management of bone infections due to their injectable properties, biocompatibility, and degradability. The interconnected porous structure of calcium phosphate particles, not only promotes osteoconductivity and osteoinductivity, but also serves as an ideal carrier for antibacterial agents. Various antimicrobial agents, including polymeric compounds, antibiotics, antimicrobial peptides, therapeutic inorganic ions (TIIs) (and their nanoparticles), graphene, and iodine, have been integrated into CPC matrices in numerous studies aimed at treating bone infections in diverse applications such as defect filling, preparation of metal implant surface coatings, and coating of implant surfaces. Additionally, for bone defects and nonunions resulting from chronic bone infections, the utilization of calcium phosphate-calcium sulfate composite multifunctional cement loaded with antibacterial agents serves to efficiently deal with infection, stimulate new bone formation, and attain an optimal degradation rate of the bone cement matrix. This review briefly delves into various antibacterial strategies based on calcium phosphate cement for the prevention and treatment of bone infections, while also discussing the application of calcium phosphate-calcium sulfate composites in the development of multifunctional bone cement against bone infections.

This case report describes a geriatric male patient with myasthenia gravis (MG) secondary to giant thymoma, presenting with progressive muscle weakness and ptosis. The diagnosis of MG was confirmed through pathology, imaging, and laboratory evaluations. Considering the significant surgical risks associated with the giant thymoma, adjuvant chemotherapy was initiated. Unfortunately, 2 weeks following chemotherapy, the patient developed acute respiratory failure and sudden loss of consciousness. Emergency endotracheal intubation was performed, and he was then transferred to the intensive care unit (ICU) and treated with immunoglobulin, plasmapheresis, prednisone, and pyridostigmine. During ICU hospitalization, the patient developed severe lower limb edema accompanied by increased skin temperature, particularly on the left side. Ultrasound imaging confirmed extensive thrombosis in the left iliac and femoral veins, with thrombosis involving 50%-67% of the venous lumen. To prevent the risk of pulmonary embolism (PE), an inferior vena cava filter was implanted, and low-molecular weight heparin (LMWH) was prescribed for anticoagulation. Unfortunately, the patient later experienced intermittent melena and heparin-induced thrombocytopenia (HIT), with hemoglobin levels decreasing to 55 g/L and platelet counts decreasing to 57 × 10⁹/L. Given the adverse events associated with LMWH, sulodexide (SDX) was substituted as a novel anticoagulant with multiple benefits, including reduced thrombosis and bleeding risk, anti-inflammatory effects, and vascular endothelium protection. SDX demonstrated excellent efficacy and safety, with no adverse effects observed during the 3-year follow-up period. In conclusion, SDX should be considered an ideal potential option for long-term anticoagulation in patients with complex conditions such as MG with both thrombotic and bleeding risks.

<h4>Introduction</h4>The increasing resistance of <i>Candida albicans</i> (<i>C. albicans</i>) to conventional antifungal drugs poses a great challenge to the clinical treatment of infections caused by this yeast. Drug combinations are a potential therapeutic approach to overcome the drug- resistance of <i>C. albicans</i>. This study explored the synergistic effects of amantadine hydrochloride (AMH) combined with azole antifungal drugs against drug-resistant <i>C. albicans in vitro</i> and <i>in vivo</i>.<h4>Methods</h4>The <i>in vitro</i> sensitivity of <i>Candida</i> spp. to drugs was determined by the microdilution method. The effect of drugs on the efflux pump activity of <i>C. albicans</i> was determined by the rhodamine 6G tracer method. The egg yolk agar plate method was used to determine the activity of extracellular phospholipase, a <i>C. albicans</i> virulence factor. The <i>Galleria mellonella</i> model of <i>C. albicans</i> infection was used to test the <i>in vivo</i> efficacy of the combination therapy.<h4>Results</h4><i>In vitro</i> experiments showed that combinations of AMH with azole antifungal drugs had synergistic antifungal effects on planktonic cells of drug-resistant <i>C. albicans</i>, with fractional inhibitory concentration index values of <0.5. The <i>in vivo</i> synergistic effects and mechanism of drug combinations with AMH were further studied using fluconazole (FLC) as a representative azole antifungal drug. <i>In vivo</i>, <i>G. mellonella</i> larvae were used to evaluate the antifungal efficacy of AMH +FLC. AMH + FLC treatment increased the survival rate of larvae infected with drug-resistant <i>C. albicans</i> and reduced tissue invasion. Studies of the mechanism of synergy showed that AMH inhibited drug efflux pump activity in drug-resistant <i>C. albicans</i>, and that AMH + FLC synergistically inhibited early biofilms and the extracellular phospholipase activity of drug-resistant <i>C. albicans</i>.<h4>Conclusion</h4>This study provides strong evidence that combinations of non-antifungal drugs and antifungal drugs can effectively overcome drug-resistant <i>C. albicans</i> infection. Both AMH and FLC are FDA-approved drugs, eliminating concerns about safety. Our findings provide a foundation for further clinical antifungal research.

Clear cell renal cell carcinoma (ccRCC) is the typical pathological subtype of renal cell carcinoma (RCC), representing about 80% of RCC. Reprogramming of lipid metabolism is one of the nonnegligible pathogeneses in ccRCC. Currently, the underlying regulatory mechanisms of lipid metabolism in ccRCC remain inadequately understood. In this study, we performed bioinformatics analyses and experiments both <i>in vivo</i> and <i>in vitro</i> to explore the biological functions and specific mechanisms of the ligand dependent nuclear receptor corepressor LCOR in ccRCC. Mechanistically, RUNX1 was a transcriptional suppressor of PLCL1, LCOR could interact with RUNX1 to relieve RUNX1-mediated repression of PLCL1, leading to increased PLCL1 expression, which, in turn, inhibited the tumor progression and lipid accumulation in ccRCC. Furthermore, PLCL1 decreased lipid accumulation through UCP1-mediated lipid browning and facilitated tumor apoptosis by activating p38 phosphorylation. In conclusion, the LCOR-RUNX1-PLCL1 axis provides a novel molecular mechanism underlying the progression and lipid storage of ccRCC. LCOR modulation represents a potential therapeutic strategy for the treatment in ccRCC.

Olfactomedin-like protein 3 (OLFML3), belonging to olfactomedin (OLF) protein family, has poorly defined functions. Recent studies have reported the functions of OLFML3 in anti-viral immunity and tumorigenesis. In this study, we investigated the roles of OLFML3 in macrophages. In LPS- or <i>Pseudomonas aeruginosa</i>-induced acute lung injury (ALI) mouse model, OLFML3 depletion exacerbated inflammatory response, leading to reduced survival. OLFML3 achieved the <i>in vivo</i> activity by regulating macrophage phagocytosis and migration. Mass spectrometry analysis revealed immunoresponsive gene 1 (IRG1) as an OLFML3-interacting protein. IRG1 is a mitochondrial decarboxylase that catalyzes the conversion of <i>cis</i>-aconitate to itaconate, a myeloid-borne mitochondrial metabolite with immunomodulatory activities. Further investigation showed that OLFML3 could prevent LPS-induced mitochondrial dysfunction in macrophages by maintaining the homeostasis of mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (mtROS) and itaconate-related metabolites. In-depth protein-protein interaction studies showed that OLFML3 could promote IRG1 mitochondrial localization via a mitochondrial transport protein, apoptosis inducing factor mitochondria associated 1 (AIFM1). In summary, our study showed that OLFML3 could facilitate IRG1 mitochondrial localization and prevent LPS-induced mitochondrial dysfunction in macrophages.

<b>Background:</b> Circular RNAs (circRNAs) exhibit differential expression in cardiac hypertrophy; however, their functions and mechanisms remain largely unexplored. This study aimed to determine the involvement of circRNAs in the pathogenesis of myocardial hypertrophy. <b>Methods:</b> A mouse model of cardiac hypertrophy was established using transverse aortic constriction (TAC) and differentially expressed circRNAs were identified via high-throughput sequencing. To facilitate gene overexpression or knockdown, related viruses were injected into myocardial tissues of the mice. Cardiomyocyte hypertrophy was assessed using quantitative real-time PCR and immunofluorescence staining. RNA immunoprecipitation, RNA pull-down assay and fluorescence <i>in situ</i> hybridization were conducted to confirm the interaction between circRNAs and proteins. Protein expression and degradation were evaluated using cycloheximide-chase assay, immunoprecipitation, and western blotting. <b>Results:</b> Cardiac hypertrophy-associated circRNA (CHACR) was significantly downregulated in myocardial tissues from TAC mice. CHACR can attenuate cardiac hypertrophy through upregulating carnitine palmitoyltransferase-1b (CPT1b) expression. Mechanistically, CHACR directly interacted with CPT1b and decreased its protein degradation by inhibiting the ubiquitin-proteasome pathway to increase its expression in cardiomyocytes. Moreover, CPT1b overexpression decreased L-carnitine levels and inhibited the Jak2/Stat3 signaling pathway, which was associated with the pathogenesis of myocardial hypertrophy. <b>Conclusions:</b> CHACR attenuated cardiomyocyte hypertrophy by facilitating the expression of CPT1b, which plays a role in regulating the Jak2/Stat3 pathway via L-carnitine. CHACR may thus be a potential therapeutic target for pathological myocardial hypertrophy.

<b>Rationale:</b> Neurons producing Gonadotropin-Releasing Hormone (GnRH) are essential for human reproduction and have to migrate from nose to brain during prenatal life. Impaired GnRH neuron biology results in alterations of the reproductive axis, including delayed puberty and infertility, with considerable effects on quality of life and metabolic health. Although various genes have been implicated, the molecular causes of these conditions remain elusive, with most patients lacking a genetic diagnosis. <b>Methods:</b> GnRH neurons and non-GnRH cells were FACS-isolated from mouse embryo microdissections to perform high-resolution transcriptomic profiling during mouse embryonic development. We analyzed our dataset to reveal GnRH neuron molecular identity, gene expression dynamics, and cell-to-cell communication. The spatial context of candidate genes was validated using <i>in situ</i> hybridization and spatial transcriptomic analysis. The possible links with human reproduction in health and disease were explored using enrichment analysis on GWAS data and analyzing the genetic burden of patients with congenital GnRH deficiency. <b>Results:</b> GnRH neurons undergo a profound transcriptional shift as they migrate from the nose to the brain and display expression trajectories associating with distinct biological processes, including cell migration, neuronal projections, and synapse formation. We revealed a timely and spatially restricted modulation of signaling pathways involving known and novel molecules, including Semaphorins and Neurexins, respectively. A particular set of genes, whose expression in GnRH neurons timely rises in late developmental stages, showed a strong association with GWAS genes linked with human reproductive onset. Finally, some of the identified trajectories harbor a diagnostic potential for congenital hypogonadism. This is supported by genetic analysis in a large cohort of patients affected by congenital GnRH deficiency, revealing a high mutation burden in patients compared to healthy controls. <b>Conclusion:</b> We charted the landscape of gene expression dynamics underlying murine GnRH neuron embryonic development. Our study highlights new genes in GnRH neuron development and provides novel insights linking those genes with human reproduction.

<h4>Background</h4>Delayed cord clamping (DCC) has the potential to alleviate respiratory distress by augmenting blood volume and oxygenation, although there is currently a lack of direct evidence to support this. Late preterm and early term infants born via elective cesarean section (CS) are known to be more vulnerable to the neonatal respiratory distress (NRD). This study was designed to examine the effect of DCC on NRD of these infants.<h4>Methods</h4>Conducted from January 1, 2019 to January 31, 2024 at Shanghai First Maternity and Infant Hospital, this single-centre, phase Ⅲ, open-label randomised controlled trial included newborns delivered via elective CS between 34⁺⁰ and 38⁺⁶ weeks of gestation. Participants were excluded if fetus had suspected or confirmed congenital malformations, metabolic diseases, intrauterine growth restriction, late fetal heart rate deceleration or fetal distress. Pregnant women and their infants were randomised into immediate cord clamping (ICC) within 10 s of birth or DCC for 60 s and stratified by late preterm or early term. The primary outcome was the incidence of NRD which was defined as requiring oxygen or airway pressure support within the first 24 h of life. This study was approved from the Ethics Committee of Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University (KS 18126, KS1947). Chinese Clinical Trial Registry (ChiCTR1800017865), registered on August 18th, 2018.<h4>Findings</h4>Of 2610 randomised women, 1418 neonates were included in the DCC group and 1419 in the ICC group. The mean maternal age for both groups was 33 (4) years, and all mothers were of Han ethnicity. The mean gestational age of the neonates was 37.9 (0.9) weeks in both groups. NRD occurred in 119 (8.4%) in DCC versus 135 (9.5%) in ICC (Adjusted Relative Risk [aRR] 0.93, 95% CI 0.75-1.14). There were no significant differences in infant and maternal adverse events such as low Apgar score (aRR 0.74, 95% CI 0.25-2.19), hypothermia (aRR 1.00, 95% CI 0.89-1.12), hypoglycemia (aRR 1.04, 95% CI 0.77-1.38), maternal intrapartum massive bleeding (aRR 0.96, 95% CI 0.76-1.19), or the requirement for transfusion (aRR 0.34, 95% CI 0.10-1.15).<h4>Interpretation</h4>Delayed cord clamping was safe for both mothers and infants in late preterm and early term delivered by elective cesarean section, while it did not reduce the risk of early respiratory diseases.<h4>Funding</h4>This trial was funded by Shanghai Municipal Health Commission, China in 2019 (201940140) and National Natural Science Foundation of China in 2022 (82204047).

<h4>Background</h4>Hepatocellular carcinoma (HCC) is a prevalent malignant tumor, and the current treatment methods exhibit various limitations. In recent years, the role of mitochondrial permeability transition-driven necrosis-related genes (MPT-DNRGs) in the pathogenesis and progression of severe diseases, particularly tumors, has garnered significant attention. This study aimed to identify new targets and concepts for MPT-DNRG-targeted therapy in HCC.<h4>Methods</h4>In this study, we utilized HCC-related datasets and MPT-DNRGs to identify differentially expressed genes (DEGs) between HCC patients and control groups. By conducting a cross-analysis of the results of DEGs and MPT-DNRGs, we screened candidate genes. Subsequently, univariate Cox regression and least absolute shrinkage and selection operator (LASSO) regression analysis methods were employed to identify prognostic genes, which were used to construct a risk model and calculate individual risk scores for HCC patients. Additionally, we performed univariate and multivariate Cox regression analyses to identify independent prognostic factors and constructed a column chart based on these factors to predict the survival probability of HCC patients. Furthermore, gene set enrichment analysis (GSEA), the immune microenvironment, chemotherapy drugs, and the expression of prognostic genes between the two groups were analyzed. Finally, the expression of these prognostic genes was further confirmed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technology.<h4>Results</h4>In this study, we identified 8,515 DEGs between HCC and control samples. By performing intersection analysis between DEGs and MPT-DNRGs, we pinpointed 15 candidate genes. Subsequently, through univariate Cox regression and LASSO regression analysis, we identified six genes (<i>LMNB2</i>, <i>LMNB1</i>, <i>BAK1</i>, <i>CASP7</i>, <i>LMNA</i>, and <i>AKT1</i>) that were significantly associated with overall survival (OS) in patients. Based on the median risk score, we categorized HCC patients into high-risk and low-risk groups. Kaplan-Meier (KM) survival analysis results demonstrated a significant difference in OS between the two groups, which was further validated through additional assessment. Furthermore, we constructed a nomogram to predict the survival probability of HCC patients. Moreover, GSEA revealed a crucial correlation between these genes and HCC, and highlighted a close association between risk scores and regulatory T cells. We also identified four chemotherapy drugs related to HCC. Finally, in both the training and validation cohorts, <i>LMNB2</i>, <i>LMNB1</i>, and <i>LMNA</i> exhibited high expression levels in tumor samples. Further validation using RT-qPCR confirmed that the expression of all prognostic genes was significantly higher in HCC group compared to the control group.<h4>Conclusions</h4>This study explored six prognostic genes (<i>LMNB2</i>, <i>LMNB1</i>, <i>BAK1</i>, <i>CASP7</i>, <i>LMNA</i> and <i>AKT1</i>) associated with MPT-DNRGs in HCC, which provides a reference for further research on HCC.

<h4>Background</h4>Pyroptosis has been recently identified as a hallmark of cancer biology; however, the potential of pyroptosis-related genes (PRGs) as prognostic markers has not been fully elucidated in hepatocellular carcinoma (HCC). The aim of this study was to develop a PRG-associated risk signature for prediction prognosis in patients with HCC.<h4>Methods</h4>We identified 35 PRGs from the published literature, and pyroptosis subtypes were identified through bioinformatics methods. The risk score model was established by applying least absolute shrinkage and selection operator (LASSO) Cox regression method in The Cancer Genome Atlas (TCGA) cohort and validated in International Cancer Genome Consortium (ICGC) datasets. Additionally, immune infiltration, enriched pathways, and genomic alterations were compared between the high- and low-risk score subgroups. Finally, a nomogram containing the pyroptosis risk score and other prognosis-related clinical factors was developed for predicting the overall survival of patients with HCC.<h4>Results</h4>Based on the expression profile of PRGs, we determined two pyroptosis-related subtypes (cluster A and cluster B) of HCC associated with different immune characteristics and significantly different prognoses. The risk score model showed that upregulation of <i>GPX4, CASP8, NOD2,</i> and <i>GSDME</i> was associated with poor overall survival (OS), while high expression of <i>NLRP6</i> was associated with good prognosis. Compared with group with a lower risk score, the group with a high risk score had worse prognosis (P<0.001) and a high level of immune cell infiltration. Functional analysis indicated that the highly expressed genes in the high-risk group were mainly enriched in various signaling pathways, while the genes with low expression in the high-risk group were mainly enriched in different biochemical metabolic translations. Genomic alterations in high-risk and low-risk populations suggested that mutations in the <i>TP53</i> gene are highly associated with pyroptosis in patients with HCC. A nomogram including risk score and TNM stage demonstrated good prognostic ability in predicting 1-year, 3-year, and 5-year OS.<h4>Conclusions</h4>We developed and verified a prognostic risk model based on PRGs for patients with HCC, which may provide a robust tool for predicting outcomes in this setting.

<h4>Background</h4>Periampullary carcinoma (PAC) is a relatively rare but highly aggressive malignancy, posing challenges to the determination of the optimal therapeutic approach. The objective of this study was to clarify the potential of histopathological typing in guiding chemotherapy selection for patients with advanced PAC and to characterize the distinct molecular features, underlying functional changes, and regulatory mechanisms associated with the different subtypes.<h4>Methods</h4>We conducted a retrospective analysis of clinical data from patients with advanced PAC admitted to the Oncology Department of Xijing Hospital between January 2015 and May 2022. These patients received first-line chemotherapy with either FOLFOX (folinic acid, fluorouracil, and oxaliplatin) or gemcitabine-based regimens. Certain patients were divided into the pathological typing group and the control group. The pathological typing group received subtype-specific chemotherapy regimens, while the control group received chemotherapy regimens based on the primary tumor site. We compared the median progression-free survival (PFS) and overall survival (OS) between the two groups. Using publicly available databases (GSE60980), we conducted differential gene screening, enrichment analysis, and immune cell infiltration assessment. A protein-protein interaction (PPI) network was constructed based on the differentially expressed genes, resulting in the identification of 60 node genes. Subsequently, a core gene selection using the least absolute shrinkage and selection operator (LASSO) regression machine learning algorithm was performed to identify the key genes specific to PAC-PB subtype.<h4>Results</h4>The pathological typing group consisted of 46 patients, with 26 classified as the PB subtype and 20 as the IN subtype, while the control group comprised 40 patients. Compared to those in the control group, patients in the pathological typing group demonstrated significant improvements in overall response rate (20.5% <i>vs</i>. 12.9%; P=0.04), median PFS (8.1 <i>vs</i>. 5.4 months; P=0.04), and median OS (34 <i>vs</i>. 25.9 months; P=0.02). Multivariate Cox regression analysis revealed that pathological typing independently influenced PFS [hazard ratio (HR) =0.20, 95% confidence interval (CI): 0.10-0.44; P=0.009] and OS (HR =0.21, 95% CI: 0.17-0.71; P=0.02). Using a publicly available PAC cohort (GSE60980), we selected 154 differentially expressed genes, which were significantly enriched in signaling pathways related to the cell cycle, fibroblasts, and epithelial-mesenchymal transition. Analysis of immune cell infiltration indicated a significant increase in the abundance of fibroblast cells and a significant decrease in that of B cells and γδ T cells in the PAC-PB subtype. Furthermore, we identified core genes specific to the PAC-PB subtype and used them to construct a PAC-PB diagnostic model.<h4>Conclusions</h4>Pathologic typing-guided individualized chemotherapy resulted in prolonged survival for patients with advanced PAC. The PB and IN subtypes of PAC exhibit distinct molecular regulatory mechanisms and immune infiltration microenvironments. These findings underscore the importance of considering subtype-specific factors in the development of a PAC-PB diagnostic model.

<i>Background and Objectives</i>: Alopecia areata (AA) is a common cell-mediated autoimmune disease of the hair follicle that results in hair loss patches, affecting males and females of all ages and ethnicities. Although its etiology is not fully understood, AA is hypothesized to have a multifactorial basis with a strong genetic association. This study aims to replicate the genetic association of several risk loci in the Jordanian population for the first time. <i>Materials and Methods</i>: Genomic DNA samples of 152 patients with AA and 150 control individuals were extracted from EDTA blood tubes collected from dermatology clinics, in addition to the clinical data of participants. Genetic sequencing of the 21 targeted risk loci was carried out using the Sequenom MassARRAY^® system (iPLEX GOLD), and the results were statistically analyzed using the Statistical Package for the Social Sciences. <i>Results</i>: The results compared the distribution of alleles and genotypes and the association between control individuals and AA patients. However, our results do not support a significant association of all of the 21 SNPs in our AA cohort (<i>p</i> > 0.05). <i>Conclusions:</i> Our data emphasize that AA has a varied genetic component between ethnic groups and suggest that other additional environmental and psychological triggers may be involved.

In recent years, ferroptosis, as an emerging modality of programmed cell death, has captured significant attention within the scientific community. This comprehensive review meticulously canvasses the pertinent literature of the past few years, spanning multiple facets. It delves into the intricate mechanisms underpinning ferroptosis, tracks the evolution of its inducers and inhibitors, and dissects its roles in a diverse array of diseases, as well as the resultant therapeutic implications. A profound exploration is conducted of the functional mechanisms of ferroptosis-related molecules, intracellular pathways, metabolic cascades, and signaling transduction routes. Novel ferroptosis inducers and inhibitors are introduced in detail, covering their design blueprints, synthetic methodologies, and bioactivity profiles. Moreover, an exhaustive account is provided regarding the involvement of ferroptosis in malignancies, neurodegenerative disorders, cardiovascular ailments, and other pathologies. By highlighting the pivotal status and potential therapeutic regimens of ferroptosis in various diseases, this review aspires to furnish a thorough and profound reference framework for future investigations and clinical translations in the ferroptosis domain.

<b>Objectives:</b> Triple-negative breast cancer (TNBC) is an aggressive cancer subtype with limited treatment options due to the absence of estrogen, progesterone receptors, and HER2 expression. This study examined the impact of DNA methylation and demethylation markers in tumor tissues on TNBC patients' response to neoadjuvant chemotherapy (NACT) and analyzed the correlation between 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) and clinicopathological characteristics, offering new insights into the predictive value of these epigenetic markers. <b>Methods:</b> The study included 53 TNBC female patients, 19 of whom received neoadjuvant chemotherapy (NACT) before surgery. Global DNA methylation and demethylation levels were quantified using an ELISA-based method to measure 5-mC and 5-hmC content in DNA isolated from pre-treatment biopsy samples (in patients undergoing NACT) and postoperative tissues (in patients without NACT). <b>Results:</b> In patients who received NACT, those with disease progression had significantly higher pretreatment levels of 5-hmC (<i>p</i> = 0.028) and a trend toward higher 5-mC levels (<i>p</i> = 0.054) compared to those with pathological complete response, partial response, or stable disease. Higher 5-mC and 5-hmC levels were significantly associated with higher tumor grade (<i>p</i> = 0.039 and <i>p</i> = 0.017, respectively). Additionally, a positive correlation was observed between the Ki-67 proliferation marker and both 5-mC (r_S = 0.340, <i>p</i> = 0.049) and 5-hmC (r_S = 0.341, <i>p</i> = 0.048) levels in postoperative tissues. <b>Conclusions:</b> Our study highlights the potential of global DNA methylation and demethylation markers as predictors of tumor aggressiveness and chemotherapy response in TNBC. Further research in larger cohorts is necessary to validate these markers' prognostic and predictive value.

<h4>Background</h4>Phenotyping and genotyping of cardiomyopathies are becoming increasingly important. We describe a rare form of syndromic dilated cardiomyopathy, years after the initial diagnosis of intellectual disability and hypergonadotropic hypogonadism, known as Najjar-Malouf syndrome or cardiogenital syndrome.<h4>Case summary</h4>A 45-year-old woman was referred for inpatient treatment because of progressive dyspnoea, chest tightness, and ankle oedema. She had a diagnosis of undefined psychomotor development delay and hypergonadotropic hypogonadism, for which she has been receiving hormone substitution treatment since she was 19 years old. The physical examination revealed signs of congestion as well as a right convex thoracic scoliosis and hyperkyphosis. Cardiac biomarkers were elevated, and echocardiography showed severe dilated cardiomyopathy with impaired systolic function, severe pulmonary hypertension, and secondary mitral regurgitation. Cardiac magnetic resonance imaging showed a severely dilated left ventricle with severely reduced left ventricular ejection fraction (20%), thinned myocardium, and late-gadolinium enhancement predominantly in the septum. Genetic screening for dilatative cardiomyopathy-associated genes revealed no mutations, in particular, no mutation of the lamin A/C (LMNA) gene. The patient progressed to heart failure with severely reduced ejection fraction 26 years after diagnosis of psychomotor development delay and hypergonadotropic hypogonadism.<h4>Discussion</h4>The triad of intellectual disability, hypergonadotropic hypogonadism, and cardiomyopathy enabled the diagnosis of Najjar-Malouf syndrome, also known as cardiogenital syndrome. This case underscores the diagnostic and therapeutic challenges of Najjar-Malouf syndrome, emphasizing the significance of thorough evaluation and genetic testing, particularly considering the association with LMNA mutations. Further research is needed to improve understanding and management strategies for this rare syndrome.

Circulating Vitamin D (Vit-D) has emerged as a potent immune modulator in asthma, yet its direct impact on T_H2 cell regulation, the central effectors of allergic inflammation, remains unclear. Preliminary transcriptomic analysis of neonatal cord blood revealed that gestational Vit-D deficiency corresponds to elevated adaptive and innate immune responses, driven by T_H2 immunity and antimicrobial responses related to asthma inflammation. To elucidate cell-specific molecular mechanisms of Vit-D, we differentiated murine T_H2 cells <i>in vitro</i> under conditions mimicking Vit-D sufficiency and deficiency. Our findings demonstrate that Vit-D exposure promotes intracellular calcium ion homeostasis while suppressing prominent inflammatory cytokines characteristic of asthma. Conversely, Vit-D deficiency reprograms T_H2 cell lineage commitment, inducing overexpression of cytolytic molecules and major histocompatibility complex (MHC) class I molecules-traits typically associated with cytotoxicity rather than the canonical helper function. Our findings underscore Vit-D's role in stabilizing T_H2 cell function and fate, offering insights into asthma and autoimmune disorders.

<h4>Aims</h4>Our aim was to investigate occurrence of senescent cells directly in tendon tissue biopsies from patients with chronic shoulder tendinopathies, and to correlate senescence with Enhancer of zeste 2 (EZH2) expression, the functional subunit of the epigenetic master regulator polycomb repressive complex.<h4>Methods</h4>Human proximal long head of biceps tendons from patients with different chronic shoulder pathologies (n = 22), and controls from patients with humerus fracture (n = 6) and pathology (n = 4), were histologically scored for degeneration and analyzed for gene and protein expression of tendon specific factors, senescence markers, and EZH2. Tissues were further exposed to senotherapeutic compounds and the USA Food and Drugs Administration-approved selective EZH2 inhibitor EPZ-6438 and their senescence-associated secretory phenotype (SASP) assessed.<h4>Results</h4>Expression of senescence markers (<i>CDKN2A</i>/p16, <i>CDKN2D</i>/p19) and <i>EZH2</i> was significantly higher in tendinopathies compared to fracture or healthy tissue controls and positively correlated with the degree of tissue degeneration. Immunofluorescent stainings demonstrated colocalization of p16 and p19 with EZH2 in tenocytes. Treatment of tendon biopsies with EPZ-6438 reduced secretion of a panel of SASP factors, including interleukin-6 (IL6), IL8, matrix metalloproteinase-3 (MMP3) or GRO1, similarly to the senotherapeutic compound AG490.<h4>Conclusion</h4>We demonstrate that senescence traits accumulate in pathological tendon tissues and positively correlate with tissue degeneration. Increased expression of <i>CDKN2A</i>/p16 and <i>CDKN2D</i>/p19 coincides with EZH2 expression, while its inhibition decreased the secretion of SASP factors, indicating a possible regulatory role of EZH2 in tenocyte senescence in tendinopathies. Reduction of cellular senescence, e.g. with EPZ-6438, opens ways to new potential therapeutic approaches for enhancing regeneration in chronic tendinopathies.

<h4>Abstract</h4>Multiple morphological abnormalities of sperm flagella (MMAF) is a severe form of asthenoteratozoospermia, characterized by morphological abnormalities and reduced motility of sperm, causing male infertility. Although approximately 60% of MMAF cases can be explained genetically, the etiology of the remaining cases is unclear. Here, we identified two novel compound heterozygous variants in the gene, dynein axonemal heavy chain 10 ( DNAH10 ), in three patients from two unrelated Pakistani families using whole-exome sequencing (WES), including one compound heterozygous mutation ( DNAH10 : c.9409C>A [p.P3137T]; c.12946G>C [p.D4316H]) in family 1 and another compound heterozygous mutation ( DNAH10 : c.8849G>A [p.G2950D]; c.11509C>T [p.R3687W]) in family 2. All the identified variants are absent or rare in public genome databases and are predicted to have deleterious effects according to multiple bioinformatic tools. Sanger sequencing revealed that these variants follow an autosomal recessive mode of inheritance. Hematoxylin and eosin (H&E) staining revealed MMAF, including sperm head abnormalities, in the patients. In addition, immunofluorescence staining revealed loss of DNAH10 protein signals along sperm flagella. These findings broaden the spectrum of DNAH10 variants and expand understanding of the genetic basis of male infertility associated with the MMAF phenotype.

Klinefelter syndrome (KS), characterized by the presence of at least one extra X-chromosome, is a common cause of male infertility. However, the mechanism underlying the failure of germline specification is not well studied. Intriguingly, the differentiation efficiency of female human pluripotent stem cells (hPSCs) is often lower than that of male. This study investigates how X-linked gene dosage affects human primordial germ cell-like cells (hPGCLCs) specification in both healthy and diseased conditions. This work reveals that X-linked genes play a multifaceted role against the fate competency to hPGCLCs, with escape genes IGSF1 and CHRDL1 inhibiting the TGF-beta/Activin A and BMP pathways, respectively. Notably, this work identifies a previously unrecognized role of SOX2, upregulated by the escape gene USP9X, elucidating a species-specific function in the mammalian germline. The USP9X-SOX2 regulatory axis profoundly influenced cellular metabolism, mitochondrial morphology, and progenitor competence in hPGCLCs specification. Furthermore, the inability to downregulate SOX2 and upregulate SOX17 in response to BMP signaling impedes downstream gene activation due to motif binding competition. These findings shed novel insights into the human germline specification by elucidating the divergent roles of SOX2 versus SOX17 in mammals, influenced by X-linked gene dosage effects. These results offer potential applications for improving the induction efficiency of hPGCLCs, facilitating disease mechanistic studies.

<i>Cryptococcus neoformans</i> is considered one of the most dangerous fungal threats to human health, and the World Health Organization recently ranked it in the critical priority group for perceived public health importance. Proliferation of <i>C. neoformans</i> within mammalian hosts is supported by its ability to overcome nutritional limitations and endure stress conditions induced by the host immune response. Previously, we reported that the Vam6/Vps39/TRAP1-domain protein Vam6 was crucial for vacuolar morphology, iron acquisition, and virulence. However, the molecular mechanisms underlying the pleiotropic phenotypes resulting from loss of Vam6 remain poorly understood. In this study, we determined that Vam6 has roles in the HOPS complex for endomembrane trafficking to the vacuole and in the vCLAMP membrane contact site between the vacuole and mitochondria. Importantly, both of these roles regulate polyphosphate (polyP) metabolism, as demonstrated by a defect in trafficking of the VTC complex subunit Vtc2 for polyphosphate synthesis and by an influence on mitochondrial functions. In the latter case, Vam6 was required for polyP accumulation in response to electron transport chain inhibition and for overcoming oxidative stress. Overall, this work establishes connections between endomembrane trafficking, mitochondrial functions, and polyP homeostasis in <i>C. neoformans</i>.IMPORTANCEA detailed understanding of stress resistance by fungal pathogens of humans may provide new opportunities to improve antifungal therapy and combat life-threatening diseases. Here, we used a <i>vam6</i> deletion mutant to investigate the role of the homotypic fusion and vacuole protein sorting (HOPS) complex in mitochondrial functions and polyphosphate homeostasis in <i>Cryptococcus neoformans</i>, an important fungal pathogen of immunocompromised people including those suffering from HIV/AIDS. Specifically, we made use of mutants defective in late endocytic trafficking steps to establish connections to oxidative stress and membrane trafficking with mitochondria. In particular, we found that mutants lacking the Vam6 protein had altered mitochondrial function, and that the mutants were perturbed for additional mitochondria and vacuole-related phenotypes (e.g., membrane composition, polyphosphate accumulation, and drug sensitivity). Overall, our study establishes connections between endomembrane trafficking components, mitochondrial functions, and polyphosphate homeostasis in an important fungal pathogen of humans.

Tumor-infiltrating myeloid cells (TIMs), which encompass tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), myeloid-derived suppressor cells (MDSCs), and tumor-associated dendritic cells (TADCs), are of great importance in tumor microenvironment (TME) and are integral to both pro- and anti-tumor immunity. Nevertheless, the phenotypic heterogeneity and functional plasticity of TIMs have posed challenges in fully understanding their complexity roles within the TME. Emerging evidence suggested that the presence of TIMs is frequently linked to prevention of cancer treatment and improvement of patient outcomes and survival. Given their pivotal function in the TME, TIMs have recently been recognized as critical targets for therapeutic approaches aimed at augmenting immunostimulatory myeloid cell populations while depleting or modifying those that are immunosuppressive. This review will explore the important properties of TIMs related to immunity, angiogenesis, and metastasis. We will also document the latest therapeutic strategies targeting TIMs in preclinical and clinical settings. Our objective is to illustrate the potential of TIMs as immunological targets that may improve the outcomes of existing cancer treatments.

The clinical application of cellular immunotherapy in hepatocellular carcinoma (HCC) is impeded by the lack of a cell surface target frequently expressed in HCC cells and with minimal presence in normal tissues to reduce on-target, off-tumor toxicity. To address this, an in silico multomics analysis was conducted to identify an optimal therapeutic target in HCC. A longlist of genes (n = 12,948) expressed in HCCs according to The Human Protein Atlas database were examined. Eight genes were shortlisted to identify one with the highest expression in HCCs, without being shed into circulation, and with restrictive expression profile in other normal human tissues. A total of eight genes were shortlisted and subsequently ranked according to the combination of their transcript and protein expression levels in HCC cases (n = 791) derived from four independent datasets. TM4SF4 was the top-ranked target with the highest expression in HCCs. TM4SF4 showed more favorable expression profile with significantly lower expression in normal human tissues but more highly expressed in HCC compared with seven other common HCC therapeutic targets. Furthermore, scRNA-seq and immunohistochemistry datasets showed that TM4SF4 was absent in immune cell populations but highly expressed in the bile duct canaliculi of hepatocytes, regions inaccessible to immune cells. In scRNA-seq dataset of HCCs, TM4SF4 expression was positively associated with mitochondrial components and oxidative phosphorylation Gene Ontologies in HCC cells (n = 15,787 cells), suggesting its potential roles in mitochondrial-mediated oncogenic effects in HCC. Taken together, TM4SF4 is proposed as a promising cell surface target in HCC due to its high expression in HCC cells with restricted expression profile in non-cancerous tissues, and association with HCC oncogenic pathways.

Repeat expansions in the <i>C9orf72</i> gene are the most common genetic cause of amyotrophic lateral sclerosis and familial frontotemporal dementia (ALS/FTD). To identify molecular defects that take place in the dorsolateral frontal cortex of patients with <i>C9orf72</i> ALS/FTD, we compared healthy controls with <i>C9orf72</i> ALS/FTD donor samples staged based on the levels of cortical phosphorylated TAR DNA binding protein (pTDP-43), a neuropathological hallmark of disease progression. We identified distinct molecular changes in different cell types that take place during FTD development. Loss of neurosurveillance microglia and activation of the complement cascade take place early, when pTDP-43 aggregates are absent or very low, and become more pronounced in late stages, suggesting an initial involvement of microglia in disease progression. Reduction of layer 2-3 cortical projection neurons with high expression of CUX2/LAMP5 also occurs early, and the reduction becomes more pronounced as pTDP-43 accumulates. Several unique features were observed only in samples with high levels of pTDP-43, including global alteration of chromatin accessibility in oligodendrocytes, microglia, and astrocytes; higher ratios of premature oligodendrocytes; increased levels of the noncoding RNA NEAT1 in astrocytes and neurons, and higher amount of phosphorylated ribosomal protein S6. Our findings reveal progressive functional changes in major cell types found in the prefrontal cortex of <i>C9orf72</i> ALS/FTD patients that shed light on the mechanisms underlying the pathology of this disease.

Although substantial advancements have been made in genetic testing, several barriers continue to limit patient access, leading to delays in diagnosis, effective treatments, and preventative measures. The NEUROMYODredger-3billion Megaproject End the Diagnostic Odyssey grant offered free whole exome sequencing (WES) to 245 patients with undiagnosed neurodevelopmental or neuromuscular disorders in seven countries: Algeria, Chile, Egypt, France, Mexico, Peru, and Romania. We found pathogenic variants in 79 patients (diagnostic yield 32.24%)-36 neurodevelopmental (43.90%) and 43 neuromuscular (26.38%). Fifty patients harboured variants of uncertain significance (VUS, 20.40%)-14 neurodevelopmental (17.07%) and 36 neuromuscular (22.08%), and 116 patients had negative results (47.34%). NEUROMYODredger helped end the diagnostic odyssey in around 30% of patients, while ongoing functional studies and reanalysis strategies are used in order to reach more diagnoses. In conclusion, a singleton WES approach is valuable in determining the genetic diagnosis of neurodevelopmental and neuromuscular diseases, especially in low and middle-income countries.

Chronic kidney disease (CKD) is a clinical syndrome of metabolic disorder caused by progressive kidney impairment for more than 3 months. CKD has become a global public health problem due to its high morbidity and mortality, which is difficult to be cured for most patients. The pathogenesis of CKD is still unclear, which is closely related to glomerulosclerosis, kidney tubular injury and kidney fibrosis. LncRNA is a non-coding RNA with a length of more than 200 nucleotides. It not only participates in intracellular transcriptional regulation, post-transcriptional regulation and epigenetic activities, but also forms a regulatory network together with miRNA and mRNA, to further conduct the reticular regulation in cells. Recently, it has been found that lncRNA participates in pathophysiological mechanism of CKD by regulating glomerulosclerosis, kidney tubular injury and kidney fibrosis. This has also become a new direction of lncRNA in early diagnosis and targeted therapy of CKD.

The muscular system plays a critical role in the human body by governing skeletal movement, cardiovascular function, and the activities of digestive organs. Additionally, muscle tissues serve an endocrine function by secreting myogenic cytokines, thereby regulating metabolism throughout the entire body. Maintaining muscle function requires iron homeostasis. Recent studies suggest that disruptions in iron metabolism and ferroptosis, a form of iron-dependent cell death, are essential contributors to the progression of a wide range of muscle diseases and disorders, including sarcopenia, cardiomyopathy, and amyotrophic lateral sclerosis. Thus, a comprehensive overview of the mechanisms regulating iron metabolism and ferroptosis in these conditions is crucial for identifying potential therapeutic targets and developing new strategies for disease treatment and/or prevention. This review aims to summarize recent advances in understanding the molecular mechanisms underlying ferroptosis in the context of muscle injury, as well as associated muscle diseases and disorders. Moreover, we discuss potential targets within the ferroptosis pathway and possible strategies for managing muscle disorders. Finally, we shed new light on current limitations and future prospects for therapeutic interventions targeting ferroptosis.

Aberrant alternative splicing and abnormal alternative splicing events (ASEs) in glioblastoma multiforme (GBM) remain largely elusive. The prognostic-associated ASEs in GBM were identified and summarized into 123 genes using GBM and LGG datasets from ASCancer Atlas and TCGA. The eleven genes (C2, COL3A1, CTSL, EIF3L, FKBP9, FN1, HPCAL1, HSPB1, IGFBP4, MANBA, PRKAR1B) were screened to develop an alternative splicing prognostic risk score (ASRS) model through machine learning algorithms. The model was trained on the TCGA-GBM cohort and validated with four external datasets from CGGA and GEO, achieving AUC values of 0.808, 0.814, 0.763, 0.859, and 0.836 for 3-year survival rates, respectively. ASRS could be an independent prognostic factor for GBM patients (HR > 1.8 across three datasets) through multivariate Cox regression analysis. The high-risk group demonstrated poorer prognosis, elevated immune scores, increased levels of immune cell infiltration, and greater differences in drug sensitivity. We found that FN1, used for model construction, contained 4 abnormal ASEs resulting in high expression of non-canonical transcripts and the presence of premature termination codon. These abnormal ASEs may be regulated by tumour-related splicing factors according to the PPI network. Furthermore, both mRNA and protein levels of FN1 were highly expressed in GBM compared to LGG, correlating with poor prognosis in GBM. In conclusion, our findings highlight the role of ASEs in affecting the progression of GBM, and the model showed a potential application for prognostic risk of patients. FN1 may serve as a promising splicing biomarker for GBM, and mechanisms of processes of aberrant splicing need to be revealed in the future.

Gastric cancer precursors demonstrate highly-variable rates of progression toward neoplasia. Certain high-risk precursors, such as gastric intestinal metaplasia with advanced histologic features, may be at up to 30-fold increased risk for progression compared to lower-risk intestinal metaplasia. The biological differences between high- and low-risk lesions have been incompletely explored. In this study, we use several clinical cohorts to characterize the microenvironment of advanced gastric cancer precursors relative to low-risk lesions using bulk, spatial, and single-cell gene expression assays. We identified a 26-gene panel which is associated with advanced lesions, localizes to metaplastic glands on histopathology, and is expressed in aberrant mature and immature intestinal cells not normally present in the healthy stomach. This gene expression signature suggests an important role of the immature intestinal lineages in promoting carcinogenesis in the metaplastic microenvironment. These findings may help to inform future biomarker development and strategies of gastric cancer prevention.

Precise spatiotemporal control of drug delivery is extremely valuable for regulating stem cell fate, particularly in stem cell differentiation. A novel near-infrared (NIR)-mediated spatiotemporal delivery system is reported combining photo-switchable arylazopyrazole (AAP)-containing DNA strands and upconversion nanoparticles (UCNPs). This nano-drug delivery system (NDDS) enables precise modulation of DNA duplex structures in response to NIR stimuli, overcoming the limitations of traditional UV-responsive systems. AAP derivatives with enhanced photoswitching efficiency (≈98%) and significantly improved cis-form stability are engineered. The successful delivery of curcumin, a neurogenic compound with an affinity for the minor groove of DNA, to human neural stem cells (NSCs) is achieved using UCNP-DNA-AAP constructs. Upon 980 nm NIR light exposure, UCNPs efficiently up-converted NIR to UV light, triggering AAP photoisomerization and DNA dissociation, thus releasing curcumin. This approach enabled efficient spatiotemporal control over NSC differentiation while facilitating neuroprotection. Immunofluorescence and gene expression analyses demonstrated enhanced neuronal mRNA levels and neurite outgrowth in treated cells. In short, the NIR-mediated photo-switchable NDDS offers a precise and innovative approach to control stem cell fate, enabling spatiotemporal regulation of cellular processes. This technology has significant potential applications in nanomedicine and neuroscience, where precise drug delivery is crucial for targeted neural interventions.

Circular RNAs (circRNAs) are differentially expressed in cardiac hypertrophy; however, the exact function and mechanisms during hypertrophy development are still unknown. Here, we explored the role of a newly discovered circRNA in the pathogenesis of myocardial hypertrophy. It was found that circ-0001283 promoted the progression of cardiac hypertrophy by interacting with myosin light chain 3 (MYL3) to inhibit the protein ubiquitination and enhance its protein expression, not by the competitive endogenous RNA mechanism. Further investigation demonstrated that the reduced hypertrophy induced by circ-0001283 knockdown was counteracted by overexpression of MYL3. Mechanistically, MYL3 facilitated myocardial hypertrophy by inducing autophagy in cells via PI3K/Akt/mTOR and ERK signaling pathways. In summary, circ-0001283 can bind directly to MYL3 and up-regulate its expression, thereby promoting autophagy to accelerate cardiac hypertrophy. Circ-0001283 may serve as a potential therapeutic target for cardiac hypertrophy.

Peroxiredoxins (PRDXs) are a highly conserved family of peroxidases that serve as the primary scavengers of peroxides. Post-translational modifications play crucial roles modulating PRDX activities, tuning the balance between reactive oxygen species (ROS) signaling and stress. We previously reported that S-acylation occurs at the "peroxidatic" cysteine (Cp) site of PRDX5 and that it inhibits PRDX5 activity. Here, we show that the PRDX family more broadly is subject to S-acylation at the Cp site of all PRDXs and that PRDX S-acylation dynamically responds to cellular ROS levels. Using activity-based fluorescent imaging with DPP-Red, a red-shifted fluorescent indicator for acyl-protein thioesterase (APT) activity, we also discover that the instigation of ROS-stress via exogenous H₂O₂ activates both the cytosolic and mitochondrial APTs, whereas epidermal growth factor (EGF)-stimulated endogenous H₂O₂ deactivates the cytosolic APTs. These results indicate that APTs help tune H₂O₂ signal transduction and ROS protection through PRDX S-deacylation.

Cell-type-specific gene activation is regulated by enhancers, sometimes located at large genomic distances from target gene promoters. Whether distal enhancers require specific factors to orchestrate gene regulation remains unclear. Here, we used enhancer distance-controlled reporter screens to find candidate factors. We depleted them and employed activity-by-contact predictions to genome-wide classify genes based on enhancer distance. Predicted distal enhancers typically control tissue-restricted genes and often are strong enhancers. We find cohesin, but also mediator, most specifically required for long-range activation, with cohesin repressing short-range gene activation and prioritizing distal over proximal HBB genes competing for shared enhancers. Long-range controlled genes are also most sensitive to perturbations of other regulatory proteins and to BET inhibitor JQ1, this being more a consequence of their distinct enhancer features than distance. Our work predicts that lengthening of intervening sequences can help limit the expression of target genes to specialized cells with optimal trans-factor environments.

Huntington's disease (HD) is a neurodegenerative disorder caused by the abnormal expansion of CAG repeats in the huntingtin (Htt) gene, leading to the aggregation of mutant huntingtin protein (mHTT) in cells, particularly in cortical and striatal neurons. This results in involuntary movements, cognitive impairment, and emotional instability. One of the critical pathogenic mechanisms in HD is impaired autophagy, which plays a vital role in cellular homeostasis by degrading damaged organelles and misfolded proteins through the formation of autophagosomes that fuse with lysosomes. However, the aggregation of mHTT disrupts autophagic function, leading to the accumulation of mHTT and exacerbating the disease's pathogenesis. Carvedilol is an established clinical medication used to treat hypertension and congestive heart failure. It exerts protective effects by blocking both β1-and β2-adrenergic receptors, reducing sympathetic nervous activity, and promoting vasodilation through α1-adrenergic blockade. Carvedilol has been shown to possess antioxidant and anti-inflammatory properties. In this study, we demonstrate that (R)-carvedilol promotes the nuclear translocation of the transcription factor binding to IGHM enhancer 3 (TFE3) by reducing glycogen synthase-3β (GSK-3β) activation, which increases the expression of autophagy-related proteins and facilitates the autophagy-lysosomal pathway (ALP), thereby enhancing mHTT degradation. Additionally, systemic administration of (R)-carvedilol improves mHTT degradation, provides neuroprotection, and inhibits gliosis, effectively ameliorating behavioral impairments and improving disease progression. Overall, these findings indicate that (R)-carvedilol has therapeutic potential for managing HD by promoting autophagy, facilitating the clearance of mHTT aggregates, and demonstrating advantageous properties in an HD transgenic mouse model, highlighting its promise as a treatment option for neurodegenerative diseases.

The protein kinase ERK2 is recurrently mutated in human squamous cell carcinomas and other tumors. ERK2 mutations cluster in an essential docking recruitment site that interacts with short linear motifs found within intrinsically disordered regions of ERK substrates and regulators. Cancer-associated mutations do not disrupt ERK2 docking interactions altogether but selectively inhibit some interactions while sparing others. However, the full scope of disrupted or maintained interactions remains unknown, limiting our understanding of how these mutations contribute to cancer. We recently defined the docking interactome of wild-type ERK2 by screening a yeast two-hybrid library of proteomic short linear motifs. Here, we apply this approach to the two most recurrent cancer-associated mutants. We find that most sequences binding to WT ERK2 also interact with both mutant forms. Analysis of differentially interacting sequences revealed that ERK2 mutants selectively lose the ability to bind sequences conforming to a specific motif. We solved the co-crystal structure of ERK2 in complex with a peptide fragment of ISG20, a screening hit that binds exclusively to the WT kinase. This structure demonstrated the mechanism by which cancer hotspot mutations at Glu81, Arg135, Asp321, and Glu322 selectively impact peptide binding. Finally, we found that cancer-associated ERK2 mutations had decreased activity in phosphorylating GEF-H1/ARHGEF2, a known ERK substrate harboring a WT-selective docking motif. Collectively, our studies provide a structural rationale for how a broad set of interactions are disrupted by ERK2 hotspot mutations, suggesting mechanisms for pathway rewiring in cancers harboring these mutations.

Neuroinflammation represents a critical pathway in the brain for the clearance of foreign bodies and the maintenance of homeostasis. When the neuroinflammatory process is dysregulate, such as the over-activation of microglia, which results in the excessive accumulation of free oxygen and inflammatory factors in the brain, among other factors, it can lead to an imbalance in homeostasis and the development of various diseases. Recent research has indicated that the development of numerous neurodegenerative diseases is closely associated with neuroinflammation. The pathogenesis of neuroinflammation in the brain is intricate, involving alterations in numerous genes and proteins, as well as the activation and inhibition of signaling pathways. Furthermore, excessive inflammation can result in neuronal cell apoptosis, which can further exacerbate the extent of the disease. This article presents a summary of recent studies on the relationship between neuronal apoptosis caused by excessive neuroinflammation and neurodegenerative diseases. The aim is to identify the link between the two and to provide new ideas and targets for exploring the pathogenesis, as well as the prevention and treatment of neurodegenerative diseases.

<h4>Background</h4>Multiple pregnancy constitutes a large metabolic expense, so women with twin pregnancies and neonates born as twins might be at risk for micronutrient deficiencies. Therefore, the aim of the study was to assess the supplementation used and supply with key micronutrients: iron, vitamin B12, folic acid and vitamin D in women with twin pregnancies and the correlations with cord blood indicators.<h4>Methods</h4>Maternal and cord blood samples were collected from 51 patients with twin pregnancies and 102 newborns born from those pregnancies between October 2020 and September 2023. Ferritin, vitamin B12, folic acid and vitamin D metabolites concentrations were measured. Additionally, the patients completed a questionnaire regarding pre-and intragestational supplementation.<h4>Results</h4>Iron, vitamin B12, and vitamin D deficiency were diagnosed in 20.8, 13.7 and 12.5% of women with twin pregnancies, respectively. No maternal folate deficiency was identified. Positive weak to moderate correlations were demonstrated between the concentrations of all studied indicators in the maternal and cord blood. Pregestational folic acid and vitamin D supplementation was associated with higher cord blood levels of folic acid and 3-epi-25(OH)D3, respectively. 25(OH)D3 and total 25(OH)D concentrations were higher in newborns whose mothers had supplemented vitamin D during pregnancy.<h4>Conclusion</h4>The problem of iron, vitamin B12 and vitamin D deficiencies in twin pregnancies is still valid. Pre-and intragestational supplementation, as well as maternal micronutrient supply affect the cord blood composition of twins.

Dipeptidyl peptidase IV (DPP-IV) is a serine protease whose inhibition has been an object of considerable interest in the context of developing novel treatments for type 2 diabetes mellitus. The development of novel DPP-IV inhibitors from natural or synthetic origin has seen a growing scientific interest in recent years, especially during the SARS-CoV-2 pandemic, when DPP-IV inhibitors were found to be of beneficial therapeutic value for COVID-19 patients. The present manuscript aims to summarize the most recent information on the synthesis of different DPP-IV inhibitors, emphasizing the various heterocyclic scaffolds that can be found in them. Special attention is devoted to DPP-IV inhibitors that are currently in clinical trials. Different synthetic approaches for the construction of DPP-IV inhibitors are discussed, as well as the most recent developments in the field.

In this work, we describe the synthesis of three new <i>meso</i>-arylporphyrins, named <i>meso</i>-tetrakis [4-(nicotinoyloxy)phenyl] porphyrin (<b>H₂TNPP</b>), <i>meso</i>-tetrakis [4-(picolinoyloxy)phenyl] porphyrin (<b>H₂TPPP</b>), and <i>meso</i>-tetrakis [4-(isonicotinoyloxy) phenyl] porphyrin (<b>H₂TIPP</b>). These new synthesized <i>meso</i>-arylporphyrins are characterized using spectroscopic analysis: Fourier Transform Infrared Spectroscopy (FTIR) and One-dimensional Nuclear Magnetic Resonance (1D NMR), and mass spectrometry (MS). The photophysical studies (UV-visible absorption, singlet oxygen (¹O₂) luminescence, and fluorescence emissions) demonstrate their potential uses as photosensitizers (PSs) in photodynamic therapy (PDT) applications. An in vitro investigation of the anti-fungal activity of <b>H₂TNPP</b>, <b>H₂TPPP</b>, and <b>H₂TIPP</b> against <i>Candida</i> (<i>C.</i>) species (<i>C. albicans</i>, <i>C. glabrata</i>, and <i>C. tropicalis</i>) reveals that their minimum inhibitory concentration (MIC) values ranged from 1.25 to 5 mg/mL. In addition, their in vitro anti-fungal susceptibilities against three dermatophyte clinical isolates (<i>Trichophyton rubrum</i>, <i>Microsporum canis</i>, and <i>Trichophyton mentagrophytes</i>) are also evaluated and they demonstrate good anti-fungal activities. A molecular docking study of these <i>meso</i>-arylporphyrins as anti-fungal agents against <i>C. tropicalis</i> extracellular aspartic proteinases, Protein data Bank in Europe (PDBe code: 1J71) and <i>Trichophyton rubrum</i> Sialidases (PDBe code: 7P1D) underlines the possible interactions of <b>H₂TNPP</b>, <b>H₂TPPP</b>, and <b>H₂TIPP</b> with the key amino acid residues of these fungal target proteins.

Radiotherapy (RT) is the primary treatment modality for hepatocellular carcinoma (HCC). Inevitably, the X-ray exposure also increases the metabolic stress and energy demands in surviving tumor cells, which leads to metabolic reprogramming that reduces the sensitivity of HCC to clinical treatments including RT. Nevertheless, the current research in tumor metabolic therapy predominantly focuses on inhibiting glycolytic pathways, and the consequent metabolic compensation behavior of tumor cells exacerbates the risks of drug resistance and recurrence. To address this challenge, we innovatively proposed a tumor-specific multi-metabolic pathway regulation strategy navigated by tumor cell surface nutrient transporter (2-DG/BP MRs), which can be triggered by X-ray radiation to achieve dual blockade of glycolysis and glutamine metabolism pathways. Thus, this nanosystem reconfigured metabolic pathways within tumor cells to counteract RT-induced metabolic reprogramming through dual metabolic inhibition (glycolysis and glutamine metabolism). This approach disrupted the essential energy supply required for cancer cell proliferation without causing metabolic disorders in normal cells, thereby sensitizing HCC to RT. This tumor cell-specific metabolic intervention strategy provides a safe and effective approach for combination therapy in clinically RT-resistant tumors.

MicroRNAs (miRNAs) are small non-coding RNAs that act as critical regulators of gene expression by repressing mRNA translation. The role of miRNAs in cell physiology spans from cell cycle control to cell proliferation and differentiation, both during development and in adult tissues. Accordingly, dysregulated expression of miRNAs has been reported in several diseases, including cancer, where miRNAs can act as oncogenes or oncosuppressors. Of note, miRNA signatures are also under investigation for classification, diagnosis, and prognosis of cancer patients. Brain tumours are primarily associated with poor prognosis and high mortality, highlighting an urgent need for novel diagnostic, prognostic, and therapeutic tools. Among miRNAs investigated in brain tumours, miR-326 has been shown to act as a tumour suppressor in adult and paediatric brain cancers. In this review, we describe the role of miR-326 in malignant as well as benign cancers originating from brain tissue. In addition, since miR-326 expression can be regulated by other non-coding RNA species, adding a further layer of regulation in the cancer-promoting axis, we discuss this miRNA's role in targeted therapy for brain cancers.

In late 2021, Eurasian-lineage highly pathogenic avian influenza (HPAI) A(H5N1) viruses from HA clade 2.3.4.4b were first detected in the United States. These viruses have caused severe morbidity and mortality in poultry and have been detected in numerous wild and domestic animals, including cows and humans. Notably, infected cows transmitted the virus to cats, causing extreme pathogenicity and death. While human-to-human spread of the virus has not been recorded, efficient transmission of the bovine-origin virus has also led to extreme pathogenicity and death in ferret models. Recently, markers in PB2 (E627K) and HA (E186D, Q222H), indicating mammalian adaptation mutations, were detected in an H5N1-infected patient manifesting critical illness in Canada. These, combined with instances of interspecies spread of the virus, have raised global public health concerns. This could highlight the potential for the virus to successfully adapt to mammals, posing a serious risk of a global outbreak. A One Health approach is, thereby, necessary to monitor and control the outbreak. This review aims to analyze the epidemiology, transmission, and ecological impacts of HPAI A(H5N1) clade 2.3.4.4b in the U.S., identify knowledge gaps, and inform strategies for effective outbreak management and mitigation.

Inflammasomes are protein complexes that trigger pro-inflammatory responses and promote many diseases, including adipose tissue dysfunction. Linagliptin (L), a DPP-4 inhibitor used for type 2 diabetes therapy, has putative anti-inflammatory effects. This work explores L effects on inflammasome regulation, inflammation, and adipose tissue dysfunction in obese mice. Male C57BL/6N mice were fed a normal chow (NC) diet, high-fat (HF) diet, or HF diet with L (HFL) for 15 weeks. Gene expression and histological examinations were performed on visceral (VAT) and subcutaneous (SAT) adipose tissue samples. Biomarkers were quantified on sera. Murine macrophages were utilized for in vitro analyses. L decreased HF-induced endotoxemia and circulating inflammatory indicators. Despite having no effect on body weight, L reduced VAT inflammation by decreasing endotoxemia-induced <i>NLRC4</i> inflammasome, inflammation severity, and fat cell hypertrophy. Although SAT response differed from VAT, inflammation was slightly reduced in this tissue too. In vitro, L modulated inflammation by directly reducing the pro-inflammatory macrophage phenotype. In obesity, increased <i>NLRC4</i> inflammasome expression links endotoxemia and VAT inflammation. L protected against endotoxemia, maybe by affecting gut permeability and VAT responses. The decreased polarization of macrophages toward a pro-inflammatory phenotype and the reduction in adipocyte hypertrophy are involved in the response to L.

<h4>Introduction</h4>Breast cancer (BC) is the leading cause of morbidity and mortality worldwide. Owing to early diagnosis and better therapeutic care, survivorship in these patients have improved tremendously. Chemotherapy, cornerstone in BC management have been associated with debilitating side effects including the effect on cognitive function, which significantly impairs the quality of life in these patients. Thus, it is imperative to understand the timeline and magnitude of the effects of chemotherapy on cognition to develop better management strategies. This is even more relevant in developing country like India, where there is inconspicuous absence of data in this regard.<h4>Aim</h4>To study the acute and long-term effects of chemotherapy on the cognitive function in BC patients compared to chemotherapy naïve (Cx naïve) BC patients (disease controls) and matched healthy controls (HC) using subjective, objective questionnaires and neuropsychological tests (NPTs).<h4>Methods</h4>The current cross-sectional study involved 120 participants, 30 each of Cx naïve BC patients, during chemotherapy BC patients (during Cx), post-chemotherapy BC patients (post Cx) and HC; all matched for age and education levels. Both subjective and objective assessments of cognitive functions were done in all the groups. Hindi Mental State Examination (HMSE) and FACT Cog questionnaire V3 were used for subjective assessment while Addenbrooke Cognitive Examination-III (ACE-III) questionnaire and domain specific computer based NPT (Wisconsin card sorting task (WCST) (learning), Flanker's (attention and interference) and <i>n</i> back task (working memory) were done for objective assessment. The data were analysed for descriptive and inferential statistics, as appropriate using GraphPad Prism V9.<h4>Results</h4>The subjective assessment using HMSE questionnaire revealed a significantly lower score in post Cx group as compared to HC (<i>p</i> < 0.001); however, it was comparable in other groups. FACT Cog V3 questionnaire revealed significantly higher cognitive impairment among those during Cx compared to Cx naïve patients (<i>p</i> < 0.001), post Cx BC patients (<i>p</i> < 0.001) and HC (<i>p</i> < 0.0001). Meanwhile, the objective assessment using ACE-III examination revealed significantly lesser scores among during Cx patients (<i>p</i> < 0.001), and post Cx BC patients (<i>p</i> < 0.0001) compared to HC group. In NPTs, WCST and <i>N</i> back working memory task revealed significantly lower accuracy in Cx naïve versus post Cx (<i>p</i> = 0.0054, <i>p</i> = 0.0068, respectively) and HC versus post Cx (<i>p</i> = 0.0054, <i>p</i> = 0.0045, respectively), while no significant difference was found in Flanker's task. Furthermore, in WCST there were significantly higher scores present in total reaction time in post Cx compared to Cx naïve: <i>p</i> = 0.00444 and HC: <i>p</i> = 0.0003). In Flanker's task reaction time was higher in all the groups (Cx naïve: <i>p</i> = 0.002, during Cx: <i>p</i> = 0.0007 and Post Cx: <i>p</i> < 0.0001) compared to HC. In addition negative correlation was found between the duration of chemotherapy with Perceived Cognitive Abilities (<i>r</i> = -0.482; <i>p</i> = 0.006), between the total number of cycles with Fact Cog Total (<i>r</i> = -0.373, <i>p</i> = 0.04) and Perceived Cognitive abilities (<i>r</i> = 0.39, <i>p</i> = 0.03), and between total dose and perceived cognitive abilities (<i>r</i> = -0.42, <i>p</i> = 0.014) also a positive correlation was seen between dose of epirubicin with reaction time of <i>n</i> back (<i>r</i> = 0.373, <i>p</i> = 0.04).<h4>Conclusion</h4>Chemotherapy can have a negative impact on the cognitive function in BC patients, manifested as both acute and long-term effects, based on patient reported/subjective and laboratory based/objective cognitive function tests. The deficits were seen mostly seen in domains of attention and working memory across groups compared to matched HC.

Trisomy of chromosome 21, the cause of Down syndrome (DS), is the most commonly occurring genetic cause of Alzheimer’s disease (AD). Here, we compare the frontal cortex proteome of people with Down syndrome-Alzheimer’s disease (DSAD) to demographically matched cases of early-onset AD and healthy ageing controls. We find wide dysregulation of the proteome, beyond proteins encoded by chromosome 21, including an increase in the abundance of the key AD-associated protein, APOE, in people with DSAD compared to matched cases of AD. To understand the cell types that may contribute to changes in protein abundance, we undertook a matched single-nuclei RNA-sequencing study, which demonstrated that APOE expression was elevated in subtypes of astrocytes, endothelial cells and pericytes in DSAD. We further investigate how trisomy 21 may cause increased APOE. Increased abundance of APOE may impact the development of, or response to, AD pathology in the brain of people with DSAD, altering disease mechanisms with clinical implications. Overall, these data highlight that trisomy 21 alters both the transcriptome and proteome of people with DS in the context of AD, and that these differences should be considered when selecting therapeutic strategies for this vulnerable group of individuals who have high-risk of early-onset dementia.

<h4>Summary</h4> Huntington’s Disease (HD) is caused by a CAG repeat expansion in the gene encoding Huntingtin (HTT ) . While normal HTT function appears impacted by the mutation, the specific pathways unique to CAG repeat expansion versus loss of normal function are unclear. To understand the impact of the CAG repeat expansion, we evaluated biological signatures of HTT knockout ( HTT KO) versus those that occur from the CAG repeat expansion by applying multi-omics, live cell imaging, survival analysis and a novel feature-based pipeline to study cortical neurons (eCNs) derived from an isogenic human embryonic stem cell series (RUES2). HTT KO and the CAG repeat expansion influence developmental trajectories of eCNs, with opposing effects on the growth. Network analyses of differentially expressed genes and proteins associated with enriched epigenetic motifs identified subnetworks common to CAG repeat expansion and HTT KO that include neuronal differentiation, cell cycle regulation, and mechanisms related to transcriptional repression and may represent gain-of-function mechanisms that cannot be explained by HTT loss of function alone. A combination of dominant and loss-of-function mechanisms are likely involved in the aberrant neurodevelopmental and neurodegenerative features of HD that can help inform therapeutic strategies.

We here report the synthesis of two types of six-membered cyclic carbonate monomers equipped with various drug molecules through ester linkages. The target compounds can be isolated in good yields and feature diagnostic IR and ¹³C NMR spectroscopic fingerprints in line with their proposed connectivities. As a potential application, we investigated their ring-opening polymerization (ROP), showing that the nature of the cyclic carbonate is crucial towards macromolecular carbonate formation. The functionalized polycarbonates have molecular weights of up to 10 kg/mol, controllable functionality and a variable drug-to-carbonate ratio. This work demonstrates the adaptive synthesis of new types of functionalized six-membered cyclic carbonates with potential as precursors to polycarbonate-drug type macromolecules.

This study investigates the energy landscapes of amyloid monomers, which are crucial for understanding protein misfolding mechanisms in Alzheimer's disease. While proteins possess inherent thermodynamic stability, environmental factors can induce deviations from native folding pathways, leading to misfolding and aggregation, phenomena closely linked to solubility. Using the UNOPTIM program, which integrates the UNRES potential into the Cambridge energy landscape framework, we conducted single-ended transition state searches and employed discrete path sampling to compute kinetic transition networks starting from PDB structures. These kinetic transition networks consist of local energy minima and the transition states that connect them, which quantify the energy landscapes of the amyloid monomers. We defined clusters within each landscape using energy thresholds and selected their lowest-energy structures for the structural analysis. Applying graph convolutional networks, we identified solubility trends and correlated them with structural features. Our findings identify specific minima with low solubility, characteristic of aggregation-prone states, highlighting the key residues that drive reduced solubility. Notably, the exposure of the hydrophobic residue Phe19 to the solvent triggers a structural collapse by disrupting the neighboring helix. Additionally, we investigated selected minima to determine the first passage times between states, thereby elucidating the kinetics of these energy landscapes. This comprehensive approach provides valuable insights into the thermodynamics and kinetics of Aβ monomers. By integration of multiple analytical techniques to explore the energy landscapes, our study investigates structural features associated with reduced solubility. These insights have the potential to inform future therapeutic strategies aimed at addressing protein misfolding and aggregation in neurodegenerative diseases.

<h4>Background</h4>Periodontitis and bone loss in the maxillofacial and dental areas pose considerable challenges for both functional and aesthetic outcomes. To date, implantable dental barrier membranes, designed to prevent epithelial migration into defects and create a favorable environment for targeted cells, have garnered significant interest from researchers. Consequently, a variety of materials and fabrication methods have been explored in extensive research on regenerative dental barrier membranes.<h4>Methods</h4>This review focuses on dental barrier membranes, summarizing the various biomaterials used in membrane manufacturing, fabrication methods, and state-of-the-art applications for dental tissue regeneration. Based on a discussion of the pros and cons of current membrane strategies, future research directions for improved membrane designs are proposed.<h4>Results and conclusion</h4>To endow dental membranes with various biological properties that accommodate different clinical situations, numerous biomaterials and manufacturing methods have been proposed. These approaches provide theoretical support and hold promise for advancements in dental tissue regeneration.

Female mosquitoes are vectors of many devastating human diseases because they require blood feeding to initiate reproduction. Thus, elucidation of molecular mechanisms managing female mosquito reproduction is essential. Although the regulation of gene expression during the mosquito gonadotrophic cycle has been studied in detail, how this process is controlled at the chromatin level remains unclear. Chromatin must be accessible for transcription factors (TFs) governing gene expression. A specialized class of TFs, called pioneer factors (PFs), binds and remodels closed chromatin, permitting other TFs to bind DNA and activate the gene expression. Here, we identified a homolog of the vertebrate PF FoxA in the mosquito <i>Aedes aegypti</i> and used the CRISPR-Cas9 system to generate mosquitoes deficient in <i>AaFoxA</i>. We found that ovary development was severely retarded in mutant females. Multiomics and molecular biology analyses have shown that AaFoxA increased histone acetylation and decreased methylation of H3K27 by controlling the chromatin accessibility of histone modification enzymes and chromatin remodelers. AaFoxA is bound to the loci of chromatin remodelers, changing their chromatin accessibility and modulating their temporal expression patterns. AaFoxA increased the accessibility of the ecdysone receptor (EcR) and E74 loci, indicating the important role of AaFoxA in the hormonal regulation of mosquito reproductive events. Further, the CUT&RUN and ATAC-seq analyses revealed that AaFoxA temporarily bound closed chromatin, making it differentially accessible during the mosquito gonadotrophic cycle. Hence, this study demonstrates that AaFoxA modulates chromatin dynamics throughout female mosquito reproduction.

Microgels offer broad applications in bioengineering due to their customizable properties, supporting innovations in mechanobiology, tissue engineering, drug delivery, and cell therapy. This study focuses on characterizing ionically cross-linked alginate microgels using a nanoindentation technique, enabling precise assessment of their mechanical properties at the microscale. We report on the microfluidic fabrication of alginate microgels with varying sizes at different cross-linker concentrations and on the mechanical characterization of the resulting microgels in terms of Young's moduli as well as viscoelastic behavior. Measurements conducted using dynamic nanoindentation reveal that microgel elasticity is strongly influenced by the ionic composition of the surrounding media, in particular, the concentration of calcium and sodium. We demonstrate that the highest Young's modulus observed for ionically cross-linked alginate microgels is in deionized water (7.2 ± 0.9 kPa). A drastic softening effect is observed when the calcium cross-linked microgels are placed into a storage buffer containing divalent ions (0.7 ± 0.1 kPa) and cell culture media consisting of Dulbecco's Modified Eagle Medium (0.2 ± 0.1 kPa) with fetal bovine serum (0.4 ± 0.1 kPa). High concentrations of sodium were found to disrupt ionic cross-links, decreasing stiffness and increasing viscosity, with reversible effects observed upon switching back to deionized water. These findings highlight the importance of media selection for applications requiring mechanical stability, and we provide guidelines for measuring the mechanical properties of microgels in a robust manner that is applicable to a wide range of different conditions.

Although immune checkpoint inhibition in particular has shown promise in cancer immunotherapy, it is not always efficient. Recent studies suggest that SMARCAL1 may play a role in tumor immune evasion, yet its pan-cancer role is unclear. We conducted a comprehensive analysis of SMARCAL1 using TCGA, GTEx, and CCLE databases, evaluating its expression, genetic alterations, epigenetic modifications, and their clinical correlations across 33 cancer types. Our findings indicate that SMARCAL1 is overexpressed in several cancers, such as Glioma, LUAD, KIRC, and LIHC, impacting prognosis. Elevated SMARCAL1 is linked to poor outcomes in Glioma, LUAD, and LIHC but correlates with better survival in KIRC. We also found significant associations between SMARCAL1 expression and DNA methylation in 13 cancers. Furthermore, SMARCAL1 expression correlates with immune infiltration, suggesting it as a potential therapeutic target in cancer immunotherapy. This study underscores the need for further research on SMARCAL1 to enhance immunotherapeutic strategies.

<h4>Background</h4>The rapid spread of covid-19 overwhelmed healthcare systems. This study aimed to investigate the impact of the covid-19 pandemic on hospitalizations and hospital deaths due to cerebrovascular diseases (CVD) in São Paulo state, Brazil.<h4>Methods</h4>This ecologic study evaluated the CVD hospitalizations and hospital deaths (2017-2021) by demographic features and CVD type. During the pandemic (2020-2021), segmented regression models were used to detect changes in CVD trends. We also evaluated the detrended cross-correlation between CVD deaths and hospitalization with the SARS-Cov-2 infection series.<h4>Results</h4>During the pandemic, there is a 35% reduction in CVD hospitalizations, mainly in elective admissions and ischemic stroke, but a 6.5% increase in deaths, especially in Black and Brown individuals, and those aged 20-29 years. From 2020 to 2021, Black and Brown individuals experience an earlier and more prolonged increase in hospital deaths. Ischemic CVD hospitalizations decrease in the first quarter of 2020. Older people exhibit a monthly increase of 2.9% in hospitalizations and 5.3% in deaths in the 2nd and 3rd quarters of 2021. SARS-Cov-2 infections are inversely correlated to CVD hospitalizations and directly correlated to CVD hospital deaths.<h4>Conclusions</h4>Covid-19 pandemic negatively affects CVD hospitalizations and deaths, particularly in Black and Brown individuals. The decrease in hospitalizations and increase in hospital deaths of ischemic CVD highlights vulnerability in accessing healthcare resources during the pandemic.

Atrial fibrillation (AF) is a predominant cardiac arrhythmia with unclear etiology. This study used bioinformatics and machine learning to explore the relationship between mitochondrial energy metabolism-related genes (MEMRGs) and immune infiltration in AF. The datasets GSE31821, GSE41177, and GSE79768 were retrieved from the Gene Expression Omnibus (GEO) database, and differential expression analysis identified 59 mitochondrial energy metabolism-related differentially expressed genes (MEMRDEGs) associated with AF. Key MEMRDEGs were selected using the least absolute shrinkage and selection operator (LASSO) and support vector machine (SVM) methods, and a diagnostic model was developed. Immune infiltration was assessed using single-sample gene set enrichment analysis (ssGSEA) and the microenvironment cell population counter (MCPcounter). The diagnostic model, based on the key genes ACAT1, ALDH1L2, HTT, OGDH, and SLC25A3, achieved an area under the curve (AUC) of 0.903. Significant differences in immune cell composition were observed between the AF and control groups. ALDH1L2 was positively correlated with most immune cells, while SLC25A3 showed a negatively correlated with the monocytic lineage. The findings indicate that MEMRGs interact with immune responses in AF, offering insights into the potential molecular mechanisms and therapeutic targets for AF.

<h4>Objective</h4>The study objective was to determine whether associations between a genetic risk score (GRS) for insulin resistance (IR) and measures of insulin sensitivity differ by race and/or BMI status in African American (AA) and European American (EA) adults without diabetes.<h4>Methods</h4>Fifty-three AA and 54 EA participants were classified into "high" or "low" BMI groups using the sample median (25.9 kg/m²) as the cut point. The GRS was derived from 52 previously identified genetic variants. Skeletal muscle insulin sensitivity was measured with the hyperinsulinemic-euglycemic clamp. The homeostasis model assessment of insulin resistance (HOMA-IR) and the Matsuda index of insulin sensitivity were calculated from oral glucose tolerance test values to determine hepatic and whole-body insulin sensitivity, respectively. Linear regression models, stratified by race, assessed interactions between BMI status and GRS on measures of insulin sensitivity.<h4>Results</h4>In EA participants, associations of GRS with HOMA-IR and the Matsuda index differed by BMI status, where the GRS was associated with IR in the high-BMI group only. In AA participants, associations from the clamp differed by BMI status, but an association was observed only in the low-BMI group.<h4>Conclusions</h4>These results highlight the heterogeneity of IR and support the hypothesis that the relationship between genetic predisposition for IR and obesity is race- and tissue-specific.

A hyperexcitability of the motor system is consistently observed in Amyotrophic Lateral Sclerosis (ALS) and has been implicated in the disease pathogenesis. What drives this hyperexcitability in the vast majority of patients is unknown. This is important to know as existing treatments simply reduce all neuronal excitability and fail to distinguish between pathological changes and important homeostatic changes. Understanding what drives the initial pathological changes could therefore provide better treatments. One challenge is that patients represent a heterogeneous population and the vast majority of cases are sporadic. One pathological feature that almost all (~97%) cases (familial and sporadic) have in common are cytoplasmic aggregates of the protein TDP-43 which is normally located in the nucleus. In our experiments we investigated whether this pathology was sufficient to increase neuronal excitability and the mechanisms by which this occurs. We used the TDP-43(ΔNLS) mouse model which successfully recapitulates this pathology in a controllable way. We used in vivo intracellular recordings in this model to demonstrate that TDP-43 pathology is sufficient to drive a severe hyper-excitability of spinal motoneurones. Reductions in soma size and a lengthening and constriction of axon initial segments were observed, which would contribute to enhanced excitability. Resuppression of the transgene resulted in a return to normal excitability parameters by 6-8 weeks. We therefore conclude that TDP-43 pathology itself is sufficient to drive a severe but reversible hyperexcitability of spinal motoneurones.

Stress granules are membraneless organelles that serve as a protective cellular response to external stressors by sequestering non-translating messenger RNAs (mRNAs) and regulating protein synthesis. Stress granules formation mechanism is conserved across species, from yeast to mammals, and they play a critical role in minimizing cellular damage during stress. Composed of heterogeneous ribonucleoprotein complexes, stress granules are enriched not only in mRNAs but also in noncoding RNAs and various proteins, including translation initiation factors and RNA-binding proteins. Genetic mutations affecting stress granule assembly and disassembly can lead to abnormal stress granule accumulation, contributing to the progression of several diseases. Recent research indicates that stress granule dynamics are pivotal in determining their physiological and pathological functions, with acute stress granule formation offering protection and chronic stress granule accumulation being detrimental. This review focuses on the multifaceted roles of stress granules under diverse physiological conditions, such as regulation of mRNA transport, mRNA translation, apoptosis, germ cell development, phase separation processes that govern stress granule formation, and their emerging implications in pathophysiological scenarios, such as viral infections, cancer, neurodevelopmental disorders, neurodegeneration, and neuronal trauma.

The nervous system processes a vast amount of information, performing computations that underlie perception, cognition, and behavior. During development, neuronal guidance genes, which encode extracellular cues, their receptors, and downstream signal transducers, organize neural wiring to generate the complex architecture of the nervous system. It is now evident that many of these neuroguidance cues and their receptors are active during development and are also expressed in the adult nervous system. This suggests that neuronal guidance pathways are critical not only for neural wiring but also for ongoing function and maintenance of the mature nervous system. Supporting this view, these pathways continue to regulate synaptic connectivity, plasticity, and remodeling, and overall brain homeostasis throughout adulthood. Genetic and transcriptomic analyses have further revealed many neuronal guidance genes to be associated with a wide range of neurodegenerative and neuropsychiatric disorders. Although the precise mechanisms by which aberrant neuronal guidance signaling drives the pathogenesis of these diseases remain to be clarified, emerging evidence points to several common themes, including dysfunction in neurons, microglia, astrocytes, and endothelial cells, along with dysregulation of neuron-microglia-astrocyte, neuroimmune, and neurovascular interactions. In this review, we explore recent advances in understanding the molecular and cellular mechanisms by which aberrant neuronal guidance signaling contributes to disease pathogenesis through altered cell-cell interactions. For instance, recent studies have unveiled two distinct semaphorin-plexin signaling pathways that affect microglial activation and neuroinflammation. We discuss the challenges ahead, along with the therapeutic potentials of targeting neuronal guidance pathways for treating neurodegenerative diseases. Particular focus is placed on how neuronal guidance mechanisms control neuron-glia and neuroimmune interactions and modulate microglial function under physiological and pathological conditions. Specifically, we examine the crosstalk between neuronal guidance signaling and TREM2, a master regulator of microglial function, in the context of pathogenic protein aggregates. It is well-established that age is a major risk factor for neurodegeneration. Future research should address how aging and neuronal guidance signaling interact to influence an individual's susceptibility to various late-onset neurological diseases and how the progression of these diseases could be therapeutically blocked by targeting neuronal guidance pathways.

While recent work has identified roles for immune mediators in regulating neural activity, how innate immune signaling within neurons influences neurotransmission remains poorly understood. Emerging evidence suggests that the modulation of neurotransmission may serve important roles in host protection during infection of the central nervous system. Here, we showed that receptor-interacting protein kinase-3 (RIPK3) preserved neuronal survival during flavivirus infection through the suppression of excitatory neurotransmission. These effects occurred independently of the traditional functions of RIPK3 in promoting necroptosis and inflammatory transcription. Instead, RIPK3 promoted phosphorylation of the neuronal regulatory kinase calcium/calmodulin-dependent protein kinase II (CaMKII), which in turn activated the transcription factor cyclic AMP response element-binding protein (CREB) to drive a neuroprotective transcriptional program and suppress deleterious glutamatergic signaling. These findings identify an unexpected function for a canonical cell death protein in promoting neuronal survival during viral infection through the modulation of neuronal activity, highlighting mechanisms of neuroimmune crosstalk.

The bespoke environments in enzyme active sites can selectively accelerate chemical reactions by as much as 10¹⁹. Macromolecular and supramolecular chemists have been inspired to understand and mimic these accelerations and selectivities for applications in catalysis for sustainable synthesis. Over the past 60+ years, mimicry strategies have evolved with changing interests, understanding, and synthetic advances but, ubiquitously, research has focused on use of a molecular "cavity". The activities of different cavities vary with the subset of features available to a particular cavity type. Unsurprisingly, without synthetic access to mimics able to encompass more/all of the functional features of enzyme active sites, examples of cavity-catalyzed processes demonstrating enzyme-like rate accelerations remain rare. This perspective will briefly highlight some of the key advances in traditional cavity catalysis, by cavity type, in order to contextualize the recent development of robust organic cage catalysts, which can exploit stability, functionality, and reduced symmetry to enable promising catalytic modes.

The late stages of mammalian pregnancy are accompanied by a mild increase in insulin resistance likely due to enhanced glucose demand of the growing fetus. Therefore, as an adaptive process to maintain euglycemia during pregnancy, maternal β-cell mass expands leading to increased insulin release. Defects in functional β-cell adaptive expansion during pregnancy can lead to gestational diabetes mellitus (GDM). While the exact mechanisms that promote GDM are poorly understood, GDM is associated with inadequate functional β-cell mass expansion and with a systematic increase of oxidative stress. Here, we show that NRF2 levels are upregulated in mouse β-cells at gestational day 15 (GD15). Inducible β-cell-specific Nrf2 deleted (βNrf2KO) mice display reduced β-cell proliferation, increased β-cell oxidative stress and lipid peroxidation, compromised β-cell function, and elevated β-cell death, leading to impaired β-cell mass expansion and dysregulated glucose homeostasis towards the end of pregnancy. Importantly, the gestational hormone 17-β-estradiol (E2) increases NRF2 levels, and downregulation of NRF2 suppresses E2-induced protection of β-cells against oxidative stress, suggesting that E2 exerts its antioxidant effects through activation of NRF2 signaling in β-cells. Collectively, these data highlight the critical role of NRF2 in regulating oxidative stress during the adaptive response of β-cells in pregnancy and identify NRF2 as a potential therapeutic target for GDM treatment.

Oxidative stress-induced cardiomyocyte apoptosis was the primary causative factor of cardiovascular disease (CVD). However, the existing therapy drugs for oxidative stress were much less investigated, which underlined the necessity for new drug discovery and development. Herein, we aimed to synthesize several novel idebenone (IDE) derivatives and investigate the protective effect and mechanism of these derivatives against H₂O₂-induced oxidative stress injury in H9C2 cells by determining cell proliferation rate, detecting the reactive oxygen species (ROS) level, and the expression of related proteins. Additionally, the study also investigated the protective effect of IDE-1 pretreatment on Balb/c mice after hypoxia-reoxygenation. <i>In vivo</i> experiments, the damage to cardiomyocytes was assessed using hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining. The results showed that IDE-1 possessed the highest antioxidant damage activity among all IDE derivatives, which could notably decrease the levels of intracellular ROS. Furthermore, the antioxidant mechanism was confirmed to be potentially linked to the expression levels of the oxidation-related pathway heme oxygenase-1 (HO-1) and the apoptosis-related pathway Bcl-2/Bax and caspase-3. Our results demonstrated that IDE derivatives could be a new research direction for the treatment of cardiovascular diseases associated with oxidative stress.

Colorectal cancer (CRC) is a common and lethal malignancy that affects millions of people worldwide. Iron is an essential micronutrient that plays a vital role in various biological processes, but also has pro-oxidant and pro-inflammatory effects that may contribute to carcinogenesis. The relationship between iron and CRC is complex and influenced by multiple factors, such as dietary intake, absorption, storage, metabolism, and excretion of iron, as well as genetic and environmental factors that modulate iron homeostasis. This review article aims to provide an overview of the current evidences on the role of iron in CRC, discussing the potential mechanisms by which iron may affect CRC development and progression, as well as the implications for prevention and treatment. This review tries to focus on the following aspects: an introduction to iron and its role in CRC, role of heme and non-heme iron in CRC, dietary patterns, nutrition, and CRC, iron overload in CRC, iron deficiency and its role in CRC especially in surgery outcome and iron therapy and blood transfusion in CRC.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that affects millions of people worldwide and is characterized by persistent inflammation, pain, and joint destruction. In RA, the dysregulation of the immune system is well documented. However, the genetic basis of the disease is not fully understood, especially when extra-articular organs are involved. Interstitial lung disease (ILD) is a major cause of morbidity and mortality in patients with RA. Notably, RA-ILD shares several risk factors with idiopathic pulmonary fibrosis (IPF), namely male gender, smoking history, usual interstitial pneumonia (UIP) pattern of fibrosis, and association with the <i>MUC5B</i> rs35705950 polymorphism. In addition, other genetic susceptibilities are reported in RA-ILD for some HLA alleles and other less studied polymorphisms. However, the pathobiology of RA-ILD, particularly whether and to what extent genetic and environmental factors interact to determine the disease, remains elusive. In this review, we summarize and critically discuss the most recent literature on the genetics and pathogenesis of RA-ILD. The main clinical aspects of RA-ILD are also discussed.

Axon guidance proteins not only play a role in the formation of proper neural circuits but also have other important functions, such as cell survival, migration, and proliferation in the brain. Therefore, mutations in the genes encoding these proteins frequently cause various types of neurological disorders, including psychiatric disorders and neurodegenerative diseases. We previously identified an axon guidance protein, draxin, that is essential for the development of several neural circuits and cell survival in the brain. Recently, the deletion of the <i>draxin</i> gene was identified in an inbred BTBR T⁺ Itpr3^tf/J (BTBR/J) mouse, which is a widely used model of Autism Spectrum Disorder (ASD), suggesting that <i>draxin</i> deletion is a genetic factor for ASD-like characteristics in BTBR/J mice. In this review, I summarize the neuroanatomical abnormalities in <i>draxin</i> knockout mice by comparing them to BTBR/J mice and discuss the possible contributions of draxin to anatomical and behavioral phenotypes in BTBR/J mice.

Atherosclerosis (AS)-related cardiovascular disease and depression are often comorbid, with patients with cardiovascular disease facing an increased risk of depression, which worsens AS. Both diseases are characterized by oxidative stress and lipid metabolism disorders. Ferroptosis, a form of cell death characterized by iron overload and harmful lipid peroxide accumulation, is found in various diseases, including AS and depression. Consistent with the iron deposition and lipid peroxidation (LPO) that characterize the ferroptosis mechanism, disturbances in iron and lipid metabolism are also crucial pathogenic mechanisms in AS and depression. The comorbid mechanisms are complex, posing challenges for clinical treatment. Chinese herbs hold significant potential owing to their multi-target pharmacological effects. Therefore, this review aims to investigate iron overload, LPO, and ferroptosis across various cell types, the shared pathogenesis of AS and depression with ferroptosis, and research on Chinese herbal medicine targeting ferroptosis in the treatment of anti-AS co-depression. This provides a comprehensive understanding of AS co-depression disease from the perspective of ferroptosis.

Nonsuicidal self-injurious behaviour (SIB) is a debilitating manifestation of physical aggression commonly observed across neurodevelopmental, psychiatric, and genetic disorders. This behaviour arises from a multifactorial aetiology involving genetic predispositions, epigenetic modifications, neurotransmitter dysregulation, and environmental stressors. Dysregulation in dopaminergic, serotonergic, glutamatergic, and GABAergic systems has been implicated in the pathophysiology of SIB, alongside structural and functional abnormalities within fronto-limbic-striatal circuits. These disruptions impair key processes, such as emotional regulation, reward processing, and behavioural inhibition, contributing to the emergence and reinforcement of SIB. Advances in preclinical research using genetic, lesion-based, pharmacological, and environmental animal models have been instrumental in elucidating the molecular and neurocircuitry underpinnings of SIB. Emerging neuromodulation therapies targeting critical nodes within the fronto-limbic-striatal network, particularly deep brain stimulation, have shown promise in treating severe, refractory SIB and improving quality of life. This review integrates current evidence from clinical studies, molecular research, and preclinical models to provide a comprehensive overview of the pathophysiology of SIB and therapeutic approaches. By focusing on the molecular mechanisms and neural circuits underlying SIB, we highlight the translational potential of emerging pharmacological and neuromodulatory therapies. A deeper understanding of these pathways will pave the way for precision-based interventions, bridging the gap between molecular research and clinical applications in SIB and related conditions.

Aberrant protein glycosylation is closely associated with a number of biological processes and diseases. However, characterizing the types of post-translational modifications (PTMs) from the complex biological samples is challenging for comprehensive glycoproteomic analysis. The development of high-performance enrichment materials and strategies during the sample pretreatment process is a prerequisite to glycoproteome research. Here in this work, a sulfonate-rich covalent organic framework (COF) called TpPa-(SO₃H)₂ (referred to as SCOF-2) was synthesized using the Schiff base reaction for the identification of glycopeptides. Benefiting from high hydrophilicity and abundant sulfonate affinity, a total of 28 and 16 glycopeptides could be efficiently detected from the standard glycoproteins of horseradish peroxidase (HRP) and immunoglobulin G (IgG) tryptic digest, respectively. Moreover, the as-prepared sulfonate-rich SCOF-2 has an ultralow detection limit (0.01 fmol μL^-1), excellent enrichment selectivity (molar ratio HRP:BSA = 1:5000), satisfactory recovery rate (89.1%), high adsorption capacity (150 mg g^-1) and good reusability in the individual enrichment. Meanwhile, by using the SCOF-2 adsorbent, 196 and 194 endogenous glycopeptides in the serum of ovarian cancer patients and healthy people among triplicates were successfully enriched and identified, respectively, using combined nanoLC-MS/MS technology. It demonstrated its great application potential in glycoproteomics research and provided a novel insight for the design of affinity materials.

Pyrethroids are one of the most commonly used insecticides worldwide in agriculture, public health, and household products. To monitor the presence of pyrethroids in the environment and in food, a broad-spectrum monoclonal antibody (mAb), CL/CN-1D2, was prepared. This mAb demonstrates a 50% inhibitory concentration (IC50) for different pyrethroids: cypermethrin (129.1 µg/L), β-cypermethrin (199.6 µg/L), cyfluthrin (215.5 µg/L), fenpropathrin (220.3 µg/L), λ-cyhalothrin (226.9 µg/L), β-cyfluthrin (241.7 µg/L), deltamethrin (591.2 µg/L), and fenvalerate (763.1 µg/L). Using the mAb CL/CN-1D2, a highly sensitive heterologous indirect competitive ELISA (ic-ELISA) was developed for the rapid detection of these pyrethroids. The limit of detection (LOD) for the eight pyrethroids in water, milk, celery, and leek matrices ranged from 24.4 to 152.2 μg/kg. The recoveries ranged from 65.1% to 112.4%, with a coefficient of variation (CV) below 15%. A robust correlation (<i>R</i>² = 0.9945) between the ic-ELISA and GC indicated that the ic-ELISA is a reliable tool for the rapid and cost-effective screening of pyrethroids residues.

Formaldehyde, a pervasive environmental toxin, has well-documented deleterious effects on critical organ systems. This review examines the therapeutic potential of <i>Ferula asafoetida</i> (FA) root extracts in protecting the testes, kidneys, and liver against formaldehyde-induced toxicity in rodent models. The literature reveals that FA's active constituents, known for their potent antioxidant and anti-inflammatory activities, may counteract oxidative stress and cellular damage caused by formaldehyde exposure. The review explores formaldehyde-induced pathophysiological mechanisms and FA's protective effects, including mitigation of oxidative damage, inflammation, and apoptosis. By analyzing empirical evidence, it compares the efficacy of various extract preparations, dosage regimens, and treatment durations. The review also addresses methodological heterogeneity and challenges in extrapolating findings to humans. It concludes with a call for rigorous, controlled clinical trials to validate FA's therapeutic viability, offering hope for those affected by formaldehyde toxicity.

Tail docking is still used in pigs to reduce the prevalence of tail biting, although it is purely symptomatic and contrary to animal welfare. Genetic selection for shorter tails might, however, help to avoid tail docking and has therefore been proposed. A genetic basis for tail length is known for many species. Variability in tail length, including moderate heritability, has also been demonstrated in pigs. The aim of the present study was to identify genetic markers for tail length and to define candidate genes. To this end, 140 piglets were phenotyped and genotyped at 3 days of age and a genome-wide association study was performed. Seven SNPs were mapped on chromosomes 1, 2, 6, 11, and 15. Two linked SNPs on chromosome 2 resulted in a functional amino acid exchange. The genotypes at the SNPs were only associated with small differences in relative tail length of up to 16.5% (short genotype versus long genotype at SSC15), but at the same time with the occurrence of malformations in the form of tail kinks. The small effect size and the association between tail length and tail kinks, together with the generally pure symptomatic effect on tail biting, argue against the applicability of selection for shorter tails in pigs.

Disrupted iron balance causes anemia and iron overload leading to hypoxia and systemic oxidative stress. Iron overload may arise from red blood cell disorders such as sickle cell disease, thalassemia major and primary hemochromatosis, or from treatment with multiple transfusions. These hematological disorders are characterized by constant red blood cell hemolysis and the release of iron. Hemolysis is a continuous source of reactive oxygen species whose accumulation changes the redox potential in the erythrocyte, the endothelium and other tissue causing damage to organ systems. Iron overload and its consequences can be treated with iron chelating therapy. We have carried out structural studies of small molecule ligands that were previously reported for their iron chelating ability. The chelators were analyzed using mass spectrometry, proton nuclear magnetic resonance and infrared spectroscopy. The iron chelators, 2-benzoylpyridine-4,4-dimethyl-3-thiosemicarbazone, 3-ethyl-1-{[2-phenyl-1-(pyridin-2-yl)ethylidene]amino}thiourea and 1-{[2-phenyl-1-(pyridin-2-yl)ethylidene]amino}-3-(prop-2-en-1-yl)thiourea in their unbound conformation were crystallized and their structures were determined. This work addresses the evolution of a thiosemicarbazone class of iron chelators by analyzing and comparing the structure and properties of a series of closely related molecules, relating these to their <i>in vitro</i> activity thus providing valuable update to the search for newer, better and more effective iron chelators and metal-based therapeutics.

B-cell acute lymphoblastic leukemia (B-ALL) is the most common hematological malignancy of childhood. The initial risk-stratification includes the genetic evaluation for common recurrent translocations and aneuploidies in B-ALL using karyotyping, fluorescence in-situ hybridization (FISH), reverse transcription polymerase chain reaction (RT-PCR), etc. However, many cases remain genetically unclassified and are labeled as B-other ALL. Recent advances in the high throughput genomics have provided new insights and many distinct genetic subtypes have been identified in B-ALL. These new subtypes have been incorporated in the latest World Health Organization (WHO)-HAEM5 classification of haematolymphoid tumors and the International Consensus Classification. Besides the driver mutations, there is also a role of secondary genetic events like copy number alterations (CNAs) in the pathogenesis of B-ALL as well as for better risk-stratification. New molecular genetic risk evaluation like Moorman's criteria, IKZF1plus and MRplus scores have been recently validated which can risk-stratify B-ALL patients at baseline based on the CNA profile. The new diagnostic armamentarium now also includes flow-based ploidy analysis, cytokine receptor like factor 2 (CRLF2/TSLPR) overexpression screening on flow cytometry, multiplex ligation dependent probe amplification (MLPA) for CNAs, targeted RNA sequencing and whole transcriptome sequencing for detection of known and novel fusions. The gene expression profiling data can also be assessed with the machine learning based genetic classifiers for prediction of the newly identified subtypes of B-ALL. This review attempts to provide updates about the latest developments in the field of B-ALL genetics with inputs of Indian data, wherever available.

<title>Abstract</title> <p><bold>Background</bold>: Ferroptosis, a novel form of regulated cell death, has emerged as a significant research focus due to its involvement in various cancers. This study combines single-cell RNA sequencing with Mendelian randomization (MR) methods to identify genetic factors that are associated with ferroptosis and explore their potential impact on the pathogenesis of glioblastoma (GBM). <bold>Methods</bold>: MR and multiple validation methods were used to identify ferroptosis-related genes in glioblastoma. Single-cell analysis was performed to evaluate the expression levels of ferroptosis markers across different glioblastoma cell types. Additionally, gene enrichment analysis was conducted to explore gene functions, while survival analysis examined the relationship between gene expression and patient prognosis. Immune cell infiltration analysis was also carried out for genes associated with prognosis. <bold>Results</bold>: MR and sensitivity analyses identified 9 ferroptosis-related genes that are associated with GBM: SIRT1, KDM3B, VCP, GPT2, PRDX6, CISD2, TP53, FLT3, and FANCD2. Co-localization analysis showed a significant association between the VCP gene and GBM. Single-cell analysis revealed that PRDX6 is highly expressed in tumor tissues. Gene enrichment analysis highlighted the biological processes such as cell metabolism, DNA repair, ubiquitination, and autophagy in the occurrence and progression of GBM. Immune infiltration and survival analyses suggest that CISD2 was related to CD8+ and CD4+ T cell infiltration, and may affect patient prognosis. <bold>Conclusions</bold>: This study combines MR analysis and single-cell analysis to reveal the crucial role of ferroptosis genes in GBM. These genes influence the occurrence and development of GBM by regulating processes such as metabolism, DNA repair, oxidative stress, and autophagy. Among them, PRDX6 plays an important role in microscopic research, while ubiquitination and autophagy are key drivers of GBM progression. The CISD2 gene may influence patient prognosis by regulating T cell infiltration, thereby promoting immune tolerance mechanisms in tumor cells.</p>

Progressive neurological decline in multiple sclerosis is associated with axonal loss and synaptic dysfunction in the non-demyelinated normal appearing gray matter (NAGM) and prominently in the cerebellum. In contrast to early disease stages, where synaptic and neuro-axonal pathology correlates with the extent of T cell infiltration, a prominent role of the innate immune system has been proposed for progressive MS. However, the specific contribution of microglia and astrocytes to synaptic cerebellar pathology in the NAGM- independent of an adaptive T cell response - remains largely unexplored. In the present study, we quantified synaptic changes in the cerebellar NAGM distant from demyelinated lesions in a mouse model of toxic demyelination. Proteomic analysis of the cerebellar cortex revealed differential regulation of synaptic and glutamate transport proteins in the absence of evident structural synaptic pathology or local gray matter demyelination. At the functional level, synaptic changes manifested as a reduction in frequency-dependent facilitation at the parallel fiber- Purkinje cell synapse. Further, deficiency of MyD88, an adaptor protein of the innate immune response, associated with a functional recovery in facilitation, reduced changes in the differential expression of synaptic and glutamate transport proteins, and reduced transcription levels of inflammatory cytokines. Nevertheless, the characteristics of demyelinating lesions and their associated cellular response were similar to wild type animals. Our work brings forward an experimental paradigm mimicking the diffuse synaptic pathology independent of demyelination in late stage MS and highlights the complex regulation of synaptic pathology in the cerebellar NAGM. Moreover, our findings suggest a role of astrocytes, in particular Bergmann glia, as key cellular determinants of cerebellar synaptic dysfunction.

We report a case series of three pediatric patients with inflammatory bowel disease (IBD) and variants in <i>HFE</i> which causes hereditary hemochromatosis (HH) type 1. Mice models suggest that these patients may be at increased risk for colitis and colon cancer. We detail the clinical course of these patients regarding IBD and iron overload and <i>HFE</i>-related comorbidities. We also review the known literature regarding HH and IBD overlap, HH detection and screening recommendations, HH management strategies, and iron management strategies in the context of both iron overload risk and IBD.

The hypoxic and high-pressure microenvironment of the intervertebral discs poses a major challenge to the survival and therapeutic efficiency of exogenous stem cells. Therefore, improving the utilization efficiency and therapeutic effect of exogenous stem cells to delay intervertebral disc degeneration (IVDD) is of great importance. Here, hypoxic induction studies are conducted in vivo and in vitro using rat costal cartilage-derived skeletal stem cells (SSCs) and find that hypoxia activates the cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)/stimulator of interferon genes (STING) signaling pathway and increased reactive oxygen species (ROS) accumulation, triggering ferroptosis in SSCs through hypoxia-inducible factor-1 alpha-dependent mitophagy. Progressive hypoxia preconditioning reduce STING expression and ROS accumulation, inducing SSCs differentiation into nucleus pulposus-like cells via the Wnt signaling pathway. Considering this, a 3-D sustained-release culture carrier is generated by mixing SSCs with methacrylated hyaluronic acid and polydopamine nanoparticles coated with the STING inhibitor C-176 and evaluated its inhibitory effect on IVDD. This carrier is demonstrated to inhibit the cGAS/STING pathway and prevent ROS accumulation by continuously releasing C-176-coated polydopamine nanoparticles, thereby reducing ferroptosis, promoting differentiation, and ultimately attenuating IVDD, suggesting its potential as a novel treatment strategy.

No abstract available.

Iron-refractory iron deficiency anaemia (IRIDA) is a rare hereditary microcytic anaemia characterized by partial or complete resistance to oral iron supplementation, caused by elevated plasma hepcidin levels resulting from pathogenic variants in the TMPRSS6 gene. Although intravenous iron supplementation is often effective, patient responses can vary significantly due to various factors, and potential side effects of this treatment remain unclear. Additionally, evidence-based international guidelines for diagnosing and managing IRIDA are lacking. This review aims to provide patient-tailored treatment strategies, informed by case studies and expert opinion, to address the specific therapeutic needs of both children and adults with IRIDA.

Movement disorders are a group of heterogeneous neurological conditions associated with alterations of tone, posture and voluntary movement. They may either occur in isolation or as part of a multisystemic condition. More recently, the advent of next generation sequencing technologies has facilitated better understanding of the underlying causative genes and molecular pathways, thereby identifying targets for genetic therapy. In this review, we summarize the advances in genetic therapy approaches for both hyperkinetic and hypokinetic movement disorders, including Parkinson's Disease, Huntington's Disease and rarer monogenic conditions of childhood onset. While there have been significant advances in the field, multiple challenges remain, related to safety, toxicity, efficacy and brain biodistribution, which will need to be addressed by the next generation of genetic therapies.

The pathology of reactive, dysplastic, and neoplastic sinonasal seromucinous glands is complex, and their contribution to tumorigenesis of sinonasal carcinomas remains controversial. In our practice, we have observed the presence of respiratory epithelial adenomatoid hamartomas (REAH) and seromucinous hamartomas (SH) associated with adenoid cystic carcinomas (AdCC) in a subset of cases. In many of these cases, genuine atypical features and dysplastic characteristics of the glands were noted at the interface of SH and AdCC. To investigate this phenomenon further, 88 sinonasal AdCC cases were selected from the authors' files and analyzed histologically, immunohistochemically, and genetically searching for MYB/MYBL1 and NFIB gene fusions. HPV testing was also performed. Univariate statistical analysis was conducted on our cohort. Thirty-one cases (35%) showed features of atypical sinonasal glands arising in SH (ASGSH) at the SH-AdCC interface, characterized by bilayered epithelium, architectural disarray, mild nuclear polymorphism, and atypia, sometimes with colloid-like material in the lumen. The MYB immunomarker was negative in 14 ASGSHs (with a positive internal control in AdCC cells), while only two cases showed faint and moderate to weak expression of the antibody in ASGSH glands. In 12 cases, the immunostaining of ASGSH could not be properly assessed, while AdCC cells were negative. The immunostaining was not performed in five cases. Our findings suggest that a subset of sinonasal AdCC may originate in a multistep dysplastic process within SH, consistent with an SH-ASGSH-AdCC progression sequence.

In male mammals, spermatogonial stem cells (SSCs) are essential for sustaining lifelong spermatogenesis within the testicular open niche, a unique environment that allows SSC migration over an extended niche area. As SSCs undergo continuous mitotic division, mutations accumulate and are transmitted to the descendant SSC clones. Therefore, SSC clonal fate behaviors, in terms of their efficiencies in completing spermatogenesis and undergoing expansion within the niche, influence sperm genomic diversity. We aimed to elucidate the effects of physiological aging on SSC clonal fate behavior within the testicular open niche. We used single-cell RNA sequencing, lineage tracing, and intravital live imaging to investigate SSC behavior in aged mouse testes, where spermatogenesis, although reduced, persists. We found that undifferentiated spermatogonia maintained gene expression heterogeneity during aging. Among these, GFRα1⁺ cells, which exhibited state heterogeneity, showed accelerated proliferation and persistent motility, continuing to function as SSCs in older mice. In contrast, a subset of SSCs characterized by low Egr4 and Cops5 expression did not contribute to spermatid formation. These non-sperm-forming SSC clones increased in proportion among the total SSC clones and expanded spatially within the testicular open niche in old mice, a phenomenon not observed in young mice. The expansion of non-sperm-forming SSC clones in aged testes suggests that they occupy a niche space, limiting the availability of functional SSCs and potentially reducing sperm production and genetic diversity. These findings highlight age-specific clonal characteristics as hallmarks of stem cell aging within the testicular open niche and provide novel insights into the mechanisms governing reproductive aging.

<h4>Background</h4>High heterogeneity in gastric cancer (GC) remains a challenge for standard treatments and prognosis prediction. Dysregulation of sialic acid metabolism (SiaM) is recognized as a key metabolic hallmark of tumor immune evasion and metastasis. Herein, we aimed to develop a SiaM-based metabolic classification in GC.<h4>Methods</h4>SiaM-related genes were obtained from the MsigDB database. Bulk and single-cell transcriptional data of 956 GC patients were acquired from the GEO, TCGA, and MEDLINE databases. Proteomic profiles of 20 GC samples were derived from our institution. The consensus clustering algorithm was applied to identify SiaM-based clusters. The SiaM-based model was established via LASSO regression and evaluated via Kaplan‒Meier curve and ROC curve analyses. In vitro and in vivo experiments were conducted to explore the function of ST3GAL1 in GC.<h4>Results</h4>Three SiaM clusters presented distinct patterns of clinicopathological features, transcriptomic alterations, and tumor immune microenvironment landscapes in GC. Compared with clusters A and B, cluster C presented elevated SiaM activity, higher metastatic potential, more abundant immunosuppressive features, and a worse prognosis. Based on the differentially expressed genes between these clusters, a risk model for six genes (ARHGAP6, ST3GAL1, ADAM28, C7, PLCL1, and TTC28) was then constructed. The model exhibited robust performance in predicting peritoneal metastasis and prognosis in four independent cohorts. As a hub gene in the model, ST3GAL1 promoted GC cell migration and invasion in vitro and in vivo.<h4>Conclusions</h4>Our study proposed a novel SiaM-based classification that identified three metabolic subtypes with distinct characteristics of tumor microenvironment and clinical outcomes in GC.

Germline mutations of YY1 cause Gabriele-de Vries syndrome (GADEVS), a neurodevelopmental disorder featuring intellectual disability and a wide range of systemic manifestations. To dissect the cellular and molecular mechanisms underlying GADEVS, we combined large-scale imaging, single-cell multiomics and gene regulatory network reconstruction in 2D and 3D patient-derived physiopathologically relevant cell lineages. YY1 haploinsufficiency causes a pervasive alteration of cell type specific transcriptional networks, disrupting corticogenesis at the level of neural progenitors and terminally differentiated neurons, including cytoarchitectural defects reminiscent of GADEVS clinical features. Transcriptional alterations in neurons propagated to neighboring astrocytes through a major non-cell autonomous pro-inflammatory effect that grounds the rationale for modulatory interventions. Together, neurodevelopmental trajectories, synaptic formation and neuronal-astrocyte cross talk emerged as salient domains of YY1 dosage-dependent vulnerability. Mechanistically, cell type resolved reconstruction of gene regulatory networks uncovered the regulatory interplay between YY1, NEUROG2 and ETV5 and its aberrant rewiring in GADEVS. Our findings underscore the reach of advanced in vitro models in capturing developmental antecedents of clinical features and exposing their underlying mechanisms to guide the search for targeted interventions.

<h4>Background</h4>Type 2 diabetes (T2D) etiology is highly complex due to its multiple roots of origin. Polygenic risk scores (PRS) based on genome-wide association studies (GWAS) can partially explain T2D risk. Asian Indian people have up to six times higher risk of developing T2D than European people, and underlying causes of this disparity are unknown.<h4>Methods</h4>We have performed targeted sequencing of ten T2D GWAS/candidate regions using endogamous Punjabi Sikh families and replication studies using unrelated Sikh people and families from three other Indian endogamous ethnic groups (EEGs).<h4>Results</h4>We detect rare and ultra-rare variants (RVs) in KCNJ11-ABCC8 and HNF4A (MODY genes) cosegregated with late-onset T2D. We also identify RV enrichment in two new genes, SLC38A11 and ANPEP, associated with T2D. Gene-burden analysis reveals the highest RV burden contributed by HNF4A (p = 0.0003), followed by KCNJ11/ABCC8 (p = 0.0061) and SLC38A11 (p = 0.03). Some RVs detected in Sikh people are also found in Agarwals from Jaipur, both from Northern India, but were monomorphic in other two EEGs from South Indian people. Despite carrying a high burden of T2D and RVs, most families have a significantly lower burden of PRS. Functional studies show that an intronic regulatory variant (RV) in ABCC8 affects the binding of Pax4 and NF-kB transcription factors, influencing downstream gene regulation.<h4>Conclusions</h4>The high burden of T2D in these families may stem from the enrichment of noncoding RVs in a small number of major known genes (including MODY genes) with oligogenic inheritance alongside RVs from genes associated with polygenic susceptibility. These findings highlight the need to conduct deeper evaluations of families from non-European ancestries to identify potential novel therapeutics and implement preventative strategies.

Peritoneal metastasis (PM) is the primary site of distant metastasis in gastric cancer (GC) and is associated with an advanced disease stage and poor prognosis. Due to its high resistance to chemotherapy, disseminated peritoneal lesions are often untreatable. A primary reason for therapy resistance in cancer cells is often their defective cell death execution mechanisms. Ferroptosis, a newly identified type of regulated cell death, is strongly linked to the emergence and formation of tumors. Earlier studies have demonstrated the significant role of RNA-binding proteins in ferroptosis. Nevertheless, the fundamental process linking Staufen Double-Stranded RNA Binding Protein 1 (STAU1) to ferroptosis in the peritoneal metastasis of gastric cancer is yet to be clarified. This study shows that the RNA-binding protein STAU1 is crucial for regulating ferroptosis in gastric cancer cells. Elevated levels of STAU1 are linked to unfavorable outcomes in individuals diagnosed with gastric cancer. STAU1 was up-regulated by PLAGL2 and decreased the stability of NCOA4 mRNA by binding to the 3'-untranslated region. Decreased NCOA4 expression inhibits the accumulation of reactive iron, the occurrence of the Fenton reaction, and cellular ROS generation in the GC cells. Additionally, we showed that NCOA4 is crucial in the process of ferritinophagy triggered by the reduction of STAU1 in gastric cancer cells. Ultimately, the process safeguards GC cells from ferroptosis. These findings elucidate the function of PLAGL2/STAU1/NCOA4 in the ferroptosis of gastric cancer cells and provide theoretical backing for possible diagnostic markers and treatment targets for peritoneal metastasis in gastric cancer.

Jiawei Kongsheng Zhenzhong Pill(JKZP) is based on Kongsheng Zhenzhong Pill contained in the Tang Dynasty's "Thousand Golden Prescriptions," which exhibited good anti-ischemic and antidepressant effects in the previous study. However, its specific effects on post-stroke depression (PSD) and the mechanism are not clear. This study aimed to investigate the effects of JKZP in the treatment of PSD and related mechanisms. The decoction of JKZP was first analyzed for its medicinal chemical composition and screened for representative components of JKZP. The Middle cerebral artery occlusion (MCAO) method combined with solitary rearing and chronic unpredictable mild stress (CUMS) was used to establish a rat model of PSD, and to observe the effects of JKZP on the behavior and synaptic plasticity of PSD rats, and to investigate the mechanism of JKZP in the treatment of PSD by detecting the mRNA level, protein expression and activity of Netrin-1/DCC signaling pathway-related proteins. The results showed that the JKZP decoction contained loganin, β-asarone and other pharmaceutical ingredients, which have been reported to protect against cerebral ischemic injury and antidepressant effects. JKZP significantly improved the depression-like behavior of PSD rats and improved the damage to pyramidal neurons in the medial prefrontal cortex (mPFC) of PSD rats. Moreover, JKZP increased the density of dendritic spines in the mPFC of PSD rats, improved synaptic gap width and thickness of the post-synaptic density, and increased the number of synaptic vesicles. The results of Real-Time quantitative reverse transcription PCR (qRT-PCR), Western blotting, and pull-down assays revealed that JKZP increased netrin-1, deleted in colorectal cancer (DCC), and focal adhesion kinase (FAK) mRNA and protein expression, elevated the p-FAK/FAK ratio, and decreased myosin II protein expression and Ras homolog gene family member A (RhoA-GTP) activity in the mPFC of PSD rats. Taken together, JKZP can affect synaptic structural remodeling and improve depressive manifestations and neuronal damage in PSD rats by regulating the expression and activity of signaling molecules related to the netrin-1/DCC signaling pathway.

Altered levels of reactive oxygen species (ROS) are recognized as one of the key factors in mediating tumor cell survival in the tissue microenvironment, where they play a role in the initiation, progression and recurrence/relapse of colorectal cancer (CRC). Tumor cells can adapt to oxidative stress (OS) using genetic or metabolic reprogramming in the long or short term. In addition, tumor cells defend themselves through positive regulation of antioxidant molecules, enhancing ROS-driven proliferation. Balanced oxidative eustress levels can influence chemotherapy resistance, allowing tumor cells to survive treatment. Secondary effects of chemotherapy include increased ROS production and redox stress, which can kill cancer cells and eliminate drug resistance. Anticancer treatments based on manipulating ROS levels could represent the gold standard in CRC therapy. Therefore, exploring the modulation of the response to OS in deregulated signaling pathways may lead to the development of new personalized CRC treatments to overcome therapy resistance. In this review, we explore the role of ROS in the initiation and progression of CRC and their diagnostic implications as biomarkers of disease. Furthermore, we focused on the involvement of ROS in different CRC therapeutic options, such as surgery, radiotherapy, theranostic imaging, chemotherapy and immunotherapy and other precision medicine approaches.

Globally, ovarian cancer (OC) is the eighth most common malignant tumour in women. Unfortunately, its symptoms-especially at the early stages-are vague and non-specific, and, thus, most patients are diagnosed at the advanced stages of the disease (stage III and IV) when treatment is not curative. The currently available approved biomarkers are not sufficient for effective screening, prognosis, or monitoring of OC. Liquid biopsy tests such as circulating tumour DNA (ctDNA) analysis has the advantage of monitoring response to treatment in real time and providing a comprehensive genotypic profile of primary, metastatic, and recurrent tumours. Thus, ctDNA analysis can be used as a complementary test for effective diagnosis and monitoring of OC. We comprehensively review current studies (2019-2024) on OC, critically highlighting recent developments and applications of ctDNA for the diagnosis and management of the disease.

<h4>Background</h4>Patients with end-stage chronic diseases, especially those undergoing hemodialysis (HD), often experience mineral bone disease (MBD), leading to hypocalcemia, hyperphosphatemia, and elevated parathyroid hormone (PTH). Vitamin D deficiency and metabolism disorders are also common, resulting from impaired conversion of 25(OH)D3 to its active form, 1,25(OH)2D3, and reduced inactivation to 24,25(OH)2D3. This study aimed to assess the levels of 25(OH)D2, 25(OH)D3, 24,25(OH)2D3, 3-epi-25(OH)D3, and the vitamin D metabolism ratio (VMR) in patients with maintenance HD.<h4>Methods</h4>A cross-sectional study was conducted on 66 HD patients (22-90 years, average 61.3 ± 16.4), with a control group of 206 adults without chronic kidney disease (CKD), both without cholecalciferol supplementation.<h4>Results</h4>the HD patients had significantly lower 25(OH)D3 levels (15 ng/mL vs. 22 ng/mL) and higher deficiency rates (69% vs. 39%) compared to the controls. However, both groups showed similarly low levels of optimal vitamin D3. The HD patients had lower 24,25(OH)D3 levels (0.1 vs. 2.1 ng/mL) and a lower VMR (0.9% vs. 9%). 3-epi-25(OH)D3 levels and its ratio to 25(OH)D3 were significantly lower in the HD group. Alphacalcidol supplementation raised 1,25(OH)2D3 levels (30.4 vs. 16.2 pg/mL) without affecting other vitamin D metabolites. The HD patients had higher levels of 25(OH)D2 compared to the controls (0.61 vs. 0.31 ng/mL).<h4>Conclusions</h4>Vitamin D3 reserves are lower, and both functional deficiency and impaired catabolism of vitamin D3 are present in HD patients compared to the general population. The VMR index is the most sensitive parameter for vitamin D3 deficiency assessment, highlighting the importance of measuring 24,25(OH)D3. Alphacalcidol supplementation increases 1,25(OH)2D3 levels without affecting other vitamin D metabolites. 25(OH)D2 is the only metabolite that was higher in HD patients than the controls.

<b>Objective(s):</b> Premature ovarian insufficiency (POI), affecting 1% of women, is characterized by the loss of ovarian activity with amenorrhea or oligomenorrhea and increased gonadotropins occurring before the age of 40 years. Iatrogenic, autoimmune, and genetic causes are known to be involved in POI, but nearly 70% of all forms remain unexplained. Recent and new genetic analyses promote the identification of new candidate genes. The aim of this study was to evaluate the contribution of array-CGH and next-generation sequencing (NGS) in the diagnosis of POI. <b>Study design:</b> Twenty-eight idiopathic POI patients with primary or secondary amenorrhea underwent genetic screening by array-CGH and NGS using a custom capture design of 163 genes known or suspected to be involved in ovarian function. The clinical, biological, and ultrasound characteristics of the patients were also recorded. <b>Results:</b> Four of the twenty-eight patients had primary amenorrhea (14.3%), and twenty-four (85.7%) had secondary amenorrhea, with an average age at diagnosis of 27.7. Eleven patients (39.3%) had a family history of POI. Our study identified a genetic anomaly in 16 of 28 patients (57.1%): one patient carried a causal copy number variation (CNV), eight patients carried a causal single nucleotide variation (SNV)/indel variation (28.6%), and seven other patients carried variants of uncertain significance. <b>Conclusions:</b> Our study was the first to combine genetic analyses by using both array-CGH and NGS in the same patients. It confirmed the usefulness of both analyses in the identification of pathogenic variations responsible for idiopathic POI. Early genetic diagnosis plays a major role in the management of complications and the screening of relatives.

In polyglutamine (polyQ) diseases, including Huntington disease (HD) and spinocerebellar ataxia type 3 (SCA3), targeting the mutant CAG tract in mRNA could be a therapeutic strategy for lowering pathogenic protein. We explored the viability of this therapeutic strategy <i>in vivo</i> at the level of the reagent design, toxicity, systemic delivery, brain regions transduction, silencing efficiency, and allele preference. We designed a series of CAG-directed short hairpin RNAs (shRNAs) based on a previous A2 reagent, allele selective <i>in vitro</i>. Humanized HD (Hu^128Q/21Q) and SCA3 (Ki^150Q/21Q) mice with mutant ∼100 CAGs and normal 21 CAGs alleles were used to simulate biallelic conditions occurring in patients. We administered AAV-PHP.eB shRNAs-encoding vectors into the blood as an equivalent of non-invasive CAG-directed brain-targeted therapy crossing the blood-brain barrier. We demonstrate that optimized CAG-targeted A4(P10) and A4(P10,11) shReagents can lower mutant huntingtin and ataxin-3 protein and its aggregates by targeting brain regions selectively and with diminished toxicity compared to other tested shRNAs. The important considerations of the approach are the silencing efficiency depending on the transduction region and careful dose adjustment. Moreover, the CAG approach could be suitable to target somatic expansion. Our work paves the way toward developing the therapy for polyQ diseases, potentially shortening drug development.

Huntington's disease (HD) and Spinocerebellar Ataxia (SCA) are debilitating neurological disorders triggered by the expansion of C<b>A</b>G sequences within the specific genes (HTT and ATXN, respectively). These are characterized as poly glutamine (polyQ) disorders, which are marked by widespread neurodegeneration and metabolic irregularities across systemic, cellular, and intracellular levels. This study aimed to identify small molecules that specifically interact with and target the toxic C<b>A</b>G repeat RNA. Here, we investigated the neuroprotective effects of Oseltamivir, an antiviral drug, against the HD and SCA-causing C<b>A</b>G repeats, through biophysical, cellular, and <i>Drosophila</i> model-based studies. Using a multidimensional approach encompassing biophysical techniques, cellular assays, and a <i>Drosophila</i> model, we explored Oseltamivir's interaction with toxic C<b>A</b>G repeat RNA. Our comprehensive analyses, including circular dichroism (CD), isothermal titration calorimetry (ITC), electrophoretic mobility shift assay (EMSA), and nuclear magnetic resonance (NMR) spectroscopy, demonstrated Oseltamivir's specific binding affinity for AA mismatches and its potential to mitigate the toxicity associated with polyQ aggregation. Moreover, the identified U.S. FDA-approved drug effectively mitigated polyQ-induced toxicity in both HD cells and the <i>Drosophila</i> model of the disease. The results obtained from this drug repurposing approach are indicative of the neuro-shielding role of Oseltamivir in HD and several SCAs, paving the way for its translation into clinical practice to benefit patients afflicted with these devastating diseases.

Cell cycle events are ordered by cyclin-dependent kinases (CDKs), which phosphorylate hundreds of substrates. Multiple phosphatases oppose CDK substrate phosphorylation, yet a systematic understanding of how these phosphatases collectively influence phosphorylation timing is lacking. Here, we show that phosphatases influence the timing of CDK substrate phosphorylation during G2 and mitosis in fission yeast. We identify substrates of four phosphatases (PP2A-B55, PP2A-B56, CDC14, and PP1), showing that each phosphatase targets a distinct subset of CDK substrate sites. On average, sites dephosphorylated by CDC14 and PP2A-B56 are phosphorylated earlier during G2, followed by sites dephosphorylated by PP1 and then PP2A-B55. This suggests that the identity of the phosphatase impacts the timing of CDK substrate phosphorylation, establishing different phosphorylation thresholds at the G2/M transition. Consistent with this, depletion of PP2A-B55 and CDC14 advances mitotic onset independently of CDK activity regulation, likely due to the earlier phosphorylation of their respective CDK substrates.

<h4>Summary</h4> Oocyte cytoplasmic lattices are critical for early embryo development but their composition and function are not fully understood. Mutations in PADI6 , an essential component of cytoplasmic lattices, lead to early embryonic developmental arrest and female infertility. To investigate PADI6 function in mRNA storage, global protein levels, and lattice composition during early mammalian development we used single cell transcriptomics and proteomics methods to study two mouse models. Padi6 null mutation resulted in inhibition of embryonic genome activation, defective maternal mRNA degradation, and disruption to protein storage on the cytoplasmic lattices. Distinct developmental phenotypes were observed with a hypomorphic Padi6 mutation. By developing a powerful single cell proteomic fractionation method, we define the cytoplasmic lattice enriched proteome in which we find essential components of another major oocyte-specific compartment, the endolysosomal vesicular assembly (ELVA), suggesting previously unknown interconnections between them. Our findings highlight a critical scaffolding function of PADI6 and implicate cytoplasmic lattices as regulatory hubs for key processes in the oocyte and early embryo, including translation, respiration and protein degradation.

West Nile virus (WNV) is a neurotropic mosquito-borne orthoflavivirus, representing a relevant public health threat. Identification of biomarkers that would predict the course of WNV infection is of interest for the early identification of patients at risk and for supporting decisions on therapeutic interventions. In this study, serum levels of glial fibrillary acidic protein (sGFAP) and neurofilament light chain (sNfL), which are markers of brain tissue damage and inflammation, were analysed in 103 subjects with laboratory-confirmed WNV infection, comprising 13 asymptomatic blood donors, 23 with WN fever (WNF), 50 with encephalitis/meningoencephalitis (E/ME) and 17 with acute flaccid paralysis (AFP). In addition, 55 WNV-negative subjects with fever, encephalitis or healthy asymptomatic were included as controls. Age-adjusted levels of both sNfL and sGFAP were significantly higher in patients with neuroinvasive disease than in those with fever or asymptomatic (both WNV-positive and WNV-negative), suggesting a broad association of these biomarkers with systemic inflammation and brain injury resulting from infection. In WNV patients, the combined analysis of sNfL and sGFAP early after symptom onset allowed discrimination between neuroinvasive disease and fever with 67.2% sensitivity and 91.3% specificity, but not between E/ME and AFP. Furthermore, high levels of sNfL and sGFAP were significantly associated with prolonged hospital stay, intensive care unit admission and the occurrence of death or severe sequelae. Detection of WNV RNA in CSF was associated with increased sGFAP. In conclusion, our study indicates the potential utility of sNfL and sGFAP as biomarkers of WNV disease severity and adverse outcome.

MicroRNAs (miRNAs/miRs) are endogenous, small non‑coding RNAs conserved across species that post‑transcriptionally regulate gene expression by both suppressing translation and inducing mRNA degradation. miRNAs are found in various tissues, exhibit variable expression and their dysregulation is implicated in numerous disease processes. Furthermore, miRNA expression levels have a key role in the normal development of kidney tissue and are key regulators of kidney function, modulating diverse biological processes across renal cell lineages. miR‑378 participates in pathological processes associated with kidney diseases, including kidney cancer, kidney transplantation and diabetic nephropathy. Despite its considerable effects on these conditions, a comprehensive summary of the roles of miR‑378 is unavailable. In the present review, the existing literature on miR‑378 in kidney diseases is consolidated, and its validated gene targets and biological effects in both malignant and non‑malignant conditions are highlighted, thereby providing a foundation for future research.

λ-type carrageenan (λ-CAR) polysaccharides remain overlooked in the preparation of medical nanoparticles (NP) due to their unsuitable rheological properties and undesired biological effects, although they can also offer advantageous properties. To overcome these obstacles, oligosaccharide derivatives (λ-COS) have been successfully applied to the synthesis of stable NP incorporating both, ferrite cores and divalent manganese (Mn²⁺) releasable ions. The acute pro-inflammatory behavior and anticoagulant activity of native λ-CAR were significantly reduced in the case of λ-COS and λ-COS NP, rendering possible their use for medical applications. In vivo MRI studies in mice showed that the λ-COS NP framework is promising for two applications. The first is partial Mn²⁺ release into the plasma to achieve intracellular Mn²⁺-based contrast of the myocardium and imaging of the hepatobiliary system. Second, it serves as a novel sugar-based coating that confers suitable pharmacokinetic properties to NP, making it promising for further targeted therapy applications.

Low nephron numbers are associated with an increased risk of developing chronic kidney disease (CKD) and hypertension, which are significant global health problems. To investigate the impact of nephron deficiency, our laboratory developed a novel inbred rat model (HSRA rat). In this model, ∼75% of offspring are born with a single kidney (HSRA-S), compared with two-kidney littermates (HSRA-C). HSRA-S rats show impaired kidney development, resulting in ∼20% fewer nephrons. Our previous data and current findings demonstrate that nephron deficit (failure of one kidney to form and altered development in the remaining kidney) predisposes HSRA-S to CKD late in life (with increased proteinuria by 18 mo of age in HSRA-S = 51 ± 3.4 vs. HSRA-C = 8 ± 1.5 mg/24 h). To understand early molecular mechanisms contributing to the increased predisposition to CKD, Methylseq using reduced representation bisulfite sequencing, single-nuclei (sn)RNAseq, and discovery proteomics were performed in kidneys of 4-wk-old HSRA rats. Methylation analysis revealed a small number of differences, including five differentially methylated cytosines and six differentially methylated regions between groups. The snRNAseq analysis identified differentially expressed genes in most kidney cell types, with several hundred genes dysregulated depending on the analysis method (Seurat vs. DESeq2). Notably, many genes are involved in kidney development. Discovery proteomic analysis identified 366 differentially expressed proteins. A key finding was dysregulation of <i>Deptor</i>/DEPTOR and <i>Amdhd2</i>/AMDHD2 across omics layers, suggesting a potential role in compensatory mechanisms or the genetic basis of altered kidney development. Further understanding of these mechanisms may guide interventions to preserve nephron health and slow kidney disease progression.<b>NEW & NOTEWORTHY</b> The HSRA rat is a novel model of nephron deficiency and provides a unique opportunity to study the association between nephron number and chronic kidney disease (CKD). Previous work characterized the impact of age, hypertension, and diabetes on the development of CKD in HSRA animals. This study examined early changes in epigenetics, cell-type specific transcriptome, and proteomic changes in the kidney that likely predispose the model to CKD with age.

<h4>Background</h4>Deep Vein Thrombosis (DVT) represents a significant complication following orthopedic injuries, particularly patellar fractures. Despite the prevalence, comprehensive studies assessing the incidence rates and identifying specific risk factors in patellar fracture patients are limited.<h4>Methods</h4>This retrospective analysis reviewed electronic medical records from 3311 patients treated for patellar fractures at two tertiary hospitals between November 2013 and January 2023. The study focused on patient demographics, fracture characteristics, comorbidities, and laboratory parameters to evaluate the incidence and predictors of DVT. DVT prophylaxis measures and diagnostic criteria, including Doppler Ultrasound Scans, were rigorously applied.<h4>Results</h4>In patients with patellar fractures, the DVT incidence was 30.8%, with 1,790 clots identified in 1,021 diagnosed individuals, predominantly on the injured side (96.7%), and a minor portion on the uninjured side (3.2%). Key risk factors included older age (P<0.001, OR = 1.038), the presence of open injuries (P = 0.002, OR = 1.521), multiple injuries (P<0.001, OR = 3.623), and prolonged time from injury to surgical treatment (P<0.001, OR = 1.097). Conversely, higher levels of albumin (ALB) (P = 0.029, OR = 0.983) and sodium (Na) (P = 0.028, OR = 0.971) were identified as protective factors against DVT. Besides, ROC curve analysis revealed that the age of 52 years and a duration of 4 days from injury to surgery serve as predictive cut-off values for assessing the risk of DVT.<h4>Conclusion</h4>Our study investigates the incidence of thrombosis in patellar fracture patients and identifies key risk factors for DVT, including age, open and multiple injuries, and the time from injury to surgery. Additionally, we highlight sodium and albumin levels as protective factors. By establishing threshold values for age and surgical delay, our findings improve DVT risk assessment, facilitating earlier and more targeted interventions.

<h4>Introduction</h4>Children with beta-thalassemia major (β-TM) commonly experience metabolic bone diseases. Understanding fibroblast growth factor 23 (FGF-23) levels in these children can shed light on phosphate dysregulation. This study aimed to assess changes in phosphate homeostasis and associated factors, including FGF-23 and explore relationships between iron overload, FGF23 levels, and phosphorus regulation for clinical management of phosphate disorders, in children with β-TM.<h4>Methods</h4>143 β-TM patients (57.3% male, median age 12 years) were recruited from Fatimid Foundation Karachi, a blood transfusion facility from January to October 2022. Clinical and biochemical evaluations were conducted at Aga Khan University Hospital, including serum ferritin, calcium (Ca), phosphate (P), vitamin D levels, and FGF-23. Descriptive and inferential statistics including multivariable analysis were applied.<h4>Results</h4>This study enrolled 143 patients, with 57.3% males. The median age was 12 years, with 53% underweight. Blood transfusion rates varied, with 66.4% receiving 2/month. Bone/joint pain was reported by 76.2%, with 60.8% requiring analgesics. Median serum ferritin was 2768.3 ng/mL. Hypophosphatemia and hyperphosphatemia were observed in 5.6% and 3.5% of participants, respectively. Vitamin D deficiency/insufficiency affected 92.3%. Plasma c-FGF23 was elevated in 60.8%, while i-FGF23 was high in 14%. A low TMP-GFR (glomerular filtration rate) was associated with high c-FGF23 and low i-FGF23. Multivariable regression revealed c-FGF23, TMP:GFR, Corrected Ca, iPTH, and an interaction term between corrected Ca and iPTH as predictors of serum P variability (~75%).<h4>Conclusion</h4>The study identified contributors to the variations observed in serum P levels in individuals with β-TM and recommends multidisciplinary care and prospective future studies to form targeted interventions for this population.

While genome-wide association studies (GWAS) have identified genetic variants associated with intelligence, their biological mechanisms remain largely unexplored. This study aimed to bridge this gap by integrating intelligence GWAS data with human brain proteomics and transcriptomics. We conducted proteome-wide (PWAS) and transcriptome-wide (TWAS) association studies, along with enrichment and protein-protein interaction (PPI) network analyses. PWAS identified 44 genes in the human brain proteome that influence intelligence through protein abundance regulation (FDR P <  0.05). Causal analysis revealed 36 genes, including GPX1, involved in the cis-regulation of protein abundance (P <  0.05). In independent PWAS analyses, 17 genes were validated, and 10 showed a positive correlation with intelligence (P <  0.05). TWAS revealed significant SNP-based heritability for mRNA in 28 proteins, and cis-regulation of mRNA levels for 20 genes was nominally associated with intelligence (FDR P <  0.05). This study identifies key genes that bridge genetic variants and protein-level mechanisms of intelligence, providing novel insights into its biological pathways and potential therapeutic targets.

Huntington's disease (HD) is a fatal neurodegenerative disease characterized by the expression of huntingtin protein (htt) that has a polyglutamine (CAG; polyQ) repeat domain consisting of 36 or more glutamines (mhtt). Historically, mhtt is more broadly associated with HD severity, as are elevated metal levels observed in HD patients. The depletion of wild-type (WT) htt (fewer than 36Qs) is also recognized as a contributing factor to HD progression; however, many questions remain about the interactions of biorelevant metals with WT htt and the impact of the interactions on protein aggregation. In the present work, we utilize a combination of biochemical assays and spectroscopic techniques to provide insights into the interaction of copper with an <i>in vitro</i> htt model (N171-17Q). Herein, we use sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and dynamic light scattering to show that the addition of equimolar or higher concentrations of Cu(II) to htt induces time- and temperature-dependent protein oligomerization/aggregation. Additionally, chelation assays, trapped ion mobility spectrometry, and mass spectrometry confirm the (i) rapid reduction of Cu(II) in the presence of N171-17Q htt, (ii) direct binding of multiple copper ions per protein, and (iii) complex Cu:htt speciation profile with a preference for three distinct Cu:htt states. These findings contribute to our molecular level understanding of copper's role in the depletion and oligomerization/aggregation of WT htt while underscoring the physiological significance of our work, its potential relevance to metal binding in mhtt, and its significance for identifying new avenues for biomarker exploration and therapeutic design strategies.

The complex relationship between frailty and schizophrenia has yet to be fully understood. This study aims to clarify their relationship by investigating their genetic links. We hypothesize a shared genetic architecture and a bidirectional causal relationship between the two conditions. Utilizing summary genetic data from European genome-wide association studies, we analyzed genetic associations through global and local correlations, shared genomic loci, tissue enrichments, and functional genes. Bidirectional Mendelian Randomization (MR) was employed to infer causality. Our findings show a positive genetic correlation between frailty and schizophrenia (LDSC: r_g = 0.117, p = 6.686 × 10^-7; HDL: r_g = 0.101, p = 5.63 × 10^-13) and local correlations in three genomic regions (chr9: 94167203-96671698, p = 2.21 × 10^-6; chr11: 112459488-114257728, p = 1.01 × 10^-5; and chr18: 77149991-78017158, p = 9.57 × 10^-6). We identified 111 genomic loci associated with both conditions and demonstrated that genetic variants for frailty and schizophrenia share tissue enrichments and functional genes in brain. MR analysis suggests that frailty increases the likelihood of schizophrenia (OR: 1.763, 95% CI: 1.259-2.468, p = 0.001) and vice versa (β: 0.012, 95% CI: 0.006-0.018, p < 0.001). Our research supports the presence of a shared genetic basis and bidirectional causality between frailty and schizophrenia. These findings necessitate further investigation in diverse populations to confirm and expand on this genetic understanding.

Structural diversity of the nucleosome affects chromatin conformations and regulates eukaryotic genome functions. Here we identify DEK, whose function is unknown, as a nucleosome-binding protein. In embryonic neural progenitor cells, DEK colocalizes with H3 K27 trimethylation (H3K27me3), the facultative heterochromatin mark. DEK stimulates the methyltransferase activity of Polycomb repressive complex 2 (PRC2), which is responsible for H3K27me3 deposition in vitro. Cryo-electron microscopy structures of the DEK-nucleosome complexes reveal that DEK binds the nucleosome by its tripartite DNA-binding mode on the dyad and linker DNAs and interacts with the nucleosomal acidic patch by its newly identified histone-binding region. The DEK-nucleosome interaction mediates linker DNA reorientation and induces chromatin compaction, which may facilitate PRC2 activation. These findings provide mechanistic insights into chromatin structure-mediated gene regulation by DEK.

<h4>Background</h4>The financial strain fertility issues cause the dairy cattle industry is substantial, with over $7 billion in lost revenue accrued annually due to a relatively low cow conception rate (CCR; 30-43%) for US dairy cows. While CCR has been improving through genomic selection, identification of causal mutations would help improve the rate of genetic progress with genomic selection and provide a better understanding of infertility. The objectives of this study were to: (1) identify genes and gene-sets associated with CCR to the first breeding (CCR1) and the number of breedings required to conceive (TBRD) in Holstein cows and (2) identify putative functional variants associated with CCR1 and TBRD through a custom genotype-by-sequencing array. The study consisted of 1,032 cows (494 pregnant to first breeding, 472 pregnant to subsequent [2-20] services, and 66 that never conceived). Cows were artificially inseminated, and pregnancy was determined 35d later by rectal palpation of uterine contents. Gene-set enrichment analyses with SNP data (GSEA-SNP) were conducted for CCR1 and TBRD with a normalized enrichment score (NES) ≥ 3.0 required for significance. Leading edge genes (LEG) and positional candidate genes from this and 26 additional studies were used to validate 100 loci associated (P < 1 × 10^- 5) with cow fertility using a custom sequencing genotyping array of putative functional variants (exons, promoters, splice sites, and conserved regions).<h4>Results</h4>GSEA-SNP identified 95 gene-sets (1,473 LEG) enriched for CCR1 and 67 gene sets enriched (1,438 LEG) for TBRD (NES ≥ 3). Thirty-four gene-sets were shared between CCR1 and TBRD along with 788 LEG. The association analysis for TBRD identified three loci: BTA1 at 83 Mb, BTA1 at 145 Mb, and BTA 20 at 46 Mb (P < 1 × 10^- 5). The loci associated with TBRD contained candidate genes with functions relating to implantation and uterine receptivity. No loci were associated with CCR1, however a single locus on BTA1 at 146 Mb trended toward significance with an FDR of 0.04.<h4>Conclusions</h4>The validation of three loci associated with CCR and TBRD in Holsteins can be used to improve fertility through genomic selection and provide insight into understanding infertility.

<h4>Objective</h4>The objective of this study was to investigate the efficacy and safety of tranexamic acid (TXA) in hepatectomy when administered as per the standardized protocol of controlled low central venous pressure (CLCVP).<h4>Methods</h4>This study was a randomized, double-blind, controlled study. Patients who fulfilled the inclusion criteria were randomly assigned to the TXA group (group T) or the placebo group (group N). The central venous pressure (CVP) was maintained at below 5 cmH2O before complete dissection of the liver parenchyma. Patients in group T received an intravenous infusion of 10 mg/kg of TXA 30 min before surgery, and it was continuously pumped intravenously at a rate of 1 mg/(kg.h) until the end of surgery. Patients in group N were infused with 1 mL/kg of normal saline 30 min before surgery, and it was continuously pumped intravenously at a rate of 0.1 mL/(kg.h) until the end of surgery. The primary outcome indicators were intraoperative blood loss, blood transfusion rate, intraperitoneal drainage at 24 h after surgery, and the occurrence of compound bleeding within 30 days.<h4>Results</h4>The baseline indicators were similar (P > 0.05), and there was no significant difference in intraoperative blood loss between the two groups, but the red blood cell transfusion rate was lower in the T group than in the N group (P < 0.05). The infusion volume, surgical field grade, and surgery duration were comparable between the two groups (P > 0.05). Patients in group T had a shorter hilar occlusion time, lower D-dimer and fibrinogen degradation products (FDPs) on the day of surgery, and significantly less intraperitoneal drainage at 24 h after surgery (all P < 0.05). There were two cases of compound bleeding and three cases of thromboembolism among patients in group N, but there were no such complications in group T.<h4>Conclusion</h4>The use of TXA in hepatectomy under CLCVP reduced the intraoperative blood transfusion rate and improved the postoperative bleeding outcome without increasing the risk of adverse events such as hepatic and renal insufficiency and thrombosis.

<h4>Introduction</h4>Preeclampsia (PE) is a severe pregnancy complication with limited early diagnostic and therapeutic options. Ferroptosis, an iron-dependent cell death pathway, has emerged as a potential mechanism in PE pathogenesis. This study investigated ferroptosis-related genes (FRGs) in PE to identify diagnostic biomarkers and therapeutic targets.<h4>Methods</h4>Differentially expressed genes were identified from GEO databases and intersected with FRGs. Hub genes were selected using RandomForest and LASSO algorithms. Their diagnostic potential was evaluated through ROC analysis. Regulatory networks were constructed using transcription factors, microRNAs and potential drug targets. Hub gene expression was validated through immunohistochemistry, Western blot, and RT-qPCR in placental tissues and hypoxic trophoblasts.<h4>Results</h4>We identified 25 ferroptosis-related differentially expressed genes enriched in ferroptosis and HIF-1 pathways. Four hub genes (NDRG1, P4HA1, LDHA, and IDO1) showed high diagnostic efficiency (AUC=0.9182). Immune cell analysis revealed altered levels of plasma cells, CD8+ T cells, Tregs, monocytes, and M2 macrophages in PE, correlating significantly with hub gene expression. We identified 84 mRNA-miRNA and 119 mRNA-TF interactions. Among 19 potential drugs, Tetrahydro-NAD showed promising targeting potential. Experimental validation confirmed elevated expression of NDRG1, P4HA1, and LDHA, and decreased IDO1 in PE tissues and hypoxic conditions.<h4>Discussion</h4>This study identified four FRGs as potential PE biomarkers and therapeutic targets, providing new insights into PE pathogenesis through integrated bioinformatics and experimental validation. These findings may facilitate early PE diagnosis and treatment development.

<h4>Background</h4>Convergent evolution is the independent evolution of similar traits in unrelated lineages across the Tree of Life. Various genomic signatures can help identify cases of convergent evolution at the molecular level, including changes in substitution rate in the same genes or gene networks. In this study, utilizing tarsus measurements of ~ 5400 species of birds, we identify independent shifts in tarsus length and use both comparative genomic and population genetic data to identify convergent evolutionary changes among focal clades with shifts to shorter optimal tarsus length.<h4>Results</h4>Using a newly generated, comprehensive and broadly accessible set of 932,467 avian conserved non-exonic elements (CNEEs) and a whole-genome alignment of 79 birds, we find strong evidence for convergent acceleration in short-tarsus clades among 14,422 elements. Analysis of 9854 protein-coding genes, however, yielded no evidence of convergent patterns of positive selection. Accelerated elements in short-tarsus clades are concentrated near genes with functions in development, with the strongest enrichment associated with skeletal system development. Analysis of gene networks supports convergent changes in regulation of broadly homologous limb developmental genes and pathways.<h4>Conclusions</h4>Our results highlight the important role of regulatory elements undergoing convergent acceleration in convergent skeletal traits and are consistent with previous studies showing the roles of regulatory elements and skeletal phenotypes.

Most GWAS loci are presumed to affect gene regulation; however, only ∼43% colocalize with expression quantitative trait loci (eQTLs). To address this colocalization gap, we map eQTLs, chromatin accessibility QTLs (caQTLs), and histone acetylation QTLs (haQTLs) using molecular samples from three early developmental-like tissues. Through colocalization, we annotate 10.4% (n = 540) of GWAS loci in 15 traits by QTL phenotype, temporal specificity, and complexity. We show that integration of chromatin QTLs results in a 2.3-fold higher annotation rate of GWAS loci because they capture distal GWAS loci missed by eQTLs, and that 5.4% (n = 13) of GWAS colocalizing eQTLs are early developmental specific. Finally, we utilize the iPSCORE multiomic QTLs to prioritize putative causal variants overlapping transcription factor motifs to elucidate the potential genetic underpinnings of 296 GWAS-QTL colocalizations.

<h4>Background</h4>Colistin resistance poses a growing global challenge, particularly in low- and middle-income countries where antibiotic misuse is prevalent. This study investigates the prevalence of colistin resistance in Klebsiella spp. and characterizes the genetic features of resistant isolates, focusing on the mcr-8.2 gene identified in a Klebsiella pneumoniae isolate from pediatric diarrheal cases in southern Vietnam.<h4>Methods</h4>Stool samples were collected from 500 pediatric patients (aged 0-5 years) hospitalized with diarrhea in two tertiary hospitals in Ho Chi Minh City between March and September 2022. Samples were cultured on Violet Red Bile Glucose Agar, then presumptive Klebsiella spp. colonies were selected, purified on nutrient agar, and identified using MALDI-TOF MS. Colistin resistance was determined via minimum inhibitory concentration testing, and the presence of mcr genes was confirmed through polymerase chain reaction. Whole-genome sequencing was performed on the Klebsiella pneumoniae strain harboring mcr-8.2 to elucidate resistance mechanisms. Strain characterization was performed using multi-locus sequence typing, while conjugation experiments assessed horizontal gene transfer potential.<h4>Results</h4>Among 121 Klebsiella spp. isolates, 49 (40.5%) were resistant to colistin. The mcr-1 gene was detected in 31 isolates (25.6%), whereas the mcr-8 was identified in a single isolate (0.8%), with a colistin MIC of 16 µg/mL. Genomic analysis revealed 34 antibiotic resistance genes, including mcr-8.2 and multiple β-lactamase genes, alongside plasmid types IncFIB and IncFII. Chromosomal mutations in phoP, phoQ, and lpxM were also implicated in colistin resistance.<h4>Conclusions</h4>This study documents the emergence of mcr-8.2-mediated colistin resistance in K. pneumoniae from pediatric diarrhea in Vietnam and highlights a high prevalence of multidrug resistance in Klebsiella spp.. Continuous surveillance of mcr genes and novel therapeutic strategies are urgently needed.

<h4>Background</h4>Autologous bone remains the gold standard for surgical bone reconstruction but presents clinical challenges like donor site complications and operational difficulties.<h4>Method</h4>We investigate the osteogenic effects of a newly designed, ceramic and collagen-based, submicron-processed Flexible Hydrated-Hardening Bone Graft (FHBG), using both murine and human mesenchymal stem cells. We also compare the efficacy and safety of FHBG with a commercially available (CA) graft in New Zealand white rabbits with cranial bone defects. Rabbits were divided into three groups: no graft, CA, and FHBG, and evaluated using Micro-CT and histological analysis at three and six weeks post-surgery. Safety was assessed through blood samples.<h4>Results</h4>In vitro, FHBG promoted osteogenesis and upregulated osteogenic-associated genes in mesenchymal stem cells. In vivo, FHBG significantly enhanced bone regeneration, showing approximately 25% and 30% more improvement than the control at three and six weeks post-surgery. FHBG also had about half the residual content compared to the CA group. Blood analysis showed no hepatotoxicity or nephrotoxicity associated with the graft.<h4>Conclusion</h4>FHBG significantly promotes bone regeneration both in vitro and in vivo. Additionally, FHBG has been demonstrated to be safe, with fewer residuals remaining in the body compared to currently in-use clinical bone grafts. This study validates the ability of the newly designed FHBG to facilitate osteogenesis in vitro and demonstrates its efficacy and safety in new bone formation in vivo. The lower residual material further suggests a reduced long-term impact and associated risk with the graft.

<h4>Objective</h4>To examine the association of a numerical Sarnat score (NSS) and modified Sarnat staging among newborn infants with moderate/severe hypoxic-ischemic encephalopathy and their neurodevelopmental outcomes at 18-22 months.<h4>Study design</h4>This secondary analysis included participants with gestational age ≥36 weeks and moderate/severe hypoxic-ischemic encephalopathy (HIE) from the Induced Hypothermia and Optimizing Cooling trials. Early (<6 hours age) neurologic examinations were performed by trained examiners to categorize HIE severity by modified Sarnat staging. The NSS was calculated by summing abnormal scores (2 for moderate, 3 for severe) in the 6 examination categories. The primary outcome was death or moderate/severe disability. Statistical analysis included logistic regression, adjusting for center, trial, and cooling group, and linear regression for continuous scales.<h4>Results</h4>The cohort (n = 528) included infants with 71% moderate and 29% severe HIE (37% Induced Hypothermia and 63% Optimizing Cooling participants). Median (IQR) NSS of infants with moderate and severe HIE were 11 (9-13) and 16 (16-17), respectively. There were significant associations among NSS, NSS tertiles, and modified Sarnat staging and death or disability, but there were no differences found in their area-under-the-curve estimates. Similar to modified Sarnat staging, NSS showed significant associations with Bayley-III cognitive, language, and motor scores and Bayley-II mental and psychomotor developmental indices.<h4>Conclusion</h4>Among infants with moderate or severe HIE, the NSS did not improve the predictive accuracy for death or disability at 18-22 months of age, compared with modified Sarnat staging performed in the initial 6 hours after birth by trained examiners.

Metformin is a well-tolerated drug frequently prescribed for managing type 2 diabetes. Extended metformin use has been linked to a significant decrease in cancer incidence across both diabetic and non-diabetic populations. Here we investigate the anti-proliferative effects of metformin on fission yeast <i>S. pombe</i>. Our findings demonstrate that metformin's inhibitory impact on cell proliferation is effective in the absence of AMP-activated protein kinase (AMPK). Using an unbiased genetic screen we identified the plasma membrane signalling scaffold Efr3, critical for phosphatidylinositol signalling and the generation of PI4Ps, as a key determinant of resistance to the anti-proliferative effect of metformin. Deletion of <i>efr3</i> resulted in both AMPK-dependent and AMPK-independent resistance to metformin. We show that Efr3 does not influence cell proliferation by controlling Ras1 activity or its cellular localization in yeast. We observe that <i>dnm1</i> (DRP1) mutants with elongated mitochondria are also resistant to the anti-proliferative effect of metformin and that metformin treatment promotes mitochondrial fusion. Metabolic measurements after prolonged metformin exposure demonstrated a reduction in respiration in both wild type and the <i>efr3</i> deletion, however, that reduction is less pronounced in the <i>efr3</i> deletion, which also contained elongated mitochondria. It is likely that mitochondrial fusion enhances yeast fitness in response to metformin exposure. Together we provide a new perspective on the cellular response to metformin.

Protein palmitoylation, a reversible post-translational lipid modification, is catalyzed by the ZDHHC family of palmitoyltransferases and reversed by several acyl protein thioesterases, regulating protein localization, accumulation, secretion, and function. Neurological disorders encompass a spectrum of diseases that affect both the central and peripheral nervous system. Recently, accumulating studies have revealed that pathological protein associated with neurological diseases, such as β-amyloid, α-synuclein, and Huntingtin, could undergo palmitoylation, highlighting the crucial roles of protein palmitoylation in the onset and development of neurological diseases. However, few preclinical studies and clinical trials focus on the interventional strategies that target protein palmitoylation. Here, we comprehensively reviewed the emerging evidence on the role of protein palmitoylation in various neurological diseases and summarized the classification, processes, and functions of protein palmitoylation, highlighting its impact on protein stability, membrane localization, protein-protein interaction, as well as signal transduction. Furthermore, we also discussed the potential interventional strategies targeting ZDHHC proteins and elucidated their underlying pathogenic mechanisms in neurological diseases. Overall, an in-depth understanding of the functions and significances of protein palmitoylation provide new avenues for investigating the mechanisms and therapeutic approaches for neurological disorders.

The integration of liquid biopsy with epigenetic markers offers significant potential for early lung cancer detection and personalized treatment. Epigenetic alterations, including DNA methylation, histone modifications, and noncoding RNA changes, often precede genetic mutations and are critical in cancer progression. In this study, we explore how liquid biopsy, combined with epigenetic markers, can provide early detection of lung cancer, potentially predicting onset up to 4 years before clinical diagnosis. We discuss the challenges of targeting epigenetic regulators, which could disrupt cellular balance if overexploited, and the need for maintaining key gene expressions in therapeutic applications. This review highlights the promise and challenges of using liquid biopsy and epigenetic markers for early-stage lung cancer diagnosis, with a focus on optimizing treatment strategies for personalized and precision medicine.

Although large-scale genetic association studies have proven useful for the delineation of neurodegenerative disease processes, we still lack a full understanding of the pathologic mechanisms of these diseases, resulting in few appropriate treatment options and diagnostic challenges. To mitigate these gaps, the Neurodegenerative Disease Knowledge Portal (NDKP) was created as an open-science initiative with the aim to aggregate, enable analysis, and display all available genomic datasets of neurodegenerative disease, while protecting the integrity and confidentiality of the underlying datasets. The portal contains 218 genomic datasets, including genotyping and sequencing studies, of individuals across 10 different phenotypic groups, including neurologic conditions such as Alzheimer disease, amyotrophic lateral sclerosis, Lewy body dementia, and Parkinson disease. In addition to securely hosting large genomic datasets, the NDKP provides accessible workflows and tools to effectively use the datasets and assist in the facilitation of customized genomic analyses. Here, we summarize the genomic datasets currently included within the portal, the bioinformatics processing of the datasets, and the variety of phenotypes captured. We also present example use cases of the various user interfaces and integrated analytic tools to demonstrate their extensive utility in enabling the extraction of high-quality results at the source, for both genomics experts and those in other disciplines. Overall, the NDKP promotes open science and collaboration, maximizing the potential for discovery from the large-scale datasets researchers and consortia are expending immense resources to produce and resulting in reproducible conclusions to improve diagnostic and therapeutic care for patients with neurodegenerative disease.

G Protein-Coupled Receptors (GPCRs) are one of the most attractive therapeutic targets due to their active role in different systems and disease types. The increasing three-dimensional structure information of GPCRs has made them interesting for Structure-Based Drug Design (SBDD) studies. There are various orphan GPCRs whose endogenous molecules have not yet been identified, although their structural information is known. The recent discovery of the three-dimensional structure of GPR52, an orphan GPCR involved in central nervous system diseases, made it stand out as a drug target. In this study, it is aimed to find a lead drug molecule candidate for GPR52 by using structure-based drug design techniques. The study comprises a set of SBDD methods, including preparation of a small molecule library, pharmacophore modeling, molecular docking, consensus scoring, molecular dynamics simulations, calculation of binding free energy, and in silico pharmacokinetic studies for GPR52. It is expected that the molecules obtained as a result of the study may be strong candidates for in vitro and in vivo experiments or could be used as lead drug molecules in new drug discovery and development studies.

Alzheimer's disease and related dementias have a multifactorial aetiology and heterogeneous biology. The current study aims to identify different biological signatures in a deeply phenotyped memory clinic patient population. In this cross-sectional study, we analysed 49 pre-specified proteins using a multiplex antibody-based suspension bead array in 278 CSF samples from the real-world research database and biobank at the Karolinska University Hospital Memory Clinic, Solna, Sweden. Patients with a clinical diagnosis of subjective cognitive decline (<i>N</i> = 151), mild cognitive impairment (<i>N</i> = 61), Alzheimer's disease (<i>N</i> = 47), or other diagnoses (<i>N</i> = 19; vascular dementias, alcohol-related dementia, unspecified dementias, or other amnesias) were included. Principal component analyses were performed, and resulting principal components (PCs) were tested for associations with clinical variables and Alzheimer's disease biomarkers (CSF biomarkers beta-amyloid 42, beta-amyloid 42/40, phosphorylated tau 181, phosphorylated tau 181/beta-amyloid 42). PC 1 (explaining 52% of the variance between patients) was associated with the clinical Alzheimer's disease CSF biomarkers beta-amyloid 42, phosphorylated tau 181, and total tau but not with Alzheimer's disease-related neurodegeneration imaging markers, cognitive performance, or clinical diagnosis. PC 2 (explaining 9% of the variance) displayed an inflammatory profile with high contributions of chitinase 3 like 1 (CHI3L1) and triggering receptor expressed on myeloid cells 2 (TREM2) and significant correlation to CSF free light chain kappa. In contrast to PC 1, PC 3 (explaining 5% of the variance) showed associations with all the clinical Alzheimer's disease CSF biomarkers, the imaging markers, cognitive impairment and clinical diagnosis. Serpin family A member 3 (SERPINA3), chitinase 1 (CHIT1), and neuronal pentraxin 2 (NPTX2) contributed most to PC 3. PC 4 (explaining 4% of the variance) exhibited an inflammatory profile distinct from PC 2, with the largest contributions from TREM2, leucine-rich alpha-2-glycoprotein 1 (LRG1) and complement C9. The component was associated with peripheral inflammation. We found that CSF protein profiles in a memory clinic cohort reflect molecular differences across diagnostic groups. Our results emphasize that real-world memory clinic patients can have different ongoing biological processes despite receiving the same diagnosis. In the future, this information could be utilized to identify patient endotypes and uncover precision biomarkers and novel therapeutic targets.

Neuropathic pain (NPP) is caused by damage to the somatosensory nervous system. Its prominent symptoms are spontaneous pain, hyperalgesia and abnormal pain. This pain is long-lasting and unbearable, seriously affecting the patient's quality of life. At present, the clinical treatment effect of painkillers to relieve NPP is still not ideal, nor can it repair damaged nerves and achieve long-term treatment results. In recent years, the application of cell therapy strategies in the field of pain has yielded encouraging results, including preclinical studies and clinical trials. Mesenchymal stem cells (MSCs) are pluripotent progenitor cells derived from mesogenesis. They have the ability to self-renew and differentiate into multiple cell types and have been widely studied and applied in the field of neuroregenerative medicine. MSCs play an important mechanism functional role in promoting injured nerve regeneration and pain relief by regulating multiple processes in target cells, including immunoregulation, anti-inflammatory properties, promoting axon regeneration and re-myelination, promoting angiogenesis, and secreting neurotrophic factors. Moreover, MSCs can also release exosomes, which may be part of their analgesic effects. Exosomes derived from MSC also have the functional properties of mother cells and have therapeutic potential for treating NPP by promoting cell proliferation, regulating inflammatory responses, reducing cell death, promoting axon regeneration and angiogenesis. Therefore, in this article, we discussed current treatment strategies for NPP and explored the functional role and mechanism of MSCs in the treatment of NPP. We also analyzed the current problems and challenges in the application of MSCs in clinical trials of NPP.

<h4>Background</h4>Accumulating evidences suggested that immune checkpoints (ICPs) played an important role in malignancies including breast cancer (BRCA). We aimed to investigate whether inhibitory-to-stimulatory immune checkpoint ratio (ISICPR) could be used as a prognostic marker for BRCA.<h4>Methods</h4>BRCA patients were enrolled from The Cancer Genome Atlas (TCGA). Survival analysis was performed with Kaplan-Meier (KM) methods. X-tile was used to calculate the optimal cut-off values of ISICPRs. Univariate and multivariate Cox regression analysis were carried out to identify prognostic factors for BRCA patients. Tissue microarray was used to validate our findings.<h4>Results</h4>In total, 586 BRCA patients were collected, including 104 cases of stage I, 330 of stage II, 139 of stage III, and 13 of stage IV. Univariate analysis showed that four ISICPRs (PDCD1/CD27 ratio, PDCD1/TNFSF4 ratio, IDO1/TMIGD2 ratio, and IDO1/TNFSF4 ratio) were significantly associated with the survival of BRCA patients. After adjusting for confounders, multivariate analysis indicated that only the IDO1/TMIGD2 ratio was an independent prognostic factor. The optimal cut-off values for the IDO1/TMIGD2 ratio were set at 4.4 and 6.3. Survival analysis indicated that the high-ratio group (ratio > 6.3) had a worse prognosis than both the low-ratio (ratio < 4.4) and medium-ratio group (4.4 < ratio < 6.3) (P < 0.001), which was further validated by BRCA tissue microarray.<h4>Conclusions</h4>We found that IDO1/TMIGD2 ratio was an independent prognostic factor for BRCA. On one hand, dual targeting of IDO1 and TMIGD2 may be a more effective therapeutic strategy for patients with a high IDO1/TMIGD2 ratio. On the other hand, ISICPR was a promising indicator with high clinical values and worthy of further promotion in other cancers.

With an ultrahigh aspect ratio and a similar chemical composition to the biomineral in bone and tooth, ultralong hydroxyapatite nanowires (UHAPNWs) exhibit a meritorious combination of high flexibility, excellent mechanical performance, high biocompatibility, and bioactivity. Despite these exciting merits, the rapid and green synthesis of UHAPNWs remains challenging. In this work, we have developed an environment-friendly, rapid, and highly efficient synthesis of ultrathin UHAPNWs by the microwave-assisted calcium oleate precursor hydrothermal method using biogenic creatine phosphate as the bio-phosphorus source. Owing to the controllable hydrolysis of bio-phosphorus-containing creatine phosphate and the highly efficient heating of microwave irradiation, ultrathin UHAPNWs with a homogeneous morphology of several nanometers in diameter (single nanowire), several hundred micrometers in length, and ultrahigh aspect ratios (>10,000) can be rapidly synthesized within 60 min. This effectively shortens the synthesis time by about two orders of magnitude compared with the traditional hydrothermal method. Furthermore, ultrathin UHAPNWs are decorated in situ with bioactive creatine and self-assembled into nanowire bundles along their longitudinal direction at the nanoscale. In addition, ultrathin UHAPNWs exhibit a relatively high specific surface area of 84.30 m² g^-1 and high ibuprofen drug loading capacity. The flexible bio-paper constructed from interwoven ibuprofen-loaded ultrathin UHAPNWs can sustainably deliver ibuprofen in phosphate-buffered saline, which is promising for various biomedical applications such as tissue regeneration with anti-inflammatory and analgesic functions.

Glioblastoma (GBM) is the most aggressive primary brain cancer, with poor prognosis due to its aggressive behavior and high heterogeneity. This study aimed to identify cellular senescence (CS) and lipid metabolism (LM)-related prognostic genes to improve GBM prognosis and treatment. Transcriptome and scRNA-seq data, CS-associated genes (CSAGs), and LM-related genes (LMRGs) were acquired from public databases. Prognostic genes were identified by intersecting CSAGs, LMRGs, and differentially expressed genes (DEGs), followed by WGCNA and univariate Cox regression. A risk model and nomogram were constructed. Analyses covered clinicopathological features, immune microenvironment, somatic mutations, and drug sensitivity. GBM scRNA-seq data identified key cells and prognostic gene expression. SOCS1 and PHB2 were identified as prognostic markers, contributing to the construction of a robust risk model with excellent predictive ability. High-risk group (HRG) patients had poorer survival, higher immune and stromal scores, and distinct somatic mutation profiles. Drug sensitivity analysis revealed significant differences in IC50 values. In microglia differentiation, SOCS1 and PHB2 showed dynamic expression patterns. These findings provide new strategies for GBM prognosis and treatment.

<h4>Background/objective</h4>Ferroptosis is an iron-dependent form of programmed cell death characterized by lipid peroxidation products (LPOs). A chemotherapeutic drug, 5-fluorouracil (5-FU), can induce epithelial mucositis and favor drug synergism with erastin in ferroptosis. <i>Camellia</i> tea saponin extract (TS) is known to exert antioxidative properties. This study aims to delineate the protective role of TS in mitigating 5-FU-induced ferroptosis and inflammation in human keratinocytes.<h4>Methods</h4>HaCaT cells were induced by 5-FU and erastin, treated with different TS doses, and their viability was then determined. Levels of cellular reactive oxygen species (ROS), LPOs, labile iron pool (LIP), glutathione (GSH), glutathione peroxidase 4 (GPX-4) activity, as well as IL-6, IL-1β, and TNF-α levels, and their wound healing properties were assessed.<h4>Results</h4>TS per se (at up to 25 µg/mL) was not toxic to HaCaT cells but was unable to restore the viability of 5-FU-induced cells up to the baseline levels. The compound significantly diminished increases in cellular ROS, LPOs, and LIP, while restoring GSH content and GPX-4 activity. Additionally, it suppressed the cytokine production of 5-FU-induced cells in a concentration-dependent manner. Moreover, TS exerted wound-healing effects against skin injuries and 5-FU damage significantly and dose dependently.<h4>Conclusions</h4>The insights of this work have identified biochemical mechanisms using tea saponin extract to protect against 5-FU-induced keratinocyte ferroptosis and inflammation. This study highlights the promising adjunctive potential of tea saponin in the mitigation and management of chemotherapy-induced mucositis.

RNA activation (RNAa), a gene regulatory mechanism mediated by small activating RNAs (saRNAs) and microRNAs (miRNAs), has significant implications for therapeutic applications. Unlike small interfering RNA (siRNA), which is known for gene silencing in RNA interference (RNAi), synthetic saRNAs can stably upregulate target gene expression at the transcriptional level through the assembly of the RNA-induced transcriptional activation (RITA) complex. Moreover, the dual functionality of endogenous miRNAs in RNAa (hereafter referred to as mi-RNAa) reveals their complex role in cellular processes and disease pathology. Emerging studies suggest saRNAs' potential as a novel therapeutic modality for diseases such as metabolic disorders, hearing loss, tumors, and Alzheimer's. Notably, MTL-CEBPA, the first saRNA drug candidate, shows promise in hepatocellular carcinoma treatment, while RAG-01 is being explored for non-muscle-invasive bladder cancer, highlighting clinical advancements in RNAa. This review synthesizes our current understanding of the mechanisms of RNAa and highlights recent advancements in the study of mi-RNAa and the therapeutic development of saRNAs.

Infection with the turkey hemorrhagic enteritis virus (THEV) can cause hemorrhagic enteritis, which affects young turkeys. This disease is characterized by bloody diarrhea and immunosuppression (IMS), which is attributed to apoptosis of infected B cells. Secondary infections due to IMS exacerbate economic losses. We performed the first transcriptomic analysis of a THEV infection to elucidate the mechanisms mediating THEV-induced IMS. After infecting and sequencing mRNAs of a turkey B-cell line, trimmed reads were mapped to the host turkey genome, and gene expression was quantified with StringTie. Differential gene expression analysis was followed by functional enrichment analyses using gprofiler2 and DAVID from NCBI. RT-qPCR of select genes was performed to validate the RNA-seq data. A total of 2343 and 3295 differentially expressed genes (DEGs) were identified at 12 hpi and 24 hpi, respectively. The DEGs correlated with multiple biological processes including apoptosis, ER unfolded protein response, and cell maintenance. Multiple pro-apoptotic genes, including APAF1, BMF, BAK1, and FAS were upregulated. Genes that play a role in ER stress-induced unfolded protein response including VCP, UFD1, EDEM1, and ATF4 were also upregulated and may contribute to apoptosis. Our data suggest that several biological processes and pathways including apoptosis and ER response to stress are important aspects of the host cell response to THEV infection. It is possible that interplay between multiple processes may mediate apoptosis of infected B-cells, leading to IMS.

The arachidonic acid (AA) pathway promotes tumor progression by modulating the complex interactions between cancer and immune cells within the microenvironment. In this Review, we summarize the knowledge acquired thus far concerning the intricate mechanisms through which eicosanoids either promote or suppress the antitumor immune response. In addition, we will discuss the impact of eicosanoids on immune cells and how they affect responsiveness to immunotherapy, as well as potential strategies for manipulating the AA pathway to improve anticancer immunotherapy. Understanding the molecular pathways and mechanisms underlying the role played by AA and its metabolites in tumor progression may contribute to the development of more effective anticancer immunotherapies.

<h4>Background</h4>Huntington's disease (HD) is a hereditary neurodegenerative disorder characterized by cognitive, neuropsychiatric and motor symptoms caused by a CAG trinucleotide repeat expansion in the huntingtin gene. Imaging techniques are crucial for understanding HD pathophysiology and monitoring disease progression.<h4>Objectives</h4>This review is targeted at general neurologists and movement disorders specialists with an interest in HD and aims to bring complex imaging, including new experimental techniques, closer to the practicing clinician.<h4>Methods</h4>We provide a summary of findings from conventional structural, diffusion and functional imaging in HD studies, together with an update on emerging novel techniques, including multiparametric mapping, multi-shell diffusion techniques, ultra-high field 7-Tesla MRI, positron emission tomography and magnetoencephalography.<h4>Results</h4>Conventional imaging techniques have deepened our understanding of neuropathological progression in HD, from striatal atrophy to widespread cortical and white matter changes. The integration of novel imaging techniques reviewed has further improved our ability to interrogate, quantify and visualize disease-specific alterations with high precision.<h4>Conclusions</h4>Novel imaging techniques have promising roles to further our understanding of HD pathology and as imaging markers for clinical trials, disease staging and therapeutic monitoring. Additionally, the synergistic potential of combining imaging modalities with molecular and genetic data, along with wet biomarkers and clinical data, will help provide a complete and comprehensive view of HD pathology and progression.

Dengue virus (DENV) is a global health threat, with approximately 390 million infections annually, ranging from mild dengue fever to severe dengue hemorrhagic fever and shock syndrome. MicroRNA (miRNA) are crucial post-transcriptional regulators which may regulate host resistance to DENV infection. This study aimed to identify miRNAs involved in natural resistance to DENV infection. Individuals from a dengue-endemic area were classified as susceptible (SD) or resistant (RD) according to their anti-DENV antibody status. RD individuals were seronegative despite high local DENV infection prevalence. Monocytes susceptibility to DENV infection was assessed in vitro. The miRNome profiles of the monocytes from 7 individuals per group were assessed upon mock or DENV-2 infection. The antiviral effect of differentially expressed miRNAs was analyzed using miRNA mimics in HeLa cells followed by infection with DENV-1, DENV-2, DENV-3, and DENV-4 serotypes. We performed RNA-seq on miRNA mimic-transfected cells to identify miRNA-targeted genes interacting with DENV proteins. Monocytes from RD individuals exhibit lower DENV-2 production in vitro. The miRNAs miR-155, miR-132-3p, miR-576-5p were overexpressed in monocytes from RD group upon DENV-2 infection. The transfection of miR-155-5p mimic reduced DENV infection and viral production in HeLa cells, regulating 18 genes interacting with DENV proteins and downregulating target genes involved in interferon response, TP53 regulation, apoptosis, and vesicle trafficking (e.g. HSD17B12, ANXA2). Therefore, we show that monocytes from RD individuals show a distinct miRNA expression profile and reduced viral production. In vitro miR-155-5p upregulation induces an antiviral state, revealing potential therapeutic targets to treat dengue.

This study investigated ERS-related gene expressions in CESC, identifying two molecular subtypes, P1 and P2, and constructing a precise prognostic model based on these subtypes. TCGA's whole-genome expression profiles were used to recognize these subtypes through the ConsensusClusterPlus method, further refining prognostic models with univariate and Lasso Cox regression analyses validated by the GSE39001 dataset. The study analyzed the expression distribution of ERS marker genes within T cell subgroups using scRNA-seq data (GSE168652), highlighting T cell diversity. The critical role of the CCL3 gene in prognostic models was examined explicitly in CD8 + T cells from healthy individuals and CESC patients. Elevated CCL3 levels were observed in patients' CD8 + T cells compared to healthy controls. Functional experiments involving CCL3 knockdown and overexpression in HeLa and SiHa CESC cell lines were conducted to investigate its impact on cell proliferation, migration, and invasion. These findings were subsequently validated in a nude mouse model. The results demonstrated that suppressing CCL3 inhibited cell proliferation, migration, and invasion significantly, while its overexpression promoted these processes. In the mouse model, CCL3 silencing reduced tumor growth and decreased Ki-67 labeling within the tumor tissues, indicating the therapeutic potential of targeting CCL3 in CESC treatment, possibly through CD8 + T cell regulation. This study contributes new prognostic assessment tools and personalized treatment options for CESC patients, paving the way for more targeted therapies in CESC by discovering the CCL3 gene, presenting significant clinical implications.

Viral proteins frequently mutate to evade host innate immune responses, yet the impact of these mutations on the molecular energy landscape remains unclear. Epistasis, the intramolecular communications between mutations, often renders the combined mutational effects unpredictable. Nonstructural protein 1 (NS1) is a major virulence factor of the influenza A virus (IAV) that activates host PI3K by binding to its p85β subunit. Here, we present a deep analysis of the impact of evolutionary mutations in NS1 that emerged between the 1918 pandemic IAV strain and its descendant PR8 strain. Our analysis reveals how the mutations rewired interresidue communications, which underlie long-range allosteric and epistatic networks in NS1. Our findings show that PR8 NS1 binds to p85β with approximately 10-fold greater affinity than 1918 NS1 due to allosteric mutational effects, which are further tuned by epistasis. NMR chemical shift perturbation and methyl-axis order parameter analyses revealed that the mutations induced long-range structural and dynamic changes in PR8 NS1, relative to 1918 NS1, enhancing its affinity to p85β. Complementary molecular dynamics simulations and graph theory-based network analysis for conformational dynamics on the submicrosecond timescales uncover how these mutations rewire the dynamic network, which underlies the allosteric epistasis. Significantly, we find that conformational dynamics of residues with high betweenness centrality play a crucial role in communications between network communities and are highly conserved across influenza A virus evolution. These findings advance our mechanistic understanding of the allosteric and epistatic communications between distant residues and provide insight into their role in the molecular evolution of NS1.

Depression is a common and severe mental disorder that affects more than 300 million people worldwide. While it is known to have a moderate genetic component, identifying specific genes that contribute to the disorder has been challenging. Previous Genome-wide association studies (GWASs) have identified over 100 genomic loci that are significantly associated with depression. But finding useful therapeutic targets and diagnostic biomarkers from this information has proven difficult. To address this challenge, I conducted a plasma protein proteome-wide association study (PWAS) for depression, using human plasma protein QTL (pQTL) and depression GWAS data. I identified four proteins that were significantly associated with depression: BTN3A3 (P value = 6.41 × 10^-06), PSMB4 (P value = 1.42 × 10^-05), TIMP4 (P value = 3.77 × 10^-05), and ITIH1 (P value = 7.86 × 10^-05). Specifically, I found that BTN3A3 and PSMB4 play a causal role in depression, as confirmed by colocalization and Mendelian Randomization (MR) analysis. Interestingly, I also discovered that PSMB4 was significantly associated with depression in both the brain proteome studies and the plasma PWAS results, which suggests that it may be a particularly promising candidate for further study. Overall, this work has identified 4 new risk proteins for depression and highlights the potential of plasma proteome data for uncovering novel therapeutic targets and diagnostic biomarkers.

Brain-machine interface performance can be affected by neuroinflammatory responses due to blood-brain barrier (BBB) damage following intracortical microelectrode implantation. Recent findings suggest that certain gut bacterial constituents might enter the brain through damaged BBB. Therefore, we hypothesized that damage to the BBB caused by microelectrode implantation could facilitate microbiome entry into the brain. In our study, we found bacterial sequences, including gut-related ones, in the brains of mice with implanted microelectrodes. These sequences changed over time. Mice treated with antibiotics showed a reduced presence of these bacteria and had a different inflammatory response, which temporarily improved microelectrode recording performance. However, long-term antibiotic use worsened performance and disrupted neurodegenerative pathways. Many bacterial sequences found were not present in the gut or in unimplanted brains. Together, the current study established a paradigm-shifting mechanism that may contribute to chronic intracortical microelectrode recording performance and affect overall brain health following intracortical microelectrode implantation.

Early detection of cartilage injuries is crucial due to their limited self-repair capacity and risk of joint dysfunction. Conventional contrast agents like gadolinium-diethylenetriamine-pentaacetic acid (Gd-DTPA) offer low specificity and T1 relaxivity (r1), limiting MRI application. This study introduces NaGdF₄ nanoparticles (NPs) modified with polyethylene glycol (PEG) and cholesterol (CLS) to enhance hydrophilicity and lipophilicity. Targeting is achieved using a matrix metalloproteinase-13 (MMP13) cartilage-binding peptide. NaGdF₄@PEG-CLS@MMP13 CBP NPs demonstrate an increased r1 value (8.07 mM^-1 s^-1) compared to NaGdF₄@PEG-CLS NPs (6.65 mM^-1 s^-1) and Gd-DTPA (3.01 mM^-1 s^-1), enabling deeper cartilage penetration and stronger cartilage affinity. Two hours post-injection, these NPs improved the signal-to-noise ratio at injury sites by 2.4-fold over pre-injection values. Biocompatibility was confirmed with no adverse effects in blood or organs, and the NPs were metabolized in kidneys and liver, with excretion via urine. This study supports NaGdF₄@PEG-CLS@MMP13 CBP NPs as an effective MRI contrast agent, enhancing early detection of cartilage injuries.

Pain is a common and complex non-motor symptom in people with Parkinson's disease (PWP). Little is known about the genetic drivers of pain in PWP, and progress in its study has been challenging. Here, we conducted two genome-wide association studies (GWAS) to identify genetic variants associated with pain experienced during the earliest stages of Parkinson's disease. The study population consisted of 4,159 PWP of European ancestry who were mapped to five previously-described, longitudinal pain trajectories. In the first GWAS, the extreme pain trajectories (highest burden versus no significant pain over time) were compared, and in the second GWAS, a multinomial approach was undertaken. While no variant reached genome-wide significance, we identified promising associations, such as rs117108018 (OR_GWAS-Extreme=8.96, p_GWAS-Extreme=2.5 × 10^- 7), a brain/nerve eQTL for L3MBTL3 and EPB41L2, and rs61881484 (p_{GWAS-Multinomial}=2 × 10^- 7), which intersects a transcription factor peak targeting CREB1, critical in sensory neuron synaptic plasticity and neuropathic pain regulation. Gene-based tests implicated CTNNB1 (p_GWAS-Extreme=3.2 × 10^- 5), KLK7 (p_GWAS-Extreme=7 × 10^- 5), and SLITRK3 (p_{GWAS-Multinomial}=3.2 × 10^- 5), which have been associated with neurodevelopment. At the pathway-level, there was an enrichment for genes involved in neurotransmitter regulation and opioid dependence. This study implicates neuropathic pain mechanisms as prominent drivers of elevated pain in PWP, suggests potential therapeutic genetic targets for further research.

High-dimensional multiplexed imaging can reveal the spatial organization of tumour tissues at the molecular level. However, owing to the scale and information complexity of the imaging data, it is challenging to discover and thoroughly characterize the heterogeneity of tumour microenvironments. Here we show that self-supervised representation learning on data from imaging mass cytometry can be leveraged to distinguish morphological differences in tumour microenvironments and to precisely characterize distinct microenvironment signatures. We used self-supervised masked image modelling to train a vision transformer that directly takes high-dimensional multiplexed mass-cytometry images. In contrast with traditional spatial analyses relying on cellular segmentation, the vision transformer is segmentation-free, uses pixel-level information, and retains information on the local morphology and biomarker distribution. By applying the vision transformer to a lung-tumour dataset, we identified and validated a monocytic signature that is associated with poor prognosis.

Ionizing radiation causes three divergent effects in the human body: On one side, tissue death (= deterministic effects) sets on, on the other side, mutations and cancer growth (= stochastic effects) can occur. In recent years, the additional phenomenon of accelerated aging has come to light. In the following, we argue that these seemingly contradictory radiation responses namely: (i) increased cancer growth, (ii) ablation of cancer tissue or (iii) deterministic senescence, share an underlying cause from damage at the lamin A C-terminus. In other words, besides the typically described genomic radiation impact, we propose an additional destabilization pathway via oxidation at the nuclear envelope. We propose five concrete hypotheses that draw a direct mechanistic model from radiation damage and cellular oxidative stress, to micronuclei and clinical symptoms. In conjunction with lamin B compensation, we might be able to explain why deterministic or stochastic responses dominate. If our model holds true, a novel target for radiotherapeutics and radiooncology arises, and a rationale to closer connect laminopathy and radioprotection research.

<h4>Introduction</h4>Thrombophilia, a blood coagulation disorder, poses risks of venous thromboembolism (VTE). Coagulation assays may not be sufficient to assess VTE risk and global assays such as Rotational Thromboelastometry (ROTEM) may add valuable information. We investigated ROTEM's capacity to detect hypercoagulability in patients undergoing thrombophilia screening, its potential impact on patient outcomes, and limitations.<h4>Methods</h4>Comprehensive clinical, laboratory, genetic tests, and ROTEM (EXTEM and INTEM) were conducted for 356 patients referred for thrombophilia screening at an academic hospital outpatient unit. Hypercoagulability was identified as a shorter clot formation time (CFT), larger alpha angle (AA), and greater maximum clot firmness (MCF), and was compared in patients with and without VTE. Statistically this was analyzed using Mann-Whitney U and Chi-square tests with p < 0.05 considered significant.<h4>Results</h4>Among 356 patients, 64.6% had previous VTE, with 76.9% experiencing one event, 14.3% recurrent (35.6% unprovoked, 64.4% provoked). 22.5% of patients were on anticoagulation. Those with VTE history exhibited significant alterations in EXTEM and INTEM parameters compared to those without (p < 0.001), showing decreased CFT and increased AA and MCF. However, receiver operating characteristic curves for these variables indicated that none were able to discriminate between those individuals with and without thromboembolic complications.<h4>Conclusion</h4>ROTEM does not appear to be a strong discriminatory test. However, it can detect hypercoagulopathy in patients referred for thrombophilia screening. Abnormal ROTEM may indicate a higher risk for recurrence. However, this can only be determined in prospective cohort studies.

Congenital heart disease (CHD) is the most common congenital anomaly and a leading cause of infant morbidity and mortality. Despite extensive exploration of the monogenic causes of CHD over the last decades, ∼55% of cases still lack a molecular diagnosis. Investigating digenic interactions, the simplest form of oligogenic interactions, using high-throughput sequencing data can elucidate additional genetic factors contributing to the disease. Here, we conducted a comprehensive analysis of digenic interactions in CHD by utilizing a large CHD trio exome sequencing cohort, comprising 3,910 CHD and 3,644 control trios. We extracted pairs of presumably deleterious rare variants observed in CHD-affected and unaffected children but not in a single parent. Burden testing of gene pairs derived from these variant pairs revealed 29 nominally significant gene pairs. These gene pairs showed a significant enrichment for known CHD genes (p < 1.0 × 10^-4) and exhibited a shorter average biological distance to known CHD genes than expected by chance (p = 3.0 × 10^-4). Utilizing three complementary biological relatedness approaches including network analyses, biological distance calculations, and candidate gene prioritization methods, we prioritized 10 final gene pairs that are likely to underlie CHD. Analysis of bulk RNA-sequencing data showed that these genes are highly expressed in the developing embryonic heart (p < 1 × 10^-4). In conclusion, our findings suggest the potential role of digenic interactions in CHD pathogenesis and provide insights into unresolved molecular diagnoses. We suggest that the application of the digenic approach to additional disease cohorts will significantly enhance genetic discovery rates.

Tumor resistance to chimeric antigen receptor T cell (CAR-T) and, in general, to adoptive cell immunotherapies (ACTs) is a major challenge in the clinic. We hypothesized that inhibiting the tumor drivers' methyltransferases EZH2 and EZH1 could enhance ACT by rewiring cancer cells to a more immunogenic state. In human B cell lymphoma, EZH2 inhibition (tazemetostat) improved the efficacy of anti-CD19 CAR-T by enhancing activation, expansion, and tumor infiltration. Mechanistically, tazemetostat-treated tumors showed upregulation of genes related to adhesion, B cell activation, and inflammatory responses, and increased avidity to CAR-T. Furthermore, tazemetostat improved CAR- and TCR-engineered T cell efficacy in multiple liquid (myeloma and acute myeloid leukemia) and solid (sarcoma, ovarian, and prostate) cancers. Lastly, combined EZH1/EZH2 inhibition (valemetostat) further boosted CAR-T efficacy and expansion in multiple cancers. This study shows that EZH1/2 inhibition reprograms tumors to a more immunogenic state and potentiates ACT in preclinical models of both liquid and solid cancers.

<h4>Background</h4>Olfactomedin 4 (OLFM4) gene polymorphisms have been associated with variations in inflammatory responses and the severity of infections. This study aimed to investigate the association between OLFM4 single nucleotide polymorphisms (SNPs) rs17552047 and rs1891944 and severe outcomes in preterm neonatal sepsis.<h4>Methods</h4>A prospective observational cohort study was conducted from April 2023 to April 2024, involving all preterm infants diagnosed with neonatal sepsis. Genotyping was performed using real-time polymerase chain reaction, and the associations with severe outcomes were analyzed using genetic models (dominant, recessive, and additive) through multivariate logistic regression and survival analysis.<h4>Results</h4>Among the 174 preterm newborns included in the study, 39 experienced severe outcomes. The AA/AG genotypes of SNP rs17552047 and TT/TC genotypes of rs1891944 were associated with a reduced risk of severe outcomes (adjusted hazard ratio: 0.271, 95 % confidence interval [CI]: 0.115-0.641, p = 0.003, and adjusted hazard ratio: 0.349, 95 % CI: 0.175-0.698, p = 0.003, respectively). The odds of severe outcomes decreased by 65 % for each additional A allele (95 % CI: 0.15-0.78, p = 0.01). The model incorporating both SNPs and clinical variables demonstrated good predictive capability (area under the receiver operating characteristic curve: 0.826, 95 % CI: 0.748-0.903, p = 0.03).<h4>Conclusions</h4>The OLFM4 rs17552047 AA/AG and rs1891944 TT/TC genotypes have been linked to favorable outcomes in neonatal sepsis. These SNPs hold promise for predicting severe outcomes in neonatal sepsis.

In recent years, gene therapy and bio-engineered hemostatic molecules have revolutionized treatment for people with hemophilia. These innovative therapies aim to decrease treatment burden and improve patient quality of life. Additional novel therapies, including next-generation mimetics and agents that rebalance hemostasis, are currently being evaluated in clinical trials. Technological advances such as point-of-care musculoskeletal ultrasound and artificial intelligence may improve patient diagnostic and treatment outcomes. However, for the majority of patients with hemophilia worldwide, diagnosis and effective treatment are inaccessible. Achieving health equity for all hemophilia patients requires improved identification of barriers to optimal care, including socioeconomic status, race/ethnicity, gender, disease severity, inhibitor status, age, and use of Hemophilia Treatment Centers. Access to novel hemophilia therapies should be ensured for all patients. Approaches to improving equity include a decision-making partnership between the patient and clinician, stakeholder engagement, and pharmaceutical industry support. The development of novel hemophilia therapies should be leveraged with a patient-centered care approach to improve health equity for all patients.

<h4>Introduction</h4>Suicide is a significant public health problem, with increased rates in low- and middle-income countries such as Mexico; therefore, suicide prevention is important. Suicide is a complex and multifactorial phenomenon in which biological and social factors are involved. Several studies on the biological mechanisms of suicide have analyzed the proteome of the dorsolateral prefrontal cortex (DLPFC) in people who have died by suicide. The aim of this work was to analyze the protein expression profile in the DLPFC of individuals who died by suicide in comparison to age-matched controls in order to gain information on the molecular basis in the brain of these individuals and the selection of potential biomarkers for the identification of individuals at risk of suicide. In addition, this information was analyzed using machine learning (ML) algorithms to propose a model for predicting suicide.<h4>Methods</h4>Brain tissue (Brodmann area 9) was sampled from male cases (n=9) and age-matched controls (n=7). We analyzed the proteomic differences between the groups using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry. Bioinformatics tools were used to clarify the biological relevance of the differentially expressed proteins. In addition, this information was analyzed using machine learning (ML) algorithms to propose a model for predicting suicide.<h4>Results</h4>Twelve differentially expressed proteins were also identified (<i>t</i> ₁₄ ≤ 0.5). Using Western blotting, we validated the decrease in expression of peroxiredoxin 2 and alpha-internexin in the suicide cases. ML models were trained using densitometry data from the 2D gel images of each selected protein and the models could differentiate between both groups (control and suicide cases).<h4>Discussion</h4>Our exploratory pathway analysis highlighted oxidative stress responses and neurodevelopmental pathways as key processes perturbed in the DLPFC of suicides. Regarding ML models, KNeighborsClassifier was the best predicting conditions. Here we show that these proteins of the DLPFC may help to identify brain processes associated with suicide and they could be validated as potential biomarkers of this outcome.

Mitochondrial integrity is fundamental to cellular function, upheld by a network of proteases that regulate proteostasis and mitochondrial dynamics. Among these proteases, AFG3L2 is critical due to its roles in maintaining mitochondrial homeostasis, regulating mitochondrial protein quality, and facilitating mitochondrial biogenesis. Mutations in AFG3L2 are implicated in a spectrum of diseases, including spinocerebellar ataxia type 28 (SCA28) and spastic ataxia 5 (SPAX5), as well as other systemic conditions. This study employs a multi-omics approach to investigate the biochemical impact of AFG3L2 mutations in immortalized lymphoblastoid cell lines derived from a patient with biallelic variants leading to spastic ataxia (SPAX5). Our proteomic analysis revealed AFG3L2 impairment, with significant dysregulation of proteins critical for mitochondrial function, cytoskeletal integrity, and cellular metabolism. Specifically, disruptions were observed in mitochondrial dynamics and calcium homeostasis, alongside downregulation of key proteins like COX11, a copper chaperone for complex IV assembly, and NFU1, an iron-sulfur cluster protein linked to spastic paraparesis and infection-related worsening. Lipidomic analysis highlighted substantial alterations in lipid composition, with significant decreases in sphingomyelins, phosphatidylethanolamine, and phosphatidylcholine, reflecting disruptions in lipid metabolism and membrane integrity. Metabolomic profiling did not reveal any significant findings. Our comprehensive investigation into loss of functional AFG3L2 elucidates a pathophysiology extending beyond mitochondrial proteostasis, implicating a wide array of cellular processes. The findings reveal substantial cellular disturbances at multiple levels, contributing to neurodegeneration through disrupted mitochondrial respiratory chain, calcium homeostasis, cytoskeletal integrity, and altered lipid homeostasis. This study underscores the complexity of SPAX5 pathophysiology and the importance of multi-omics approaches in developing effective strategies to address the impact of loss of functional AFG3L2. Our data also highlight the value of immortalized lymphoblastoid cells as a tool for pre-clinical testing and research, offering a detailed biochemical fingerprint that enhances our understanding of SPAX5 and identifies potential areas for further investigation.

Tendon and enthesis injuries are a global health problem affecting millions of people, causing huge medical expenditure and labor loss every year. However, due to their intricate tissue architecture, unique mechanical properties, and especially their sluggish and limited innate regenerative capacity, repairing these injuries remains a formidable clinical challenge. Here, we present a comprehensive review of biomaterials advances in tendon and enthesis repair recently. These biomaterials are categorized into two primary groups based on their potential clinical application conditions: biomaterials for T/E repairing and biomaterials for T/E replacement. The T/E repairing biomaterials were further divided into two groups: mechanical-enhanced biomaterials and bioactive biomaterials, according to the approaches they used to improve sutured tendon healing. We delve into the characteristics and underlying mechanisms of these various biomaterials to gain a deeper understanding of the current landscape in tendon and enthesis repair biomaterials. This review aims to highlight the prominent advancements while identifying the remaining gaps, ultimately inspiring future biomaterial design strategies.

In the context of global warming, heat tolerance is becoming a crucial physiological trait influencing fish species' distribution and survival. While our understanding of fish heat tolerance and stress has expanded from behavioral studies to transcriptomic analyses, knowledge at the transcriptomic level is still limited. Recently, the highly conserved microRNAs (miRNAs) have provided new insights into the molecular mechanisms of heat stress in fish. This review systematically examines current research across three main reference databases to elucidate the universal responses and mechanisms of fish miRNAs under heat stress. Our initial screening of 569 articles identified 13 target papers for comprehensive analysis. Among these, at least 214 differentially expressed miRNAs (DEMs) were found, with 15 DEMs appearing in at least two studies (12 were upregulated and 13 were downregulated). The 15 recurrent DEMs were analyzed using DIANA mirPath v.3 and the microT-CDS v5.0 database to identify potential target genes. The results suggest that multiple miRNAs target various genes, forming a complex network that regulates glucose and energy metabolism, maintains homeostasis, and modulates inflammation and immune responses. Significantly, <i>miR-1</i>, <i>miR-122</i>, <i>let-7a</i>, and <i>miR-30b</i> were consistently differentially expressed in multiple studies, indicating their potential relevance in heat stress responses. However, these miRNAs should not be considered definitive biomarkers without further validation. Future research should focus on experimentally confirming their regulatory roles through functional assays, conducting transcriptomic comparisons across different species, and performing target validation studies. These miRNAs, conserved across species, could be valuable for monitoring wild fish health, enhancing aquaculture breeding, and guiding conservation strategies. However, the specific regulatory mechanisms of these miRNAs need clarification to confirm their reliability as biomarkers for thermal stress.

In ruminants, the survival and development of the conceptus are heavily dependent on the composition of the uterine lumen fluid (ULF), which is influenced by prostaglandins (PGs). However, the variations in underlying PG-mediated ULF remain unclear. Herein, cycling heifers received an intrauterine infusion of vehicle as a control (CON) or meloxicam (MEL) on days 12-14 of the estrous cycle. Then, the ULF was collected on day 15 and alternations in its protein and lipid levels were analyzed. The suppression of prostaglandins induced by meloxicam resulted in 1343 differentially abundant proteins (DAPs) and 59 differentially altered lipids. These DAPs were primarily associated with vesicle-mediated transport, immune response, and actin filament organization, and were mainly concentrated on the ribosome, complement and coagulation cascades, cholesterol metabolism, chemokine signal pathway, regulation of actin cytoskeleton and starch and sucrose metabolism. These differential lipids reflected a physiological metabolic shift as the abundance of cell membrane-related lipids was modulated, including an accumulation of triacylglycerols and reductions in lysophosphatidylcholines, hexosyl ceramides, ceramides, and sphingomyelins species. Integration analysis of the DAPs and differentially altered lipid metabolites revealed that glycerophospholipid metabolism and choline metabolism were the core pathways. These findings highlight the potential roles of prostaglandins in ULF, providing new insights into the contributions of prostaglandins in the development of the conceptus.

<b>Background:</b> Patients with psoriasis often experience psychiatric comorbidities, such as depression and anxiety. These comorbidities can lead to poorer adherence to treatment regimens, reduced effectiveness of therapies, and a heightened disease burden. This study aims to explore the scientific output related to psoriasis, depression, and anxiety using a comprehensive analysis combining bibliometric statistical methods. <b>Methods:</b> The study performed a bibliometric analysis of publications related to psoriasis, depression, and anxiety between 1974 and December 2023. This study employed the Latent Dirichlet Allocation (LDA) algorithm to identify key research topics and used the HJ-Biplot technique to visualize the relationships between publications and research indicators. The inclusion criteria were limited to English-language research articles. <b>Results:</b> Over 49 years, the analysis identified 5059 documents published across 1151 sources. The annual growth rate for research was 12.26%. <i>The Journal of the European Academy of Dermatology and Venereology</i> and <i>The British Journal of Dermatology</i> were found to be the leading journals in this field. The United States emerged as the top contributor, followed by China, Italy, and Germany. The most prevalent research topics were inflammation and cellular function, with a significant focus on patient treatment and the impact of depression and anxiety. <b>Conclusions:</b> This bibliometric analysis underscores the increasing of studies on the comorbidities of depression and anxiety in patients with psoriasis. This study provides a comprehensive overview of research trends and emerging topics in this field, offering valuable insights for future investigations.

<h4>Background</h4>The literature on the relationships among blood iron levels, cognitive performance, and brain iron levels specific to women at the menopausal transition is ambiguous at best. The need to better understand these potential relationships in women for whom monthly blood loss (and thus iron loss) is ceasing is highlighted by iron's accumulation in brain tissue over time, thought to be a factor in the development of neurodegenerative disease.<h4>Methods</h4>Non-anemic women who were either low in iron or had normal iron levels for their age and race/ethnicity provided blood samples, underwent MRI scans to estimate brain iron levels, and performed a set of cognitive tasks with concurrent EEG.<h4>Results</h4>Cognitive performance and brain dynamics were positively related to iron levels, including measures associated with oxygen transport. There were no relationships between any of the blood measures of iron and brain iron.<h4>Conclusions</h4>Higher iron status was associated with better cognitive performance in a sample of women who were neither iron deficient nor anemic, without there being any indication that higher levels of systemic iron were related to higher levels of brain iron. Consequently, addressing low iron levels at the menopausal transition may be a candidate approach for alleviating the "brain fog" commonly experienced at menopause.

Immune cells within the tumor microenvironment impact cancer progression, resistance, response to treatments. Despite remarkable outcomes for some cancer patients, immunotherapies remain unsatisfactory for others. Here, we designed an experimental setting using the <i>Alb-R26</i> ^<i>Met</i> "inside-out" mouse model, faithfully recapitulating molecular features of liver cancer patients, to explore the effects of distinct anticancer targeted therapies on the tumor immune landscape. Using two treatments in clinical trials for different cancer types, Decitabine and MEK+BCL-XL blockage, we show their capability to trigger tumor regression in <i>Alb-R26</i> ^<i>Met</i> mice and to superimpose distinct profiles of immune cell types and immune-checkpoints, impacting immunotherapy response. A machine learning approach processing tumor imaging and immune profile data identified a putative signature predicting tumor treatment response in mice and patients. Outcomes exemplify how the tumor immune microenvironment is differentially reshaped by distinct anticancer agents and highlight the importance of measuring its modulation during treatment to optimize oncotherapy and immunotherapy combinations.