Stroke Research and Treatment

The TRAF family of molecules are intracellular signaling adaptors that regulate various signaling pathways. These pathways are not only mediated by the TNFR superfamily and the Toll-like receptor/IL-1 receptor superfamily but also by unconventional cytokine receptors like IL-6 and IL-17 receptors. Overactive immune responses caused by TRAF signaling following the activation of these receptors frequently result in inflammatory and autoimmune diseases such as rheumatoid arthritis, inflammatory bowel disease, psoriasis, and autoinflammatory syndromes. Therefore, it is crucial to comprehend the signaling processes controlled by TRAFs, which have a significant influence on the determination of cell fate (life or death) and the functioning, specialization, and endurance of cells in the innate and adaptive immune systems. Our data indicate that the dysregulation of cellular expression and/or signaling of TRAFs leads to the excessive production of pro-inflammatory cytokines, hence promoting abnormal activation of immune cells. The objective of our investigation was to comprehend the function of these molecules in SARS-CoV-2 infection both prior to and during SARS-CoV-2 vaccination. Our results demonstrate a clear inactivation of the TRAF5 and TRAF6 genes when infection occurs after immunization, in contrast to infection without prior vaccination. This can bolster the belief that immunization is essential while also demonstrating the involvement of these molecules in the pathogenesis of SARS-CoV-2.

Background: The etiology of obesity has been associated with genetic and epigenetic factors, hormonal changes, unhealthy lifestyle habits, and infectious agents such as human adenovirus-36 (HAdV-36). Viral infections induce reactive oxygen species, and the imbalance between oxidative stress/antioxidant results in fat accumulation. In the Mexican population, little is known about the frequency of HAdV-36 and its effect on the balance between antioxidants and oxidants, inflammation, and metabolic markers. The purpose of our study was to evaluate the frequency of HAdV-36 seroprevalence and its relation to body mass index (BMI), lipid profiles, glucose levels, inflammation, and levels of antioxidants and oxidative stress in a representative sample. A cross-sectional study was carried out on 112 healthy adults between 18 and 28 years old, who were divided into four groups according to their BMI: underweight (BMI < 18.5); normal weight (BMI 18.5–24.9); overweight (BMI ≥ 25); and obese (BMI ≥ 30). Blood samples were taken to evaluate lipid and glucose profiles, as well as antioxidant and oxidative stress status, using colorimetric techniques. Seropositivity for HAdV-36 and levels of TNF-α, IL-6, and cortisol were determined using an enzyme-linked immunosorbent assay. The HAdV-36 frequency was 15.6% in underweight subjects, 18.7% in the normal-weight subjects, 34.37% in the overweight subjects, and 31.24% in the obese subjects. The subjects who were positive for HAdV-36 seroprevalence had increased levels of IL-6, cortisol, and oxidative stress, independently of BMI. The HAdV-36-positive subjects had reduced LDL-C and HDL-C levels only in the low-weight groups. Glutathione and SOD levels increased in the underweight and normal-weight subjects with positive HAdV-36 seroprevalence, while catalase levels decreased in the normal-weight, overweight, and obese subjects. In conclusion, for the first time, an HAdV-36 seroprevalence in the adult Mexican population is reported which was higher and had a relation with the presence of inflammation, alterations in the lipid profile, and imbalance between oxidative stress and antioxidant status, regardless of BMI. The oxidative stress/antioxidant imbalance could be participating in the stimulation of white adipose tissue deposition.

Bone morphogenetic proteins (BMPs) belong to the TGF-β family. They perform diverse roles in development, osteogenesis, and vasculogenesis. BMPs have crucial functions in embryonic development and regulate the specialization of various cell types. The dysregulation of BMP activity at various stages in signal transduction is associated with a diverse range of human diseases. It is not surprising that BMPs also have a role in tumor formation and control the progression of cancer through different phases. Nevertheless, their specific roles remain ambiguous and the findings regarding this have been inconsistent. The objective of this review is to highlight the important functions of BMP ligands, receptors, and signaling mediators and the subsequent effects on final cellular responses resulting from these signaling modalities. This review elucidates the dysregulation of BMPs identified in various cancer types, which serves as a predictive sign for favorable results in cancer therapy. Alterations in the BMP pathway can represent a crucial milestone in the genetic and molecular mechanisms that facilitate cancer formation. This review has shown that alterations in certain components of the BMP pathway are evident in various tumor forms, including breast, gastric, colorectal, and myeloma cancer. This review reinforces the conclusion that BMPs exert both beneficial and detrimental effects on cancer biology. Collectively, these findings indicate that BMPs serve multiple functions in cancer; therefore, directing therapeutic efforts to focus on BMP may be a highly effective method for treating several cancers.

Metabolic dysfunction-associated steatotic liver disease (MASLD) and its progressive form, metabolic dysfunction-associated steatohepatitis (MASH), represent a growing global health problem linked to obesity, insulin resistance, and dyslipidemia. MASLD often leads to fibrosis, cirrhosis, and hepatocellular carcinoma. Currently, therapeutic options are limited, emphasizing the need for novel, targeted pharmacological interventions. Resmetirom, a selective thyroid hormone receptor beta (THR-β) agonist, offers a promising approach by specifically enhancing hepatic metabolism while minimizing systemic effects. Clinical trials have demonstrated its capacity to reduce hepatic triglyceride accumulation and improve lipid profiles. Early- and advanced-phase studies, including the MAESTRO program, highlight significant reductions in hepatic fat content and favorable impacts on noninvasive biomarkers of fibrosis with minimal side effects. This review highlights evidence from pivotal studies, explores resmetirom’s mechanism of action, and compares its efficacy and safety with other emerging therapeutic agents. While resmetirom marks a breakthrough in non-cirrhotic MASH management, further long-term studies are essential to fully evaluate its clinical benefits and potential regulatory approval for broader use in MASLD and MASH.

Zearalenone (ZEN), a toxic estrogenic mycotoxin in cereals, threatens human and animal health through reproductive, immune, and cytotoxic effects, necessitating sensitive detection methods. While nanobodies offer advantages over conventional antibodies for on-site ZEN detection, their application remains unexplored. This study aimed to develop an anti-ZEN nanobody derived from an anti-ZEN phage display nanobody library. An alpaca was immunized with a ZEN-bovine serum albumin (ZEN-BSA) antigen, achieving peak serum antibody titers (1:25,600) following four immunizations. A high-capacity phage display nanobody library (1.0 × 1011 plaque-forming units/mL) was constructed. Following four rounds of biopanning, an enrichment factor of 479 was achieved. Phage ELISA screening identified six phage display nanobodies with specific ZEN-binding activity, and multiple sequence alignment revealed four unique nanobody sequences. The selected phage display nanobody, designated phage-V44, was expressed and purified, and its presence was validated by SDS-PAGE and western blotting, which detected a single approximately 17 kDa band consistent with the expected nanobody size. We established a working curve for an indirect competitive enzyme-linked immunoassay (ELISA) for ZEN, which showed an IC50 value of 7.55 ng/mL. The specificity and affinity of the V44 were also verified. Collectively, the study successfully constructed an anti-ZEN phage display nanobody library, screened four specific ZEN-binding phage display nanobodies, and prepared the anti-ZEN nanobody V44. Thereby establishing a foundation for the nanobody’s future integration into rapid on-site detection methods for ZEN in both animal feed and human food products.

Objective: Non-small cell lung cancer (NSCLC) remains one of the most significant contributors to cancer-related mortality. This investigation explores the influence and underlying mechanisms of the USP1 inhibitor SJB2-043 on A549 cells, with the aim of advancing the development of anti-NSCLC therapeutics. Methods: Publicly available databases were utilized to assess USP1 expression and its association with the progression of NSCLC. Gene expression variations were ascertained through RNA sequencing, followed by the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology pathway enrichment evaluations. Various doses of SJB2-043 were administered to A549 cells to evaluate its impact on cell multiplication, motility, apoptosis, and the cell cycle using CCK-8 assays, colony formation, wound healing, flow cytometry, and Western blotting (WB). Results: USP1 was found to be overexpressed in NSCLC specimens and linked to adverse prognosis. Treatment with SJB2-043 markedly inhibited A549 cell proliferation and migration, diminished clonogenic potential, and triggered apoptosis in a dose-dependent manner. Modifications in the cell cycle were observed, showing an elevated percentage of cells in the G2 phase while exhibiting a parallel decline in the G1 phase. WB examination demonstrated diminished protein levels of N-cadherin, CyclinB1, CDK1, C-myc, Bcl-2, p-ERK/ERK, p-p38/p38, p-JNK/JNK, p-AKT/AKT, and p-mTOR/mTOR, alongside an upregulation of E-cadherin, ZO-1, occludin, p53, Bax, p-β-catenin/β-catenin, and GSK3β. Conclusions: SJB2-043 exerts a suppressive effect on A549 cell proliferation, migration, and epithelial–mesenchymal transition while enhancing apoptosis. These cellular effects appear to be mediated through the inhibition of the MAPK, Wnt/β-catenin, and PI3K/AKT/mTOR signaling cascades, in addition to modulation of the cell cycle.

D-galactose (d-gal) plays numerous roles in the organism as an energy-providing nutrient and also an important constituent of the complex glycoconjugates. However, excessive amounts of d-gal activate alternative metabolic pathways that can lead to the development of a pro-oxidative environment. This feature is used in numerous aging studies which implied intraperitoneal (i.p.) or subcutaneous (s.c.) administration of d-gal for a prolonged time. The present study aims to investigate the systemic effects of orally administered d-gal (200 mg/kg and 500 mg/kg, dissolved in tap water, for 6 weeks) by analyzing oxidative stress parameters in the liver, kidney, and heart. For comparison with natural aging, the effects were studied in rats aged 12, 18, 24, and 30 months. In addition, histopathologic analyzes and serum biochemical measurements were performed to investigate the potential structural and functional organ damage induced by d-gal administration. Our findings show that chronic oral administration of d-gal induces oxidative stress in rat organs and mimics some aspects of natural aging similar to those of 30-month-old rats. Consistent with its primary role in galactose metabolism, the liver exhibited the most pronounced oxidative damage. However, despite the increased oxidative stress, only minor histopathological changes were observed, while organ function remained largely unaffected. Oral intake of d-gal was found to have milder effects compared to i.p. or s.c. injections, suggesting that this model may induce some features of natural aging but without overt organ dysfunction.

Guar [Cyamopsis tetragonoloba (L.) Taub] is a diploid legume crop cultivated for galactomannan (guar gum) extracted from the endosperm of the seed. Previous studies have suggested that nodulation of guar can be poor in field conditions; however, solid proof has yet to be provided. The objectives of this study were to conduct a genome-wide association study (GWAS) and to identify single nucleotide polymorphism (SNP) markers associated with nodules in guar. GWAS was performed on a total of 225 guar genotypes using 19,007 filtered SNPs. Tassel 5 was used to run five models: single marker regression (SMR), generalized mixed linear model with PCA as a covariate (GLM_PCA), generalized mixed linear model with Q matrix as a covariate (GLM_Q), mixed linear model with PCA and Kinship (K) as covariates (GLM_PCA + K), and mixed linear model with Q and K as covariates (MLM_Q + K). Across all statistical models, the results showed a total of 3, 2, 25, 7, 2, and 3 SNPs were associated with plant height, nodule number per plant, fresh nodule weight, dry nodule weight, fresh aboveground plant biomass, and dry aboveground plant biomass. These SNPs could be used as a tool to select for better nodule traits in guar.

Alternative splicing (AS) plays a crucial role in regulating gene expression and protein diversity, influencing both normal cellular function and pathological conditions, including cancer. Protein pre-mRNA processing factor 8 (PRPF8), a core component of the spliceosome, is integral to the splicing process, ensuring accurate gene transcription and spliceosome assembly. Disruptions in PRPF8 function are linked to a variety of cancers, as mutations in this gene can induce abnormal splicing events that contribute to tumorigenesis, metastasis, and drug resistance. This review provides an in-depth analysis of the mechanisms by which PRPF8 regulates tumorigenesis through AS, exploring its role in diverse cancer types, including breast, liver, myeloid, and colorectal cancers. Furthermore, we examine the molecular pathways associated with PRPF8 dysregulation and their impact on cancer progression. We also discuss the emerging potential of targeting PRPF8 in cancer therapy, highlighting challenges in drug development.

Metastatic endometrial cancer continues to be a common cause of death as of 2024, even after maximal use of all currently available standard treatments. To address this problem of metastatic cancer generally in 2025, the drug repurposing movement within oncology identifies medicines in common general medical use that have clinical or preclinical experimental data indicating that they interfere with or inhibit a specific growth driving element identified in a given cancer. The drug repurposing movement within oncology also uses data from large scale in vitro screens of thousands of drugs, looking for simple empirical growth inhibition in a given cancer type. This paper outlines the data showing that five drugs from general medical practice meet these evidence criteria for inhibition of endometrial cancer growth, the EC5 regimen. The EC5 regimen uses the osteoporosis treatment drug, alendronate; the analgesic drug, celecoxib; the antifungal drug, itraconazole; the sleep aid, ramelteon; and the cholesterol lowering drug, simvastatin. Side effects seen with these drugs are usually minimal and easily tolerated by patients.

Ergosterol is a component of fungal cell membranes that has physiological functions and applications in drugs, such as anti-inflammatory, anti-tumor, anti-fungal, and other immunosuppressive activities. The fungus Athelia termitophila, also known as the termite ball fungus, primarily contains secondary metabolites (like active ingredients) that are similar to ergosterol. To enhance the synthesis of ergosterol and mycelial biomass in termite ball fungus, methyl jasmonate and salicylic acid were used to stimulate the biosynthesis of ergosterol compounds during the growth of TMB mycelium and relative quantitative levels of gene transcripts. The conditions of the inducers were optimized. Under 80 µmol/L MJ incubation conditions, the content of ergosterol compounds in TMB was increased by 2.23-fold compared with the wild-type strain. RT-qPCR results at the transcriptional level of ergosterol synthesis pathway genes showed that MJ significantly induced the expression of HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase), HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase), SE (Squalene Epoxidase), and FPS (Farnesyl Diphosphate Synthase) genes in the ergosterol synthesis pathway. For expression levels at different induction days, we collected 7/10 d and 4/6/8 d samples with similar expression patterns, as well as SS (Squalene Synthase)/FPS (Farnesyl Diphosphate Synthase), SE (Squalene Epoxidase)/MVD (Mevalonate Diphosphate Decarboxylase), and HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase)/HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase) genes with similar expression patterns, which resulted in gene transcription data during ergosterol content synthesis. The current study provides an effective method to increase the ergosterol contents in termite ball fungus and a new idea for the mechanism of MJ-induced ergosterol compound biosynthesis.

Muscle atrophy, through several pathways including increased protein catabolism, leads to adverse effects in cachexia induced by cancer and chemotherapy. Perch essence (PE) is a perch extract rich in branched-chain amino acids and peptides. The present study initially investigated the effects of PE supplementation on muscle atrophy in a mouse model of cancer cachexia induced by C26 cancer cells and compared these effects with those of tryptone. Compared with the tumor-only group, we found that PE supplementation significantly improved body weight, muscle mass, maximum limb grip strength (MLGS), and myosin heavy chain expression in the muscles of tumor-bearing mice. PE also significantly inhibited the expression of factors related to protein degradation, oxidative stress, and inflammation, while enhancing the expression of antioxidant enzymes in tumor-bearing mice. These effects of PE were associated with an increased expression of phosphorylated Akt and forkhead box protein O1, along with a reduced expression of phosphorylated nuclear factor-κB p65 in the muscles of tumor-bearing mice. Furthermore, PE similarly increased MLGS and attenuated muscle atrophy in mice exposed to cisplatin by inhibiting protein degradation. All the therapeutic effects of PE supplementation mentioned above were generally greater than those of tryptone supplementation. These results suggest the potential of PE in protecting against muscle atrophy induced by tumors or chemotherapy.

Glaucoma is a leading cause of irreversible blindness, with elevated intraocular pressure (IOP) as the most important risk factor. The trabecular meshwork (TM) and Schlemm’s canal are the main components of the proximal aqueous humor outflow pathway. Their dysfunction is a major contributor to IOP elevation. This study aims to identify and validate key biomarkers for TM and Schlemm’s canal endothelial (SCE) cells. A Microarray was performed on characterized human TM and SCE cells to analyze their transcriptome profiling. Differentially expressed genes (DEGs) were identified and cross-referenced with published single-cell RNA sequencing (scRNA-Seq) datasets to ensure cell-specific relevance. Further validation was performed using qPCR and re-confirmed on the scRNA-seq datasets. One-way ANOVA was used for statistical analysis, and p < 0.05 was considered significant. The Microarray revealed 341 DEGs, with TM cells enriched in metabolic and signaling pathways and SCE cells enriched in adhesion, immune, and morphogenesis-related processes. Cross-referencing with scRNA-Seq data refined the list of candidate biomarkers, and qPCR confirmed the significant gene expression differences between TM and SCE cells. CTTNBP2 and MGARP were identified as TM cell markers. JAM2, PODXL, and IFI27 are new SCE cell biomarkers. The validated biomarkers offer insights into glaucoma pathophysiology and lay the groundwork for targeted therapies.

(1) Background: Hepatic lipid accumulation is the initial factor in metabolic-associated fatty liver disease (MAFLD) in type 2 diabetics, leading to accelerated liver damage. The NOD-like receptor protein 3 (NLRP3) inflammasome plays a critical role in this process. Dapansutrile (DAPA) is a novel NLRP3 inflammasome inhibitor; however, its effect on ectopic lipid accumulation in the liver remains unclear. This study aimed to investigate the therapeutic effect of DAPA on hepatic lipid accumulation in a diabetic mouse model and its potential mechanisms. (2) Methods: The effects of DAPA on hepatic ectopic lipid deposition and liver function under metabolic stress were evaluated in vivo using db/db and high-fat diet (HFD) + streptozotocin (STZ) mouse models. Additionally, the role and mechanism of DAPA in cellular lipid deposition, mitochondrial oxidative stress, and inflammation were assessed in HepG2 cells treated with free fatty acids (FFA) and DAPA. (3) Results: Our findings indicated that DAPA treatment improved glucose and lipid metabolism in diabetic mice, particularly addressing liver heterotopic lipid deposition and insulin resistance. DAPA treatment also ameliorated lipid accumulation and mitochondrial-related functions and inflammation in HepG2 cells through the NLRP3-Caspase-1 signaling axis. (4) Conclusions: Targeting NLRP3 with DAPA may represent a novel therapeutic approach for diabetes-related fatty liver diseases.

Isatis indigotica Fort. is a traditional medicinal plant, which has anti-inflammatory, antioxidant, and antiviral properties. Despite the development and licensing of several cultivars in recent years, morphological similarity among cultivars complicates their identification. The genetic diversity within I. indigotica significantly impacts the biosynthesis of bioactive substances. To elucidate genetic relationships and evaluate bioactive compounds, I. indigotica cultivars were analyzed using SSR markers. A total of 109 alleles were identified across 29 cultivars at 20 SSR loci, exhibiting a genetic diversity with an average polymorphic information content (PIC) of 0.46. Phylogenetic, principal coordinate analysis (PCoA), and Bayesian clustering revealed that genetic relationships were largely independent of geographic origin, potentially due to regional transplantations. Notably, some cultivars with distinct leaf sizes showed clear genetic differentiation, highlighting their potential as candidates for quality evaluation. A fingerprint was successfully constructed using five SSR markers. These findings provide technical support for cultivar identification, quality evaluation, and intellectual property protection of I. indigotica cultivars.