Benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) are quaternary ammonium compound (QAC)-based biocides co-formulated into a wide range of commercial disinfectants, resulting in frequent, simultaneous exposure in humans. Despite their ubiquitous use, the toxicity caused by combined BAC and DDAC exposure remains poorly characterized. In this study, we investigated these effects in human lung adenocarcinoma A549 cells, employing real-time stress granule (SG) formation as a sensitive early-stage biomarker of sub-lethal cell-level stress. Using A549 G3BP1-eGFP knock-in cells, the two compounds induced time- and concentration-dependent SG formation individually and eIF2α phosphorylation, accompanied by a reduction in cell viability and elevation in H₂O₂ levels. Combined exposure at sub-lethal concentrations markedly potentiated SG assembly and synergistically suppressed cell viability beyond levels predicted by the Bliss independence model. At the molecular level, co-exposure robustly amplified activation of multiple unfolded protein response (UPR) branches, including the PERK/eIF2α and IRE1α/XBP1 signaling pathways, whereas individual treatments only partially engaged UPR. Collectively, these findings demonstrate that BAC and DDAC co-exposure synergistically induced toxicity through the cooperative amplification of endoplasmic reticulum stress and oxidative stress signaling, and establish SG formation as a sensitive biomarker for detecting cell-level sub-lethal perturbations not captured by conventional assays. They underscore the need for mixture-based toxicological evaluations of co-formulated disinfectants and highlight the limitations of current frameworks for assessing the safety of single compounds.

Bronopol and benzisothiazolinone (BIT) constitute antimicrobial-preservative co-formulations in consumer products, creating mixture exposure scenarios. Current risk assessments evaluate individual compounds, potentially underestimating mixture toxicity. This study investigated individual and combined toxicological effects using stress granule formation as an early biomarker. A549 lung epithelial cells were exposed to Bronopol, BIT, or combinations, assessed through real-time stress granule monitoring, viability assays, oxidative stress measurements, and integrated stress response pathway analysis. Mixture interactions were evaluated using the Bliss Independence Model. Both preservatives induced concentration-dependent stress granule formation and PERK-eIF2α activation. Critically, co-exposure produced pronounced synergism exceeding additive predictions. Sub-threshold concentrations potentiated toxicity, enhancing stress responses at lower concentrations. Combined treatments accelerated signaling kinetics and uniquely activated JNK phosphorylation. Most notably, co-exposure amplified reactive oxygen species 9-18-fold above individual compounds, demonstrating substantial synergy. These findings indicate that mixture exposures pose greater health risks than individual compound predictions, supporting mixture-based safety evaluation frameworks.

Motivation:Accurate identification of DNA-binding proteins (DBPs) and RNA-binding proteins (RBPs) is critical for elucidating transcriptional and post-transcriptional regulatory mechanisms. However, existing computational approaches often rely on inferred labels or domain-specific annotations, which limit the subsequent generalizability. Results:This study aimed to introduce transformer-based classifiers for human DBPs and RBPs that rely solely on protein sequence information without engineered features or domain constraints. The models were implemented using ESM-2 with low-rank adaptation (LoRA) fine-tuning and trained on experimentally validated datasets, including chromatin immunoprecipitation sequencing (ChIP-seq) annotations for DBPs and eCLIP annotations for RBPs. Next, to evaluate biological relevance, we computed value-aware attention (VAT) scores aggregated across transformer layers to interpret model focus. In 20-fold cross-validation, the DBP model achieved an area under the receiver operating characteristic curve (AUROC) of 0.84 with a Matthews correlation coefficient (MCC) of 0.40, while the RBP model achieved an AUROC of 0.92 with an MCC of 0.46. Proteins predicted as nucleic acid-binding were enriched for known binding domains, and inspection of attention distributions revealed preferential focus on annotated functional regions rather than non-binding segments. These results demonstrate that attention-based protein language models can accurately identify nucleic acid-binding proteins directly from sequence data. Moreover, these models reveal biologically meaningful sequence determinants of binding, establishing an interpretable and scalable framework for proteome-wide characterization of protein-nucleic acid interactions. Availability and implementation:Code is available on GitHub (https://github.com/CSB-hub/DRBP).

BackgroundNatural killer (NK) cells are promising candidates for cancer immunotherapy due to their safety and potent anti-tumor activity. However, their therapeutic efficacy is often limited by poor persistence and activity within the tumor microenvironment (TME) caused by a lack of essential cytokines.MethodsTo overcome cytokine dependence, we engineered NK92, primary NK (pNK), and chimeric antigen receptor (CAR)-NK92 cells to express an interleukin-7 receptor with an insertion mutation (IL-7R-IM), which induces constitutive signaling. We evaluated the proliferation, viability, and cytotoxicity of these cells in vitro and analyzed downstream signaling pathways using RNA sequencing and western blotting. The in vivo anti-tumor efficacy was assessed using a metastatic leukemia xenograft mouse model.ResultsNK92-IL-7R-IM cells exhibited sustained proliferation and high viability independent of exogenous cytokines, superior to IL-2-activated NK cells. This enhanced functionality was driven by the constitutive activation of the JAK/STAT, AKT, and ERK signaling pathways, leading to the upregulation of cytotoxicity-related genes (GZMA, GZMB) and anti-apoptotic genes (BCL2L1). In vivo, NK92-IL-7R-IM cells demonstrated significantly potent anti-tumor activity and extended survival compared to control groups. Furthermore, the IL-7R-IM strategy successfully enhanced the function of CAR-NK cells targeting EphA2, EGFR, and CD5 antigens.ConclusionsThe expression of IL-7R-IM confers cytokine independence and robust anti-tumor activity to NK and CAR-NK cells. This strategy offers a practical solution to improve the persistence and efficacy of off-the-shelf NK cell therapeutics for clinical application.

Understanding how chemical stress perturbs human lung physiology requires models that capture dynamic molecular responses in real time. Here, we established a CRISPR/Cas9-engineered human induced pluripotent stem cell (hiPSC)-derived lung organoid expressing endogenous G3BP1-mCherry, enabling live, non-destructive visualization of stress granule (SG) formation under toxicant exposure. The organoids recapitulated airway and alveolar epithelial diversity and displayed lamellar body-like ultrastructures, indicating advanced maturation. Time-lapse imaging revealed rapid and reversible SG dynamics across chemically distinct stressors, while cytotoxicity assays showed that these organoids are significantly more sensitive than conventional 2D or cancerderived lung models. Importantly, SG dynamics were linked to exposure duration-dependent changes in epithelial barrier integrity, indicating that SG formation precedes overt epithelial injury and serves as an early indicator of toxicant-induced cellular stress. Integration with high-content screening enabled quantitative, image-based analysis of cellular stress phenotypes, greatly enhancing throughput and mechanistic insight, thereby provided next-generation New Approach Methodologies for lung toxicity assessment. Together, this hiPSC-derived lung organoid SG reporter platform links early molecular stress adaptation to tissue-level responses, offering a predictive and mechanistically informative framework for human-relevant lung toxicity evaluation.

Alternative splicing quantification is critical for understanding disease mechanisms and developing precision medicine approaches. Conventional gel-based methods suffer from poor resolution and limited quantitative precision, especially for similar-sized exons. Here, we present multiplex alternative splicing quantification (MASQ), a dual TaqMan probe qPCR platform enabling precise quantification of alternative splicing events. MASQ employs FAM-labeled probes targeting the alternative exon and HEX-labeled probes for constitutive exon normalization, providing internal controls and eliminating variability. Using PBRM1 exon 27 as a model system, where inclusion promotes immune checkpoint resistance through enhanced PBAF complex recruitment to the PD-L1 promoter, we demonstrate superior analytical performance. MASQ exhibits exceptional linearity (R 2 > 0.98), high precision (intra-assay CV < 9%), and femtogram-level sensitivity (LOD 195 fg). CRISPR-engineered cellular validation confirmed probe specificity, while analysis of uterine corpus endometrial carcinoma specimens revealed significantly elevated exon27 inclusion in cancer tissues (61.2% ± 14.5%) compared to normal endometrium (12.7% ± 4.0%, p < 0.01). Platform generalizability was demonstrated through adaptation to HTRA2 exon 7 quantification and monitoring splice-switching oligonucleotide effects (44.6% reduction at 24 h, p < 0.0001). MASQ represents a broadly applicable platform for alternative splicing analysis, enabling applications including biomarker discovery, diagnostic development, and monitoring.

Mitochondrial redox homeostasis depends on respiratory chain integrity, but whether environmental stress alters this system through alternative splicing remains poorly understood. Here, we identify aberrant TIMM8B alternative splicing as a post-transcriptional mechanism that compromises mitochondrial respiratory chain function and redox homeostasis. Using perfluoroundecanoic acid (PFUnDA) as an environmental stressor, transcriptomic profiling of HaCaT cells revealed suppression of mitochondrial bioenergetic programs, including oxidative phosphorylation, the tricarboxylic acid cycle, mitochondrial central dogma, and protein import. Splicing analysis identified TIMM8B exon 1a inclusion as a prominent stress-associated event, which was validated by RT-PCR. The exon 1a-included isoform showed reduced transcript stability and markedly reduced detectable protein abundance. Functionally, PFUnDA impaired mitochondrial respiration in HaCaT and HDF cells. TIMM8B isoform-function analysis further showed that the exon 1a-included isoform failed to preserve ETC activity, mitochondrial membrane potential, and MitoSOX-associated redox signal, whereas the exon 1a-excluded isoform partially restored these mitochondrial readouts toward basal levels. In zebrafish, PFUnDA reduced mitochondrial fluorescence and induced NAC-sensitive oxidant accumulation. Collectively, these findings identify aberrant TIMM8B alternative splicing as a mechanism linking environmental stress to mitochondrial respiratory chain dysfunction and redox dysregulation.

Tetramethylthiuram disulfide (TMTD), widely used in rubber manufacturing and agriculture, presents occupational inhalation hazards, yet its effects on human lung epithelial cells remain poorly characterized. Here, we investigated TMTD-induced cellular stress responses in A549 lung epithelial cells, focusing on stress granule formation, oxidative stress, and DNA damage. TMTD induced concentration-dependent cytotoxicity, with brief exposure producing effects comparable to continuous exposure, indicating persistent cellular damage. Using live-cell imaging with A549 G3BP1-GFP knock-in cells, we demonstrated that TMTD rapidly triggered SG formation within minutes, accompanied by marked eIF2α phosphorylation. TMTD exposure caused dramatic intracellular ROS accumulation and robust γ-H2AX phosphorylation. Antioxidant rescue experiments using N-acetylcysteine confirmed that oxidative stress directly drives SG formation and DNA damage. Repeated TMTD exposure significantly increased apoptotic cell populations, demonstrating that cells cannot recover from recurrent exposure. Our findings reveal a mechanistic cascade whereby TMTD induces oxidative stress, triggers SG formation as an adaptive response, causes DNA damage, and ultimately leads to apoptosis when cellular stress overwhelms protective mechanisms. This study establishes stress granule formation as a sensitive early biomarker for TMTD exposure and highlights significant respiratory health risks for workers in rubber and agricultural industries, supporting the need for re-evaluation of occupational exposure limits and implementation of stringent protective measures.

Mesenchymal stem cells (MSCs) and their derived exosomes have gained significant attention in regenerative medicine due to their unique therapeutic properties, including immunomodulatory and regenerative capabilities. However, the development of a simple, scalable, and reproducible method for isolating exosomes from MSC-conditioned media remains a challenge. We optimized a polyethylene glycol (PEG)-based precipitation method for exosome isolation by initially evaluating three different PEG molecular weights (1140, 3350, and 8000 Da). Protein quantification and nanoparticle tracking analysis (NTA) were performed for all three PEG types to assess yield and particle size. Based on these results, PEG 3350 and PEG 8000 were selected for further characterization by scanning electron microscopy (SEM), size exclusion chromatography (SEC), and Western blotting. Subsequently, proteomic and metabolomic analyses were conducted using exosomes isolated with PEG 3350. For functional assays, HeLa cells were exposed to increasing concentrations of MSC-derived exosomes under either 1

Particulate matter (PM) poses significant adverse impacts on respiratory health, yet most studies investigating PM effects have relied on cancer-derived cell lines or animal models with limited physiological relevance to human lung tissue. To address this limitation, we developed functional lung organoids (hLOs) from human pluripotent stem cells that recapitulate the cellular complexity of human lungs for evaluating transcriptomic and toxicological responses to PM exposure. Transcriptome profiling of hLOs exposed to European Reference Material (ERM) identified 283 differentially expressed genes, predominantly enriched in xenobiotic metabolism and oxidative stress-related pathways. ERM exposure dose-dependently upregulated CYP1A1 expression and enzymatic activity while enhancing reactive oxygen species production. Comparative analysis with diesel particulate matter (DPM) treatment revealed distinct gene expression profiles but identified a conserved subset of commonly upregulated genes including CYP1A1, CYP1B1, and metallothionein family members (MT2A, MT1E, MT1G)-key regulators of xenobiotic metabolism and oxidative stress defense. Network analysis demonstrated molecular connectivity between these gene families, confirming that hLOs recapitulate metabolic and oxidative stress responses to compositionally distinct PMs. This study establishes a physiologically relevant in vitro platform for assessing pulmonary responses to environmental pollutants and identifies conserved molecular pathways activated upon PM exposure.

Liquid–liquid phase separation (LLPS) segregates the eukaryotic nucleus into membraneless ribonucleoprotein (RNP) condensates that orchestrate multiple stages of gene expression. In contrast to cytoplasmic granules, these nuclear assemblies lie in direct contact with chromatin and nascent pre‑mRNA, granting first‑order control over transcriptional initiation, co‑transcriptional RNA processing, and mRNA export. Consequently, alterations in their biochemical properties can propagate transcriptome‑wide disturbances and increase disease susceptibility. This review synthesizes current knowledge of the molecular composition, architectural scaffolds, and regulatory roles of the four canonical nuclear condensates—nuclear speckles, paraspeckles, Cajal bodies, and histone locus bodies. We discuss how these dynamic hubs accelerate spliceosome assembly, enforce RNA quality control, and reprogram transcription under stress, and we compile evidence that condensate hardening, mislocalization, or compositional rewiring contributes to diverse pathologies. Finally, we evaluate emerging therapeutic strategies that reengineer condensate phase behavior and outline future directions for biophysical and multi-omics approaches needed to translate condensate biology into precision medicine.

Stress granules (SGs) are cytoplasmic condensates that regulate mRNA translation and signaling in response to stress. Although SGs have been widely studied in antiviral responses, their function in bacterial infections is not well understood. Here, we demonstrate that SGs promote Mycobacterium tuberculosis (Mtb) pathogenesis by suppressing mitochondrial metabolism and innate immunity. Quantitative proteomics revealed that Mtb-induced SGs sequester mTORC1 and suppress cap-dependent mRNA translation. This leads to decreased expression of proteins necessary for mitochondrial respiration and immune activation in bone marrow-derived macrophages (BMDMs). Disrupting SG assembly restored mTORC1 signaling, enhanced oxidative phosphorylation, and increased the production of antimicrobial mediators, such as reactive oxygen species, nitric oxide, and proinflammatory cytokines. This restricted intracellular Mtb growth in vitro and in vivo. Mechanistically, intracellular ATP depletion triggered by Mtb phagocytosis was sufficient to drive SG formation, linking energy stress to translational repression. Furthermore, SGs captured Ndufa12, a complex I subunit, thereby impairing mitochondrial electron transport and ATP production. These findings identify SGs as key regulators that couple translational arrest to metabolic and immune suppression, enabling Mtb persistence. Targeting SG formation or function could be a host-directed strategy to restore mitochondrial activity and strengthen immune responses against Mtb infection.

BACKGROUND:Alternative splicing (AS) plays a crucial role in regulating protein function through the generation of structurally distinct isoforms. OBJECTIVE:We identify a novel splicing event in Chitinase 3-like 1 (CHI3L1) that modulates its secretion through conformational changes. METHODS:CHI3L1 alternative splicing was analyzed using the GTEx dataset. The regulation of CHI3L1 splicing was examined in response to THP-1 and BEAS-2B cells using RT-PCR. Structural modeling of CHI3L1 isoforms was conducted with AlphaFold to predict conformational changes caused by exon 8 exclusion. Protein expression and secretion levels of CHI3L1 isoforms were analyzed by Western blotting. RESULTS:Analysis of the GTEx dataset revealed tissue-specific regulation of CHI3L1 exon 8, with pronounced exclusion in lung tissue. The splicing pattern of CHI3L1 was dynamically regulated during THP-1 macrophage differentiation and by cell density in lung-derived epithelial BEAS-2B cells, suggesting its responsiveness to cellular context. While both full-length and exon 8-excluded CHI3L1 proteins showed cytoplasmic localization, structural analysis using AlphaFold revealed that exon 8 exclusion significantly altered the orientation of the signal peptide. Consequently, exon 8-excluded CHI3L1 exhibited minimal secretion into the culture medium compared to the full-length protein. CONCLUSION:These findings demonstrate that alternative splicing-mediated exclusion of exon 8 serves as a novel regulatory mechanism controlling CHI3L1 secretion through conformational changes, providing new insights into the post-transcriptional regulation of secreted proteins.

Perfluorooctanoic acid (PFOA), a perfluoroalkyl acid, induces neuroinflammation. However, present understanding regarding its fundamental role in neuroinflammation remains limited. Therefore, in this study, we aimed to clarify the potential association between PFOA and astrocyte activation via the modulation of the endoplasmic reticulum (ER) stress-autophagy axis. The results obtained revealed that PFOA activated astroglia in A-172 astrocytoma cells and primary astrocytes by upregulating the expression levels of autophagy-related proteins (ATG5, BECN1, SQSTM1, and MAP1LC3B-II). It also activated autophagy in A-172 astrocytoma cells and primary astrocytes via the upstream activation of ER stress-related proteins, such as ATF4, GRP78, and CHOP. Further, the pharmacological inhibition of ER stress as well as autophagy prevented PFOA-induced activation of astrogliosis in PFOA-treated A-172 cells and primary astrocytes. We also observed that PFOA-mediated activation of GFAP upregulated the transcription of pro-inflammatory cytokines, such as IL-1β, TNF-α, and IL-6. These findings confirmed the existence of a relationship between ER stress-induced autophagy and astrogliosis in PFOA-treated astrocytes, suggesting that targeting the ER stress-autophagy axis may be a potential therapeutic strategy for reducing PFOA-induced neuroinflammation.

Objective This study investigates the potential of grape tendril extracts (G.T.) derived from Shine Muscat variety in mitigating oxidative stress, inflammation, and allergic reactions, focusing on stress granule (SG) dynamics, inflammatory pathways, and mast cell responses.Methods G.T. was extracted using 50% ethanol and assessed for antioxidant activity via ABTS radical scavenging assay. Its anti-inflammatory effects were evaluated in RAW 264.7 cells by measuring inflammatory cytokine expression (TNF-alpha, IL-6, IL-1 beta) using qRT-PCR and NF-kappa B pathway components by immunoblotting. SG formation was visualized in AGS cells under arsenite-induced oxidative stress through G3BP1 immunofluorescence staining. Anti-allergic properties were determined through beta-hexosaminidase release assay in RBL-2H3 mast cells.Results G.T. demonstrated potent antioxidant activity with 88% radical scavenging at 4 mg/mL. It significantly suppressed LPS-induced pro-inflammatory cytokine expression and I kappa B alpha phosphorylation in RAW 264.7 cells. Under oxidative stress, G.T. (0.5 mg/mL) reduced both the number of SG-positive cells and SGs per cell in AGS cells. Moreover, G.T. inhibited compound 48/80-induced beta-hexosaminidase release in RBL-2H3 cells dose-dependently.Conclusions These findings establish G.T. as a promising natural therapeutic agent with significant antioxidant, anti-inflammatory, and anti-allergic properties. The demonstrated effects on SG formation provide new insights into its cellular protective mechanisms.

KRAS is a well-established oncogene that exhibits high-frequency mutations at cancer-driver hotspot loci across human cancers. While the oncogenic roles of mutant KRAS have been extensively investigated, the functional significance of KRAS splicing isoforms generated by exon 4 (E4) alternative splicing remains poorly understood. Here, we analyzed the expression patterns of KRAS E4 splicing variants in cancer tissues using The Cancer Genome Atlas (TCGA) data and found that certain cancer types exhibit relatively higher expression of the E4-included KRAS (KRAS4A) splicing variant compared to the E4-excluded KRAS (KRAS4B) splicing variant. Functional analysis revealed that the oncogenic properties of KRAS4A were significantly enhanced compared to those of KRAS4B. Furthermore, we identified RBM47 and PTBP1 as key regulators that promote KRAS E4 inclusion. Our findings demonstrate that RBM47- and PTBP1-mediated alternative splicing of KRAS contributes to enhanced tumor progression, highlighting KRAS alternative splicing as a promising therapeutic target for cancer treatment.

Bavachin, a bioactive phytoestrogen from Psoralea corylifolia, has shown promising therapeutic potential in various cancers, but its effects on ovarian cancer remain unexplored. In this study, we investigate the anti-cancer mechanisms of bavachin in ES2 and OV90 ovarian cancer cells and its potential to enhance paclitaxel sensitivity. Bavachin significantly inhibited cell proliferation and spheroid formation while inducing caspase-dependent apoptosis through modulation of Bcl-2 family proteins. Mechanistically, bavachin disrupted mitochondrial function by inducing membrane depolarization and calcium dysregulation, leading to comprehensive impairment of cellular bioenergetics including both oxidative phosphorylation and glycolysis. Furthermore, bavachin activated the endoplasmic reticulum stress pathway as evidenced by upregulation of GRP78, ATF4, PERK, and CHOP, and notably inhibited phosphorylation of ERK1/2 and p38 MAPK signaling pathways essential for tumor cells survival. Combination treatment with bavachin enhanced the cytotoxic effects of paclitaxel, showing synergistic anti-cancer activity in both cell lines. These findings demonstrate that bavachin effectively suppresses ovarian cancer growth through multiple mechanisms and may serve as a promising therapeutic agent, particularly in combination with conventional chemotherapy.

Purpose Perivascular adipose tissue (PVAT) dysfunction contributes to vascular impairment in obesity, primarily through altered lipid accumulation, inflammatory cytokine imbalance, and disrupted endothelial signaling. This study aimed to investigate whether aerobic exercise can ameliorate PVAT-induced vascular dysfunction in mice with obesity induced by a high-fat diet. Methods Male C57BL/6J mice were fed either a chow or a high-fat diet for 12 weeks, followed by 8 weeks of aerobic treadmill training or sedentary control. Body weight, PVAT morphology, and levels of inflammatory cytokines (tumor necrosis factor alpha, interleukin 1β, monocyte chemoattractant protein 1, and intercellular adhesion molecule 1), adipokines (leptin and adiponectin), and phosphorylation of endothelial signaling proteins (Akt, AMP-activated protein kinase [AMPK], and endothelial nitric oxide synthase [eNOS]) were evaluated in the thoracic aorta and PVAT. Endothelium-dependent relaxation (EDR) was assessed using acetylcholine-induced vasodilation in aortic rings with or without PVAT. Results High-fat diet-induced obesity led to increased adipocyte size in PVAT, while it also impaired vascular relaxation, elevated levels of proinflammatory cytokines, and reduced phosphorylation of Akt, AMPK, and eNOS in both PVAT and the aorta. Aerobic exercise training significantly reduced PVAT adipocyte size, restored EDR, suppressed inflammatory cytokine levels, increased adiponectin expression, and promoted phosphorylation of vascular signaling molecules in both PVAT and the aorta. Conclusions Aerobic exercise training restores PVAT homeostasis and endothelial function in obese mice by modulating inflammation, adipokines, and vascular signaling. These findings suggest aerobic exercise as a nonpharmacological approach to improve vascular function in obesity by targeting PVAT dysfunction.

Skeletal muscle health is crucial for maintaining physical function and metabolic homeostasis. This study investigated the effects of gallic acid, a naturally occurring phenolic compound, on myogenesis and muscle function. In vitro experiments using mouse primary myoblasts demonstrated that gallic acid (10 μg/mL) enhanced myogenic differentiation, evidenced by increased Myh3 protein expression (3.3-fold under growth conditions and 1.3-fold under differentiation conditions) and enhanced myotube formation in both conditions. In vivo studies were conducted using C57BL/6N mice fed either a control diet or a 0.2% gallic acid-supplemented diet for 8 weeks. While gallic acid supplementation did not affect body weight, food intake, or general metabolic parameters, it significantly increased soleus muscle mass (10.56 ± 1.35 mg vs. 8.28 ± 0.68 mg in controls). Furthermore, gallic acid-fed mice showed improved muscle function, with increased running distance (345.0 ± 28.8 m vs. 262.2 ± 58.6 m), extended time to exhaustion (24.2 ± 1.2 min vs. 20.1 ± 3.0 min), and enhanced grip strength (182.2 ± 19.4 N vs. 157.3 ± 11.5 N). These findings suggest that gallic acid could serve as a promising natural supplement for improving muscle function and exercise performance.