Search Results (5,261)

In this study, a mitochondria-targeting near-infrared (NIR) fluorescent probe, Mito-V, based on an oxanthrene-like hybrid structure was designed and synthesized, for detecting viscosity with pH-cooperative response characteristics. Upon increasing viscosity, Mito-V exhibits a significant NIR fluorescence enhancement with a high signal-to-noise ratio and excellent selectivity. Under weakly acidic conditions, the viscosity response is further enhanced, demonstrating a pH-cooperative effect that improves detection sensitivity. Cellular imaging experiments confirmed that Mito-V can effectively monitor fluctuations in intracellular viscosity, and co-localization studies revealed its high specificity for mitochondria, with a Pearson’s coefficient of 0.89. Furthermore, the pH-cooperative effect on viscosity detection was verified at the cellular level. In vivo imaging in zebrafish successfully visualized viscosity variations, demonstrating the probe’s applicability and biosafety for monitoring viscosity in biological systems. These findings indicate that Mito-V holds great potential for studying viscosity-related processes in live cells and organisms. Full article

Background/Objectives: Gamma-aminobutyric acid (GABA) is increasingly recognized as an important modulator of pancreatic beta-cell function, but the mechanisms by which it regulates intracellular Ca²⁺ oscillations and coordinated beta-cell activity remain insufficiently understood. The aim of this study was to investigate how GABA influences the amplitude, frequency, phase adjustment, entrainment, and synchronization of beta-cell Ca²⁺ oscillations. Methods: We developed a reduced ATP–Ca²⁺ oscillation model, based on established beta-cell oscillatory frameworks, and coupled it to the GABA-shunt subsystem derived from our previously established Dual Anaplerotic Model. The model incorporates explicit dynamics of cytosolic Ca²⁺, endoplasmic reticulum Ca²⁺, ATP, and a regulatory variable controlling Ca²⁺ influx, while the interstitial GABA signal is represented as a delayed feedback signal acting on cellular excitability. Single-cell and two-cell simulations were performed to analyze GABA-dependent oscillatory regulation and intercellular coupling. Results: The model reproduced key experimental observations under both control and GABA-deficient conditions, including reduced Ca²⁺-oscillation amplitude and a prolonged oscillation period when GABA production was suppressed. Mechanistically, GABA affected single-cell oscillations through two complementary pathways: metabolically, by modulating ATP production through PEP-related and TCA-related contributions linked to the GABA shunt, and as an interstitial/paracrine signal, by adjusting the phase of Ca²⁺ influx through fast and delayed inhibitory feedback. In the reduced two-cell model, delayed interstitial GABA signaling could phase-lock non-identical oscillators over finite ranges of parameter mismatch. When included as an additional weak effective term, electrical coupling broadened these ranges, consistent with a complementary interaction between GABA-mediated phase adjustment and established electrical coupling. Conclusions: GABA acts as a dual regulator of beta-cell dynamics, linking intracellular metabolism to Ca²⁺-oscillation patterning and promoting coordinated activity through intercellular phase adjustment. The model provides a mechanistic framework connecting GABA metabolism, ATP dynamics, Ca²⁺ signaling, and beta-cell synchronization in pancreatic islets. Full article

Snake venom is a remarkably complex cocktail of bioactive molecules capable of hijacking diverse host physiological processes, yet how individual venom components drive these cellular responses remains a bit of a black box. To map these dynamics, we ran a comparative transcriptomic analysis on human osteoclastogenic cultures, exposing them continuously to crude Bothrops moojeni venom and its high (HMM) and low (LMM) molecular mass fractions throughout differentiation. This allowed us to capture the cumulative transcriptional shifts that unfold across the entire lifecycle of osteoclast development. The crude venom triggered a sweeping response, deeply impacting neuroimmune, extracellular matrix remodeling, inflammatory, and apoptotic pathways—reflecting a massive reshuffling of cellular regulatory networks. When we looked at the fractions, clear dividing lines emerged. The HMM fraction, packed with metalloproteinases and serine proteases, mostly drove pathways tied to cytoskeletal remodeling, intracellular trafficking, and osteoclast-associated signaling. In contrast, the LMM fraction—home to phospholipases A₂, disintegrins, and small peptides—steered a much more targeted course, influencing immune regulation, proliferative signaling, and metabolic homeostasis while noticeably turning down catalytic and binding functions. Interestingly, all venom-treated groups shared a drop-off in ATP-dependent and ligand-binding categories, pointing to a common disruption in core metabolic and signaling processes. Taken together, these findings offer a clearer mechanistic look at how different B. moojeni venom components target bone remodeling pathways, highlighting the power of transcriptomics for untangling complex venom–host interactions. Full article

Pancreatic cancer remains a highly lethal malignancy, with standard therapies offering limited benefits in advanced stages; thus, novel strategies that exploit specific cancer cell vulnerabilities are urgently needed. Building on our previous findings that nanosecond pulsed electric fields (nsPEF) combined with cold atmospheric plasma (CAP) produce enhanced cytotoxicity, this study investigates the molecular mechanisms underlying this synergy. Pan02 pancreatic cancer cells were subjected to nsPEF, CAP, or a combination of both. We assessed cell viability, reactive oxygen species (ROS) production, and mitochondrial integrity using metabolic assays, flow cytometry, and fluorescence microscopy. Apoptotic markers were evaluated via Western blotting and caspase activity assays. Combined nsPEF–CAP treatment significantly outperformed either modality alone in inducing cell death. Mechanistically, dual treatment triggered a surge in intracellular ROS, particularly mitochondrial superoxide, indicating severe oxidative stress. Distinct mitochondrial responses were observed: nsPEF reduced mitochondrial membrane potential, whereas CAP alone caused a slight elevation. Notably, while CAP induced apoptosis (evidenced by increased cleaved caspase-3 and caspase-3/7 activity), lethal nsPEF (100 pulses) caused cell death without triggering apoptotic signaling. However, mild nsPEF (20 pulses) significantly potentiated CAP-induced apoptosis. These findings suggest that nsPEF sensitizes cells to CAP treatment by amplifying oxidative stress and mitochondrial dysfunction. This synergistic combination represents a promising therapeutic approach for managing pancreatic cancer cells resistant to conventional therapies. Full article

Marine medicinal algae represent a valuable reservoir of bioactive metabolites for anticancer drug discovery, yet the efficient identification of target-relevant compounds from chemically complex marine matrices remains challenging. In this study, an integrated cathepsin B-oriented strategy was developed to discover, prioritize, isolate, and validate antitumor metabolites from the brown alga Sargassum polycystum. Affinity ultrafiltration LC-MS was first applied to screen CTSB-binding constituents from the crude extract, followed by molecular docking, molecular dynamics simulation, and gray relational analysis for multidimensional candidate prioritization. Seven CTSB-binding metabolites were characterized, including chlorogenic acid, caffeic acid, cynarin, loliolide, taxifolin, senkyunolide H, and dihydroactinidiolide, with binding degrees of 73.99–85.61% at 2.5 U/mL CTSB. Molecular docking showed predicted binding affinities ranging from −6.3 to −9.4 kcal/mol, compared with −10.2 kcal/mol for the positive control CA-074Me. Integrated computational and biological evaluation identified caffeic acid, cynarin, and taxifolin as the top-ranked candidates. Preparative recovery was then achieved using counter-current chromatography combined with semi-preparative HPLC, and the isolated compounds were structurally identified by LC-MS/MS and NMR. Cellular assays in NCI-H1975 cells suggested that these metabolites reduced CTSB-associated enzymatic activity and intracellular CTSB-related fluorescence signals to different extents, with phenolic acid-type compounds exhibiting comparatively stronger effects. At the extract level, S. polycystum dose-dependently suppressed NCI-H1975 xenograft tumor growth, with inhibition rates of 48.78%, 36.58%, and 22.86% in the high-, middle-, and low-dose groups, respectively, without evident hepatorenal histopathological toxicity. This effect was associated with reduced CTSB, Ki-67, and Bcl-2 staining, increased Bax staining, enhanced apoptosis, and ultrastructural alterations in tumor tissues. Overall, this study provides a practical CTSB-oriented workflow for discovering antitumor metabolites from marine medicinal algae and supports further investigation of S. polycystum as a potential source of anti-NSCLC candidates. Full article

Ischaemic stroke induces a dynamic neuroimmune response in which microglia act as central regulators of both secondary injury and tissue repair. In the acute phase, microglial activation amplifies neuronal damage through inflammatory signalling and vascular dysfunction; over subsequent days, these cells undergo coordinated transcriptional and metabolic reprogramming toward reparative states. The repeated failure of immunomodulatory therapies in clinical translation, however, suggests that current approaches fundamentally mischaracterise the underlying biology. We propose that microglial state transitions are governed not by discrete linear pathways but by a coupled regulatory network integrating proteostatic clearance, receptor-mediated signalling, inflammasome activation, and intracellular metabolism. Within this network, impaired clearance of cellular debris sustains exposure to damage-associated molecular patterns, perpetuating inflammasome activity and a pro-inflammatory metabolic programme; conversely, restoration of clearance capacity shifts network equilibrium toward resolution and repair. Microglial phenotypes therefore emerge from dynamic shifts in network state rather than progression through fixed activation stages. This framework accounts for the limited efficacy of non-selective or temporally misaligned interventions and identifies the post-acute transitional phase as a window of maximal network plasticity. Aligning therapy with the temporal and functional dynamics of this network—guided by phase-specific biomarkers—provides a mechanistic basis for precision immunomodulation and improved clinical translation in ischaemic stroke. Full article

Muscle wasting associated with gastric cancer represents a complex, multifactorial systems disorder involving inflammatory, anabolic, mechanosensory, calcium-regulatory, mitochondrial, and proteostatic disruption. This review synthesizes current evidence regarding the cellular and physiological mechanisms involved in skeletal muscle dysfunction in gastric cancer and provides a unifying framework centered on loss of signaling coherence. Specifically, it examines IL-6/STAT3 and NF-κB inflammatory signaling, the myostatin–activin–ACVR2B–SMAD pathway, PI3K/Akt/mTOR signaling, mechanotransduction, excitation–metabolism coupling, calcium homeostasis, mitochondrial function, and proteostasis. Although individual components of these pathways have been implicated in muscle wasting associated with chronic disease, current evidence suggests that they interact through positive feedback loops. Inflammation, anabolic resistance, impaired force-to-signal conversion, mitochondrial stress, altered intracellular calcium homeostasis, and disrupted protein quality control may reinforce one another, contributing to metabolic, structural, and transcriptional instability. Within this context, muscle wasting reflects not only loss of muscle mass or strength, but also loss of functional integrity resulting from disrupted integration of mechanical, metabolic, inflammatory, and anabolic signals. Given the systemic nature of these effects, this review proposes kinesitherapy as a potentially useful nonpharmacological adjunctive strategy that may modulate inflammation, restore responsiveness to mechanical stimuli, support calcium homeostasis and mitochondrial function, improve anabolic sensitivity, and maintain protein quality control. Overall, this review presents a systems-biology model of gastric cancer-associated muscle wasting and supports further investigation of exercise-based therapies for this condition. Full article

Background: Cancer-associated cachexia (CAC) can affect up to 80% of patients with late-stage cancer and is characterized by depletion of skeletal muscle mass with or without loss of fat tissue. No effective treatments are currently available, and reversing CAC requires understanding the intracellular processes of muscle atrophy and its cancer-related extracellular triggers. In this study, we aimed to disentangle tumor- and host-driven mechanisms in CAC muscle wasting. Methods: In skeletal muscle tissue obtained from control non-tumor-bearing mice and cachectic mice resulting from orthotopically implanted 344P lung adenocarcinoma cells, transcriptomic analyses were performed to identify muscle wasting-associated processes. To explore whether these reflected direct tumor-induced effects, 344P tumor-conditioned medium (tCM) was applied to in vitro cultured C2C12 skeletal muscle cells to investigate the impact on muscle proteolysis, myogenesis and mitochondrial function. Results: RNAseq data revealed increased proteolysis along with decreased myogenesis-related processes, and prominent downregulation of genes encoding mitochondrial OXPHOS complexes, in cachectic mouse muscle. Exposure of cultured skeletal muscle cells to tCM reduced mitochondrial respiration and induced changes in mitochondrial mass and mitochondrial DNA copy number. tCM did not induce myotube atrophy, or activation of proteolysis-related signaling, in fully differentiated myotubes. In contrast, tCM reversibly inhibited myoblast–myotube fusion, and reduced myogenic and muscle-specific gene expression in differentiating myoblasts. Application of CCCP to simulate muscle mitochondrial dysfunction reproduced the myogenesis-impairing phenotype caused by tCM. Conclusions: Our results show that factors present in the cachexia-inducing lung tumor secretome directly impair myogenesis and muscle mitochondrial function, whereas activation of muscle catabolic processes requires host-dependent mechanisms. Full article

Xanthotoxin (8-methoxypsoralen) belongs to the group of naturally occurring furanocoumarin (furocoumarin) compounds and is a product of plant secondary metabolism. Analysis of the available literature indicates that xanthotoxin exhibits a broad spectrum of pharmacological activities, including anti-inflammatory, antioxidant, immunomodulatory, and antibacterial effects. Xanthotoxin has been shown to stimulate autophagy via inhibition of the AKT/mTOR pathway, as well as to block cell migration by modulating RIG-1 and NF-κB signaling. Moreover, its effects on JNK/MAPK, PI3K/AKT, Calcium–CaMYK/PYK2, and other signaling cascades have been confirmed. Among its most promising properties is the ability to inhibit ABC transporters, thereby preventing the reduction of chemotherapeutic agent concentrations within tumor cells and enhancing their intracellular accumulation. Thus, the aim of this study was to evaluate xanthotoxin as a potential anticancer agent. The literature review was based on publications indexed in Google Scholar, Scopus, Web of Science, and PubMed and published between 2010 and 2026. Studies describing the biological properties of xanthotoxin, its toxicity, anticancer mechanisms of action, and modulation of ABC transporters were included. This literature review summarizes the pharmacological profile of xanthotoxin, and its biological activities and therapeutic potential, as well as its antitumor effects in various cancer cell lines. The available evidence may provide a foundation for the future development of xanthotoxin as a lead compound for anticancer drug discovery. Full article

Glutamine-Fructose-6-Phosphate Transaminase 1 (GFPT1), the rate-limiting enzyme of the hexosamine biosynthetic pathway (HBP), provides the UDP-N-acetylglucosamine (UDP-GlcNAc) required for protein glycosylation. Biallelic mutations in GFPT1 cause congenital myasthenic syndromes (GFPT1-CMS), yet the molecular mechanisms linking impaired glycosylation to skeletal muscle dysfunction remain incompletely understood. Here, we combine cellular models of inducible Gfpt1 knockdown and a skeletal muscle-specific Gfpt1 knockout mouse (Gfpt1^Tm1d/Tm1d) with whole-cell proteomics, immunoblot studies and secretomics to define glycosylation-dependent defects in intracellular trafficking, ER stress signaling and autophagy. Global proteomic profiling of Gfpt1-deficient myoblasts revealed marked downregulation of protein trafficking pathways and impaired secretion of key muscle cargo proteins, including serglycin (Srgn). Loss of GFPT1 reduced both high-molecular-weight glycosylated serglycin and its core protein, accompanied by intracellular retention and decreased secretion. These trafficking defects coincide with robust activation of the unfolded protein response (UPR), evidenced by increased Xbp1 expression and accumulation of spliced Xbp1s across pharmacologic, cellular, and mouse models of GFPT1 deficiency. Converging evidence from proteomics, immunoblotting, and immunofluorescence demonstrated impaired autophagy, including increased LC3-II accumulation, elevated p62/Sqstm1 levels, and enhanced p62-positive puncta in both Gfpt1-deficient C2C12 myoblasts and skeletal muscle. Soluble/insoluble fractionation further confirmed p62 accumulation, indicating defective autophagic flux and buildup of aggregated cargo. Together, these findings identify a glycosylation-dependent failure in protein trafficking that triggers ER stress, UPR activation, and autophagy impairment in Gfpt1-deficient skeletal muscle. This mechanistic cascade provides a unifying explanation for muscle pathology in GFPT1-CMS and suggests that restoring glycosylation or improving proteostasis may represent viable therapeutic approaches. Full article

Background/Objective: Accumulation of amyloid-beta (Aβ) senile plaques in the human brain is a major hallmark of Alzheimer’s disease (AD), which manifests as progressive decline in memory and cognitive functions and currently lacks effective disease-modifying therapies. Emerging evidence demonstrates that polyphenol-rich plant foods are potential complementary therapies for AD. Methods: In this study, we investigated crude polyphenol extracts (CPEs) and purified polyphenol extracts (PPEs) from three sorghum genotypes for their ability to inhibit Aβ₄₂-induced toxicity in MC-65 cells. Thioflavin T fluorescence, cell viability, mitochondrial function, oxidative stress assays, and Western blotting, along with RNA sequencing and computational analyses, were used to characterise both functional and transcriptomic responses of the cells to polyphenol treatments. Results: CPEs and PPEs inhibited Aβ₄₂ aggregation by 67–76% and significantly reduced Aβ oligomer species. The extracts increased cell viability against Aβ-induced toxicity by more than 70%, decreased intracellular oxidative stress, and enhanced mitochondrial activity by over 80%. Transcriptomic profiling revealed differential modulation of genes associated with ferroptosis and MAPK/NF- κB signalling pathways, indicating regulation of inflammatory and oxidative-stress responses are mechanisms underlying the observed neuroprotection. Conclusions: This study demonstrates that polyphenol extracts from black and red sorghum genotypes exert strong multitarget neuroprotection against Aβ₄₂ toxicity in MC-65 cells. These findings support further evaluation of sorghum-derived polyphenols as complementary therapeutic candidates for AD, with in vivo studies required to establish efficacy and translational potential. Full article

Hair loss is closely associated with oxidative stress, which impairs the function of human follicle dermal papilla cells (HFDPCs) and disrupts hair follicle homeostasis. Current pharmacological treatments, such as minoxidil and finasteride, are effective but may cause adverse effects, highlighting the need for safer alternatives. In this study, we utilized a patented high-pressure processing method to produce black ginseng concentrate (BGC), which is significantly enriched with rare bioactive ginsenosides, including Rg3, Rg5, and Rk1, through optimized chemical transformation. We aimed to elucidate the protective effects of BGC against oxidative stress-induced damage in HFDPCs. BGC significantly reduced intracellular reactive oxygen species (ROS) levels. BGC also improved mitochondrial function, including an increased oxygen consumption rate (OCR). In addition, BGC activated hair growth-related signaling pathways by upregulating Wnt/β-catenin and increasing the phosphorylation levels of ERK and AKT. Collectively, these findings demonstrate that BGC protects HFDPCs from oxidative stress, improves mitochondrial function, and supports key signaling pathways associated with hair growth. This study suggests that BGC has potential as a natural agent for preventing oxidative stress-induced cellular dysfunction related to hair loss. Full article

The widespread application of per- and polyfluoroalkyl substances (PFASs) has established perfluorooctanesulfonic acid (PFOS), a representative PFAS, as a critical environmental pollutant. Although PFOS exposure causes significant bioaccumulation and potential myelotoxicity, its specific impact on the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) remains to be elucidated. In this study, we established a murine model of PFOS exposure to isolate primary BMSCs and investigated this issue through in vitro differentiation assays, cellular senescence evaluations, and an in vivo subcutaneous implantation model using gelatin methacryloyl (GelMA) hydrogel scaffolds. Our results demonstrated that PFOS exposure triggered intracellular reactive oxygen species (ROS) accumulation and induced a senescent phenotype in BMSCs, characterized by restricted cellular proliferation and the release of senescence-associated secretory phenotype (SASP) factors, thereby markedly suppressing their chondrogenic capacity. Mechanistically, the inhibition of the Nrf2 signaling pathway by PFOS was identified as the principal driver of this process. Furthermore, both in vitro and in vivo assays confirmed that pharmacological activation using the Nrf2 agonist sulforaphane (SFN) effectively mitigated the senescent phenotype and restored the chondrogenic potential of PFOS-exposed BMSCs. Altogether, these findings elucidate the specific mechanisms of PFOS-induced stem cell toxicity and offer a potential strategy to overcome the resulting limitations in BMSC-based cartilage regeneration. Full article

Human spaceflight in microgravity induces profound physiological adaptations, yet its effects on the sensory system remain comparatively underexplored. While musculoskeletal and cardiovascular changes are well documented, sensory alterations pose equally important challenges to astronaut safety, performance, and post-mission recovery. Calcium ions (Ca²⁺), as universal intracellular messengers, play central roles in sensory transduction, neurotransmitter release, and adaptive signaling across all sensory modalities. Emerging evidence suggests that microgravity may influence Ca²⁺ homeostasis and Ca²⁺-dependent cellular processes, potentially affecting the functional integrity of sensory pathways. In this review, we synthesize current findings on the impact of microgravity on Ca²⁺-dependent processes in the five classical senses. Evidence from spaceflight studies, ground-based analogs, and related physiological models suggests possible alterations in taste receptor signaling, Ca²⁺-binding protein expression, mechanotransduction pathways, and vestibular function. However, direct evidence for microgravity-induced disruption of Ca²⁺ signaling remains limited for several sensory modalities. Collectively, these changes are associated with altered taste and smell perception, visual disturbances, reduced tactile sensitivity, and vestibular imbalance. By integrating both direct evidence and mechanistic hypotheses across sensory systems, this review highlights Ca²⁺ signaling as a potential unifying mechanism underlying sensory adaptation to microgravity. We further identify key knowledge gaps and discuss potential directions for developing targeted countermeasures aimed at preserving sensory function during long-duration missions. Beyond spaceflight, these insights contribute to a broader understanding of Ca²⁺-mediated sensory physiology under extreme environmental conditions. Full article

Virus infection requires coordinated activation of pathogen-sensing, innate immune, and cellular stress response pathways to mount an effective antiviral defense. Recognition of nucleic acid pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs) initiates signaling cascades that drive the production of type I interferons (IFNs) and proinflammatory cytokines. These responses are often accompanied by the activation of integrated stress response pathways that help optimize host defense. Cytosolic double-stranded dsDNA, generated during viral infection or released from damaged mitochondria, is sensed by cyclic GMP-AMP synthase (cGAS), which generates 2′3′-cGAMP to activate stimulator of interferon genes (STING). Activated STING translocates from the endoplasmic reticulum to the Golgi, where it drives TBK1-dependent IFN and cytokine production. Previous reports show that cGAS activity is enhanced by Ras-GAP SH3 domain binding protein 1 (G3BP1), a key nucleator of stress granules (SGs), independent of its role in SG assembly. Here, we identify a non-canonical role of G3BP1 as a regulator of DNA sensing responses at multiple levels, including STING intracellular trafficking, in addition to potentiating cGAS activity. Loss of G3BP1 impaired STING-dependent IFN and cytokine responses to HSV-1 infection and viral DNA. G3BP1-deficient cells showed reduced cGAMP-induced STING translocation to the Golgi, induction of type I IFN and proinflammatory cytokines, and activation of the ER stress kinase PERK and stress granule formation. Together, these findings demonstrate G3BP1-STING as a node linking DNA sensing, innate immunity, and stress signaling with broad implications for antiviral defense and diseases characterized by aberrant DNA sensing and stress responses, including neurodegeneration, fibrosis, and autoimmunity. Full article