Search Results (44,268)

The intestinal barrier is a key interface maintaining host–microbial segregation and systemic homeostasis. A broad range of viruses, including enteric, respiratory, and systemic pathogens, can disrupt this barrier through effects on epithelial integrity, vesicular transport, immune responses, and microbial ecology. Whether these diverse insults converge on shared regulatory nodes or act through distinct virus-specific pathways that ultimately result in barrier failure remains unclear. Building on this premise, this review systematically delineates the molecular and cellular mechanisms underlying virus-induced disruption of the intestinal barrier. Viral infection disrupts epithelial integrity through multiple converging processes, including disassembly of tight junction architecture, activation of programmed cell death pathways, degradation of the mucus layer, impaired regeneration driven by intestinal stem cells, and dysregulation of transcellular transport. These processes are interconnected and collectively drive epithelial dysfunction and barrier breakdown. Beyond epithelial damage, we further highlight the pivotal contribution of host immune responses to barrier breakdown. Viral infection induces dysregulated cytokine production and aberrant immune activation, which amplify epithelial damage and further increase barrier permeability. In parallel, increasing evidence supports a bidirectional interaction between viral infection and gut microbiota dysbiosis, in which each process reinforces the other to accelerate barrier disruption and disease progression. We also discuss emerging therapeutic strategies aimed at restoring intestinal homeostasis, including antiviral therapies, host-targeted interventions, and microbiota modulation. Despite recent progress, key questions remain, particularly regarding mechanisms of failed barrier repair after viral clearance and the multilayered regulatory networks linking viruses, immunity, and the microbiota. Together, this review provides a framework for understanding virus-induced intestinal barrier dysfunction and identifies potential therapeutic nodes for intervention. Full article

First-generation H₂S donors fail in pulmonary arterial hypertension (PAH) not due to lack of efficacy, but because they release H₂S indiscriminately. This review articulates a conceptual framework for advancing H₂S donor therapy along three complementary directions. Donor 2.0 for precision delivery: Donors remain inert in normal tissues, but release H₂S upon sensing PAH microenvironment signals [reactive oxygen species (ROS), hypoxia, esterases, matrix metalloproteinases (MMPs)], combined with lesion-selective enrichment and organelle targeting. Donor–endogenous synergy: Move from chronic exogenous supplementation to restoring the patient’s own H₂S synthesis via epigenetic derepression of CSE, oxidative reactivation of CBS, and substrate support for 3-MST. Systemic sensitization: Redefine H₂S donors as combination enhancers that reverse acquired insensitivity to ERAs, PDE5i, and prostacyclin analogues through protein S-sulfhydration. These three mutually reinforcing dimensions transform H₂S donors from passive releasers into programmable, context-sensitive therapeutic platforms, addressing the fundamental limitations of current PAH therapies. Full article

Oral delivery of bioactive compounds in veterinary medicine offers important opportunities to improve animal health, productivity, and welfare, but its effectiveness is often limited by species-specific gastrointestinal physiology. This review aims to provide a comprehensive overview of microencapsulation strategies for oral veterinary delivery, focusing on species-specific gastrointestinal barriers and the formulation approaches developed to overcome them. Monogastric and ruminant animals present distinct gastrointestinal environments that compromise the stability, bioavailability, and therapeutic performance of orally administered compounds. In monogastrics, gastric acidity, digestive enzymes, gastrointestinal transit, and microbiota-mediated interactions represent major barriers, whereas in ruminants, ruminal fermentation, prolonged retention, and physicochemical conditions may cause premature degradation of bioactive compounds. These barriers significantly hinder the successful use of probiotics, enzymes, essential oils, nutrients, vaccines, and veterinary drugs. However, microencapsulation has emerged as a promising solution, providing a protective barrier that improves compound stability, enhances gastrointestinal survival, and enables controlled or site-specific release. By preserving bioactivity and modulating release kinetics, microencapsulation contributes to improved delivery efficiency and functional performance. A distinctive feature of this review is the integration of species-specific gastrointestinal physiology with microencapsulation technologies to provide a rational framework for designing oral delivery systems in veterinary medicine, rather than focusing solely on individual encapsulation technologies. Current challenges related to material selection, formulation optimisation, and industrial scalability are discussed. Overall, this review highlights that integrating gastrointestinal physiology with advanced microencapsulation technologies is essential for developing effective, targeted, and sustainable oral delivery systems, ultimately supporting improved animal health, productivity, and welfare. Full article

Glioblastoma (GB), the most prevalent and aggressive brain tumor, remains one of the most lethal challenges in modern oncology. Standard therapy remains largely ineffective, mainly due to limited therapeutic penetration across the blood–brain barrier (BBB) and adaptive tumor resistance. Consequently, there is an urgent need for innovative strategies to enhance therapeutic precision and efficacy. To address these limitations, we engineered a targeted peptide-based co-delivery system using the WRAP5 cell-penetrating peptide functionalized with a transferrin receptor (TfR)-targeting T7 peptide (sequence: HAIYPRH), enabling simultaneous delivery of temozolomide (TMZ) and a p53-encoding plasmid DNA. The resulting peptide-based TMZ/p53 nanocomplexes exhibited favorable physicochemical properties, enhanced TfR-mediated cellular uptake, and targeted antitumor activity mediated through p53-induced apoptosis in two-dimensional (2D) U87 MG cell cultures. To better reproduce the structural and cellular complexity of the tumor microenvironment, a three-dimensional U87 MG spheroid model was established and optimized using a Design of Experiments (DoE) approach to improve reproducibility and physiological relevance. The developed WRAP5-based nanocomplexes induced a significant dose-dependent inhibition of growth and morphological alterations in the U87 MG spheroid model, accompanied by deep penetration and cell death throughout the spheroid. Collectively, these findings highlight the potential of this targeted and tailored nanosystem to enhance cellular transfection, enable drug/gene co-delivery, restore p53 function, and promote apoptosis, representing a promising therapeutic strategy for GB treatment. Full article

The study of body composition (BC) is of increasing clinical and research relevance, as it provides a detailed characterization of metabolic status beyond traditional anthropometric indices such as body mass index (BMI). BC describes the quantitative ratio of fat mass (FM), lean mass (LM)—including skeletal muscle and internal organs—and bone mineral density (BMD). The pathogenetic mechanisms underlying specific BC phenotypes arise from a complex interplay of endocrine, metabolic, inflammatory, and genetic factors. Given the shared pathways involved in phenotype formation, the study of BC alterations across different diseases holds promise for predicting complications and identifying common therapeutic targets. This review aims to systematize current knowledge on combined BC phenotypes—sarcopenic obesity, osteosarcopenia, osteopenic obesity, and osteosarcopenic obesity—and to explore their pathogenetic unity, with particular emphasis on type 1 diabetes mellitus as a dynamic model of sequential phenotypic shifts. Full article

Plasma biochemistry often presents significant limitations in diagnosing borderline metabolic disorders, particularly within complex neurodevelopmental phenotypes. Here, we present the clinical genomic evaluation of a six-year patient presenting with early-onset hypotonia and severe gastrointestinal complications whose newborn screening panel did not evaluate biotinidase (BTD) activity. While initial baseline plasma biochemistry yielded borderline residual BTD function (46% of the population mean), targeted sequencing identified a novel, compound heterozygous hypomorphic variant (p.Thr459Met) in trans with the common p.Asp424His allele. In vitro functional validation confirmed that p.Thr459Met induces severe protein misfolding and intracellular retention, impairing enzyme secretion. Biotin supplementation triggered a documented and favorable therapeutic improvement, establishing this borderline enzymatic background as an actionable metabolic vulnerability unmasked by chronic gastrointestinal stressors. This study underscores the critical value of functional genomic characterization over static enzymatic biomarkers to identify highly treatable metabolic components within heterogeneous clinical landscapes. Full article

Influenza remains a significant national health security threat, particularly for vulnerable populations, as existing control measures do not fully mitigate its impact. This gap in protection is especially pronounced early in a pandemic when a well-matched vaccine may not yet be available, as well as in populations unable to mount an optimal vaccine response. Clinical studies support pre-exposure prophylaxis (PrEP) therapeutics as a promising complementary strategy to reduce influenza transmission and disease severity, potentially easing the strain on healthcare systems during outbreaks. This manuscript outlines the Biomedical Advanced Research and Development Authority’s (BARDA’s) target product profile (TPP) for a long-acting influenza PrEP product, reviews the current development landscape, and models the potential impact of early PrEP product deployment using agent-based modeling in a synthetic population of 19.5 million people across pandemic scenarios resembling the 1918, 1968, and 2009 influenza pandemics. Simulations showed that early deployment of a 70%–effective PrEP reduced cumulative and peak infections, delayed the epidemic peak, and provided the greatest benefit in less transmissible pandemics; at 40–50% coverage, PrEP fully mitigated a 2009-like pandemic and substantially reduced transmission in 1918- and 1968-like scenarios. The TPP defines key characteristics of an effective PrEP option for seasonal and pandemic influenza that (1) demonstrates a strong safety and tolerability profile across all populations; (2) targets a direct-acting antiviral mechanism of action; (3) reduces the relative risk of symptomatic influenza infection by at least 70% in an unvaccinated population and (4) provides single-dose, season-long protection to optimize patient adherence. In this context, integrating PrEP into influenza prevention strategies could improve control of virus spread, strengthen protection for high-risk groups, and significantly reduce the overall public health impact of seasonal and pandemic influenza. Full article

Background: Regional heterogeneity of the macula and peripheral retinas contributes to the differences in retinal physiology, metabolic demand and susceptibility to macular diseases such as age-related macular degeneration (AMD), diabetic macular oedema (DME) and Macular telangiectasia type 2 (MacTel). Extracellular vesicles (EVs) released from the retinal pigment epithelium (RPE)-choroid are increasingly recognised as mediators of extracellular communication and correlate with the molecular biological states of their tissue of origin. However, regional variation in EV composition in human RPE-Choroid tissues remains poorly characterised. Methods: Human macular and peripheral RPE-choroid explants from non-diseased donor eyes (n = 4) were cultured ex vivo using a transwell system. EV and EV-depleted conditioned media were collected by differential ultracentrifugation, respectively. EVs were characterised by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). Proteomic profiling was performed using LC-MS/MS followed by multivariate, pathway, and differential expression analyses. Results: TEM and NTA confirmed the presence of vesicle-like particles in a 119–140 nm size range, indicating small EVs in both regions. Proteomic analysis demonstrated a region-associated separation between the macular and peripheral samples in both EV proteome and EV-depleted soluble secretome datasets. GO enrichment analysis revealed that macular EV proteomes were enriched for wound healing, cell–substrate adhesion, and focal adhesion-related terms with a high abundance of integrin and annexin family members, whereas peripheral EV proteomes were enriched for retinoid- and vitamin-binding terms. In the EV-depleted soluble secretome, macular-enriched proteins were associated with actin binding and extracellular matrix-related terms, while peripheral-enriched proteins were correlated with RNA localisation and nuclear compartment terms. Comparative analysis identified EV-specific, secretome-specific, and shared extracellular protein pools. Several EV-specific markers and membrane proteins such as annexins and integrins, showed a relatively high enrichment in macular EV proteomes, whereas CD63 is more abundant in peripheral EV proteomes. In contrast, EV-depleted soluble secretome contains cytokine- and ligand-associated proteins, including MIF, SPP1, and IL6, which may suggest that EVs and soluble secreted proteins represent partially distinct extracellular signalling compartments. Conclusions: Human macular and peripheral RPE-choroid explants released secretory proteins in a regional differentiated manner, supported by PLS-DA and GO enrichment analyses of both EV proteomes and EV-depleted soluble secretome. Further analysis of the EV proteomes may show that such differences were also able to be reflected in protein categories such as EV-specific markers, mitochondrial and membrane proteins. These findings provide a foundation for future investigations into the role of EV-mediated communication in macular diseases and may support the development of region-specific extracellular biomarkers and therapeutic targets. Full article

Alzheimer’s disease (AD) is characterized by neurite degeneration and neuronal death. Extracellular amyloid-β 1-42 (Aβ42) oligomers (EAO) not only disrupt the homeostasis and function of the extracellular matrix (ECM) but also damage neural cells through direct binding. Previous studies have demonstrated that EAO binding to membrane integrins reduces neuronal motility, adhesion, and neuritogenesis. To identify the key molecular switch(es) responsible for these actin cytoskeleton dysfunction-associated events, this study utilized neuronal and glial cell lines as well as AD model mice to investigate the cascade underlying EAO-induced actin cytoskeleton dysfunction. This study revealed that EAO induce the dual activation of ROCK2 through RhoA and granzyme B (GzmB) mediation, with GzmB-mediated ROCK2 activation constituting a significant component of this process. ROCK2 hyperactivation in response to EAO causes dynamic dysregulation of the actin cytoskeleton, defective neuritogenesis, and ultimately reduced cell survival, leading to disturbances in brain cell populations. However, the excessive inhibition of ROCK2 activity might cause excessive neurite outgrowth, which may disrupt intrinsic neuronal networks or normal neural transmission. Thus, the disruption of ROCK2 activity might lead to impaired neuritogenesis and disturbances in brain cell populations. The findings of this study may provide important insights into AD pathogenesis and feasible therapeutic targets. Full article

The growing interest in natural bioactive compounds has positioned sea buckthorn (Hippophae rhamnoides L.) as a source of high-value phytochemicals for pharmaceutical and functional applications. Renowned for its complex biochemical profile, this species synthesizes significant levels of lipophilic and hydrophilic antioxidants, notably carotenoids and ascorbic acid (vitamin C). These constituents provide extensive therapeutic and pharmacological benefits, including strong antioxidant, anti-inflammatory, and tissue-regenerative properties. Consequently, characterizing the exact bioactive composition of sea-buckthorn is essential for identifying cultivars with exceptional multi-functional potential. However, a major challenge in exploiting this botanical resource lies in the significant variability of its chemical profile. While the fundamental pharmacological attributes of the plant are genetically determined, the absolute concentration of its bioactive compounds can vary. Factors such as specific cultivar traits and inter-annual climatic conditions, including severe seasonal droughts, are known to influence secondary metabolite accumulation. Although individual varieties exhibit distinct nutritional profiles, systematic data tracking these fluctuations over consecutive years remain limited, particularly regarding certified cultivars adapted to specific regional ecosystems. To extend current understandings, this study evaluates the multi-functional characteristics and inter-annual stability of 17 distinct sea buckthorn varieties over a continuous three-year monitoring period. Specifically, this research focuses on four select varieties officially registered in the Republic of Moldova Official Catalogue of Plant Varieties and Species. The primary aim of this investigation is twofold: first, to quantify key quality parameters—including carotenoid levels, vitamin C content, total acidity, pH, and dry matter—across the different cultivars; and second, to determine whether genetic variety or inter-annual climatic variations exert the dominant influence on these bioactive levels over the medium term. By evaluating these parameters, this study establishes reliable baselines for selecting stable, high-yield cultivars optimized for targeted pharmacological applications. Full article

Marrubium vulgare L. aerial parts are a rich source of polyphenols with recognized antioxidant and anti-inflammatory properties; however, their therapeutic potential is limited due to poor stability and bioavailability. To enhance its pharmacological efficacy, a liposomal formulation of M. vulgare polyphenolic extract (MV-Lipos) was developed in this study by employing the thin-film hydration method. Before encapsulating, the free-extract (MV-Ext) was analyzed using LC-MS, and the resultant nanoliposomes were tested for physicochemical qualities, biological activity, and safety. MV-Lipos exhibited particle sizes ranging from 127 to 200 nm, an 84% encapsulation efficiency, and high colloidal stability (zeta potential −29.58 ± 0.40 mV). In vitro evaluations revealed anti-inflammatory and antioxidant activities without cytotoxic effects. In vivo, MV-Lipos significantly improved analgesic and anti-inflammatory responses. Specifically, a dose of 100 mg/kg lowered acetic acid induced writhing by up to 75.9% and carrageenan induced paw edema by 72%, with efficacy comparable to ibuprofen. A 28-day subacute toxicity assessment found no treatment-related adverse effects. Furthermore, molecular docking analyses validated the experimental results by revealing possible interactions with inflammation-related targets. Overall, liposomal encapsulation improved the biological efficacy and safety profile of M. vulgare polyphenols, highlighting their potential as natural agents for the management of pain and inflammatory conditions. Full article

Background/Objectives: The NLRP3 inflammasome–Caspase-1–IL-1β pyroptotic axis participates in peripheral inflammatory responses, yet its function in peripheral–central neuroimmune crosstalk underlying endometriosis (EM)-associated pain remains unclear. This study aimed to clarify whether NLRP3-mediated pyroptosis establishes a brain–lesion neuroimmune axis connecting ectopic lesion inflammation with central neuroimmune remodeling and to explore the therapeutic mechanism of transcranial direct current stimulation (tDCS). Methods: An EM rat model was established to detect NLRP3 pathway expression in ectopic lesions and anterior cingulate cortex (ACC), together with central nervous system pathological alterations. Animals received tDCS intervention to evaluate inflammatory, neuropathological and pain behavioral changes. The closed-loop brain–lesion regulatory circuit was further interpreted. In a clinical cohort including 40 EM patients, pain and quality-of-life scores were compared between active and sham tDCS groups. Results: NLRP3, Caspase-1 and IL-1β were upregulated in ectopic lesions and ACC of EM rats, accompanied by ACC mitochondrial injury, microglial activation and thalamic demyelination. tDCS inhibited pyroptosis-related molecules, decreased systemic proinflammatory cytokines, improved central pathological lesions and relieved pain hypersensitivity. Mechanically, top-down descending pain inhibitory pathways, vagal cholinergic anti-inflammatory pathway and the HPA axis jointly mediate therapeutic effects, whereas circulating cytokines and visceral afferents transmit peripheral inflammatory signals to the brain. Clinical data demonstrated that active tDCS effectively alleviated EM-related pain and improved patients’ quality of life. Conclusions: NLRP3-mediated pyroptosis acts as a key mediator linking peripheral and central neuroimmune communication. Targeting this pathway via tDCS interrupts the inflammation–pain vicious cycle through multiple neuroregulatory pathways and remodels the central neuroimmune microenvironment in endometriosis. Full article

Cancer is the second leading cause of death worldwide, and tumor heterogeneity remains a major obstacle to effective therapy. In vitro reconstruction of the tumor microenvironment (TME) is particularly challenging because of its complexity, dynamic nature, and spatial heterogeneity, which limits the predictive value of conventional models for anticancer drug evaluation. Traditional Chinese medicine (TCM) has attracted increasing attention in cancer therapy owing to its multi-component, multi-target, and multi-pathway therapeutic characteristics. However, the complexity of TCM formulations and the diversity of their bioactive constituents make their pharmacological mechanisms difficult to elucidate using conventional experimental models. Microfluidic tumor microenvironment-on-a-chip (TME-on-a-chip) platforms integrate engineered cell culture with dynamic perfusion systems to recapitulate key structural, biochemical, and cellular features of the TME, thus providing a more physiologically relevant platform for anticancer research. With advantages such as low sample consumption, precise spatiotemporal control, multicellular co-culture, and real-time monitoring, these platforms provide a promising strategy for evaluating the efficacy and microenvironment-dependent effects of TCM-derived compounds and formulations. In this review, we summarize recent advances in the construction of TME-on-a-chip models, including multicellular organization, extracellular matrix simulation, vascularization, and immune microenvironment reconstruction, and discuss how these systems can be applied to key questions in TCM-based anticancer research. We further analyze current technical and methodological challenges that limit their broader adoption in TCM research and highlight future directions for promoting mechanism-driven and precision-oriented development of TCM in cancer therapy. Full article

Colorectal cancer (CRC) is a major cause of cancer morbidity and mortality worldwide, and metabolic reprogramming is increasingly recognized as an important feature of its progression. Among these changes, fatty acid metabolism (FAM) has drawn growing attention because it supports energy supply, membrane synthesis, redox balance, and stress adaptation in tumor cells. CRC cells can increase fatty acid uptake, activate de novo synthesis, adjust fatty acid oxidation (FAO), and alter lipid droplet (LD) dynamics according to metabolic demand. These processes are strongly influenced by the hypoxic tumor microenvironment. Under hypoxic conditions, signaling pathways centered on hypoxia-inducible factors (HIFs) reshape lipid uptake, synthesis, oxidation, and storage, allowing CRC cells to maintain survival and adapt to limited oxygen and nutrient availability. Increasing evidence suggests that this metabolic shift is closely linked to invasion, metastasis, stem-like behavior, and resistance to therapy. In this review, we provide an integrated overview of the hypoxia–FAM axis in CRC. We first summarize the major steps of FAM reprogramming, then highlight how hypoxia reshapes these processes through HIF-dependent and related pathways. We also discuss FAM crosstalk with stromal and immune cells, experimental models, and metabolic heterogeneity between primary CRC and liver metastases. Finally, we discuss therapeutic strategies targeting FAM and hypoxia-associated signaling in CRC. Full article

Background: Miscarriage is defined as the spontaneous loss of a pregnancy before fetal viability and includes all pregnancy losses occurring from conception until 24 weeks of gestation. Recurrent pregnancy loss (RPL) is traditionally defined as three or more consecutive miscarriages. Emerging evidence suggests that both miR-99a and miR-143 may serve as promising biomarkers for recurrent pregnancy loss. Methods: A total of 42 women of reproductive age (18–45 years) were enrolled in this study. The control group consisted of 13 women with no history of recurrent miscarriage, whereas the study group included 29 women diagnosed with spontaneous miscarriages/recurrent implantation failure. Results: Both miR-99a and miR-143 showed significantly lower relative expression in women with recurrent miscarriage/recurrent implantation failure compared with the control group. Conclusions: Despite the relatively small sample size, statistically significant differences in the expression of miR-99a and miR-143 were observed between the case and control groups, with lower relative expression of both miRNAs in the case group. These findings suggest that both microRNAs may be associated with molecular pathways involved in reproductive failure. Furthermore, investigating the expression patterns of miR-99a and miR-143 may contribute to a better understanding of the molecular mechanisms underlying recurrent miscarriage/recurrent implantation failure. Such insights may facilitate the development of genetic risk profiles for women with unexplained pregnancy loss and implantation failure, as well as support the identification of potential therapeutic targets. Further studies are required to determine their potential value as molecular markers and to clarify their functional relevance in reproductive failure. Full article