Search Results (54,547)

Chronic exposure to even low levels of pesticides is a serious public health issue, mainly due to the role of oxidative stress in damaging the liver and promoting cancer. This has driven interest in finding natural, plant-based antioxidants that can counteract this kind of chemical injury. In this study, we tested whether a methanol extract from the leaves of Pteropyrum scoparium (PSE) could protect the liver against oxidative harm caused by four common pesticides: acetochlor, deltamethrin, thiamethoxam, and rotenone. Chemical analysis showed that the extract contains high levels of phenolics (345.1 ± 7.6 mg GAE/g) and flavonoids (17.3 ± 1.3 mg CAE/g). GC–MS profiling revealed a diverse set of compounds, including fat-soluble antioxidants like squalene, α-tocopherol, and γ-sitosterol, and water-soluble phenolics like pyrogallol and catechol, suggesting PSE is equipped with a multi-layered antioxidant defence. In the animal experiment, rats were given each pesticide for 30 days, with or without PSE. All four pesticides caused clear oxidative stress in the liver: glutathione (GSH), total antioxidant capacity (TAC), antioxidant enzymes activities dropped, while markers of lipid damage (MDA) and free radical activity (DPPH) rose. Co-administration of PSE significantly restored GSH, TAC and antioxidant enzymes levels and reduced MDA and residual DPPH values compared to pesticide-only groups; these parameters were statistically comparable to the controls (p > 0.05), indicating a substantial recovery of hepatic redox balance. Histopathological examination of liver tissues confirmed these findings, as pesticide treatment caused visible liver injury; deltamethrin and thiamethoxam led to congestion in central veins, while rotenone and acetochlor triggered clusters of inflammatory Kupffer cells. In animals that also received PSE, liver structure remained largely normal, with much less congestion and inflammation. These results show that the combination of antioxidant constituents in PSE might contribute to hepatoprotection through redox modulation and preservation of endogenous antioxidant balance, as suggested by the observed biochemical and histological improvements. Full article

Seed coating technology is regarded as one of the optimal strategies to promote sustainable agricultural development. It can effectively optimize the physical and physiological characteristics of seeds, improve germplasm quality, and enhance crop resistance to abiotic and biotic stresses. Saline–alkali soils, characterized by high salinity and alkalinity, severely restrict plant growth and development. However, alfalfa, a high-quality leguminous forage, faces substantial challenges in large-scale popularization and cultivation in saline–alkali regions. At present, research on the application of microbial seed coating technology in alfalfa production under saline–alkali conditions remains insufficient, and relevant techniques and formulations still require optimization. Under field conditions, this study used a randomized complete block design with alfalfa as the research material. Different coating treatments combining plant growth-promoting rhizobacteria (PGPR), rhizobia, and extracellular polysaccharides (EPSs) were established to systematically investigate the effects of various coating formulations on alfalfa yield, nutritional quality, root system architecture, and rhizosphere soil properties. Meanwhile, high-throughput sequencing was employed to analyze shifts in rhizosphere soil microbial community structure. The results demonstrated that all microbial coating treatments exerted significant growth-promoting effects on alfalfa grown in saline–alkali soils, among which the T8 treatment (combined coating of rhizobia + PGPR + EPS) performed the best. This treatment not only significantly improved alfalfa yield and nutritional quality but also modified root system architecture and enhanced soil enzyme activities, soil nutrient contents, and soil physical structure, thereby creating a favorable growth environment for plants. Among the single microbial coating treatments, the combined coating of rhizobia and EPS outperformed other single treatments and exhibited favorable application potential. Sequencing results revealed that microbial seed coating treatments significantly increased the relative abundance of beneficial soil bacteria, decreased the abundance of harmful fungi, regulated rhizosphere microbial community structure, and consequently promoted improvements in alfalfa yield and quality by optimizing the plant growth microenvironment. The findings of this study provide important theoretical support for the popularization and application of microbial seed coating technology in crop cultivation in saline–alkali soils, offer a key reference for optimizing alfalfa-specific seed coating formulations for saline–alkali conditions, and are of great significance for promoting the efficient utilization of saline–alkali land resources and the development of ecological agriculture. Full article

Hypoxia-dependent microRNAs play an important role in orchestrating a plant’s response to low-oxygen stress. To assess the regulatory mechanisms of the adaptive response of maize (Zea mays L.) to hypoxia, an antisense sequence was developed, and the short tandem target mimic (STTM) system was used to induce the loss of function of the mature microRNA775A (miR775a) in maize. A recombinant binary vector pBI121 cloned in E. coli cells containing the antisense sequence anti-miR775A to maize miR775A was acquired to create a line of modified A. tumefaciens EHA105. Using the puncturing method on soaked seeds, maize plants with an active anti-miR775A construct were obtained, as evidenced by a decrease of more than 10-fold in mature miR775A content and by developmental changes in the seedlings. The size of seedlings of the maize knockdown line was almost twice smaller than that of the wild-type (WT) plants. An assessment of the effects of hypoxic conditions induced by flooding of 14-day-old maize plants revealed differences in the expression and activity of several enzymes between WT and knockdown plants. The reduced miR775A levels led to a 2.1-fold drop in pyruvate levels, which resulted in decreased pyruvate kinase, pyruvate dehydrogenase, and lactate dehydrogenase activities as compared to WT plants. A decrease in miR775A content in the maize knockdown cell line also affected the function of mitochondrial and extramitochondrial isoenzymes of citrate synthase, aconitase, and fumarase under hypoxic conditions. Full article

Snakebite envenoming remains a major global health challenge. Naja atra (N. atra) envenomation induces severe acute kidney injury (AKI), largely driven by snake venom phospholipase A₂ (SVPLA₂). Increasing evidence suggests that immune dysregulation, in addition to direct cytotoxicity, contributes to delayed renal injury. Here, we investigated whether N. atra SVPLA₂ exposure is associated with macrophage immunometabolic remodeling and functional changes relevant to AKI progression. In vivo, AKI was induced in C57BL/6J mice by intraperitoneal administration of N. atra venom, followed by treatment with the SVPLA₂ inhibitor varespladib. In vitro, bone marrow–derived macrophages were exposed to venom with or without varespladib. N. atra venom exposure was associated with extensive tubular apoptosis, increased renal macrophage abundance, and elevated kidney injury biomarkers. Macrophages exhibited a shift toward a pro-inflammatory polarization signature accompanied by reduced efferocytic capacity. Targeted metabolomics revealed coordinated increases in glycolytic intermediates together with upregulation of key glycolytic enzymes. Pharmacological inhibition of SVPLA₂ partially restored macrophage metabolic features and efferocytic capacity and was accompanied by attenuation of renal injury. Together, these findings support a model in which SVPLA₂ exposure is associated with macrophage immunometabolic remodeling and impaired apoptotic cell clearance during venom-induced AKI. Full article

Microbial cutinases are promising biocatalysts for polymer recycling. Here, we investigated the structural basis of catalytic activation in a thermophilic cutinase from Chaetomium thermophilum (CtCut). Differential scanning calorimetry revealed a three-state thermal unfolding pathway (Tm = 66.4 °C and 69.5 °C), indicating hierarchical stability. To capture distinct conformational states while avoiding affinity-tag artifacts, we employed both tag-free and tagged constructs. We determined apo-structures of wild-type and S136A mutant CtCut at 1.7 Å resolution and a complementary inhibitor complex at 2.65 Å. In the apo-state, a chloride ion coordinated the electrostatically pre-organized active site, while the catalytic H204 adopted a solvent-exposed, inactive loop conformation. In the inhibitor complex, p-nitrophenol displaced the chloride, establishing a characteristic oxyanion hole network. Concomitantly, the “lid” loop transitioned to an open state, with H204 exhibiting pronounced conformational heterogeneity across eight independent molecules. These complementary structures provide structural evidence for conformational dynamics of the catalytic lid loop, consistent with the conformational cycling model previously proposed for a mesophilic homolog. Full article

To address the need for enhanced geotechnical performance in gravelly soil stabilization, this study investigated the synergistic effects of guar gum as an additive in enzyme-induced calcium carbonate precipitation (EICP) treatment. Through systematic experimentation combining unconfined compressive strength (UCS) tests, carbonate content quantification, and triaxial analysis, the mechanical behavior of treated soils was evaluated under varying EICP solution concentrations (0–2 mol/L) and curing durations. Results demonstrated that a 1.5 mol/L EICP solution achieved peak strength and carbonate precipitation before subsequent decline, while a 1% guar gum dosage optimized mechanical properties by balancing initial strength enhancement and precipitation efficiency. Scanning electron microscopy revealed microstructural mechanisms wherein guar gum provided heterogeneous nucleation sites for calcite crystals, while its interaction with EICP enabled dual-phase pore filling and interparticle bonding. This synergistic effect created a three-dimensionally reinforced matrix, significantly improving both UCS and unconsolidated undrained shear strength compared to native and EICP-only specimens. The findings establish a theoretical framework for regulating calcite precipitation patterns and enhancing cementation mechanisms in gravelly soil improvement, offering practical guidelines for foundation engineering applications through the combined use of guar gum and EICP. Full article

Metabolic dysfunction-associated steatotic liver disease (MASLD, introduced in 2023), formerly termed non-alcoholic fatty liver disease (NAFLD), is a leading cause of chronic liver disease worldwide and is closely linked to obesity, insulin resistance, and cardiometabolic dysfunction. Exercise is widely recommended as a cornerstone of MASLD management; however, the magnitude of its hepatic and metabolic benefits and the underlying molecular mechanisms remain incompletely defined. We aim to synthesize evidence from randomized controlled trials assessing how structured exercise interventions influence hepatic steatosis and metabolic dysfunction in adults with MASLD. A targeted search of PubMed from database inception to February 2025 identified eligible trials, of which eleven were included in the qualitative synthesis. Across studies, aerobic and resistance exercise interventions were consistently associated with reductions in hepatic fat content, improvements in plasma lipid profiles and liver enzyme concentrations, and enhanced indices of insulin sensitivity, frequently occurring independently of substantial weight loss. Mechanistically, exercise-induced activation of pathways related to mitochondrial function, lipid oxidation, inflammation modulation, and insulin signaling likely contributes to these benefits. Despite heterogeneity in intervention design, duration, and outcome assessment, the collective evidence supports structured exercise as an effective non-pharmacological strategy for improving hepatic steatosis and metabolic dysfunction in MASLD. Future studies integrating molecular biomarkers with clinical endpoints are warranted to refine exercise prescriptions and elucidate mechanisms of therapeutic response. Full article

The Black Sea waters and sediments accumulate a wide range of pollutants, which, together with natural factors, can have diverse and combined effects on marine organisms. Cellular oxidative stress (OS) is a common response to environmental stressors and thus an indicator of the physiological resilience of organisms. The present study analyzed OS in fish species from the Bulgarian part of the Black Sea. Samples of 18 fish species were obtained during monitoring trawling in the northern and southern regions of the Bulgarian Black Sea and were analyzed. The OS-specific biomarkers (lipid peroxidation, glutathione, and antioxidant enzymes) were spectrophotometrically measured in the liver and gills. The obtained results revealed pronounced variability in pro- and antioxidant responses, shaped by sampling location, species identity, and tissue type. In fish sampled from the northern part, the liver showed stronger antioxidant activation, while the gills exhibited higher lipid peroxidation. Benthic fish exhibited pronounced hepatic antioxidant responses. Pelagic species had higher gill lipid peroxidation. Generalist feeders displayed variable stress responses associated with a diverse diet and the presence of multiple contaminants in it. In conclusion, OS was most pronounced in benthopelagic and pelagic species, which were more stressed in the northern Bulgarian part of the Black Sea, while benthic or coastal fish species experienced greater environmental pressure in the southern part. Full article

Large-scale expansion of human mesenchymal stem cells (hMSCs) remains a major challenge due to the intrinsic trade-off between cell proliferation and the maintenance of multipotency in conventional culture systems. Stiff substrates, such as tissue culture polystyrene or rigid hydrogels, promote rapid proliferation but induce progressive loss of stemness, whereas very soft matrices preserve multipotency at the expense of cell growth. To overcome this limitation, we developed mechanically soft, phase-separated gelatin–phenol/hyaluronic acid–phenol (Gtn-Ph/HA-Ph) hydrogels with precisely controlled microstructures via enzyme-mediated crosslinking. These hydrogels consist of HA-rich, dot-like domains embedded within a continuous Gtn-rich network, allowing for independent tuning of stiffness and domain architecture. On single-component Gtn-Ph hydrogels, hMSC proliferation increased with substrate stiffness, whereas soft hydrogels with a storage modulus (G′) of approximately 0.6 kPa markedly suppressed proliferation while preserving stemness marker expression, confirming the stiffness-dependent trade-off. In contrast, phase-separated Gtn-Ph/HA-Ph hydrogels supported robust hMSC proliferation even under soft mechanical conditions while maintaining high expression of stemness-associated markers. During long-term culture, hMSCs achieved a 68- to 195-fold increase in cumulative cell yield on soft Gtn-Ph/HA-Ph hydrogels (G′ = 0.5 kPa) compared with tissue culture polystyrene. Expression of α-smooth muscle actin (α-SMA) mRNA, encoded by the ACTA2 gene and associated with cellular senescence and fibrotic activation, was completely suppressed, while hMSCs retained robust adipogenic, osteogenic, and chondrogenic differentiation capacities. These results demonstrate that phase-separated Gtn-Ph/HA-Ph hydrogels effectively resolve the proliferation–multipotency dilemma in hMSC expansion and provide a promising platform for scalable manufacturing of therapeutic stem cells. Full article

The effects of static magnetic field (SMF) treatment on the solid-state culture of Sanghuangporus vaninii (SV) were investigated to enhance metabolite production and bioactivity. SMF parameters including intensity, exposure duration, and temperature were optimized, and treatment at 4 mT for 2 h per day produced the most pronounced effects, increasing total flavonoid (TFC), polyphenol (TPC), and triterpenoid (TTC) contents by 61–438% compared with the control. Ultrasonic extraction and semi-preparative chromatography enabled the isolation of three key compounds: D-(+)-trehalose (1), 5,7-dihydroxy-3,4′-dimethoxyflavone (2), and pinolenic acid (3), all of which were elevated following SMF treatment. Importantly, SMF exposure was associated with enhanced inhibitory activities against enzymes relevant to chronic metabolic disorders. The overall inhibitory activities against α-amylase, α-glucosidase, pancreatic lipase, and xanthine oxidase increased by 6–28% compared with the control, reaching a maximum inhibition of 97.60 ± 0.17%. Preliminary in vitro screening at 100 μg/mL showed that compounds 1 and 2 inhibited both α-amylase and α-glucosidase, whereas compound 3 selectively inhibited pancreatic lipase. Subsequent IC₅₀ analysis confirmed that compound 2 under SMF treatment exhibited inhibitory activity comparable to acarbose against α-amylase (45.62 μg/mL vs. 52.18 μg/mL) and α-glucosidase (38.74 μg/mL vs. 35.42 μg/mL). In addition, compound 3 showed moderate inhibition of pancreatic lipase with an IC₅₀ value of 42.15 μg/mL. These findings suggest that SMF treatment may enhance metabolite production and in vitro enzyme inhibitory activity in S. vaninii. However, these results are limited to in vitro assays, and further studies including cellular and in vivo validation, toxicity assessment, and pharmacokinetic evaluation, are required before any therapeutic or industrial applications can be considered. Full article

Plant growth regulation is orchestrated by complex hormonal networks involving gibberellin and auxin signaling pathways. In this study, a comprehensive in silico approach was employed to comparatively evaluate the plant growth regulators (PGRs) forchlorfenuron (CPPU) and strigol (STG) against two key proteins from Arabidopsis thaliana: Gibberellin 3-beta-dioxygenase 2 (GA3Ox2), a rate-limiting enzyme in the biosynthesis of bioactive gibberellins, and the auxin signaling repressor IAA7. These targets were specifically selected because they represent critical regulatory nodes in two major hormonal pathways controlling plant growth: GA3Ox2 governs the final steps of gibberellin activation, while IAA7 modulates auxin-responsive gene expression through its interaction with Auxin Response Factors. Therefore, their combined analysis enables the evaluation of potential regulatory effects of PGRs on both gibberellin biosynthesis and auxin-mediated transcriptional control. Molecular docking analyses revealed that both ligands exhibited higher binding affinity toward GA3Ox2 than IAA7, with STG showing slightly more favorable binding energies (−7.91 kcal/mol for GA3Ox2 and −5.43 kcal/mol for IAA7) compared to CPPU (−7.18 and −4.79 kcal/mol, respectively). These results suggest a structural preference of both PGRs toward the gibberellin biosynthetic pathway. To further assess complex stability under near-physiological conditions, 100 ns molecular dynamics (MD) simulations were conducted using the CHARMM36m force field. Despite its slightly lower docking scores, CPPU demonstrated greater conformational stability, lower RMSD fluctuations, and more persistent hydrogen bonding patterns, particularly in complexes with IAA7. In contrast, STG induced more pronounced conformational rearrangements, although it promoted slightly more compact protein conformations in certain systems. Fourier-transform infrared (FTIR) spectroscopy supported the computational findings by confirming the presence of key functional groups responsible for hydrogen bonding and hydrophobic interactions. Collectively, the results indicate that although STG exhibits higher initial binding affinity, CPPU forms more dynamically stable complexes with both proteins. These findings suggest that CPPU may represent a more robust candidate for sustained modulation of auxin and gibberellin signaling pathways in plant growth regulation. Full article

Mechanical pretreatment techniques are essential for overcoming lignocellulosic biomass recalcitrance in emerging biorefineries. This review critically synthesizes advances from 2020 to 2025 across fundamental mechanisms, hybrid technologies, energy efficiency, Life Cycle Assessment, and industrial scalability. The analysis reveals that effective pretreatment targets supramolecular modification—defect generation in cellulose crystallites and the creation of reactive sites—beyond simple particle size reduction. Impact–shear regimes prove most effective for fibrous materials. Hybrid approaches are examined: mechanocatalysis enables solvent-free depolymerization, while mechanoenzymatic technologies achieve hydrolysis without bulk water, though enzyme denaturation under mechanical stress remains unresolved. Energy consumption is the primary upscaling barrier, with Life Cycle Assessment identifying electricity use as the dominant environmental hotspot and emphasizing burden per unit of final product as the critical metric. Technology Readiness Level assessment provides a strategic framework: continuous extruders and mills are industrially mature for bulk applications, while high-intensity batch devices are suited for high-value coproducts. A research agenda prioritizing mechanistic understanding, hybrid process engineering, feedstock diversification, and embedded sustainability assessment is proposed. Full article

In situ cancer vaccines activate antitumor immune responses by locally capturing and presenting tumor-derived antigens, in which polymers play a key role as antigen-capturing materials. However, the influence of polymer composition and degree of polymerization (DP) on antigen capture efficiency and protection mechanisms remains insufficiently understood. In this study, the tumor-specific antigen MAGE-A3, highly expressed in esophageal squamous cell carcinoma (ESCC), was employed to investigate antigen capture and stabilization by five representative polymers—chitosan, polyethyleneimine (PEI), alginate, polycaprolactone (PCL), and poly (lactic-co-glycolic acid) (PLGA)—with different DPs, using molecular dynamics simulations and in vitro experiments. All-atom simulations revealed that hydrophobic interactions dominate polymer–antigen binding, while electrostatic interactions from cationic polymers synergistically enhance binding affinity and capture efficiency. Binding free energy analysis showed that van der Waals and electrostatic contributions stabilize the complexes, whereas polar solvation partially counteracts these effects. Experimentally, low-DP chitosan exhibited the highest antigen-capture efficiency (38.9%), attributed to its small molecular size, enabling multipoint binding across the antigen surface. In contrast, high-DP polymers generated pronounced steric hindrance that suppressed antigen–enzyme interactions and inhibited hydrolysis. These findings clarify how polymer composition and chain length jointly regulate antigen capture and protection, providing mechanistic guidance for the rational design of polymer-based in situ cancer vaccines. Full article

The prolonged low temperature in cold regions significantly inhibits the initiation of straw composting and lignocellulose degradation, thereby restricting straw resource utilization. In this study, 24 cellulose-degrading strains capable of stable growth under low-temperature conditions were screened. Based on multiple indicators, including carboxymethyl cellulase (CMCase) activity, strain LDT1 was identified as the best-performing isolate under low-temperature conditions and as Paenarthrobacter nitroguajacolicus. Subsequently, an efficient mutant strain, LDT1-8, was obtained through atmospheric and room-temperature plasma mutagenesis. The CMCase activity of LDT1-8 at 10 °C increased to 74.25 U/mL, representing a 21.72% increase compared to the wild-type strain. In a straw degradation system at 10 °C, LDT1-8 significantly accelerated early-stage degradation kinetics, with straw degradation rates at 3 and 6 d being 72.72% and 38.15% higher than those of the wild-type strain, respectively. Multi-enzyme profiling further indicated enhanced activities of multiple lignocellulose-degrading enzymes at low temperatures, accompanied by a partial shift in the optimal temperature of some enzymes (e.g., laccase) toward lower temperatures. Whole-genome sequencing revealed increased gene numbers related to energy, amino acid, and lipid metabolism in LDT1-8. Comparative genomic analysis suggested that mutations were mainly enriched in regulatory regions, accompanied by local structural variations. Transcriptional analyses further verified the coordinated upregulation of genes involved in cellulose and hemicellulose degradation, cold adaptation, and transcriptional and protein homeostasis processes in LDT1-8. Overall, this study provides an efficient microbial resource and a mechanistic basis for straw bioconversion in cold regions. Full article

Background: Fecal calprotectin (FC) is a well-established, non-invasive biomarker of intestinal inflammation and is widely used to differentiate inflammatory bowel disease (IBD) from functional gastrointestinal disorders. Although enzyme-linked immunosorbent assays (ELISA) remain the reference method, rapid immunochromatographic tests (ICTs) offer important operational advantages for point-of-care (POC) diagnostics. However, variability in analytical performance among available ICTs remains a concern. Objective: This study aimed to evaluate the diagnostic accuracy of the CerTest Calprotectin one-step card (CerTest Biotec S.L., Zaragoza, Spain) in comparison with the Actim^® Calprotectin lateral flow assay and the reference Calprest^® ELISA (Eurospital Diagnostics, Italy). Methods: A total of 128 fresh stool samples from patients clinically suspected of IBD were analyzed in parallel using all three assays. For the reference ELISA (Calprest^®), a cutoff value of >40 µg/g was applied according to the manufacturer’s instructions. For discrepant results between assays, a cutoff of 200 ng/mL (equivalent to 200 µg hCp/g stool) was employed for ELISA Calprest^® to resolve inconsistencies. The results of the lateral flow assays (CerTest^® Calprotectin ICT and Actim^® Calprotectin) were interpreted using their respective manufacturer-recommended thresholds. Diagnostic sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated using ELISA as the reference standard. Agreement between methods was assessed using Cohen’s κ coefficient. Results: Using ELISA, 47 of 128 samples (36.7%) exceeded the 40 µg/g cutoff. Compared with the Actim^® assay, the CerTest card demonstrated a sensitivity of 88.0% (95% CI: 75.7–95.5), a specificity of 100.0% (95% CI: 95.4–100), and a strong agreement (κ = 0.90). When compared with ELISA, the CerTest assay showed a sensitivity of 87.2% (95% CI: 74.3–95.2), a specificity of 96.3% (95% CI: 89.6–99.2), a PPV of 93.2%, an NPV of 93.2%, and a strong agreement (κ = 0.85). Conclusions: The CerTest Calprotectin one-step card provides a rapid and reliable detection of fecal calprotectin, demonstrating a high sensitivity and specificity that are comparable to both other lateral flow assays and the ELISA reference method. These findings support the use of rapid immunochromatographic testing as a valuable tool for preliminary screening and clinical decision-making in patients suspected of IBD, while acknowledging that histology remains the gold standard for definitive diagnosis. Full article