Search Results (168)

Mycotoxin contamination in agricultural products poses a serious challenge to food safety, severely threatening human and animal health and causing significant economic losses. This study aimed to investigate the degradation and detoxification capabilities of Trichoderma reesei GG-T40 against two representative mycotoxins—aflatoxin B₁ (AFB₁) and zearalenone (ZEN). The results showed that the degradation rates of AFB₁ and ZEN by this strain reached 98.6% and 88.4%, respectively. Using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF MS), the degradation products were systematically characterized, leading to the identification of six AFB₁ degradation products (C₁₇H₁₄O₇, AFD₁: C₁₆H₁₄O₅, C₁₁H₁₀O₄, C₁₄H₁₆O₄, C₁₅H₁₀O₄, and C₁₇H₁₄O₅) and two ZEN degradation products (α-ZOL and β-ZOL). Toxicity evaluation revealed that the key toxic structures of AFB₁ were disrupted, significantly reducing or even eliminating the toxicity of its degradation products; ZEN was mainly converted into β-ZOL (accounting for 91.5%), which has lower estrogenic activity. Further toxicological experiments in mice confirmed that the degradation products were non-toxic and non-pathogenic under actual testing conditions, demonstrating systematic verification of their safety. In conclusion, T. reesei GG-T40 can efficiently and safely degrade AFB₁ and ZEN, showing great potential for developing green control technologies for mycotoxin contamination in food and feed raw materials, with important application value for ensuring food safety. Full article

Cerebral ischemia–reperfusion injury triggers mitochondrial dysfunction and oxidative stress, exacerbating neuronal apoptosis. Emerging evidence highlights hydrogen sulfide (H₂S) as a gasotransmitter modulating redox balance, autophagy, and apoptosis. This study investigates the neuroprotective mechanisms of Enriched Environment (EE) against ischemic injury, focusing on mitochondrial dynamics and H₂S-mediated pathways. Using MCAO mice and OGD/R-treated SH-SY5Y neurons, interventions targeting H₂S synthesis, hypoxia-inducible factor 1-alpha (HIF-1α), and mitophagy were implemented. Behavioral, histological, and molecular analyses demonstrated EE significantly improved neurological outcomes, suppressed apoptosis, and attenuated oxidative damage (reduced MDA, elevated MnSOD/glutathione). Mechanistically, EE enhanced mitophagy via dual pathways: canonical PINK1/parkin-mediated mitochondrial clearance, corroborated by transmission electron microscope and LC3B/parkin colocalization, and non-canonical HIF-1α/BNIP3L axis activation. Transcriptomic and Co-immunoprecipitation (Co-IP) data revealed EE upregulated endogenous H₂S biosynthesis post-injury by promoting dopamine-induced calcium influx, which activated calmodulin-dependent signaling to stimulate cystathionine β-synthase/γ-lyase expression. Pharmacological blockade of H₂S synthesis or HIF-1α abolished mitochondrial protection, confirming H₂S as a central mediator. Notably, H₂S exerted antiapoptotic effects by restoring mitochondrial integrity through synergistic mitophagy activation and oxidative stress mitigation. These findings propose a novel neuroprotective cascade: EE-induced dopaminergic signaling potentiates H₂S production, which coordinates PINK1/parkin and HIF-1α/BNIP3L pathways to eliminate dysfunctional mitochondria, thereby preserving neuronal homeostasis. This study elucidates therapeutic potential of EE via H₂S-driven mitochondrial quality control, offering insights for ischemic brain injury intervention. Full article

Anaerobic digestion of sewage sludge generates large volumes of liquid digestate, which is often returned to wastewater treatment plants (WWTPs) due to the presence of pathogens and pollutants, limiting its safe reuse in agriculture. This study evaluated plasma-based post-treatment as a method to improve the sanitary quality of digestate. The liquid phase from mesophilic digesters at WWTP “Ravda” was treated for 5 min using two plasma sources, the β-device and the Surfaguide WR340 (SAIREM, Décines-Charpieu, France). Disinfection effectiveness was assessed for aerobic and anaerobic heterotrophs, fecal and total coliforms, Escherichia coli, Salmonella sp., and Clostridium sp. Physicochemical parameters measured included pH, COD, NH₄⁺, NO₂⁻, NO₃⁻, and PO₄³⁻. The β-device achieved partial disinfection, with reductions ranging from 16.3% to 89.8% for different microbial groups, whereas coliforms persisted and Clostridium sp. reappeared. The Surfaguide produced near-complete disinfection, eliminating coliforms, E. coli, Salmonella sp., and Clostridium sp., and markedly reduced microbial diversity. Both treatments caused slight pH increases, COD decreases, release of NH₄⁺ and PO₄³⁻, and rises in NO₂⁻ and NO₃⁻. Plasma-based disinfection, particularly with the Surfaguide, effectively improves the sanitary quality of the digestate and modifies its chemical properties, supporting the potential for sustainable digestate valorization and its safe reuse in agriculture. Full article

In agricultural production, Deoxynivalenol (DON) generally exists and contaminates wheat, corn, and other grains, causing intestinal damage and immunotoxicity. Lycopene (LYC), an antioxidant, anti-inflammatory carotenoid, is mainly found in red fruits such as tomatoes and has been investigated for its great medicinal advantages. This study aimed to investigate the protective effect of LYC against DON-induced enterotoxicity. Our findings demonstrated that incubation of IPEC-J2 cells with 0.5 μM DON for 24 h caused intestinal barrier impairment and oxidative stress induction, which subsequently led to increased secretion of pro-inflammatory factors (TNF-α, IL-1β, IL-18, and IL-6) and decreased secretion of the counterregulatory factor (IL-10). Furthermore, DON ultimately induced NLRP3 inflammasome activation through the stimulation of the MAPK/NF-κB pathway. It is worth mentioning that the above changes were reversed after adding 30 μg/mL of LYC to DON-exposed IPEC-J2 cells. In addition, further experiments confirmed that ERK activator (4-Methylbenzylidene camphor, 4-MBC) eliminated the positive effect of LYC on alleviating enterotoxicity induced by DON in IPEC-J2 cells. In addition, further experiments confirmed that 4-MBC eliminated the positive effect of LYC on alleviating enterotoxicity induced by DON. In general, our study certified that ERK is a therapeutic target for LYC protecting DON-induced intestinal barrier dysfunction and NLRP3 inflammasome activation. Full article

Biodegradable magnesium (Mg) alloys are promising materials for temporary orthopaedic implants, combining favourable mechanical properties and superplastic behaviour with in vivo resorption. This enables (i) prolonged implant duration, (ii) fabrication of complex-shaped prostheses via superplastic forming (SPF), (iii) elimination of removal surgery, and (iv) reduced risk of long-term complications. However, rapid corrosion under physiological conditions remains a major limitation, highlighting the need for surface treatments that slow degradation while preserving implant integrity. This study investigates the effects of hydrothermal surface treatment on MgAZ31-SPF alloys, focusing on immunomodulatory responses, antibacterial potential, and degradation behaviour. Hydrothermally treated MgAZ31-SPF (MgAZ31-SPF-HT) extracts released lower Mg²⁺ concentrations (29.2 mg/dL) compared to untreated MgAZ31-SPF (47.5 mg/dL) while maintaining slightly alkaline pH (7–8.7), indicating improved control of early degradation. In vitro assays with human peripheral blood mononuclear cells (hPBMCs) and normal human dermal cells (NHDCs) showed that MgAZ31-SPF-HT extracts maintained higher cell viability over 24–72 h. Gene expression analysis revealed significant downregulation of pro-inflammatory markers CTSE and TNF-α, while protein quantification via ELISA and BioPlex confirmed reduced secretion of TNF-α, TGF-β1, TGF-β2, IL-6, and IL-8, suggesting mitigation of early immune activation. Antibacterial assays demonstrated limited Staphylococcus aureus colonisation on both MgAZ31-SPF and MgAZ31-SPF-HT scaffolds, with CFU counts (~10⁵–10⁶) well below the threshold for mature biofilm formation (~10⁸), and SEM analysis confirmed sparse bacterial distribution without dense EPS-rich layers. Overall, hydrothermal treatment improves Mg alloy biocompatibility by controlling Mg²⁺ release, modulating early immune responses, and limiting bacterial adhesion, highlighting its potential to enhance clinical performance of Mg-based implants. Full article

Background/Objectives: Hip osteoarthritis (HOA) is a progressive joint disease characterized by cartilage loss, subchondral bone changes, and synovial inflammation. While tumor necrosis factor receptor 1 (TNFR1), interleukin-6 (IL-6), and transforming growth factor-beta 1 (TGF-β1) are recognized as key mediators of joint pathology, their compartment-specific expression in the human hip synovium remains insufficiently characterized. Therefore, we aimed to investigate their localization and expression in the intimal and subintimal compartments of synovial tissue in patients with HOA compared to controls (CTRL). Methods: Synovial membrane samples were obtained from 19 patients with primary HOA undergoing total hip arthroplasty and 10 CTRL subjects undergoing arthroplasty for acute femoral neck fracture without HOA. Specimens were processed for hematoxylin and eosin (H&E) and immunofluorescence staining. Expression of TNFR1, IL-6, and TGF-β1 was quantified in the intima and subintima using ImageJ analysis. Group differences were assessed using two-way Analysis of variance (ANOVA) with Tukey’s test when assumptions were met; for heteroscedastic outcomes we applied Brown–Forsythe ANOVA with Dunnett’s T3 multiple comparisons. Results: Histological analysis confirmed synovitis in HOA samples, with intimal hyperplasia and mononuclear infiltration. IL-6 was significantly upregulated in the intima of HOA synovium compared with CTRLs, while subintimal expression remained unchanged. In contrast, TGF-β1 expression was reduced in the HOA intima, eliminating the normal intima–subintima gradient. For TNFR1, the within-HOA contrast (int > sub) was significant, whereas the intimal HOA vs. CTRL comparison showed a non-significant trend. Transcriptomic analysis supported IL-6 upregulation, while TNFR1 and TGF-β1 did not reach statistical significance at the mRNA level in an orthogonal, non-hip (knee-predominant) dataset. Conclusions: These findings demonstrate compartment-specific cytokine dysregulation in HOA, with increased intimal TNFR1 and IL-6 alongside reduced intimal TGF-β1. The synovial lining emerges as a dominant site of inflammatory signaling, underscoring its importance in disease progression. Full article

Background: This study evaluates the impact of fibrinogen enrichment on the structural, mechanical, and bioactive properties of fibrin scaffold derived from balanced protein-concentrate plasma (BPCP), an autologous platelet-rich plasma (PRP) formulation with elevated extraplatelet content. Methods: A novel high-fibrinogen BPCP (HF-BPCP) scaffold was produced by combining BPCP platelet lysate with a concentrated fibrinogen solution at a 1:1 ratio, yielding nearly four-fold physiological fibrinogen levels. Comparative analyses between HF-BPCP and standard BPCP included platelet and fibrinogen quantification, scanning electron microscopy (SEM), rheology, indentation, adhesion testing, coagulation kinetics, retraction assays, biodegradation profiling, and growth factor (GF) release kinetics. Results: HF-BPCP displayed significantly denser fibrin networks with thinner fibers, higher porosity, and markedly faster coagulation times compared to BPCP. Mechanically, HF-BPCP exhibited greater stiffness, higher energy dissipation, and more stable adhesion, while almost eliminating scaffold retraction at 24 h. Despite improved early handling and structural integrity, HF-BPCP degraded more rapidly in vitro under tissue plasminogen activator exposure. GF release analysis showed reduced early peaks of platelet-derived factors (TGF-β1, PDGF-AB, VEGF) but sustained release thereafter, while extraplatelet factors (IGF-1, HGF) exhibited similar profiles between scaffolds. Conclusions: These results indicate that fibrinogen enrichment synergizes with the elevated extraplatelet protein profile of BPCP to enhance scaffold mechanical stability, handling properties, and controlled GF delivery. HF-BPCP combines the adhesive, structural, and bioactive features of fibrin sealants with the regenerative potential of PRP, offering a fully autologous alternative for clinical applications requiring rapid coagulation, high mechanical support, and sustained GF availability. Further preclinical and clinical studies are needed to evaluate therapeutic efficacy in the regenerative medicine field. Full article

In this study, two γ-carbon-substituted dialkylphosphinic acids—symmetrical di-(3,5,5-trimethylhexyl)phosphinic acid (P355) and unsymmetrical (2-ethylhexyl)(3,5,5-trimethylhexyl)phosphinic acid (P227-355)—were synthesized via a precise free radical addition method. Their chemical structures were fully characterized using ESI-HRMS, ¹H NMR, ³¹P NMR, and FT-IR. Their middle REE extraction/separation performance, anti-emulsification behavior, and underlying mechanisms were investigated. Key results showed that P355 had better Dy saturation capacity (357.51 mg/L) and good selectivity for middle REEs (their average value of β_{N + 1/N} = 3.18), while P227-355 showed higher back-extraction efficiency (≈90% Dy stripping at ≥0.02 mol/L H₂SO₄). Methyl n-pentyl ketone (MNPK) eliminated emulsification and boosted saturation capacity (324.18 mg/L Sm and 357.51 mg/L Dy for P355). Mechanistically, the extraction followed cation exchange (Sm³⁺ + 2(HL)₂ ↔ Sm·L₃·(HL) + 3H⁺); MNPK formed hydrogen-bonded associates (HL·MNPK) with free extractants, slightly reducing the effective concentration of (HL)₂ but not altering the core cation exchange mechanism. Full article

Lactic acid bacteria (LAB) play a crucial role in acid-curd cheese production by driving milk protein coagulation and forming metabolites that determine texture, safety, and flavor. This study investigated the effect of enzymatic lactose hydrolysis using β-D-galactosidase (Maxilact LX5000) on the quality of full-fat curd cheeses (16.5% and 20.8% dry matter) produced without whey separation. Cheeses were manufactured with or without prior lactose hydrolysis, inoculated with a mesophilic Flora Danica starter culture, and stored for 28 days at 4 °C. Chemical composition, sugar profile (HPLC), pH, LAB viability, textural properties (hardness, adhesiveness, and water-holding capacity), and sensory attributes were determined. Lactose hydrolysis completely eliminated lactose and increased glucose and galactose concentrations, without significant changes in protein, fat, or pH level. In our data, lactose was undetectable in hydrolyzed samples across storage, glucose/galactose exhibited only minor fluctuations, and LAB counts and pH remained stable, indicating a largely stable sugar profile and limited microbial activity under refrigeration. Hydrolyzed samples showed improved texture, especially higher hardness and moisture retention in low-dry-matter variants, while sensory characteristics were comparable to the control and free from excessive sweetness. These results demonstrate that enzymatic lactose hydrolysis is an effective tool for producing lactose-free curd cheese without compromising quality. This process can be recommended for sustainable whey-free cheese manufacture aimed at lactose-intolerant consumers. Full article

During metastasis, cancer cells detach from the primary tumor, and the floating cells enter the circulation and reattach in distant organs. Floating cells are highly chemoresistant to anticancer drugs, but the underlying mechanisms are poorly understood. We hypothesized that floating cells transition into a quiescent/senescent (Q/S) state. Using human lung carcinoma H460 and H23, human prostate adenocarcinoma PC3, and human breast adenocarcinoma MDA-MB-231 cells, we found (1) a progressive increase in activity of β-galactosidase (β-Gal), a marker associated with Q/S cells, (2) a transition to a non-proliferative state while growing under anchorage-independent conditions, and (3) upon reattachment, the β-Gal activity returned to the basal level and cells resumed proliferation. Similar experiments were performed in parallel with cells treated with etoposide (Eto), a well-known inductor of senescence. Eto-untreated floating cells resumed proliferation faster and showed a quicker decrease in β-Gal activity compared to Eto-induced senescent cells. We conclude that cell detachment per se triggers a reversible (plastic) increase in β-Gal. Our findings provide a partial explanation for chemoresistance under anchorage-independent conditions and a new target to eliminate highly resistant floating cells. Ultimately, eliminating Q/S floating cells may prevent or reduce metastasis. Full article

Cellular senescence is a state of irreversible growth arrest characterized by a pro-inflammatory phenotype, playing dual roles in development. In the fetal intestine, the regulation of senescent cells is critical for maintaining tissue homeostasis. Human breast milk (HBM), known for its rich composition of bioactive molecules, may play a role in modulating senescence, although its effects on senescent intestinal cells remain unexplored. This study investigated whether HBM selectively eliminates senescent cells in the FHs74Int fetal intestinal epithelial cell line. Senescence was assessed via β-galactosidase activity and expression of p16 and p21. The model cell line was treated with HBM, infant formula, and milk fractions, and outcomes included cell recovery, senescence markers, apoptosis, and mitochondrial potential. Total free fatty acids (FFA) were quantified and correlated with senolytic activity. HBM reduced senescent cell recovery without affecting non-senescent cells, correlating with decreased β-galactosidase activity, reduced phospho-p38 and γH2AX expression, mitochondrial depolarization, and caspase activation. Only the lipid fraction retained senolytic activity, which was associated with elevated FFA levels. Incubation of HBM at 37 °C increased FFA content and conferred senolytic activity. These findings are consistent with the idea that HBM exerts selective senolytic effects via FFA, revealing a novel mechanism by which breast milk could contribute to intestinal homeostasis. Full article

To express and purify staphylococcal enterotoxin M (SEM) using immobilized metal affinity chromatography (IMAC), a signal peptide-truncated (ΔNsp) wild-type SEM (SEM_WT) was N-terminally fused in pET-28a(+) to a polyhistidine tag (His_6×-) and thrombin cleavage site (TCS; LVPR↓GS), generating His_6×-TCS-ΔNspSEM_WT. Unexpectedly, 4 °C desalting reduced the fusion protein’s molecular weight by ~2.0 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). N-terminal sequencing and mass spectrometry identified cleavage specifically at the arginine (R) and glycine (G) peptide bond (R–G bond) within the TCS motif. AlphaFold 3 revealed an exposed serine protease catalytic triad: histidine 172, serine 178, and aspartic acid 212 (H₁₇₂/S₁₇₈/D₂₁₂) in the β-grasp domain, suggesting intrinsic thrombin-like activity (TLA). Sequential IMAC and size-exclusion high-performance liquid chromatography (SE-HPLC) purification eliminated contaminant concerns, while chromogenic substrate S-2238 (S-2238) assays demonstrated increasing specific activity and purification fold, supporting intrinsic TLA. Critically, the mutation of serine at position 178 to alanine (His_6×-TCS-ΔNspSEM_S178A) abolished TLA but preserved the secondary/tertiary structure, confirming the activity’s origin within the wild-type construct. Molecular dynamics (MD) simulations probed the atomistic mechanism for specific R–G bond cleavage. This work establishes a foundation for understanding ΔNspSEM_WT’s TLA. Full article

Background/Objectives: Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli poses a significant global health challenge, as it leads to antimicrobial treatment failure and is associated with elevated mortality rates. The use of β-lactam/β-lactamase inhibitor combinations offers an alternative approach for combating ESBL-producing bacteria. Ceftriaxone (CRO) and sulbactam have been coadministered in the clinical settings; however, discrepancies in their pharmacokinetics raise concerns regarding the rationality of this combination. Methods: This study was designed to investigate the postβ-lactamase inhibitor effect (PLIE) under both static and dynamic conditions, with the aim of supporting the clinical application of this combination. Results: The minimum inhibitory concentration (MIC) of CRO/SBT (2:1 ratio) against E. coli NCTC 13353 was determined to be 32/16 μg/mL. The PLIEs were determined to be −1.26, −0.57, and 0.37 h at CRO/SBT concentrations ranging from 1/2 MIC to 2 MIC, respectively. The results of CRO concentration, β-lactamase activity, bla_CTX-M-15 expression, and cell morphology collectively support that SBT exerts PLIEs and protects against the antibacterial activity of CRO. In the dynamic hollow-fiber infection model, CRO monotherapy showed no inhibitory effect on E. coli, whereas CRO/SBT combination therapy rapidly eliminated SBT, achieved comparable bactericidal effects, prolonged CRO exposure, and maintained low β-lactamase activity levels. Conclusions: In conclusion, CRO/SBT exerts an inhibitory effect on enzyme-producing strains by being able to produce PLIE to maintain the inhibition of β-lactamase. Full article

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) encompasses a spectrum of liver conditions and involves gut–liver axis crosstalk. We aimed to evaluate whether oral vancomycin modifies liver injury and the cecal microbiota in a methionine–choline-deficient (MCD) diet model of NASH. Male C57BL/6J mice (n = 28) were block-randomized to four groups (n = 7 each) for 10 weeks: standard diet (STD); MCD diet; STD + vancomycin (VANC); and MCD + VANC (2 mg/mouse ≈ 50 mg/kg, every 72 h). After 10 weeks, liver tissues were analyzed for histological changes, cytokine levels [interleukin-6 (IL-6), interleukin-8 (IL-8), transforming growth factor beta 1 (TGF-β1)], and immunohistochemical markers [ubiquitin and cytokeratin 18 (CK18)]. Cecal microbiota composition was evaluated with 16S ribosomal RNA (rRNA) sequencing. The MCD reproduced key NASH features (macrovesicular steatosis, lobular inflammation). Vancomycin shifted steatosis toward a microvesicular pattern and reduced hepatocyte injury: CK18 and ubiquitin immunoreactivity were decreased in MCD + VANC vs. MCD, and hepatic IL-8 and TGF-β1 levels were lower in MCD + VANC vs. STD. Taxonomically, STD mice had Lactobacillus-rich microbiota. The MCD diet alone reduced alpha diversity (α-diversity), modestly lowered Firmicutes and increased Desulfobacterota/Fusobacteriota. Vancomycin alone caused a much larger collapse in richness, depleting Gram-positive commensals and promoting blooms of Escherichia–Shigella, Klebsiella, Parabacteroides, and Akkermansia. In the MCD + VANC group, vancomycin profoundly remodeled the microbiota, eliminating key commensals (e.g., Lactobacillus) and enriching Desulfobacterota, Fusobacteriota, and Campylobacterota. Oral vancomycin in the MCD model of NASH improved liver injury markers and altered steatosis morphology, but concurrently reprogrammed the gut into a low-diversity, pathobiont-enriched ecosystem with near-loss of Lactobacillus. These findings highlight a therapeutic trade-off—hepatic benefit accompanied by microbiome cost—that should guide microbiota-targeted strategies for NAFLD/NASH. Full article

Phyllostachys edulis is a vital bamboo resource in China, known for its economic benefits and ecological functions. However, under natural conditions, its seed germination rate is very low. Exogenous gibberellin (GA) directly supplements endogenous GA levels, while paclobutrazol (PAC) is an inhibitor of GA biosynthesis that can prevent seed germination. Preliminary experiment indicated that a treatment of 50 mg/L GA₃ markedly enhanced the germination rate of P. edulis seeds, whereas 50 μmol/L PAC had an opposite function. To study the exogenous GA₃ effects on the seed germination of P. edulis, seeds were soaked in ddH₂O (CK), Gibberellic acid 3 (GA₃), and PAC solutions for 24 h, respectively. Then, we analyzed and compared the physiology, biochemistry, and transcriptome at different germination stages. The results demonstrated that exogenous GA₃ treatment significantly reduced the contents of starch and soluble protein while increasing the levels of soluble sugar by inducing the activities of β-amylase and protease, respectively. In addition, the activities of superoxide dismutase (SOD), polyphenol (PPO), and ascorbate peroxidase (APX) were enhanced to eliminate ROS during seed germination under exogenous GA₃ treatment compared to CK and PAC treatments. Moreover, the endogenous levels of GA₃ and JA were found to be higher in exogenous GA₃-treated seeds than those in CK and PAC-treated seeds. Furthermore, RNA-seq results revealed that the expressions of 10 related genes are consistent with the observed physiological changes. In summary, exogenous GA₃ effectively accelerated the seed germination of P. edulis by influencing storage reserves, antioxidant enzymes activity, and endogenous hormone through the coordinated transcriptional regulation of related genes. These findings provide novel insights into the regulation mechanisms of exogenous GA₃ on the seed germination of P. edulis. Full article