Springer最新文献

Diabetic nephropathy (DN) is a major complication of diabetes characterized by progressive renal dysfunction driven by oxidative stress and chronic inflammation. Although Withaferin A (WA) possesses antioxidant and anti-inflammatory properties, its clinical application is limited by poor aqueous solubility and rapid systemic clearance. This study developed a thermosensitive chitosan/β-glycerophosphate (CS/β-GP) hydrogel as a sustained-release platform for WA delivery and evaluated its renoprotective potential using high glucose-treated renal cells. An early streptozotocin-induced diabetes-associated renal injury animal model following 4 days of treatment and a 28-day observation period. The CS/β-GP-WA hydrogel demonstrated favorable physicochemical properties, including efficient drug encapsulation, thermoresponsive gelation, injectability, sustained drug release, and good batch-to-batch reproducibility. Under high-glucose conditions, the hydrogel improved cell viability, reduced intracellular reactive oxygen species generation, preserved mitochondrial membrane potential, restored antioxidant enzyme activity, and decreased pro-inflammatory cytokine production. In diabetic rats, CS/β-GP-WA treatment was associated with improved glycemic control and renal function, reflected by reduced serum creatinine, blood urea nitrogen, urinary protein, and oxidative stress markers. Histological analyses further showed preservation of renal structure and reduced tissue injury. These improvements were accompanied by increased Nrf2, HO-1, and NQO1 mRNA expression and reduced Keap1 expression, suggesting an association with enhanced antioxidant responses. Overall, sustained delivery of WA using the CS/β-GP hydrogel was associated with attenuation of diabetes-associated renal injury and improved renal functional and histological outcomes in this early experimental model. However, the absence of protein-level mechanistic validation and comprehensive pharmacokinetic and tissue-distribution analyses limits definitive conclusions regarding pathway activation and renal targeting. Further studies are warranted to confirm the underlying mechanisms and evaluate long-term therapeutic potential.

Liver cancer is a leading cause of cancer-related mortality worldwide due to its abrupt proliferation and inadequate treatment. Developing novel and effective therapeutics against liver cancer has become a formidable challenge in the arsenal of biomedical research due to the limitations of conventional treatment strategies. Recently, natural product-based metal nanocomposites have emerged as promising candidates for targeted and biocompatible drug delivery. Herein, a Triphala-functionalized gold nanocomposite (t-AuNC) was synthesized employing Triphala extract and gold nanoparticles (AuNPs) as the precursors to enhance targeted delivery against liver cancer. Different analytical techniques, including TEM, DLS, UV-visible spectroscopy, XRD, zeta potential, FT-IR, and EDX analysis, were used to evaluate the synthesized nanocomposite, confirming its successful formation and stability. The MTT and SRB assays demonstrated the anti-proliferative implications of t-AuNC against the HepG2 cell line. Flow cytometry analysis confirms the induction of apoptosis in the HepG2 cell line by the t-AuNC. Antioxidant analysis using DPPH and ABTS assays showed significant radical scavenging activity of the t-AuNC. Furthermore, the t-AuNC also demonstrated notable anti-inflammatory properties validated by its protein denaturation inhibition. Collectively, these findings highlight that t-AuNC could be a contemporary and efficient medication for liver cancer therapy with minimized side effects.

Madin-Darby canine kidney (MDCK) cells are important hosts for cell culture-based viral vaccine production, but fetal bovine serum (FBS) supplementation increases cost, batch variability, adventitious-agent risk, and regulatory burden. This study developed a 2% FBS low-serum medium optimization workflow for MDCK cells by integrating design of experiments (DoE) with interpretable machine learning. A Plackett-Burman (PB) design screened 27 nutritional components and identified seven factors that significantly affected proliferation. L-glutamine, L-asparagine, and L-tyrosine showed positive effects and were further optimized using a three-factor Box-Behnken design (BBD). A Kolmogorov-Arnold network (KAN) modeled the nonlinear proliferation response, and its response function served as the objective function for Bayesian optimization (BO). After two KAN-BO validation cycles, the optimized formulation contained 292.30 mg/L L-glutamine, 54.36 mg/L L-tyrosine, and 11.34 mg/L L-asparagine. The predicted relative proliferation rate was 159.38%, in close agreement with the experimental value of 158.63 ± 1.70%. Compared with Baseline medium, Optimized medium enhanced MDCK cell proliferation while maintaining typical adherent epithelial-like morphology and high viability. In H1N1 BVR-26 production assays, Optimized medium achieved a peak titer of 3.50 ± 0.05 log₁₀ 50% tissue culture infectious dose (TCID₅₀)/mL at 48 h post-infection, outperforming both Baseline medium and Commercial medium. Metabolic analysis showed higher apparent glucose consumption and lower lactate yield, indicating improved glucose utilization and reduced metabolic burden. These results demonstrate that the PB-BBD-KAN-BO workflow is an efficient, interpretable, and experiment-saving strategy for low-serum medium development.

To enhance lignocellulose degradation in tobacco stem fibers and improve the quality of reconstituted tobacco, this study optimized xylanase-mediated hydrolysis. Xylanase OKBK was identified as the most effective enzyme, producing a reducing sugar yield of 2.29%, significantly higher than those obtained with other tested enzymes (p< 0.05). Single-factor experiments were performed to evaluate the effects of six key parameters, including enzyme concentration, hydrolysis time, pH, solid-to-liquid ratio, temperature, and shaking speed. Plackett-Burman design identified five significant factors (p< 0.05), and the steepest ascent method was subsequently used to determine the optimal region for response surface optimization. A Box-Behnken design was then applied to develop a quadratic regression model involving temperature, hydrolysis time, and enzyme concentration, which exhibited high predictive accuracy (R²= 0.9950). The optimized conditions predicted by the model were 50 ℃, 4.5 h, and 400 U/mL, resulting in a reducing sugar yield of 21.1%, closely matching the predicted value of 21.5%. Component analysis, FTIR, and SEM characterization demonstrated that xylanase effectively disrupted the lignocellulosic structure and facilitated component degradation. Furthermore, sensory evaluation of reconstituted tobacco sheets prepared from enzymatically treated stems showed improved sensory properties, particularly reduced woody odor and smoke harshness. Overall, the optimized xylanase hydrolysis process enhanced lignocellulose degradation and sensory quality, providing a promising strategy for the high-value utilization of tobacco stem waste.

Cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) is a common female malignancy. Gut microbiota and metabolites are critical regulators of tumor immunity and therapy, but their roles in CESC remain unclear. This study integrated TCGA and GEO datasets with gut microbiota information to construct a protein-protein interaction network. These targets were prioritized using machine learning, SHAP interpretation, and Mendelian randomization analysis. Immune-related pathways were explored using single-cell and spatial transcriptomic analyses. Molecular docking and molecular dynamics simulations were performed to evaluate potential interactions between key metabolites and their targets. A total of 136 gut microbiota-related DEGs were identified, which may be associated with intercellular immune interactions. KDR was selected as a core target and may exert protective effects. Single-cell and spatial transcriptomic analyses suggested that specific metabolites may be associated with changes in the tumor microenvironment potentially involving the MIF signaling pathway. Network analysis of microbes, metabolites, and targets suggested that metabolites such as 5-(3,4-dihydroxyphenyl) pentanoic acid may serve as key mediators linking gut microbiota and KDR signaling. Additionally, eight non-toxic metabolites with favorable drug-likeness were identified, molecular docking and molecular dynamics simulation demonstrated stable binding to KDR with potential bioactivity, providing a theoretical basis for developing microbiota-related therapeutic strategies. The gut microbiota and its metabolites may be associated with the immune microenvironment and tumor progression in CESC, potentially involving the MIF signaling axis. This study provides a computational framework and preliminary evidence supporting microbiota-related hypotheses, and may inform future experimental investigations and therapeutic strategy development.

A total of twenty pyrazole derivatives were synthesized using alkoxy or halo acetophenones in reaction with N, N-dimethylformamide dimethylacetal (DMF-DMA). These compounds were characterized through ¹H NMR, ¹³C NMR, and LC-HRMS techniques. Their herbicidal efficacy against barnyard grass (Echinochloa crus-galli) was evaluated via in vitro bioassays. Notably, compound 6a (5-Methoxy-2-(1H-pyrazol-3-yl) phenol) and compound 6s (3-(4-Fluoro-2-hydroxyphenyl) pyrazole) demonstrated inhibition rates of 30.6% to 54.1% and 26.9% to 50.6%, respectively, across concentrations ranging from 62.5 to 1000 µg mL^- 1. In in vivo assessments, compound 6a yielded the highest weed control index (WCI) of 51.32%, followed by compound 6s with a WCI of 50.24%. These findings indicate that compound 6a represents a promising herbicidal agent, effectively managing barnyard grass while exhibiting tolerance toward rice seedlings following both post-emergence and soil application. Toxicity predictions for the synthesized compounds were evaluated for fish (LC₅₀: 0.0001 to 38.1 mg/L), daphnids (LC₅₀: 0.0001 to 27.7 mg/L), and green algae (EC₅₀: 0.0001 to 1.70 mg/L), including chronic toxicity (ChV). Using the ECOSAR model, the assessment found that shorter chain compounds are within acceptable limits. The binding free energy of compound 6a with HPPD was relatively low (-8.3), which was consistent with the findings of bioassay activity studies. As a result, compound 6a could serve as the starting point for the development of novel HPPD inhibitors.

Rapid and accurate detection of methicillin-resistant Staphylococcus aureus (MRSA) is crucial for guiding antimicrobial therapy and improving outcomes in infection management, including postoperative care and surgical site infections. Conventional detection methods often suffer from lengthy turnaround times, operational complexity, or limited specificity. To address these challenges, we developed a novel, integrated biosensing platform termed the Cas12a/crRNA-initiated triple self-priming cascade amplification assay for ultrasensitive and specific analysis of the mecA resistance gene. The innovation of this strategy lies in the synergistic combination of CRISPR-Cas12a for high-fidelity target recognition and an elegantly engineered isothermal amplification circuit. Target-specific activation of Cas12a triggers the trans-cleavage of a designed probe, remaining an RNA initiator that sets off a triple self-priming cascade amplification within a single, multifunctional Y-shaped probe (Y-probe). This integrated probe architecture eliminates the need for multiple independent components, significantly simplifying the workflow and enhancing assay stability. Amplification culminates in the generation of G-quadruplex (G4) structures, which are detected through a label-free turn-on fluorescence signal using thioflavin T (ThT), thereby avoiding the instability associated with covalently labeled probes. The assay demonstrated exceptional analytical performance, achieving a detection limit of 0.41 fM for synthetic mecA DNA and 5.2 CFU/mL for viable MRSA cells, with a linear range spanning five to six orders of magnitude. It exhibited high specificity, successfully discriminating MRSA from methicillin-susceptible Staphylococcus aureus and distinguishing single-base mismatches. By integrating precise CRISPR recognition, a streamlined and stable all-in-one probe design, and a robust label-free readout, the platform presents a powerful, user-friendly molecular tool for resistance genotyping, with potential for future application in clinical settings, such as postoperative infection management.

The growing burden of antimicrobial resistance (AMR) and the persistence of biofilm-associated and quorum-sensing-mediated infections represent major public health challenges, highlighting the need for new anti-infective agents from natural sources. In this context, Allium atroviolaceum is recognized for its culinary and medicinal uses, remains relatively underexplored with respect to its anti-virulence potential, despite biological activities previously attributed to its organosulfur and phenolic constituents. In this study, ethanolic Allium atroviolaceum bulb extract (EAABE) was investigated for its antimicrobial, antibiofilm, and anti-quorum sensing (QS) activities. HR-LCMS analysis tentatively annotated 19 metabolites, including organic acids, phenolics, triterpenoids, fatty acids, glycosides, alkaloids, sphingolipids, steroid derivatives, and peptide-type compounds. EAABE demonstrated moderate antibacterial effects, with inhibition zones ranging from 6.00 to 7.33 mm and MIC values between 0.292 and 2.343 mg/mL. In contrast, a relatively enhanced antifungal activity was observed, with inhibition zones of 12.67-15.00 mm and MIC values of 0.292-0.585 mg/mL. The extract also inhibited biofilm formation in a dose-dependent manner, achieving up to a 56.91% reduction, while swarming motility was reduced by up to 64.13%. In addition, EAABE significantly attenuated QS-regulated virulence traits in a concentration-dependent manner. Violacein production was reduced by 67-86% at concentrations of 250-1000 µg/mL, whereas swarming motility was inhibited by 29 to 64% at 50-100 µg/mL. Similarly, pyocyanin production and extracellular proteases activity were reduced by 67-90% and 29-46%, respectively, across the concentration range of 250-1000 µg/mL. Molecular docking analysis predicted favorable binding affinities for several tentatively annotated metabolites, ranging from - 7.9 to - 12.4 kcal/mol, with Gravelliferone, Americine, and (2'S)-Deoxymyxol 2'-α-L-fucoside as compounds with favorable predicted binding affinities toward the selected targets. These interactions were further supported by 100 ns molecular dynamics simulations. ADMET predictions and DFT calculations provided further insights into the physicochemical, predicted pharmacokinetic, and electronic properties of the tentatively annotated metabolites. Overall, the findings indicate that EAABE exhibits promising in vitro antimicrobial and anti-virulence activities; however, further isolation of individual constituents, experimental validation, and mechanistic investigations are needed to clarify the specific contribution of each metabolite to the observed effects.

Polyphenols are secondary metabolites of plants. They have potential health benefits as antioxidants and protection against the development of cancers, cardiovascular disease, and diabetes. Herein, the inhibitory potential of a series of 35 simple phenols, flavone, flavonol, flavanone, dihyrdoflavanol, flavan-3-ol, anthocyanidin, isoflavone, chalcone, silybins polyphenolics against (i) the main protease of (M^pro) of SARS-CoV-2, the enzyme involved in COVID-19 viral replication and infection, and (ii) the human peroxiredoxin 5 the antioxidant enzyme has been investigated by estimating their binding affinities by means molecular docking, molecular dynamics simulation, and by evaluating their pharmacokinetic, and the drug-likeness properties through the ADMET study. Further, the structure-activity relationship of the effects of the number of hydroxyl groups, the presence of unsaturated double bonds, the substitution of hydroxyl groups, the opening of the C-ring, and the position of the B-ring on the main protease (M^pro) and the human peroxiredoxin 5 inhibition has been discussed.

Garlic (Allium sativum) contains bioactive components with antioxidants and anti-inflammatory constituents with potential anticancer activity. We aimed to compare a petroleum-ether garlic extract (PEGE) versus an ethanolic garlic extract (EGE) in a DMBA-induced murine hepatic carcinogenesis model. Fresh garlic was extracted with petroleum ether or ethanol and chemically characterized by GC-MS. PEGE was enriched in long-chain fatty acids/derivatives, while EGE contained both lipids and more polar constituents. Male Swiss albino mice received DMBA and were assigned to concurrent (DMBA + extract) or post-DMBA (DMBA → extract) regimens; extract-only and vehicle groups served as toxicity controls. Biochemical parameters (ALT, AST, AFP), oxidative stress markers (MDA, GSH), inflammatory signaling (NF-κB by ELISA), apoptosis (caspase-3 gene expression by qPCR), proliferation (PCNA by flow cytometry), and histopathological changes (H&E staining) were evaluated. Extract-only showed no significant differences compared to control groups, indicating no overt toxicity. DMBA administration resulted in significant hepatic injury, evidenced by elevated liver enzymes and AFP, increased lipid peroxidation and NF-κB levels, depleted GSH, enhanced cellular proliferation, and severe histopathological alterations. Across endpoints, the post-DMBA PEGE regimen produced the largest overall normalization, followed by EGE. Post-treatment outperforming concurrent dosing for both extracts. Histology aligned the biochemical trends, showing reduced necrosis, inflammation, vascular congestion, and dysplastic changes in treated groups. Both garlic extracts showed hepatoprotective effects in this preclinical model; they were well tolerated and mitigated DMBA-induced injury, with PEGE given after DMBA yielding the strongest biochemical, molecular, and histopathological improvements. Chemical differences between the lipid-rich PEGE and the more polar EGE likely contribute to their differential efficacy and merit further standardization and mechanistic study.