Acetic Acid

Abstract

To investigate the species distribution and antifungal susceptibility profiles of clinical mould isolates from China to olorofim, manogepix, amphotericin B, triazoles and echinocandins.

Abstract

Amyloid-β (Aβ) peptides are a defining feature of Alzheimer's disease (AD). These peptides are produced by the proteolytic processing of the amyloid precursor protein (APP), which can occur through the synaptic vesicle (SV) cycle. However, how amyloidogenic APP processing alters SV composition and presynaptic function is poorly understood. Using App knock-in mouse models of amyloid pathology, we found that proteins with impaired degradation accumulate at presynaptic sites together with Aβ42 in the SV lumen. Levetiracetam (Lev) is a US Food and Drug Administration-approved antiepileptic that targets SVs and has shown therapeutic potential to reduce AD phenotypes through an undefined mechanism. We found that Lev lowers Aβ42 levels by reducing amyloidogenic APP processing in an SV2a-dependent manner. Lev modified SV cycling and increased APP cell surface expression, which promoted its preferential processing through the nonamyloidogenic pathway. Stable isotope labeling combined with mass spectrometry confirmed that Lev prevents Aβ42 production in vivo. In transgenic mice with aggressive amyloid pathology, electrophysiology and immunofluorescence confirmed that Lev restores SV cycling abnormalities and reduces synapse loss. Last, early Aβ pathology in brains from donors with Down syndrome was characterized by elevated presynaptic proteins. Together, these findings highlight the potential to prevent Aβ pathology before irreversible damage occurs.

Abstract

Optimal purification, high purity and robust purity control for synthetic peptides are critical, as even minor impurities can alter biological activity, distort analytical results and compromise downstream applications. Reliable translation of analytical HPLC results to preparative FPLC conditions remains challenging due to system-specific differences in column geometry, gradient formation and mobile-phase chemistry. This study addresses these limitations through a systematic evaluation of chromatographic parameters influencing peptide selectivity, resolution and transferability. Using a defined peptide impurity library, the effects of gradient steepness, flow rate, temperature and mobile-phase modifier were quantified, revealing that flow-rate optimization and modifier choice have the greatest impact on separation quality. A correction equation was developed to compensate for system-dependent deviations, reducing transfer errors in elution percentage from approximately 17% to less than 5%. The optimized workflow enabled initial preparative purifications with purities above 90% and yields exceeding 30%. Additionally, substitution of trifluoroacetic acid with formic acid was explored as a greener modifier, providing selective improvements in separation performance. The approach establishes a practical and sustainable workflow for the transfer of HPLC-to-FPLC methods for peptide purification.

Abstract

To address the pressing need for improving the silage quality of sweet sorghum, this study aims to establish aPichia anomala-Lactic acid bacteria (LAB) consortium suitable for the sweet sorghum silage system and to elucidate its synergistic mechanism through untargeted metabolomics. The optimal P. anomala-Leuconostoc citreum consortium was obtained through a membership function analysis based on indicators of organic acids and antifungal metabolites. Compared to L. citreum monoculture, the co-culture enhanced L. citreum viable cell count and lactic acid (LA) production by 35.80% and161.34%, respectively. Untargeted metabolomics analysis of differential metabolites related to L. citreum growth and LA synthesis revealed upregulation of amino acids and their derivatives (10), bioactive peptides (4), vitamins (4), and organic acids (2). This metabolic profilereveals the role of P. anomala in enhancing L. citreum growth and LA synthesis by providing key growth factors and LA precursors. During ensiling, the P. anomala-L. citreum consortium significantlyslowedthe loss of dry matter (DM) and water-soluble carbohydrate (WSC), and reducedthe content of fiber components and structural polysaccharides (P < 0.05). The consortium also effectively suppressed the accumulation of ammonia nitrogen (AN) and propionic acid (PPA), while significantly increasing the contents of crude protein (CP), LA, and acetic acid (AA) (P < 0.05). These findings provide a novel insight for developing high-performance yeast-LAB consortia for the improvement of biomass silage quality.

Abstract

To evaluate the synergistic protective effect of linezolid (LZD) combined with Xuebijing Injection (XBJ, a traditional Chinese medicine injection) against pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA) in mice and explore the underlying mechanism.

Abstract

Ecological dysregulation leads to the progression of inflammatory bowel disease (IBD). The present study was designed to evaluate whether ginsenoside Rb3 (GR3) ameliorates dextran sulfate sodium (DSS)-induced colitis by modifying the microbiota. The results revealed that GR3 treatment with oral doses of 5 mg/kg suppressed DSS-induced colitis in mice, and its effects were evaluated using a combination of histological analysis, enzyme-linked immunosorbent assays (ELISA), and Western blotting. This was evidenced by a significant attenuation of symptoms such as weight loss, diarrhea, hematochezia, and colonic shortening in the DSS-induced colitis mice. Furthermore, GR3 treatment remarkably elevated the expression of tight junction proteins (occludin and zonula occludens-1) while reducing both inflammatory cell infiltration and inflammatory cytokine concentrations (TNF-α, IL-1β, IL-15, IL-17A, and IL-6). Intriguingly, GR3 treatment also mitigated DSS-induced intestinal dysbiosis by prominently increasing the proliferation of Lactobacillus and decreasing the relative abundance of Bacillus. Additionally, GR3 treatment significantly modified the metabolism of short-chain fatty acids in colitis mice, especially elevating the levels of acetic acid and butyric acid. These findings suggest that GR3 ameliorates colitis by reshaping the gut microbiota and improving the intestinal barrier and inflammation.

Abstract

Maize (Zea mays L.), a globally cultivated staple grain and economically significant crop, is highly susceptible to Southern corn leaf blight (SCLB), a disease caused by Cochliobolus heterostrophus. This pathogen can lead to substantial losses in both yield and grain quality during the growing season. In this study, Bacillus velezensis strain JLU-53 was identified as a promising biocontrol agent against C. heterostrophus. JLU-53 exhibited broad-spectrum antifungal activity against several phytopathogens, including C. heterostrophus, Setosphaeria turcica, Rhizoctonia solani, Fusarium graminearum, Botrytis cinerea and Sclerotinia sclerotiorum. Genomic analysis revealed the presence of genes encoding diverse carbohydrate-active enzymes (CAZymes), and phenotypic assays confirmed the strain's capacity to produce hydrolytic enzymes such as cellulase, protease, and amylase, as well as indole-3-acetic acid (IAA). Whole-genome sequencing of JLU-53 identified 13 putative biosynthetic gene clusters responsible for the synthesis of antimicrobial peptides (AMPs). Further analysis of secondary metabolites confirmed the production of bacillomycin, macrolactin, and amicoumacin. In vitro experiments demonstrated that JLU-53 significantly inhibited mycelial growth, conidial germination, and appressorium formation of C. heterostrophus, thereby suppressing lesion development of maize leaves. In planta application of JLU-53 activated key defense-related genes in maize, enhanced host resistance, and markedly reduced disease symptoms. These findings indicate that B. velezensis JLU-53 exerts its biocontrol activity through dual mechanisms-direct antagonism and induction of systemic resistance-highlighting its potential as a sustainable and effective biocontrol agent for managing SCLB.

Abstract

Hypercholesterolemia is a major risk factor for atherosclerotic cardiovascular disease; however, current therapeutic options such as statins are limited by issues including hepatotoxicity and patient intolerance. Probiotics and their metabolites show promise in modulating cholesterol metabolism through the gut‒liver axis, yet the specific commensal bacteria and molecular mechanisms underlying these effects remain poorly understood. In this study, we isolated and characterized EPS-D1, a novel exopolysaccharide (15.003 kDa) derived from Lactiplantibacillus plantarum H6, which is composed primarily of mannose (46.10%) and glucose (33.98%) and features a highly branched structure (branching degree of 29.5%). The administration of EPS-D1 significantly reduced the serum total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) by 40.31%, 37.55%, and 43.15%, respectively, in high-cholesterol diet (HCD) mice. Additionally, it improved hepatic steatosis and reduced markers of liver injury. Through 16S rRNA sequencing and fecal microbiota transplantation (FMT), we identified Muribaculum as the key commensal bacterium enriched by EPS-D1. Direct administration of Muribaculum (Muribaculum intestinale) replicated the cholesterol-lowering effects, decreasing ileal and fecal cholesterol levels by 74.79% and 53.16%, respectively. Mechanistically, both EPS-D1 and M. intestinale activated the enterohepatic FXR‒FGF15 axis, which resulted in the upregulation of hepatic cholesterol 7α-hydroxylase (CYP7A1) expression and the downregulation of ileal ASBT and NPC1L1, thereby promoting bile acid synthesis and inhibiting cholesterol absorption. Furthermore, M. intestinale increased intestinal short-chain fatty acids (SCFAs), particularly acetic acid and caproic acid, by 37.88% while also modulating the composition of the bile acid pool. These findings establish M. intestinale as a precise microbial target for cholesterol management and demonstrate that EPS-D1 from L. plantarum H6 enhances cholesterol metabolism through microbiota-mediated activation of the enterohepatic FXR‒FGF15 axis, providing a novel therapeutic strategy for managing hypercholesterolemia.

Abstract

Background: Estrogen deficiency following human menopause or rodent ovariectomy (OVX) induces adverse alterations in body composition and metabolic function. This study investigated the combined effects of acetic acid supplementation and voluntary exercise on metabolic health and skeletal muscle mitochondrial function using an OVX mouse model. Methods: Forty female C57BL/6J mice (8 weeks old) were randomly assigned to 5 groups: sham (SHM), ovariectomized control (OVX), OVX with exercise (OVX-E), OVX with acetic acid (OVX-A), and OVX with both interventions (OVX-AE). Following a 1-week recovery from OVX, a 13-week intervention was initiated: 5% sodium acetate-supplemented chow and/or voluntary wheel running. Body composition, glucose tolerance, total energy expenditure, skeletal muscle mitochondrial function, and the contents of AMPKα, PGC-1α, and carbonyl protein were assessed. Results: OVX impaired whole-body metabolism and skeletal muscle mitochondrial function, specifically in the gastrocnemius muscle. While the exercise alone failed to mitigate the OVX-induced mitochondrial dysfunction, the combined treatment of exercise and acetic acid supplementation significantly rescued from the OVX-induced mitochondrial dysfunction. Conclusions: OVX resulted in detrimental changes in whole-body metabolism, but voluntary exercise and/or acetic acid supplementation had no rescuing effects on those parameters. In gastrocnemius muscle, acetic acid supplementation during exercise enhanced mitochondrial function in OVX mice.

Abstract

Plant-based fermented foods are increasingly promoted for glycemic control, yet their mechanisms and clinical impact remain incompletely defined. This narrative review synthesizes mechanistic, preclinical, and human data for key matrices-kimchi and other fermented vegetables, tempeh/miso/natto, and related legume ferments, kombucha and fermented teas, plant-based kefir, and cereal/pulse sourdoughs. Across these systems, microbial β-glucosidases, esterases, tannases, and phenolic-acid decarboxylases remodel polyphenols toward more bioaccessible aglycones and phenolic acids, while lactic and acetic fermentations generate organic acids, exopolysaccharides, bacterial cellulose, γ-polyglutamic acid, γ-aminobutyric acid, and bioactive peptides. We map these postbiotic signatures onto proximal mechanisms-α-amylase/α-glucosidase inhibition, viscosity-driven slowing of starch digestion, gastric emptying and incretin signaling, intestinal-barrier reinforcement, and microbiota-dependent short-chain-fatty-acid and bile-acid pathways-and their downstream effects on AMPK/Nrf2 signaling and the gut-liver axis. Animal models consistently show improved glucose tolerance, insulin sensitivity, and hepatic steatosis under fermented vs. non-fermented diets. In humans, however, glycemic effects are modest and highly context-dependent: The most robust signal is early postprandial attenuation with γ-PGA-rich natto, strongly acidified or low-glycemic sourdough breads, and selected kombucha formulations, particularly in individuals with impaired glucose regulation. We identify major sources of heterogeneity (starters, process parameters, substrates, background diet) and safety considerations (sodium, ethanol, gastrointestinal symptoms) and propose minimum reporting standards and trial designs integrating metabolomics, microbiome, and host-omics. Overall, plant-based ferments appear best positioned as adjuncts within cardiometabolic dietary patterns and as candidates for "purpose-built" postbiotic products targeting early glycemic excursions and broader metabolic risk.

Abstract

This study aimed to determine whether degeneration of intervertebral discs and subchondral bone correlates with the development of persistent low back pain (LBP) in a rat model with intradiscal monosodium iodoacetate (MIA), focusing on structural alterations, neuronal sensitization, and pain-related behavior. Male Sprague Dawley rats received MIA injections (2 mg in 2 μl) into L4/5 and L5/6 lumbar discs to induce degeneration. LBP-related behaviors were assessed over 42 days via dynamic weight-bearing and hindpaw withdrawal thresholds. Neuronal sensitization was evaluated via electrophysiological recordings of mechanosensitive afferent nerves (MSAN) and calcium imaging of labeled dorsal root ganglia (DRG) neurons innervating discs. Structural alterations in subchondral bone and disc were analyzed using in vivo/ex vivo µCT imaging, along with immunohistochemistry for TRPV1 and CGRP expression. Intradiscal MIA induced long-term LBP behaviors, characterized by forelimb weight-bearing shifts and reduced hindpaw withdrawal thresholds. MSAN exhibited hyperexcitability and lowered firing thresholds to intradiscal pressure, while DRG neurons showed enhanced TRPV1- and TRPA1-mediated calcium influx. µCT imaging revealed decreased disc volume and deteriorated trabecular bone quality. Histology confirmed disc and subchondral bone degeneration, with upregulated TRPV1 and CGRP expression in subchondral bone, indicating nociceptive fiber ingrowth. Linear discriminant analysis identified weight-bearing asymmetry (HindFore AUC), trabecular thickness, connectivity, and trabecular pattern factor as strong predictors of LBP development. Pathological degeneration of disc and bone is associated with chronic LBP through structural remodeling and peripheral neuronal sensitization. These findings suggest that therapeutic strategies targeting both tissues can alleviate chronic pain in degenerative spine conditions.

Abstract

Glyphosate (GLY) and its main degradation product aminomethylphosphonic acid (AMPA), together with the chemically related phosphinic acid herbicide glufosinate (GLU), are widely used agrochemicals. Their reliable quantification in groundwater remains analytically challenging due to complexation with divalent cations and matrix-dependent ionization. This work developed and validated a groundwater-adapted LC-MS/MS method using FMOC derivatization and isotopically labeled glyphosate (GLY*) as internal standard. The new protocol (SUBTE, from SUBTERRANEAN) introduces an acidification in the presence of excess EDTA to disrupt metal complexes prior to derivatization and was benchmarked against a conventional borate-buffer workflow (BORAX). Normalized calibration (analyte/GLY*) was linear from 0.5 to 20 µg·L-1 (R2 ≥ 0.99) and origin-compatible. Operational LOD/LOQ in matrix were 0.003/0.01 µg·L-1 for GLY, 0.01/0.04 for GLU, and 0.02/0.06 for AMPA, meeting precision criteria (≤20 % RSD at the lowest level). Across real groundwater samples spanning wide conductivity and Ca+2 + Mg+2 ranges (n = 50), SUBTE achieved mean recoveries of ∼96 % compared with ∼70 % for BORAX, demonstrating effective mitigation of divalent-cation artefacts and enhanced robustness in hard waters. No analyte exceeded the detection limit in any well at the time of sampling, consistent with the low-mobility, Ca-rich geo-pedologic setting of the Pampean plain. The SUBTE workflow thus provides a validated, matrix-resilient approach for routine monitoring of GLY, AMPA, and GLU in groundwater, ensuring reliable quantification under variable hydrogeochemical conditions.

Abstract

Hexadimethrine Bromide (HxB) has been reported to trigger marked eosinophilia even under conditions that typically cause eosinopenia. We investigated how HxB modulates eosinopoiesis and eosinophil accumulation in mice. Swiss Webster, C57BL/6, and BALB/c mice received HxB (1 or 10 mg/kg, intraperitoneally). Eosinophils were quantified in bone marrow, blood, and spleen by cytology and flow cytometry; serum cytokines/chemokines were profiled; serum transfer and montelukast pre-treatment tested mechanisms. HxB increased circulating and tissue eosinophils in a dose-dependent manner, peaking at 10 mg/kg, without neutrophilia. Bone marrow eosinopoietic potential rose, with higher frequencies of eosinophil-committed progenitors and mature eosinophils; effects generalized across strains. Serum from HxB-treated donors transferred eosinophilia to naïve recipients. A Th2-skewed burst of mediators (IL-3, IL-4, IL-5, IL-9, IL-13, eotaxin, RANTES, MIP-1α/β, and TNF-α) peaked at 24 h and returned to baseline by day 3. Montelukast did not blunt responses, indicating CysLT-independence. A second HxB dose on day 3 sustained eosinophilia through day 6. HxB elicits a rapid, cytokine-mediated, CysLT-independent pro-eosinophilic state that can be maintained with dosing every three days. This robust model yields abundant, mature eosinophils, supports mechanistic studies, and may inform future translational investigations of pharmacologically induced eosinophilia.

Abstract

We investigated whether plasma cytomegalovirus (CMV) IL-10 (cmvIL-10) levels could serve as a biomarker of active CMV replication in allogeneic hematopoietic transplant recipients (allo-HCT) in the presence or absence of letermovir (LMV) prophylaxis. A total of 189 leftover plasma samples that tested positive for CMV DNA (Alinity m CMV assay), representing 33 episodes of CMV DNAemia were run on a laboratory-developed enzyme-linked immunosorbent assay for cmvIL-10 quantification. Eighteen episodes developed during LMV prophylaxis. Overall, 16 episodes of CMV DNAemia were classified as clinically significant (CsCMVi). There was an overall very weak correlation between the two biomarkers (Rho = 0.10; p = 0.16). Overall, the median cmvIL-10 area under the curve (AUC) until CMV DNA levels reached their peak was significantly higher (p < 0.001) in CsCMVi episodes than in non-CsCMVi episodes. cmvIL-10 AUC between Days 14 and 23 after allo-HCT (AUC₁₄₋₂₃) values were significantly higher in CsCMVi episodes compared with non-CsCMVi episodes among patients receiving LMV therapy (p = 0.008). An AUC₁₄₋₂₃ cutoff value of log10 3.06 discriminated anticipately between CsCMVi and non-CsCMVi with a sensitivity and specificity of 100%. Plasma cmvIL-10 levels may reflect true CMV replication and thus provide a unique perspective on viral dynamics, serving as an ancillary marker to CMV DNA monitoring.

Abstract

Dendrobium officinale Kimura et Migo (D. officinale), a rare and precious orchid (Orchidaceae) in traditional Chinese medicine, exhibits a sweet flavor and a cold nature, and acts on lung, stomach, and kidney meridians with marked efficacy in nourishing lung yin. When applied medicinally and in functional foods, it demonstrates antioxidant, immunomodulatory, anti-inflammatory, and anti-inflammatory, and antitumor bioactivities.

Abstract

High-fat diet (HFD) is a key contributor in exacerbating food allergy, but there is little in-depth research on the mechanisms by which HFD affects allergic responses induced by α-lactalbumin (ALA). The mechanism of HFD exacerbating ALA-induced allergic responses (H-ALA) was studied via microbiome and metabolomics. HFD induced alterations in body weight and temperature in ALA-sensitized mice, while exacerbating allergic responses by increasing ALA-specific IgG/IgE levels and disrupting Th1/Th2 cytokine balance. In H-ALA, 16S rRNA gene sequencing revealed that Staphylococcus and Bilophila were the dominant genera, and untargeted metabolomics demonstrated an upregulation of histidine accompanied by reduced indole-3-acetic acid. Correlation analysis and subsequent mechanistic validation suggested that HFD might exacerbate ALA-induced allergic responses through promoting the expansion of Staphylococcus and elevating histidine levels. These findings will provide a theoretical foundation for the prevention and intervention of HFD-exacerbated food allergy.

Abstract

This study aims to explore the synergistic effect of triptolide(TP) combined with methotrexate(MTX) on lipopolysaccharide(LPS)-induced RAW264.7 macrophages and its potential mechanism based on the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING) signaling pathway, in order to infer the therapeutic effect of the combination of the two drugs on rheumatoid arthritis. RAW264.7 cells were cultured in vitro, and cell counting kit-8(CCK-8) assay was used to detect cell survival rates under the intervention of different concentrations of drugs and STING activator 5,6-dimethylxanthenone-4-acetic acid(DMXAA) and to evaluate the synergistic effect of the combination of TP and MTX by the combination index(CI). RAW264.7 cells were divided into a control group(normal cells), a model group(1 μg·mL~(-1) LPS), a TP group(0.025 μmol·L~(-1)), a MTX group(0.1 μmol·L~(-1)), combination groups(TP+MTX), and an activator group(TP+MTX+DMXAA). Cell migration and invasion abilities were detected by cell scratch and Transwell invasion assays. Levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in cell supernatants were determined by enzyme-linked immunosorbent assay(ELISA). cGAS and STING protein expression levels were analyzed in cells by Western blot. The results showed that the expression levels of IL-6, TNF-α, cGAS, and STING increased, and the cell migration and invasion abilities were enhanced in the model group compared with the control group. CCK-8 results showed that TP and MTX had a certain inhibitory effect on the proliferation of RAW264.7 cells, and there was a trend of concentration-and time-dependence. The six combination groups had a synergistic inhibitory effect on the proliferation of RAW264.7 cells(CI<1). The results of cell scratch and Transwell invasion assays showed that the migration and invasion of cells were decreased in each drug-treated group compared with the model group, and the decrease was more obvious in the combination groups(P<0.05). The results of ELISA and Western blot showed that the expression of IL-6, TNF-α, cGAS, and STING was down-regulated in the cells of each drug-treated group compared with the model group, and the effect of the combination groups was more favorable(P<0.05). In addition, the STING activator DMXAA attenuated the inhibitory effect of TP combined with MTX on LPS-induced RAW264.7 cell viability and inflammatory response(P<0.05). In conclusion, TP combined with MTX synergistically inhibited LPS-induced proliferation of RAW264.7 cells, and its mechanism of action may be related to the down-regulation of the cGAS-STING pathway and the influence on macrophage migration and invasion, which provides further evidence for the treatment of rheumatoid arthritis with TP combined with MTX.

Abstract

Intestinal inflammation, often driven by microbial dysbiosis and infections, remains a significant health challenge with limited effective treatments. Identifying probiotic strains with anti-inflammatory properties and elucidating their mechanisms is essential for developing novel therapeutic strategies. This study investigates the molecular mechanisms by which E. hirae-a lactic acid bacterium (LAB) isolated from Ningxiang piglets with low diarrhea incidence-alleviates E. coli-induced intestinal inflammation.

Abstract

Ulcerative colitis (UC) is colonic inflammation associated with increased production of pro-inflammatory cytokines, oxidative stress, and disturbances of immune responses. Mitragynine is the most abundant active alkaloid in Mitragyna speciosa (kratom) and may have anti-inflammatory, antioxidant, and antispasmodic properties. In this study, we investigated the palliative effects of mitragynine in kratom leaf extract on the symptoms of UC.

Abstract

Inflammatory bowel disease (IBD) is a chronic inflammatory condition of the intestine, for which no cure currently exists. The gut microbiota play a critical role in ameliorating IBD, and Eubacterium limosum has emerged as a potential probiotic with anti-inflammatory properties. However, the specific anti-inflammatory effects of E. limosum and the underlying mechanisms remain largely unexplored.

Abstract

Peripartum dairy cows are highly susceptible to metabolic disorders, with ketosis being the most prevalent postpartum disease associated with rumen microbial dysbiosis and systemic inflammation. However, the mechanisms by which microbial alterations compromise rumen epithelial integrity remain poorly understood. Using peripartum cows with ketosis as a model, we demonstrated that perturbations of rumen microbiota disrupt tryptophan metabolism, resulting in pronounced depletion of indole-3-acetic acid (IAA). The loss of IAA-producing taxa (Lactobacillus and Bifidobacterium) contributed to reduced IAA levels and epithelial barrier dysfunction, whereas enrichment of proinflammatory taxa (Candidatus Saccharimonas and Mycoplasma) was associated with exacerbated epithelial inflammation. In vitro, IAA supplementation activated the AhR/IL-22 signaling pathway, promoting bovine rumen epithelial cells (BRECs) regeneration and restoring barrier integrity. These findings identify the microbiota-IAA-AhR/IL-22 axis as a key regulator of rumen epithelial homeostasis and suggest that targeting this pathway represents a promising strategy to prevent metabolic disorders in dairy cows.

Abstract

Plant-derived proteolytic enzymes are widely used in biochemistry and food processing. For example, bromelain, ficin, and papain serve as meat tenderizers, while cardosin A is used as a plant-based rennet in cheese production. A cysteine protease has been identified in grapevine products such as fresh juice, wine, and wine vinegar. The enzyme (CYSP) shares sequence similarity with RD21A from Arabidopsis and other plant cysteine endopeptidases. This mass spectrometry-based study investigated the proteolytic activity of wine and wine vinegars, including both commercial and laboratory-prepared samples. Vinegar types examined included white wine vinegar, balsamic vinegar, and red wine vinegar produced through grape juice fermentation and spontaneous acetification. Protease activity and specificity were assessed using substrates such as pure protein standards, casein, and minced beef proteins. The activity assay also included spectrophotometry with azocasein and electrophoresis followed by gelatin zymography. Results confirmed the presence of CYSP and indicated aspartic protease involvement. Digestion experiments coupled with mass spectrometry identified peptide cleavage sites, with C-terminal residues frequently being L, F, R, Y, K, and D/E. This pattern reflects the combined specificity of CYSP and pepsin-like proteases. Notably, CYSP activity was higher in wine, whereas aspartic protease activity predominated in vinegar.

Abstract

Ulcerative colitis represents a chronic inflammatory bowel disease with limited therapeutic options due to inadequate efficacy and adverse effects of current treatments. This study investigated the therapeutic potential of Beaucarnea recurvata leaf extract (BRLE) against ulcerative colitis using integrated computational and experimental approaches to address the need for safer, multi-targeted interventions. Phytochemical profiling was performed using UPLC-ESI-MS/MS analysis. Network pharmacology and molecular docking predicted therapeutic targets and mechanisms. In vivo validation employed an acetic acid-induced ulcerative colitis rat model with BRLE treatment at 100, 200, and 400 mg/kg doses, evaluating clinical parameters, histopathology, oxidative stress markers, inflammatory cytokines, and protein expression. UPLC-ESI-MS/MS revealed diverse bioactive compounds including steroidal saponins, triterpenes, and flavonoids. Network pharmacology identified 24 hub targets, and molecular docking revealed strong binding affinities (-6.5 to -9.1 kcal/mol) between BRLE compounds and inflammatory proteins including EGFR, SRC, STAT3, and AKT1. BRLE at 200 mg/kg significantly improved disease activity, restored glutathione levels, reduced malondialdehyde, normalized IL-10 and TNF-α levels, downregulated EGFR, SRC, STAT3, and AKT1 expression, and enhanced mucosal healing with reduced inflammatory infiltration. BRLE demonstrates significant anti-inflammatory, antioxidant, and tissue-protection effects through multi-target mechanisms, representing a promising therapeutic intervention for ulcerative colitis treatment. Further studies in chronic models, pharmacokinetic assessments, and clinical trials are needed to support its translation into therapeutic use.

Abstract

The pro-apoptotic protein Bax is a key apoptosis regulator, as its activity is the main driver of mitochondrial outer membrane permeabilization. Bax is therefore tightly regulated, both by protein-protein interactions and post-translational modifications, such as phosphorylation. Although less studied, N-terminal acetylation has also been implicated in Bax regulation: disruption of the NatB N-terminal acetyl transferase complex in both yeast and MEFs increases Bax mitochondrial localization, although increased translocation is not sufficient to trigger its activation. Using the well-established model of heterologous expression of human Bax in yeast, we further investigated its regulation by N-terminal acetylation. We found that the sensitivity of Bax-expressing cells to acetic acid is greatly enhanced in a strain lacking the yeast NatB catalytic subunit (Nat3p). We propose that the Bax-induced cell death process shifts to a regulated necrosis in this strain due to autophagy inhibition. Furthermore, we show that the protective role of Bcl-xL against acetic acid-induced cell death of Bax-expressing yeast cells requires Nat3p. We speculate that Nat3p modulates the function of pro-death and pro-survival proteins, ultimately affecting both the levels and mode of cell death. These findings may have implications for the development of novel therapeutic strategies targeting human diseases associated with cell death dysfunction.

Abstract

Acute hepatic injury (AHI) is a sudden onset of hepatic inflammation, a key contributor to the progression of diabetes and other disorders. Diabetes mellitus also increases the risk of liver illnesses associated with inflammatory disorders. According to recent studies, Saroglitazar (SAR), originally developed for the treatment of hyperglycemia and dyslipidemia, has also demonstrated notable anti-inflammatory properties. In our search for a prime therapeutic approach for inflammatory liver disorders in diabetic patients, we investigated the effects of SAR on thioacetamide (TAA)-induced AHI in rats. In order to investigate possible interactions between SAR and thioredoxin-interacting protein (TXNIP), this research utilized a multistep methodology that included prediction of computational targets, network analysis, molecular docking, and experimental verification. Findings revealed the anti-inflammatory potential of SAR, presumably ascribed to its inhibition of the NLRP3 signaling pathway by inhibiting TXNIP, an NLRP3 inflammasome upstream regulator. Furthermore, SAR inhibited the priming signal brought on by NFκB stimulation and the succeeding inflammasome components, cleaved caspase-1, GSDMD, IL-1β, and IL-18. As a result, SAR demonstrated anti-pyroptotic properties in the injured liver. Moreover, SAR exhibited potential antiapoptotic effects, as indicated by decreased Bax levels, decreased tissue expression of cleaved caspase-3, and increased BCL2 levels. Improvements in liver function, oxidative stress markers, liver histology, and the liver weight-to-body weight ratio all supported these findings. In conclusion, SAR demonstrates potential as a preventive treatment for inflammatory liver disorders. To render these preclinical findings into efficient techniques for enhancing hepatic function, more research is required, particularly in the context of diabetes.

Abstract

This study aimed to investigate the relationship between texture analysis parameters of lacrimal gland activity and dry eye disease (DED) in participants undergoing Gallium-68 labelled Prostate-Specific Membrane Antigen ligand Positron Emission Tomography / Computed Tomography ([68 Ga]Ga-PSMA-11 PET/CT) imaging for prostate cancer (PCa).

Abstract

Escherichia coli strains are widely utilized as cell factories for recombinant protein production. However, acetate overflow remains a significant challenge that negatively impacts both biomass yield and protein expression. Here, we evaluated a previously engineered E. coli K 12 BW25113 strain with pka and arcA deletions (RV04) for the expression of a single-chain variable fragments (scFv) derived from 4D5MOC-B, a monoclonal antibody that binds to epithelial cell adhesion molecule (EpCAM) as a biologically important marker for tumor immunotherapy. According to our results, RV04 strain demonstrated a significant growth advantage over both BW25113 and BL21 strains. In minimal M9 medium, RV04 exhibited a maximum cell density that was 44% higher than wild‑type and 11% higher than BL21. In enriched M9 medium, RV04 achieved a remarkable maximum specific growth rate (µ_max) of 0.775 ± 0.003 h⁻¹ and a maximum cell density of 2.1095 ± 0.0205, even under metabolic load. Regarding acetate accumulation, RV04 fully eliminated acetate accumulation within 24 h, whereas BW25113 accumulated acetate up to 0.521 g L- 1 under the same minimal medium conditions. Similarly, in enriched M9 medium, RV04 maintained significantly lower acetate levels (1.65 g L- 1 at 24 h) compared to BW25113 (3.99 g L- 1), despite increased biomass and protein production. These results confirm that RV04 can control acetate overflow more efficiently than the wild type under both minimal and enriched conditions. The combination of using the genetically modified strain and medium enrichment strategy resulted in significantly increased recombinant protein production. In LB medium, RV04 produced 5% more protein than BL21 and 44.8% more than the wild‑type, while in enriched synthetic M9 medium, it outperformed BL21 and BW25113 by 7.1% and 59.5%, respectively. Furthermore, RV04 demonstrated markedly enhanced protein expression compared to other commercial strains; it produced approximately 33.8% more protein than SHuffle, 145.7% more than Rosetta, and over sevenfold more than Origami B. Our findings demonstrate that RV04 effectively mitigates acetate overflow, enhances growth, and substantially increases the recombinant protein titer under both minimal and enriched culture conditions. These features make RV04 a strong candidate for large-scale industrial bioprocessing operations.

Abstract

Temporomandibular joint osteoarthritis (TMJOA) is a pathological condition marked by subchondral bone remodeling. Osteoarthritis can lead to TMJ pain, nevertheless, the relationship between nociceptive mechanisms and subchondral bone in TMJOA still unclear.

Abstract

The anti-inflammatory ability of a novel peptide-compound conjugate, 2IP5MP-CW, synthesized by fusing 2-isopropyl-5-methylphenoxy acetic acid with cysteine and tryptophan, was investigated in this study. 2IP5MP-CW markedly inhibited the growth of Escherichia coli and Staphylococcus aureus and exhibited potent antioxidative activity by suppressing reactive oxygen species production and mitigating oxidative DNA damage induced by lipopolysaccharide (LPS). In RAW 264.7 macrophages, 2IP5MP-CW significantly reduced LPS-induced TNF-α production and suppressed nitric oxide (NO) generation as well as the expression of inflammation-related proteins iNOS and COX-2. The compound also attenuated the phosphorylation of ERK, JNK, NF-κB p65, IKKβ, and IκBα, thereby blocking the nuclear translocation of phosphorylated NF-κB p65. In an LPS-induced mouse model of pulmonary inflammation, 2IP5MP-CW treatment reduced the phosphorylation of ERK, JNK, NF-κB p65, IKKβ, and IκBα in LPS-challenged lung tissue. Furthermore, it markedly attenuated TNF-α production and downregulated iNOS and COX-2 expression. Histological examination further confirmed that 2IP5MP-CW alleviated inflammatory cell infiltration and tissue injury. Collectively, these findings demonstrate that 2IP5MP-CW exerts potent anti-bacterial, anti-oxidative, and anti-inflammatory effects by suppressing LPS-induced TNF-α and NF-κB/MAPK signaling, highlighting its therapeutic potential against LPS-mediated inflammatory lung injury. KEY MESSAGES: A novel peptide-compound conjugate, 2IP5MP-CW, was designed and synthesized. 2IP5MP-CW exhibited in vitro and in vivo anti-bacterial activity against E. coli and S. aureus. 2IP5MP-CW showed anti-oxidative activity by suppressing ROS production. 2IP5MP-CW demonstrated anti-inflammatory activity in LPS-stimulated RAW 264.7 cells. 2IP5MP-CW exhibited anti-inflammatory efficacy in a pulmonary inflammation mouse model.

Abstract

Castleman's disease is a rare lymphoproliferative disorder that can mimic malignancy in cancer patients. This study presents a rare case of Castleman disease coexisting with prostate cancer, highlighting the potential value of gallium 68-labelled prostate-specific membrane antigen-11 ([⁶⁸Ga]Ga-PSMA-11) positron emission tomography/computed tomography (PET/CT) in distinguishing Castleman disease from prostate cancer metastasis.