This study evaluates the antimicrobial properties of Juniperus phoenicea L. against multidrug-resistant microbes, highlighting potential synergistic effects of combining the J. phoenicea L. newly synthesized nanoparticles with conventional antibiotics and anticancer agents. Juniperus phoenicea L. ethanolic and ethyl acetate extracts were prepared. Antibacterial activity against a panel of multidrug-resistant microbes showed the ethanolic extract as superior to the ethyl acetate extract, with Acinetobacter baumannii being the most susceptible organism (minimum inhibitory MIC = 12.5 µg/mL). Phytochemical profiling of the ethanolic extract revealed predominant components, namely: Apigenin-7-glucoside, α-pinene, and Cedrol. Network pharmacology-based analysis suggested the potent effect of these compounds as anticancer and antibacterial agents. J. phoenicea L. nanoparticles were synthesized by a combined hot-melt dispersion and homogenization approach. Characterization yielded a zeta vesicle size of 98.6 nm, PDI 0.23, zeta potential + 48.5 mV, EE
Alfalfa biomass contains significant carbohydrate fractions underutilized in animal feed. This research optimized the enzymatic hydrolysis of Alfalfa biomass and evaluated single-cell protein (SCP) production using Candida utilis and Komagataella pastoris. Hydrolysis kinetics in this study showed biphasic sugar release, with enzymatic optimization increasing monomeric sugar yield from 18.5
Salmonella Typhimurium and Salmonella Choleraesuis display distinct clinical manifestations, yet the molecular basis for their differential inflammatory potential, particularly via outer membrane vesicles (OMV), are not fully understood. Here, we examined the role of FliC in the differential inflammatory responses induced by S. Typhimurium UK-1 and S. Choleraesuis C78-3 and their corresponding OMV. In vivo, UK-1 infection triggered significantly elevated serum levels of IL-1β and IL-6 compared with C78-3 infection, a difference that was also observed following OMV administration. Isogenic fliC deletion mutants, which lacked both flagella and OMV-associated flagellin, were successfully generated. While wild-type bacteria and their OMV robustly activated the NLRC4 inflammasome pathway in bone marrow-derived macrophages (BMDMs), resulting in caspase-1 cleavage and IL-1β maturation, the ΔfliC mutants and their OMV exhibited markedly diminished activity. Notably, recombinant FliC proteins from the two serovars, despite sharing only 77
The NLRP3 inflammasome is an important susceptibility factor in inflammatory bowel disease (IBD), yet its specific mechanisms in modulating gut microbiota during intestinal inflammation remain unclear. This study observed significant upregulation of NLRP3 inflammasome in colonic tissues from patients with active ulcerative colitis and a murine dextran sulfate sodium (DSS)-induced colitis model. Nlrp3 deletion markedly attenuated disease severity, evidenced by improved histopathology, reduced pro-inflammatory cytokines, and increased microbial alpha diversity. Notably, Nlrp3 deficiency mitigated the depletion of beneficial SCFA-producing taxa, specifically enriching Dubosiella and Rikenellaceae_RC9_gut_group, which was associated with the restoration of fecal propionate and butyrate levels. Furthermore, treatment with the SCFA-producing probiotic Clostridium butyricum alleviated colitis in wild-type mice but provided no additional benefit in Nlrp3 - deficient mice and failed to reduce inflammation in mice treated with the NLRP3 activator nigericin. Collectively, these results suggest that the therapeutic efficacy of C. butyricum may be influenced by the activation status of host NLRP3, underscoring the complex relationship between NLRP3 signaling and microbiota-mediated regulation in IBD pathogenesis.
The pig gut microbiota plays an important role in host nutrition, intestinal physiology, and health, and cultivation remains essential for isolating viable bacteria for functional studies and microbial resource development. However, practical guidance for preserving region-specific porcine intestinal contents prior to cultivation remains limited, and most previous evidence has been derived from human stool samples or DNA-focused preservation studies. Here, we compared fresh processing under three cryopreservation conditions to assess the recovery of the culturable bacterial fraction and the composition of cultured bacterial communities from the ileum, cecum, colon, and rectum of a healthy Berkshire boar after one month of storage at − 80 °C. Combining anaerobic cultivation, colony counting, and 16 S rRNA gene sequencing of cultured biomass, we found that fresh samples consistently yielded the highest recovery of culturable bacteria across intestinal regions. Cryopreservation reduced colony counts across intestinal regions, with unprotected freezing (P0) yielding the lowest recovery. Counts decreased from 1.19 × 109 to 2.73 × 108 CFU/g in the ileum, from 8.26 × 108 to 3.33 × 108 CFU/g in the cecum, from 4.87 × 108 to 2.10 × 108 CFU/g in the colon, and from 4.37 × 108 to 1.24 × 108 CFU/g in the rectum, with the largest absolute reduction observed in the ileum. Compared with freezing without cryoprotection, glycerol-containing formulations generally retained broader cultured communities and better preserved taxonomic representation. These results suggest that immediate processing is the most favorable approach for culture-based studies of the pig gut microbiota, while cryoprotectant-assisted freezing represents a practical alternative for short-term storage when fresh cultivation is not possible. Overall, this pilot study provides region-specific evidence to guide short-term preservation of porcine intestinal samples for bacterial isolation and future culture-based microbiome research.
The dengue virus (DENV) remains a significant global health threat, and the currently available vaccines exhibit concerns regarding safety, efficacy, and cost-effectiveness. Microalgae represent an innovative method for producing proteins that do not occur naturally. They are Generally Recognized As Safe (GRAS), easily scalable, and capable of inducing post-translational modifications in eukaryotes. This study illustrates the effective heterologous expression of recombinant dengue virus capsid–premembrane–envelope (C-prM-E) structural antigen constructs representing all four DENV serotypes (DV1–DV4) in the red microalga Porphyridium purpureum through Agrobacterium tumefaciens-mediated transformation. Gene constructs containing capsid-premembrane-envelope (CprME) polyprotein fragments with C-terminal epitope tags were cloned into pCAMBIA1301 vectors and subsequently introduced into P. purpureum. PCR amplification confirmed the successful chromosomal integration of all four serotype constructs, yielding the anticipated 2.5-kb amplicons. The SDS-PAGE analysis revealed protein bands at around 85 kDa, consistent with the expected molecular weight of the processed CprME-derived proteins. We employed both anti-DENV-2 and anti-FLAG antibodies in a western blot assay to demonstrate that the recombinant antigens were identical and interacted with the immune system across all serotypes. These findings indicate that P. purpureum is an effective system for expressing DENV antigens, paving the path for the development of cost-effective, plant-derived dengue vaccine candidates. This microalgal platform may facilitate the large-scale production of tetravalent dengue vaccines, particularly in regions where dengue is prevalent and resources are scarce.
Astragalus membranaceus is a widely utilized medicinal and edible herb, serving as immunostimulant, metabolic regulator, anticancer agent, and hepatoprotectant agent. However, its protective effect against chemotherapy-induced intestinal mucositis (CIM) remains poorly elucidated. The study aimed to investigate the therapeutic efficacy and mechanism of Astragalus membranaceus extract (AME) in alleviating CIM using Drosophila melanogaster and C57BL/6J mouse models. The mechanisms of AME were evaluated via multi-omics approaches, including transcriptomic profiling, 16 S rRNA sequencing, and targeted metabolomics. The expression levels of the TLR4/NF-κB pathway and glutathione metabolic pathway were analyzed by RT-qPCR and immunofluorescence. AME supplementation significantly mitigated CPT-11-induced systemic and intestinal damage in adult flies and mice. Mechanistically, AME markedly down-regulated the TLR4/NF-κB pathway and up-regulated glutathione metabolism. AME also modulated gut microbiota composition, increased relative abundance of Lactobacillus, unidentified_Lachnospiraceae, and Ruminococcus, and decreased abundance of Streptococcus, Bacteroides, and Alistipes. Meanwhile, AME decreased the levels of glutamine (Gln), lysine (Lys), phenylalanine (Phe), proline (Pro), and valine (Val), while increasing threonine (Thr). In conclusion, AME alleviates CIM via regulating gut microbiota-related innate immunity and amino acid metabolism.
Yeast surface display is a versatile microbial engineering strategy for recombinant protein presentation and oral antigen delivery. In this study, a conserved influenza hemagglutinin stem antigen (mini-HA) was displayed on the surface of Saccharomyces cerevisiae to evaluate the feasibility and stability of a yeast-based oral delivery system. A rationally engineered mini-HA derived from influenza A H1N1 was anchored to the yeast cell surface using the Aga1p-Aga2p display system. Efficient surface localization was confirmed by flow cytometry and confocal microscopy. The displayed antigen remained stably anchored following exposure to simulated gastric fluid and after heat inactivation at 60 °C, indicating favorable stability of the yeast surface display system under acidic and thermal stress conditions. Oral administration of the engineered yeast induced antigen-specific systemic IgG and mucosal secretory IgA responses in mice. In addition, antigen-specific lymphocyte proliferation and increased expression of IFN-γ and T-bet were observed, suggesting activation of both humoral and cellular immune responses. No significant adverse effects on body weight, serum biochemical parameters, or tissue histopathology were detected. These findings support the feasibility of using S. cerevisiae surface display for stable presentation and oral delivery of a conserved influenza antigen. The favorable stability and immunogenicity of the system support further development of yeast-based recombinant antigen delivery platforms.
Cervical cancer remains a leading cause of cancer-related death. Integrating TCGA/GTEx and GWAS, we identified PRKCZ as a risk factor (OR = 1.26, p = 0.008). PRKCZ was upregulated in tumors, and its knockdown inhibited SiHa and HeLa proliferation. Using gutMGene, we identified Escherichia coli as a microbe computationally predicted to be associated with PRKCZ based on literature-curated data. Public 16S rRNA data (45 patients vs. 30 controls) showed reduced gut microbial diversity and higher abundance of the genus Escherichia-Shigella (which includes Escherichia coli) in patients. Parallel trends of higher Escherichia-Shigella abundance and PRKCZ expression were observed in separate cohorts. Importantly, because the microbiome and transcriptomic data derive from independent, non‑paired cohorts, no direct microbiota–gene association was measured, and no causal or mechanistic relationship can be inferred. These hypothesis-generating observations suggest a hypothetical association among gut Escherichia coli, PRKCZ, and isoquercetin, warranting direct validation in paired multi‑omics cohorts and functional studies.
The increasing antimicrobial resistance of Staphylococcus epidermidis, a leading opportunistic pathogen causing device‑related and nosocomial infections, underscores an urgent need for novel anti‑staphylococcal agents. Nifuratel, a nitrofuran derivative currently used for urogenital tract infections, represents a promising candidate for drug repurposing. In this study, nifuratel showed minimal inhibitory concentrations (MICs) ranging from 0.5 to 2 μg/mL against S. epidermidis, and at 1 μg/mL in Mueller‑Hinton broth reduced the growth turbidity of RP62A and ATCC 12228. At 2×MIC, nifuratel reduced viable counts of S. epidermidis to the detection limit within 12 h. No significant resistance was observed after 15 serial passages at subinhibitory concentrations, and the single‑step induction assay also indicated extremely low resistance potential by nifuratel. Nifuratel exhibited a post‑antibiotic effect similar to ciprofloxacin and showed synergistic or partially synergistic effects with oxacillin or gentamicin. Moreover, nifuratel significantly reduced the fluorescence of the proton motive force ‑related probes DiSC3(5) and BCECF‑AM. In vivo, nifuratel significantly reduced bacterial load and inflammatory exudate in abscesses and wounds. Pathological staining further confirmed reduced inflammatory cell infiltration and accelerated wound healing. Collectively, nifuratel exerts potent bactericidal activity against S. epidermidis by disrupting the transmembrane potential and transmembrane proton gradient of the proton motive force.
Non-cystic fibrosis bronchiectasis (NCFB) is characterized by persistent Pseudomonas aeruginosa (P. aeruginosa) colonization and uncontrolled pulmonary inflammation, while the underlying metabolic regulatory mechanism remains poorly understood. This study aimed to explore the potential analyze correlations of the HSD17B8 metabolic axis in lung inflammatory injury related to P. aeruginosa in NCFB. Transcriptome sequencing was performed on peripheral blood and tissue samples from NCFB patients and healthy controls to screen differentially expressed genes. Two-sample Mendelian randomization integrated with protein quantitative trait locus data was applied to evaluate the potential association between candidate genes and NCFB susceptibility. The expression of HSD17B8 in clinical samples was further validated by RT-qPCR. Meanwhile, a rat model of P. aeruginosa-infected pulmonary inflammation was established to investigate the influences of estrone intervention on lung pathology, pro-inflammatory cytokines (IL1β, IL6), and steroid metabolite homeostasis. A total of 249 overlapping differentially expressed genes were identified. Mendelian randomization analysis indicated that downregulated HSD17B8 expression was nominally correlated with elevated NCFB risk (OR 0.76, 95
The presence of carbaryl in the environment and agricultural products poses uncertain risks to both ecosystems and human health. In the present study, a carbaryl-degrading Serratia marcescens strain A4 was isolated from orchard soil, demonstrating a degradation rate of 83.16
Human immunodeficiency virus type 1 (HIV-1) remains a major global public health challenge due to lifelong persistence, latency, and the emergence of drug resistance. Natural bioactive compounds are increasingly being explored as alternative or adjunct antiviral strategies. In this study, we evaluated the antiviral efficacy and cytotoxicity of a synergistic pharmaceutical composition comprising three phycobiliproteins: C-phycocyanin (C-PC), B-phycoerythrin (B-PE), and allophycocyanin (APC), derived from cyanobacteria and marine algae. Individual and combinatorial effects were assessed against HIV-1 using TZM-bl cells and peripheral blood mononuclear cells (PBMCs). A Box–Behnken experimental design was employed to identify optimal synergistic ratios. Among fifteen combinations initially screened at a fixed concentration of 50 µg/mL, five exhibited strong antiviral activity. The optimized formulation (CBA: C2; 73.28
Pseudomonas aeruginosa is a pathogen responsible for healthcare-associated infections, and the emergence of carbapenem-resistant strains (CRPA) poses significant clinical challenges. Phage therapy is increasingly recognized as a promising strategy against drug-resistant bacteria. This study aimed to identify a phage capable of effectively lysing CRPA. Using CRPA clinical isolates (PA19N) as hosts, phages were isolated and purified from water samples via the double-layer agar plate assay. A P. aeruginosa phage, designated Henuyfy19N, was successfully isolated. Transmission electron microscopy revealed that it possesses a head with diameter of approximately 87.94 ± 1.32 nm and a tail length of about 130.25 ± 3.65 nm, and it forms plaques with clear, distinct margins. The phage exhibited an optimal multiplicity of infection (MOI) of 0.1 against PA19N, with a latent period of 10 min, and a rise period of 30 min. It demonstrated strong stability under various temperature and pH conditions but was sensitive to UV irradiation. Whole-genome sequencing showed that the phage has a 93,413 bp linear dsDNA genome with a G + C content of 49.42
Chronic obstructive pulmonary disease (COPD) is a progressive inflammatory respiratory disorder characterized by persistent airflow limitation and systemic inflammation. Increasing evidence suggests that intestinal microbiota influence respiratory diseases through the gut–lung axis. Probiotics have emerged as potential therapeutic agents capable of modulating immune responses and restoring microbial balance. This study investigated the protective effects of Lactobacillus rhamnosus on pulmonary function, inflammatory responses, and gut microbiota composition in a cigarette smoke and lipopolysaccharide-induced rat model of COPD. Male Wistar rats were randomly assigned to Control, COPD model, Low-dose L. rhamnosus, High-dose L. rhamnosus, and Aminophylline groups. COPD was induced through chronic cigarette smoke exposure combined with intratracheal lipopolysaccharide administration. Pulmonary function parameters, including forced expiratory volume in 0.3 s (FEV₀.₃) and forced vital capacity (FVC), were measured. Lung and colon tissues were examined histopathologically, and serum concentrations of TNF-α, IL-6, IL-1β, and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA). Gut microbiota composition was analyzed using 16 S rRNA gene sequencing. COPD model rats showed significant impairment of pulmonary function, marked increases in serum TNF-α, IL-6, and IL-1β, reduced IL-10 levels, and pronounced lung and colon tissue injury compared with Control animals. Treatment with L. rhamnosus significantly improved pulmonary function, reduced pro-inflammatory cytokine levels while restoring IL-10, and attenuated histopathological damage, with the High-dose group exhibiting greater protective effects than the Low-dose group. Microbiome analysis demonstrated modest alterations in bacterial community composition, although most diversity indices and bacterial taxa did not differ significantly among groups. These findings suggest that L. rhamnosus attenuates COPD-associated inflammation and tissue injury and may exert beneficial effects through modulation of host inflammatory responses and gut microbial composition. Further studies are required to clarify the underlying mechanisms linking probiotic supplementation and the gut–lung axis in COPD.
Dental caries is a prevalent microbial disease, with Streptococcus mutans as a major pathogen. Increasing antibiotic resistance highlights the need for alternative, non-antibiotic strategies. This study evaluated the antimicrobial and anti-virulence potential of nano-hypericin (nHyp)-mediated antimicrobial photodynamic therapy (aPDT) combined with Lactobacillus casei-derived postbiotic (PSLC). nHyp was synthesized and characterized by transmission electron microscopy, UV–Vis spectroscopy, and dynamic light scattering, confirming quasi-spherical nanoparticles ( 47 nm). PSLC was purified via acid precipitation and chemically profiled by gas chromatography–mass spectrometry, revealing diketopiperazines, fatty acids, hydrocarbons, and phenolic compounds. Planktonic S. mutans were treated with nHyp (1.9–1000 µg/mL) ± blue laser irradiation (450 ± 5 nm, 1–5 min), alone or combined with PSLC at 1/2 × MIC (32 µg/mL). Bacterial viability was measured by colony forming unit (CFU)/mL, and gtfB expression quantified by qRT-PCR. nHyp-mediated aPDT induced dose- and time-dependent reductions in bacterial viability (up to 81
In recent years, probiotics have attracted increasing attention due to their roles in gut health regulation, antimicrobial activity, and metabolic benefits. Given the high-fat dietary characteristics of mink, their intestinal microbiota represents a potential source of functionally valuable lactic acid bacteria. In this study, we evaluated the probiotic properties, safety profile, and functional potential of Lactiplantibacillus plantarum ZZM01 isolated from mink intestine. ZZM01 exhibited good tolerance to simulated gastrointestinal conditions, with survival rates of 71.92
The increasing emergence of multidrug-resistant (MDR) pathogens poses a significant challenge to global healthcare, necessitating the development of alternative antimicrobial strategies. In the present study, cranberry extract was evaluated for its antimicrobial, antibiofilm, antioxidant, and anti-inflammatory activities against a panel of clinically relevant microorganisms, including Escherichia coli, Acinetobacter baumannii, Klebsiella pneumoniae, Pseudomonas aeruginosa, and methicillin-resistant Staphylococcus aureus (MRSA). The extract demonstrated notable antimicrobial activity, with minimum inhibitory concentration (MIC) values ranging from 32 to 512 µg/mL, exhibiting relatively higher efficacy against Gram-positive bacteria. Significant antibiofilm activity was observed at sub-inhibitory concentrations, where cranberry extract reduced biofilm formation by up to 75–80
Inflammatory skin diseases (ISDs) represent a significant global health burden, yet the role of the human microbiome in their pathogenesis remains unclear. This study employed a comprehensive two-sample Mendelian randomization (MR) framework to investigate potential causal associations between multi-site microbiota (3117 oral, 412 gut, and 150 skin microbial features) and five major ISDs: psoriasis, atopic dermatitis, acne, seborrheic dermatitis, and urticaria. Genetic instruments were selected from large-scale genome-wide association studies (GWAS) of microbiota composition and ISD outcomes. The inverse variance weighted (IVW) method served as the primary analytical approach, supplemented by MR-Egger regression, weighted median, and MR-PRESSO for sensitivity analyses. Multiple testing was controlled using the false discovery rate (FDR) method. Our analyses identified numerous suggestive associations between specific microbial taxa, metabolic pathways, and ISD risk. Notably, shared microbial signatures were observed across multiple ISDs, suggesting potentially shared microbiome-associated pathogenic pathways. These results provide genetic evidence supporting a role for the microbiome in ISD susceptibility and highlight potential microbial targets for future therapeutic development.
The gut–joint axis has been increasingly implicated in osteoarthritis (OA), yet the causal contribution of specific gut microbes and their downstream molecular mechanisms remain unclear. We employed a multi-stage, two-sample Mendelian randomization (MR) framework. This approach utilized summary-level data from large-scale genome-wide association studies (GWAS) for gut microbiota, plasma proteins, and OA. The analysis involved three steps: (1) identifying gut microbial taxa with potential causal associations with OA risk; (2) screening of pyroptosis-related proteins using pQTL data; and (3) performing mediation analysis to evaluate potential intermediate mechanisms. We also performed transcriptomic analysis of human cartilage and in vitro experiments in chondrocytes to support the biological relevance of our findings. Our MR analysis identified Gordonibacter pamelaeae as a suggestive microbial taxon inversely associated with OA risk. Among 11 candidate pyroptosis-related proteins, only Phospholipase C Gamma 1 (PLCG1) showed a significant inverse association with OA. MR analysis further suggested that G. pamelaeae was positively associated with genetically predicted PLCG1 levels. Mediation analysis indicated that PLCG1 partially mediated the association between G. pamelaeae and OA. Consistent with these findings, PLCG1 mRNA levels were reduced in human OA cartilage. Furthermore, our in vitro experiments demonstrated that PLCG1 knockdown enhanced IL-1β-induced inflammatory and pyroptotic responses in chondrocytes. This study suggests a potential link involving gut microbiota and OA through PLCG1-related signaling. PLCG1 may act as a context-dependent regulator that limits excessive inflammatory responses under stress conditions. These findings refine the current understanding of the gut–joint axis and may help identify potential targets for OA intervention.
