The changes in viscosity and the concentration of ONOO- in mitochondria can effectively reflect the physiological and pathological status of cells. Therefore, the development of effective fluorescent probes for the sensing of viscosity and the concentration of ONOO- in mitochondria has great significance. In this article, a mitochondrial targeted fluorescent probe named Mito-RP was synthesized for the dual responsive sensing of viscosity and ONOO- by introducing pyridine ring and phenylboronic acid ester structure into 4-dimethylamino-cinnamaldehyde with long conjugated chain structure as the parent material. Mito-RP exhibits 600 folds fluorescence enhancement of viscosity in the red-light channel at 700 nm, with pyridine cation as the mitochondrial anchoring group. Simultaneously, Mito-RP appears excellent selectivity towards ONOO- using boronic acid esters as response sites. A new ratio fluorescence analysis method was constructed based on the linear correlation between the emission intensity ratio of Mito-RP at 616 nm/700 nm and the concentration of ONOO-. The linear range is 0.05-33 μM and the detection limit is 9.2 nM. Meanwhile, Mito-RP successfully monitored the changes in viscosity during lipopolysaccharide induced inflammation and rapamycin induced mitochondrial autophagy in HeLa cells. In addition, Mito-RP has also achieved visual imaging of intracellular exogenous/endogenous ONOO-. These studies provide a novel method for in-depth investigation of mitochondrial function and its role in diseases.

Tire wear particles (TWPs), as a pervasive environmental pollutant, pose significant risks to aquatic ecosystems. This study investigates the effects of small (HS) and large (HL) TWPs produced by heavy vehicles on zebrafish, focusing on physiological, microbial, and transcriptomic levels, as well as their intergenerational consequences, under short-term (15 days) and long-term (90 days) exposure. Short-term exposure to small particles (HS15) significantly reduced body width and triggered widespread oxidative stress, while long-term exposure to large particles (HL90) increased gut weight and decreased gill weight, reflecting respiratory and digestive disruptions. Tissue-level analyses revealed that smaller particles accumulated more readily in internal organs, whereas larger particles caused localized physiological stress. Gut microbiota profiling indicated a marked decline in microbial diversity, compositional shifts, and network simplification, with HL15 enriched in Acinetobacter and xenobiotic metabolism pathways, and HS15 exhibiting Proteobacteria-dominated dysbiosis and enrichment of LPS biosynthesis genes. Liver transcriptomics revealed group-specific responses: HL15 exposure activated innate immunity via the NOD-MAPK axis, while HS15 induced atypical PI3K-NF-κB signaling, potentially linked to microbial LPS. Notably, all TWP-exposed groups showed enrichment of the herpes simplex virus 1 (HSV-1) infection pathway, suggesting a conserved antiviral-like host response. Transgenerational effects were evidenced by impaired growth and significant downregulation of GH/IGF signaling and upregulation of apoptotic genes in offspring, despite only subtle transcriptomic changes in long-term exposed parents. These findings underscore the importance of particle size, exposure duration, and microbiota-gut-liver axis interactions in mediating TWP toxicity and highlight potential transgenerational risks associated with environmental microplastic exposure.

SALMONELLA: as an important food-borne zoonotic pathogen, is found in soil and processing environment by human or animal feces, causing serious public health problems. Quaternary ammonium compounds (QACs) disinfectants are widely used in hospitals, livestock farms and food processing sites because of their low toxicity and broad-spectrum disinfection. However, sub-lethal levels of QACs disinfectants can induce bacteria to develop tolerance to disinfectants and cross-resistance to other antimicrobial agents. The acquired resistance will undoubtedly pose a threat to the prevention of antimicrobial resistance. In this study, Salmonella enterica SE211 was induced by the sub-inhibitory concentration and sub-lethal concentration of dodecyl dimethyl ammonium bromide (DDAB) in vitro. Following exposure to DDAB, the strains showed increased resistance to DDAB, doxycycline, amphenicols and fluoroquinolones, and increased sensitivity to colistin drugs. Phenotypic experiments showed that the induced strains exhibited changes in efflux pump activity, biofilm formation ability, motility and membrane characterization. Next-generation sequencing revealed mutations in induced strains involved in LPS-related genes (msbA, lptDE) and cationic antimicrobial peptide (CAMP) resistance-related genes (phoQ, pmrD). Transcriptome sequencing (RNA-seq) analysis revealed up-regulation of efflux pump genes and down-regulation of CAMP resistance, LPS and peptidoglycan related genes. Our study provided a theoretical basis for the potential consequences of disinfection failures and environmental residues of QACs disinfectants on the evolution of antibiotic resistance in salmonella. Furthermore, the induction of colistin sensitivity in salmonella by DDBA resulted in the emergence of collateral sensitivity, which offered a new strategy for drug combination applications to prevent the rise of colistin-resistant superbugs.

Acute lung injury (ALI) is associated with a high mortality rate and requires effective treatment. Tanshinone IIA (T) and Osthole (O) exhibit anti-inflammatory effects and have been used to protect against lipopolysaccharide (LPS)-induced lung injury in mice. However, the combined effects of T and O on lung injury protection and their potential protective mechanisms have not been studied.

To assess the protective effects of TO on LPS-induced ALI in mice and BEAS-2B cell injury and to investigate the potential mechanisms underlying these protective effects.

Models of ALI induced by LPS were established. The assessment encompassed the viability of BEAS-2B cells, cell count, myeloperoxidase (MPO) activity, protein content, as well as IL-6 and TNF-a levels in bronchoalveolar lavage fluid (BALF). Additionally, malondialdehyde (MDA), reactive oxygen species (ROS), and glutathione (GSH) levels in mouse lung tissue were measured. The effects of TO were assessed using immunofluorescence (IF), immunohistochemistry (IHC), Western Blot (WB), RT-PCR, and ELISA. Statistical analysis involved one-way ANOVA and t-test.

TO administration led to a significant reduction in lung edema (W/D), MDA, ROS, GSH, and superoxide dismutase (SOD) levels compared to the individual T or O groups, alleviating LPS-induced ALI. TO also significantly attenuated lung tissue damage, reduced inflammatory response, decreased Fe2+ and 4-HNE levels, and increased GPX4, SLC7A11, and Nrf2 gene expression in mice. Ultimately, TO alleviated ferroptosis in LPS-induced ALI by activating Nrf2 expression, and no markedly adverse reactions were observed.

TO alleviates LPS-induced ALI and effectively treats against LPS-induced ALI.

ADP-heptose-LPS heptosyltransferase I (encoded by waaC gene) is a crucial enzyme in the lipopolysaccharide (LPS) synthesis, maintaining the stability of LPS and cell walls in numerous Gram-negative bacteria. Vibrio mimicus is an epidemic pathogen that threatens aquatic animals and human health, resulting in high morbidity and mortality after infecting the fish. Currently, the role of waaC gene in V. mimicus is still unclear. In this study, waaC gene deletion and complementation strains of V. mimicus were constructed. Our results show that ΔwaaC exhibited a rough phenotype on LB agar, with elevated exocrine protein and a disordered cell wall observed by the electron microscope. Transcriptomic analysis revealed that following the deletion of waaC gene, the expression of 224 genes was drastically upregulated, while the expression of 229 genes was significantly downregulated. These genes involve various pathways, including material transport, metabolism, and environmental adaptation. The tricarboxylic acid cycle (TCA cycle) and pyruvate metabolism are the primary pathways affected by waaC gene. Our phenotypic analysis is consistent with transcriptomic findings, indicating that the decreased pathogenicity of ΔwaaC is related to the effect of waaC gene on these genes, which negatively impacts V. mimicus growth, motility, adhesion, and biofilm formation while enhancing its self-aggregation. The virulence of the ΔwaaC was 103-fold lower than that of the wild strain. Moreover, the expression levels of inflammatory cytokine and pro-apoptotic genes in epithelioma papulosum cyprini (EPC) cells were dramatically upregulated during the ΔwaaC infection. These results provide valuable insights into revealing the pathogenic mechanism of V. mimicus, further bringing more options for the candidate deletion targets of the V. mimicus attenuated vaccine.

This study synthesizes carbon dots derived from crude extracts of purple sweet potato (CPP-CDs) and evaluates its anti-inflammatory effects in a lipopolysaccharide (LPS) -induced acute inflammation model. Characterization revealed that CPP-CDs possess a uniform spherical structure and excellent photoluminescent properties. In vitro, CPP-CDs significantly inhibited the expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), reduced the accumulation of reactive oxygen species (ROS), suppressed pyroptosis, and facilitated the polarization of macrophages from the M1 phenotype to the M2 phenotype. In vivo, CPP-CDs significantly improved the survival rates of LPS-treated mice, mitigated tissue damage, and suppressed the levels of pro-inflammatory cytokines. Mechanistic studies indicated that CPP-CDs exert anti-inflammatory effects through the inhibition of the TLR4/NF-κB signaling pathway and the modulation of the NLRP3 inflammasome. Additionally, CPP-CDs exhibited excellent biocompatibility, with no significant toxicity observed in mice. This study provides strong evidence supporting the application of CPP-CDs as a novel anti-inflammatory material, highlighting their potential for acute inflammation treatment and expanding the possibilities for the development of carbon-dot-based anti-inflammatory therapies.

Lipopolysaccharide (LPS)-induced inflammation of lung tissues triggers irreversible alterations in the lung parenchyma, leading to fibrosis and pulmonary dysfunction. While the molecular and cellular responses of immune and connective tissue cells in the lungs are well characterized, the specific epithelial response remains unclear due to the lack of representative cell models. Recently, we introduced human embryonic stem cell-derived expandable lung epithelial (ELEP) cells as a novel model for studying lung injury and regeneration.

ELEPs were derived from the CCTL 14 human embryonic stem cell line through activin A-mediated endoderm specification, followed by further induction toward pulmonary epithelium using FGF2 and EGF. ELEPs exhibit a high proliferation rate and express key structural and molecular markers of alveolar progenitors, such as NKX2-1. The effects of Escherichia coli LPS serotype O55:B5 on the phenotype and molecular signaling of ELEPs were analyzed using viability and migration assays, mRNA and protein levels were determined by qRT-PCR, western blotting, and immunofluorescent microscopy.

We demonstrated that purified LPS induces features of a hybrid epithelial-to-mesenchymal transition in pluripotent stem cell-derived ELEPs, triggers the unfolded protein response, and upregulates intracellular β-catenin level through retention of E-cadherin within the endoplasmic reticulum.

Human embryonic stem cell-derived ELEPs provide a biologically relevant, non-cancerous lung cell model to investigate molecular responses to inflammatory stimuli and address epithelial plasticity. This approach offers novel insights into the fine molecular processes underlying lung injury and repair.

Gram-negative bacterial cell envelope consists of a surface-exposed lipid bilayer (outer membrane or OM) that serves as a permeability barrier to maintain the cellular integrity. Beneath the OM is the periplasmic space that harbours peptidoglycan (PG), a highly cross-linked mesh-like glycan polymer closely encasing the inner membrane (IM). During growth of a bacterium balanced synthesis of the envelope components is required to maintain the cellular integrity, of which little is known. In this study, we identify sanA, an ORF of unknown function encoding a predicted IM-anchored protein as a factor contributing to balanced synthesis of PG in E. coli. Absence of SanA increased the rate of nascent PG strand incorporation, and restored growth and viability to several mutants defective in either cell division or cell elongation. Detailed mutant analysis of sanA showed that it is defective in the envelope barrier properties. Interestingly, overexpression of the periplasmic endopeptidases that cleave the cross-links of the PG mesh was able to alleviate the phenotypes of sanA mutant implying the envelope defects are due to alterations in the PG sacculus. Additionally, a SanA variant (SSDsbA-SanA) targeted to the periplasm, complemented the SanA- phenotypes suggesting it functions in the periplasmic phase of the PG synthesis. Further, we find that SanA functions independently of its paralog, ElyC, known to regulate the synthesis of enterobacterial common antigen (ECA), a surface polysaccharide found in the cell envelopes of most enteric bacteria. Overall, our results suggest a role for SanA in the maintenance of optimal PG synthesis, providing evidence for the existence of an additional layer of regulation in Gram-negative cell envelope biogenesis.

Elastin and elastin-derived compounds are promising biomaterials due to their biological activity, unique natural crosslinks, and ability to mimic native tissue properties. Solubilized elastin peptides retain the bioactivity of elastin and are more suitable for wound care applications than the insoluble form. Chemically solubilized elastins have shown advantageous effects in skin regeneration in humans. Here, five solubilized elastins were prepared via chemical (stepwise and continuously hydrolyzed with oxalic acid - OxA-st-ELN and OxA-ELN, or with potassium hydroxide - KOH-ELN), enzymatic (Enz-ELN), or combined (Combi-ELN) methods. OxA-st-ELN had the largest molecular weights (MWs) fragments, while Enz-ELN and Combi-ELN yielded the smallest. The effects of elastin preparations were evaluated on primary human cells - dermal fibroblasts and macrophages. In fibroblast assays, Enz-ELN induced elastin, collagen, and fibrillin-2 protein deposition, while other preparations exhibited levels comparable to the control. α-smooth muscle actin (SMA) expression remained low across all conditions. Continuous oxalic acid hydrolysis simplified the traditional stepwise approach while maintaining bioactivity. Macrophage studies showed chemical hydrolysates preserved the M0-like subtype, while Enz-ELN promoted a pro-inflammatory M1-like phenotype, and Combi-ELN had mixed effects. OxA-ELN and KOH-ELN appeared to be the most promising options for developing biomaterial dermal scaffolds that support tissue regeneration in vivo.

It has been established that inflammatory factors are involved in the formation of pathological scars. Therefore, pathological scars are regarded to be highly associated with chronic inflammation, whereas what factors contribute to this inflammation remains unclear.

To confirm that bacterial colonization is involved in the formation of pathological scars, and to reveal that the persistent inflammatory response mediated by macrophages due to bacterial colonization promotes scar formation.

This study included 23 normal skin controls and 58 untreated pathological scar samples. To detect the presence of bacteria in surgically-excised scar samples and alterations of histology, as well as bacteria-associated gene levels, histological staining, immunoelectron microscopy, microbiological and cell culture and molecular biology detection methods were employed. The PICRUSt2 tool and BugBase were employed to identify pathways, genes, and phenotypic differences.

We found that in pathological scars, bacteria were widely distributed both extracellularly and intracellularly, with intracellular bacteria primarily located in the cytoplasm of macrophages and fibroblasts. A total of 2,260 bacterial species were detected in pathological scars, primarily from the Clostridiales, Burkholderiales, Actinomycetales, and Bacteroidales orders. Moreover, the pathogenicity and motility of colonizing bacteria were positively correlated with the degree of scar hyperplasia and invasiveness. The lysates of four clinically-relevant bacterial species had differential effects on the secretion of inflammatory cytokines from macrophages. When treated macrophage supernatant was added to fibroblasts, collagen secretion was dysregulated, and fibroblast differentiation into myofibroblasts prominently increased. In rat scar model, the expression of inflammatory factors and growth factors in the scar tissue was increased, which activated the TGF-β/Smad signaling pathway, resulting in the increasing of α-SMA.

Persistent activation of macrophages by tissue-colonizing bacteria may be a key factor in promoting inflammatory response and dysregulated collagen deposition in pathological scars, offering a potential new strategy for preventing and treating pathological scars.

Here, astaxanthin production in Escherichia coli was systematically improved step by step. By introducing the additional copy of CrtZ and fusion complex of CrtZ and CrtW, astaxanthin content in cells increased from 0.10 mg/g to 0.16 mg/g and 0.63 mg/g DCW, respectively. Remolding the astaxanthin gene cluster by replacing the PanCrtE by HpGGPPS3-1 and the fusion of CrtZ and CrtW increased astaxanthin content to 1.98 mg/g DCW. Further selecting the productive host and optimizing culture conditions dramatically increased astaxanthin content to 3.61 mg/g DCW. Subsequently, the fed-batch fermentation achieved the maximum yield of astaxanthin at 509.58 mg/L with the productivity of 7.72 mg/L/h and 5.91 mg/g DCW, covering 98.17 % of detected carotenoids. The chirality analysis assigned the same isomer of astaxanthin extracted from our fermentation system and Haematococcus pluvialis. Moreover, the radical and superoxide anion scavenging activity analysis revealed that astaxanthin achieved in this study performed better than natural astaxanthin extracted from H. pluvialis and chemical synthetic astaxanthin. This study provides a step-by-step example for bioengineering improvement of natural products in E. coli with high purity and activity.

Chronic inflammation plays a central role in the pathogenesis of lung diseases including asthma, long COVID, chronic obstructive pulmonary disease (COPD), and lung cancer. Lipopolysaccharide (LPS) is a potent inflammatory agent produced by Gram-negative bacteria and also found in cigarette smoke. Our earlier study revealed that the intranasal exposure of A/J mice to LPS for 7 days altered gene expression levels in alveolar Type II epithelial cells (AECIIs), which serve as precursors to lung adenocarcinoma and are also preferentially targeted by SARS-CoV-2. In the present work, we employed a comprehensive multi-omics approach to characterize changes in DNA methylation/hydroxymethylation, gene expression, and global protein abundances in the AECIIs of A/J mice following the sub-chronic exposure to LPS and after a 4-week recovery period. Exposure to LPS led to hypermethylation at regulatory elements within the genome such as enhancer regions and expression changes in genes known to play a role in lung cancer tumorigenesis. Changes in protein abundance were consistent with an inflammatory phenotype and also included tumor suppressor proteins. Integration of the multi-omics data resulted in a model where LPS-driven inflammation in AECIIs triggers epigenetic changes that, along with genetic mutations, may contribute to lung cancer development.

The rising demand for noninvasive portable point-of-care (POC) sensors for on-site detection of biomarkers reflects a shift toward personalized healthcare and real-time diagnostics. Lipopolysaccharide (LPS) is a key sepsis biomarker, a bacteria-borne endotoxin that can induce fatal conditions, such as septic shock, if left undetected. Timely bedside detection of LPS is critical for the appropriate intervention and treatment of sepsis. In this work, we report a highly sensitive electrochemical sensor chip designed for the selective detection of LPS, which is compatible with a portable analyzer for on-site detection. The sensor is fabricated using functionalized CNT (fCNT) and copper(I) oxide nanoparticles (Cu2O). Functionalized with the LPS-specific aptamer, the sensor demonstrated remarkable selectivity and sensitivity toward LPS. It achieved a limit of detection (LOD) of 10 ag mL-1 and exhibited a linear detection range from 10 ag mL-1 to 10 ng mL-1. The device detected LPS across various samples, including food samples such as mayonnaise and fruit juices, as well as a clinical sample, whole blood. These results underscore its potential for practical application in food quality assurance and clinical diagnostics, offering real-time analysis capabilities.

Needle-based blood draws or phlebotomy practice in clinics for centuries, often causing pain, discomfort, and inconvenience. Here, a wearable photonic device is presented by integrating a microlens array (MLA) and an optic microneedle array (OMNA) functionalized with immunobinding for safe and needle-free biomarker sampling and detection. The MLA-integrated OMNA amplifies and transmits LED light at 595 nm into skin through the OMNA, bypassing the light-absorbing melanin in the epidermal layer, and evenly distributing it in the capillary-enriched dermis independent of the skin colors. The 595 nm light is absorbed by hemoglobin (Hb) and oxygen-Hb within the capillaries, triggering thermal dilation of capillaries without damaging them or causing petechiae. The light illumination remarkably increases in the concentrations of various blood biomarkers in the skin through biomarker extravasation. These biomarkers bound specifically to the capture antibodies on OMNA with each microneedle covalently immobilized with one specific antibody. The OMNA is extensively modified to amplify the immunobinding signals and achieve sensitivity superior to that of enzyme-linked immunosorbent assay (ELISA) kits. As proof of concept, the functionality of the prototype for minimally invasive sampling and precise multiplexed blood biomarker detection in two mouse models is validated to quantify acute inflammation and specific antibody production.

The immune system orchestrates the hypothalamus-pituitary-adrenal (HPA) axis response to stress. However, the impact of invariant natural killer T (iNKT) cell activation on stress-induced glucocorticoid levels remains poorly understood. Alpha-galactosylceramide (α-GalCer), a specific agonist for iNKT cells, activates iNKT cells to produce inflammatory cytokines including interleukin (IL)-4 and interferon (IFN)-γ. Our findings indicate that treatment with α-GalCer 3 hours before acute restraint stress suppressed the elevation of adrenocorticotropic hormone (ACTH) but did not affect the increase in corticosterone (CORT) in mice. However, treatment with α-GalCer 24 hours prior to restraint stress did not alter the rise in ACTH but reduced the increase in CORT by about half. This dissociation between stress-induced ACTH and CORT levels suggests an intra-adrenal regulation of HPA axis responses to acute stress following α-GalCer treatment. We further found that administration of α-GalCer enhances lipid utilization within adrenocortical cells and elicits a hyperresponsive reaction to ACTH stimulation. Mechanistically, IL-4 elevates the expression of type II 3β-hydroxysteroid dehydrogenase/isomerase (HSD3B2) and scavenger receptor class B type I (SRBI) protein in adrenocortical cells, thereby facilitating ACTH-induced glucocorticoid release. Additionally, we observed that acute stress amplifies both α-GalCer-induced IL-4 and IFN-γ production as well as liver injury. Our findings not only elucidate the mechanistic basis underlying interactions between immunity and stress but also highlight potential targets for therapeutic intervention.

Nontyphoidal and enteric fever serovars of Salmonella enterica display distinctive interactions with serum antibodies and the complement system, which initiate the host immune response to invading microbes. This study examines the contributions of lipopolysaccharide O-antigen (O-ag) and the S. Typhi Vi polysaccharide capsule to serum resistance, complement activation and deposition, and immunoglobulin (Ig) binding in nontyphoidal S. enterica serovar Typhimurium and the enteric fever serovars S. Typhi and S. Paratyphi A. Although all three serovars are resistant to serum killing, S. Typhi and S. Paratyphi A exhibit lower levels of Ig binding, complement binding and complement activation compared to S. Typhimurium. In S. Typhimurium, WzzB-dependent long O-antigen (L O-ag) production with 16-to-35 repeating O-ag units, and FepE-dependent very long O-antigen (VL O-ag) production with over 100 repeating O-ag units, are required for serum resistance but do not prevent IgM binding or complement deposition. S. Typhi lacks VL O-ag, but its production of Vi capsule inhibits IgM binding and complement deposition, while acting in concert with L O-ag to resist serum killing. In S. Paratyphi A, L O-ag production is deficient due to a hypofunctional WzzB protein, but this is compensated by greater quantities of VL O-ag, which are required for serum resistance. Restoration of WzzB function by exchange with the S. Typhimurium or S. Typhi wzzB alleles can restore L O-ag production in S. Paratyphi A but decreases VL O-ag production, resulting in increased IgM binding. Replacement of the S. Paratyphi A O2-type polysaccharide with the S. Typhi O9 polysaccharide further increases IgM binding of S. Paratyphi A, which enhances complement activation but not complement deposition. Lastly, a gene duplication of rfbV in S. Paratyphi A is necessary for higher levels of VL O-ag and resistance to complement deposition and antibody binding. Collectively, these observations demonstrate fundamental differences between nontyphoidal and enteric fever Salmonella serovars in their interactions with innate immune effectors. Whereas nontyphoidal S. Typhimurium elicits, exploits and withstands the host acute inflammatory response, the enteric fever serovars S. Typhi and S. Paratyphi A evade it by limiting antibody recognition and complement activation and deposition.

Amylase trypsin inhibitors (ATIs) potentially play a role in irritable bowel syndrome (IBS) and non-celiac wheat sensitivity (NCWS). These cereal-derived inhibitors are suspected to bind to the TLR4-MD2-CD14 complex and trigger intestinal pro-inflammatory responses, but confirmation through more extensive cell line studies is required. In this study, an amylase trypsin inhibitors enriched fraction (AEF) was prepared and characterized. Then, AEF binding potential to TLR4-MD2-CD14 was investigated using the human TLR4 reporter cell line HEK-BlueTM. The method took into account the presence of lipopolysaccharides (LPS) using Polymyxin B (PMB) to block LPS binding to TLR4. Proteinase K was also used to hydrolyze AEF proteins and eliminate their induced response. The cell line experiments showed that PMB treatment of AEF reduced the binding signal by 92 %. Complete hydrolysis of the protein by Proteinase K doubled the TLR4 activation signal and might indicate that protein-LPS complexation reduced LPS's ability to activate the TLR4-receptor. These finding underline the need for future work to consider the non-protein part in cell assays, especially the LPS bias. Altogether, these results indicated that LPS activates TLR4-MD2-CD14 and challenges ATIs' intestinal inflammation capacity contributing in irritable bowel syndrome and non-celiac wheat sensitivity.

Bacteriophage (phage) are promising novel antimicrobials but a key challenge to their effective implementation is the rapid emergence of phage resistance. An improved understanding of phage-host interactions is therefore needed. The Anderson phage typing scheme differentiates closely related strains of Salmonella enterica serovar Typhimurium (S. Typhimurium) based on sensitivity to a panel of phage preparations. Switches in phage type are indicative of changes in phage sensitivity and inform on the dynamics of phage interaction with their host bacteria. We investigated the molecular basis of switches between the relatively phage sensitive S. Typhimurium DT8 and phage resistant DT30 strains that are present in the same phylogenetic clade. DT30 strains emerged from DT8 strains predominantly by deletion of a genomic region affecting the wzy locus encoding an O-antigen polymerase. The deletion site was flanked by two perfect direct repeats designated attL and attR. During broth culture in the presence of a typing phage that used O-antigen as primary receptor the Δwzy genotype increased in frequency compared with culture in the absence of phage and removal of attL prevented deletion of the wzy locus. Co-culture of S. Typhimurium DT8 with a strain lacking wzy resulted in reversion of the latter to wild type. We propose a model in which reversible deletion of the wzy locus enables recovery of S. Typhimurium DT8 following predation by phage that use O-antigen as their primary receptor. This was consistent with ancestral state reconstruction of DT8 and DT30 phylogeny that supported a model of reversible transition from DT8 to DT30 in natural populations.

Bovine mastitis remains a major concern for dairy farmers, mainly because of its effect on the economy of their activity and on animal welfare. Because Escherichia coli is considered a major mastitis pathogen, the diversity of E. coli strains isolated from mastitis cases has been studied for decades, with the aim to discover new ways to fight this infection. With the recent advances in whole-genome sequencing, a detailed view of the peculiarities of mastitis E. coli strains has emerged. This review aims to bring together the knowledge garnered over the years with the more recent results of whole-genome analyses. Whereas the concept of a mammary pathogenic E. coli has been proposed, because a common set of virulence genes cannot be identified among mastitis E. coli strains, we prefer the use of mastitis-associated E. coli (MAEC), with MAEC being more an "ecotype" rather than a "pathotype." Indeed, data available so far suggest that a common feature of MAEC would rather be an enrichment in fitness capabilities that makes them well-suited for survival and rapid adaptation to changing biotopes in the mammary gland, which we qualify as intramammary ecotopes.

This systematic review investigates gut bacterial diversity and composition in patients with Obstructive Sleep Apnea-Hypopnea Syndrome (OSAHS) and examines how these changes may contribute to cardiovascular complications.

A comprehensive search was conducted in PubMed, Web of Science, and Scopus up to March 2025. After removing duplicates, titles and abstracts were screened by two reviewers, and full texts were assessed for inclusion. Data extraction on study characteristics and outcomes was performed. Methodological quality was evaluated using the Joanna Briggs Institute checklist. α-diversity was assessed using richness and diversity indices, while β-diversity examined community structure differences. Meta-analysis was conducted using standardized mean differences (SMD) and confidence intervals (CIs), and heterogeneity was assessed with the Cochrane I2 test.

The review included 18 studies (16 adults, 2 pediatrics) examining the gut microbiome in OSAHS. Meta-analysis revealed significant reductions in α-diversity indices (Shannon, Chao1, observed species, ACE) in OSAHS patients, while Simpson's index showed no difference. β-diversity analyses showed distinct gut microbiome differences in OSA. Key differential bacteria included Bacteroides, Proteobacteria, Faecalibacterium, Ruminococcaceae, Megamonas, Oscillibacter, Dialister, Roseburia, and Lachnospira. Study quality was medium to high.

OSAHS is associated with significant gut microbiome alterations, including a reduction in beneficial bacteria and an increase in LPS-producing bacteria, leading to intestinal barrier dysfunction. These changes may contribute to systemic inflammation and elevate the risk of cardiovascular diseases.

Lipopolysaccharide (LPS) is an essential component of the cellular envelope of Gram-negative bacteria and contributes to antibiotic resistance and pathogenesis. Proper localization of LPS at the outer membrane is facilitated via seven distinct LPS transport (Lpt) proteins that bridge the inner and outer membranes. Mature LPS diffuses into the membrane cavity of the inner membrane ABC transporter LptB2FGC through a lateral gate formed by the LptF and LptG transmembrane (TM) helices. The TM helix of LptC intercalates within the LPS entry point and has been shown to regulate the ATPase activity of LptB2FG and contribute to thermal stability. Determination of the LptB2FGC open state structure revealed the location of the LptC TM helix within the membrane complex. However, in the closed state structure, the LptC TM helix is unresolved, suggesting the helix may be displaced from the lateral gate prior to or upon closure of the cavity. To determine the conformational states of the LptC TM helix in the open and closed LptB2FGC conformations, we utilized site-directed spin labeling in combination with both continuous wave electron paramagnetic resonance (EPR) and double electron electron resonance (DEER) spectroscopies to investigate the LptC TM helix and linker region. These data indicate that the LptC TM helix undergoes a rigid body movement away from the central LptB2FG cavity upon cavity closure. The findings presented here will support structure-based drug design optimization of recently discovered antibiotics that bind LptB2FG and occlude the LptC TM helix from the lateral gate.

Endotoxin contamination, as one of the most significant challenges in recombinant protein production by Escherichia coli, represents a critical biosafety concern and greatly hinders the biomedical application of recombinant proteins. Conventional methods, such as extreme-condition inactivation and chromatography-based separation, are plagued by issues including protein denaturation, low efficiency, and operational complexity in endotoxin removal. In this work, we developed a novel magnetic bifunctional endotoxin removal nano-agent (MagBER) with a multi-layered structure, consisting of a superparamagnetic Fe3O4 core, a mesoporous TiO2 intermediary layer, and an outer shell functionalized with boronic acid groups. This dual-functional design significantly enhances endotoxin removal efficiency through the TiO2 layer and boronic acid groups, ensuring stable endotoxin clearance performance even in high-salt environments and complex biological matrices. MagBER exhibits reusability while maintaining protein structural integrity. Moreover, MagBER has been successfully employed for endotoxin removal in various proteins, establishing it as a promising and sustainable solution for endotoxin clearance in biopharmaceutical applications.

Osteomyelitis poses substantial therapeutic challenges due to the prevalence of multidrug-resistant bacterial infections and associated inflammation. Current treatment regimens often rely on a combination of corticosteroids and antibiotics, which can lead to complications and impede effective bacterial clearance. In this study, we present CADP-10, a Cecropin A-derived peptide, designed to target methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Escherichia coli (MRE), while simultaneously addressing inflammatory responses. CADP-10 self-assembles into nanobacterial net (NBacN) that selectively identify and bind to bacterial endotoxins (LPS and LTA), disrupting membrane integrity and depolarizing membrane potential, which culminates in bacterial death. Importantly, these NBacN are bound to LPS and LTA from dead bacteria, preventing their engagement with TLR receptors and effectively blocking downstream inflammatory pathways. Our assessments of CADP-10 demonstrate good biosafety in both in vitro and in vivo models. Notably, in a rabbit osteomyelitis model, CADP-10 eliminated MRSA-induced bone infections, mitigated inflammation, and promoted bone tissue regeneration. This research highlights the potential of CADP-10 as a multifunctional antimicrobial agent for the management of infectious inflammatory diseases.

UDP-glucuronic acid (UDP-GlcUA) is a nucleotide sugar essential for various biological processes in many organisms, and its excess within the cell can disrupt cellular functions. In Cryptococcus, mutations in the UXS1 gene which encodes an enzyme responsible for converting UDP-GlcUA into UDP-xylose, result in excessive accumulation of UDP-GlcUA and confer resistance to the antifungal drug 5-fluorocytosine. Here, we demonstrate that elevation of UDP-GlcUA affects several cellular processes in Cryptococcus gattii, including growth rate, ability to grow under various stress conditions and resistance to fluorinated pyrimidine analogs. RNA-seq analyses of the uxs1Δ mutant identify three acid protease genes, notably PEP401, that are differentially expressed. The absence of PEP401 in the uxs1Δ background significantly reduces UDP-GlcUA levels and reverts all the phenotypes of the uxs1Δ mutant to the wild-type characteristics. High levels of UDP-GlcUA not only regulate expression of PEP401 at RNA and protein levels but also enhance the proteolytic activity of total protein extracts in a PEP401-dependent manner, establishing a functional link between nucleotide sugar metabolism and proteolytic regulation. Moreover, the UDP-GlcUA transporter gene, UUT1, can further modulate the levels of UDP-GlcUA in the uxs1Δ pep401Δ double mutant and manifests drug resistance phenotypes observed in the uxs1Δ mutant. Collectively, these findings reveal a previously unrecognized regulatory network that links UDP-GlcUA metabolism to protease-mediated cellular processes and the transport of UDP-GlcUA. This interaction provides a foundation for targeting nucleotide sugar metabolism and protease regulation in the development of enhanced therapeutic strategies against cryptococcosis.

Although toxicity of alcohol toward the intestines and immunity is mentioned, there might be different effect of alcohol in a low and a high dose and the rodent model development using a simple SHIRPA binary score night be useful. Hence, a low and high dose of alcohol (6.30 and 1.26 g/kg/day) were administered in might for 16 weeks before determination of several parameters. As such, the peak blood alcohol concentration (BAC) of low and high dose of alcohol were approximately at 0.05 and 0.15%, respectively, at 1 h post-administration, which correlated with SHIRPA score at 1.8 ± 0.8 and 7.2 ± 0.6, respectively. After 16 wk of administration, a significant liver injury in high-dose alcohol was indicated by liver enzymes, liver weight, histology score, apoptosis, and hepatic accumulation of triglyceride (TG) and oxidative stress (malondialdehyde; MDA) with reduced anti-oxidant (glutathione). Meanwhile, low-dose alcohol demonstrated only elevated apoptosis with increased TG and MDA in liver tissue. Leaky gut from both dose of alcohol was also demonstrated by FITC-dextran, endotoxemia, serum beta glucan, and reduced occludin. However, bacterial abundance (microbiome analysis) of the feces from small bowel of high-dose alcohol, but not the low dose, was different from the control (increased Alitipes spp. with reduced Lachnospiraceae). In conclusion, both low- and high-dose alcohol induced leaky gut, while only the high-dose caused gut dysbiosis and alcohol damaged mitochondria but enhanced glycolysis in enterocytes and macrophages. Leaky gut might be more sensitive than dysbiosis to determine alcohol-induced intestinal injury.

Bacterial lysate proteins (BLPs) serve as potential immunostimulants, recognized by pattern recognition receptors (PRRs) on immune cells, eliciting a robust immune response. In this study, THP-1 macrophages were treated with varying doses of BLPs derived from Streptococcus pyogenes (SP), Streptococcus agalactiae (SA), and Serratia marcescens (SM). The results showed significant increases (p  < 0.05) in pro-inflammatory cytokines such as TNF-α, IL-1β, IL-6, IL-12, granulocyte macrophage-colony stimulating factor (GM-CSF), eotaxin, and macrophage inflammatory protein (MIP)-1α, except for 5 µg of all BLPs for TNF-α and eotaxin, and 5 µg of SP for IL-12 production. No significant differences were found between the corresponding doses of SP and SA or SP and SM, except for GM-CSF in all doses, while SA and SM only showed a difference at the 5 µg dose for GM-CSF. Furthermore, there were no significant differences between the 10 and 20 µg doses of all BLPs, indicating that doses higher than 10 µg do not significantly enhance the pro-inflammatory response. Combination doses of SP + SM and SA + SM did not show significant differences, except for IL-1β, suggesting no synergistic effect. Cytotoxicity was observed to increase with higher BLP concentrations in a dose-dependent manner, with combinations of SP + SM and SA + SM exhibiting greater cytotoxicity than the individual BLPs. Proteomic analysis of BLPs identified immunostimulatory proteins, including heat shock proteins (HSPs; ClpB, DnaK, and GroEL), metabolic enzymes (glyceraldehyde 3-phosphate dehydrogenase (GAPDH), enolase, and arginine deiminase (ADI)), and surface and secreted proteins (ESAT-6-like protein, CRISPR-associated endonuclease Cas9, OmpA, porin OmpC, and serralysin), which are involved in immune modulation, bacterial clearance, and immune evasion. This study underscores the potential of bacterial proteins as vaccine adjuvants or supplementary therapies; however, further research is essential to find a balance between immune activation and inflammation reduction to develop safer and more effective immunostimulants.

The genus Alistipes consists of anaerobic, Gram-negative bacteria with 13 species that colonize the entire gastrointestinal tract and are a serious health concern. They contribute to gut dysbiosis, intestinal inflammation, colorectal cancer, and depression.

To explore potential therapeutic targets and inhibitors, we filtered the core genome of Alistipes strains through subtractive genomics for non-host homology, gene essentiality, PPI, KEGG pathways, virulence, cellular localization, and druggability. The potential targets were docked against two drug-like libraries (ZINC, n = 11,993) and TCM (n = 36,043). ADMET profiling for best hits and MD simulation for apo/complex structures were performed, followed by physicochemical and pharmacokinetic evaluation and complex stabilities.

A set of 39 potential proteins was drastically reduced to only two targets after sequential data mining. The 3D structures of the selected targets (LpxA and KdsB) revealed good druggability scores. The top hits (ZINC85530940, ZINC05161112, ZINC95911713, and ZINC05566415) for both targets showed maximum H-bond interactions. The RMSD and RMSF values exhibited compactness with minimum fluctuation in ligand-bound complexes. The β-factor of ZINC05161112 at 327th residue and 352nd residue exhibited higher thermal instability, consistent with the RMSF results. The globularity of the complexes and apo structures remained consistent, whereas the LpxA complexes exhibited lower solvent-accessible surface area. For the KdsB, the surface area for ZINC5566415 increased significantly, with a steep decrease for ZINC95911713, establishing rather stable protein-ligand complexes. The results highlight the importance of identifying novel inhibitors and therapeutic targets. They are crucial for establishing better treatment regimes for human health and to aid in controlling the pathogenicity of Alistipes species.

Bacteria synthesize a plethora of complex surface-associated polysaccharides which enable them to persist and thrive in distinct niches. These glycans serve an array of purposes pertaining to virulence, colonization, antimicrobial resistance, stealth, and biofilm formation. The Wzx/Wzy-dependent pathway is universally the predominant system for bacterial polysaccharide synthesis. This system is responsible for the production of lipopolysaccharide (LPS) O-antigen (Oag), enterobacterial common antigen, capsule, and exopolysaccharides, with orthologs present in both Gram-negative and Gram-positive microbes. Studies focusing principally on Pseudomonas, Shigella, and Salmonella LPS Oag synthesis have provided much of the framework underpinning the biochemical and molecular mechanism behind polysaccharide synthesis via this pathway. LPS Oag production via the Wzx/Wzy-dependent pathway occurs through the stepwise activity of multiple key biosynthetic enzymes, including primarily the polymerase, Wzy, which is responsible for the Oag assembly, and the polysaccharide co-polymerase, Wzz, which effectively modulates the length of the glycan produced. In this review, we provide a comprehensive summary of the latest genetic, structural, and mechanistic data for the main protein candidates of the Wzx/Wzy-dependent pathway, in addition to an examination of their substrate specificities. Furthermore, we have reviewed recent insights pertaining to the dynamics/kinetics of glycan synthesis by this mechanism, including the interplay of the key proteins among themselves and in complex with their substrate. Lastly, we outline key gaps in the literature and suggest future research avenues, with the aim to stimulate ongoing research into this critical pathway responsible for the production of key virulence factors for numerous debilitating and lethal pathogens.

Zosurabalpin, a novel tethered macrocyclic peptide antibiotic, exhibits potent activity against Acinetobacter spp., particularly carbapenem-resistant Acinetobacter baumannii (CRAB). Zosurabalpin inhibits lipopolysaccharide (LPS) transport by targeting the LptB2FG protein complex, resulting in toxic LPS accumulation and bacterialdeath. This study investigates zosurabalpin's molecular specificity against Acinetobacter spp., its ineffectiveness against Klebsiella pneumoniae, and its potential efficacy against Shigella flexneri. Comparative analysis of LptB2FG sequences and structures, revealed significant differences in LptB2FG protein conformations, pocket geometry and electrostatic surface surrounding the binding pocket among the three species, which may influence zosurabalpin binding. Docking results for zosurabalpin showed lower binding affinities for K. pneumoniae and S. flexneri compared to Acinetobacter baylyi. Additionally, other zosurabalpin derivatives were tested showing improved binding affinities for K. pneumoniae but not for S. flexneri. These findings underscore the need for tailored zosurabalpin derivatives to enhance efficacy against a broader spectrum of Gram-negative bacteria.

The online version contains supplementary material available at 10.1007/s40203-025-00343-3.

The hybrid FowlTα1 peptide represents a promising biomolecule synthesized from two naturally occurring peptides, namely Fowlicidins (Fowl) and Thymosin α1 (Tα1). This particular peptide exhibits remarkable anti-inflammatory and antimicrobial properties and demonstrates the capacity to effectively interact with lipopolysaccharide (LPS), while simultaneously inducing minimal cytotoxicity and hemolytic repercussions. Despite its potential, the high cost of this peptide has limited its use. To overcome this limitation, the present study developed a cost-effective and biocompatible method for expressing the FowlTα1 peptide in Pichia pastoris (P. pastoris). We obtained a transgenic strain of the hybrid FowlTα1 peptide with a predicted molecular weight of 3.1 kDa. The FowlTα1 peptide was purified followed by reverse-phase high-performance liquid chromatography (RP-HPLC), yielding 7.2 mg with a purity of 98.2%. Furthermore, physiochemical and structural analysis revealed an amphipathic helical configuration that enhances bioactivity. Moreover, in LPS-stimulated HD11 macrophages, the hybrid FowlTα1 peptide significantly reduced the release of nitric oxide (NO), TNF-α, IL-6, and IL-1β in a dose-dependent manner (p < 0.05) and displayed robust antimicrobial activity against Escherichia coli (E. coli) compared to conventional antibiotic. Overall, the results of this study highlighted the production method and potential of the FowlTα1 peptide as a novel therapeutic agent for antimicrobial, anti-inflammatory, and anti-endotoxin applications.

Antibiotic-resistant Pseudomonas aeruginosa is a pathogen notorious for its resilience in clinical settings due to biofilm formation, efflux pumps, and the rapid acquisition of resistance genes. With traditional antibiotic therapy rendered ineffective against Pseudomonas aeruginosa infections, we explore alternative therapies that have shown promise, including antimicrobial peptides, nanoparticles and quorum sensing inhibitors. While these approaches offer potential, they each face challenges, such as specificity, stability, and delivery, which require careful consideration and further study. We also delve into emerging alternative strategies, such as bacteriophage therapy and CRISPR-Cas gene editing that could enhance targeted treatment for personalized medicine. As most of them are currently in experimental stages, we highlight the need for clinical trials and additional research to confirm their feasibility. Hence, we offer insights into new therapeutic avenues that could help address the pressing issue of antibiotic-resistant Pseudomonas aeruginosa, with an eye toward practical applications in future healthcare.

Central melanocortin signaling plays a critical role in maintaining energy homeostasis by regulating energy intake and expenditure, with impairment of this system closely related to metabolic diseases such as obesity. Among melanocortin receptor subtypes, melanocortin receptor 4 (MC4R) is the primary mediator of these effects within the central nervous system. Accumulating evidence suggests that MC4R contributes to stress-induced disruptions in feeding behavior and energy homeostasis. However, the precise neural mechanisms by which stress alters MC4R activity remain incompletely understood. In this study, we compared brain-wide c-Fos expression patterns induced by two distinct stress paradigms: lipopolysaccharide (LPS)-induced inflammatory stress and restraint stress in male mice, and further examined the involvement of MC4R-expressing (MC4R+) neurons in these stress conditions. We found that both stressors elicited c-Fos activation in brain areas associated with stress responses as well as feeding regulation. Notably, LPS-induced stress, but not restraint stress, selectively activated MC4R+ neurons in the central amygdala (CeA) and oval nucleus of the bed nucleus of stria terminalis (ovBNST). These results highlight the distinct recruitment of MC4R+ neurons during acute inflammatory stress in male mice, offering novel insights into the role of MC4R in the stress-induced imbalance of energy homeostasis depending on stressor types.

Kdo (3-Deoxy-d-manno-oct-2-ulosonic acid) is an essential sugar found in bacterial lipopolysaccharides with significant biomedical relevance. This study introduces an orthogonally protected 3-iodo-Kdo fluoride donor and demonstrates its coupling to a pre-synthesized β-(1→6)-linked N-acetylglucosamine disaccharide acceptor as an example. Nuclear magnetic resonance data indicates the presence of an intraresidue hydrogen bond in the distal glucosamine unit. Two complementary glycosylation approaches are explored with an emphasis on achieving high stereoselectivity and minimizing protecting-group manipulation. The orthogonal protection of 3-iodo Kdo fluoride donor offers insights into tailoring Kdo-based donors for specific biomedical applications. While yields vary depending on the approach, they are sufficient to demonstrate the donor's applicability. These findings enable the design of advanced glycomimetic constructs for therapeutic and vaccine research.

Malus hupehensis leaves (MHL), consumed as a daily beverage in Chinese folk tradition and recently recognized as a new food material, are abundant in polyphenols and bioactive compounds that demonstrate hypoglycemic, lipid-lowering, and anti-inflammatory effects. However, the antidiabetic mechanisms have not been fully elucidated. This study aimed to investigate the protective mechanisms of Malus hupehensis leaves' extract (MHLE) against type 2 diabetes mellitus (T2DM). The results showed that MHLE effectively ameliorated glucose and lipid metabolic abnormalities in db/db mice, and attenuated hepatic macrophage activation. Transcriptomic analysis of the liver revealed that MHLE primarily affects genes involved in inflammatory responses and inhibited the TLR4/MAPK pathway to reduce hepatic inflammation. Metagenomic sequencing identified changes in gut microbiota composition and showed that MHLE restored the abundance of Lachnospiraceae bacterium, Oscillospiraceae bacterium, and Clostridia bacterium while reducing the abundance of Escherichia coli, thereby ameliorating gut dysbiosis. The integrated regulation of metabolism, immune response, and the microbial environment by MHLE significantly alleviated symptoms of T2DM. This study offers strong scientific evidence for the potential use of MHL as a functional food.

The purpose of this study was to investigate the anti-inflammatory effects of Perilla Seed Extract (PSE) and its active ingredient on Inflammatory Bowel Disease (IBD) in vitro and in vivo. Thirty-two C57/BL mice were randomly divided into four groups (n = 8): control group (CON), PBS group, LPS group (LPS 3.5 mg/kg given intraperitoneally [ip] on day 7 of the study only), and PSE group (100 mg/kg orally daily + LPS ip at 3.5 mg/kg on day 7). Mice were euthanized 24 h after LPS administration. MODE-K cells were divided into five groups: control group (CON), LPS group (50 μg/mL LPS for 2 h), and PSE group (low dose, 25 μg/mL PSE + LPS; middle dose, 50 μg/mL PSE + LPS; high dose, 100 μg/mL PSE + LPS). In vivo, compared with the CON group, LPS revealed a significant decrease in the villus length-to-crypt depth ratio (p < 0.01) and goblet cell density per unit area (p < 0.01). Conversely, PSE administration resulted in a significant increase in the villus length-to-crypt depth ratio (p < 0.01) and goblet cell density (p < 0.01). LPS significantly increased the ROS content (p < 0.01), the secretion of inflammatory cytokines of IL-6 (p < 0.01), TNF-α (p < 0.01), and the mRNA expressions of HO-1 (p < 0.01). LPS significantly decreased the mRNA expressions of Occludin (p < 0.01) and Claudin1 (p < 0.01). In contrast, PSE treatment led to a marked decrease in ROS levels (p < 0.01), along with a reduction in the secretion of inflammatory factors IL-6 (p < 0.01) and TNF-α(p < 0.05), as well as the mRNA expressions of HO-1 (p < 0.01). Concurrently, PSE significantly increased the mRNA expressions of Occludin (p < 0.05) and Claudin1 (p < 0.01). In vitro, PSE treatment also significantly reversed LPS-induced inflammation, oxidation and tight junction-related factors. Network pharmacology identified 97 potential targets for PSE in treating IBD, while transcriptomics analysis revealed 342 differentially expressed genes (DEGs). Network pharmacology and transcriptomics analysis indicated that significant pathways included the PI3K-Akt signaling pathway, MAPK signaling pathway, and TNF signaling pathway, of which the PI3K-AKT pathway may represent the primary mechanism. In an in vivo setting, compared with the CON group, LPS led to a significant increase in the protein expression of p-PI3K/PI3K (p < 0.01) and p-AKT1/AKT1 (p < 0.01). Conversely, PSE resulted in a significant decrease in the protein expression of p-PI3K/PI3K (p < 0.01) and p-AKT1/AKT1 (p < 0.01). In vitro, compared with the LPS group, PSE also significantly blocked the protein expression of p-PI3K/PI3K (p < 0.01) and p-AKT1/AKT1 (p < 0.01). The chemical composition of PSE was analyzed using UPLC-MS/MS, which identified six components including luteolin (content 0.41%), rosmarinic acid (content 0.27%), α-linolenic acid (content 1.2%), and oleic acid (content 0.2%). Molecular docking found that luteolin could establish stable binding with eight targets, and luteolin significantly decreased the p-AKT1/AKT1 ratio (p < 0.01) compared to the LPS group in MODE-K cells. In summary, PSE demonstrates efficacy against IBD progression by enhancing intestinal barrier function and inhibiting inflammatory responses and oxidative stress via the PI3K/AKT signaling pathway, and luteolin's inhibition of AKT1 protein phosphorylation appears to play a particularly crucial role in this therapeutic mechanism.

Glucocorticoid-induced osteoporosis (GIOP) is the leading cause of secondary osteoporosis. Recently, the "bone-gut axis" theory has linked bone development with gut microbial diversity, community composition, and metabolites. Curcumin, a well-studied polyphenol, shows potential in mitigating bone loss and osteoporosis. Alendronate, a standard therapeutic agent for osteoporosis, serves as a positive control in this investigation. The study demonstrates the potency of curcumin in reducing bone loss and restoring bone mineral density, enhancing trabecular parameters notably through increased trabecular number, volume, and thickness and reduced bone marrow cavity size. Gut microbiome sequencing revealed that both curcumin and alendronate treatments similarly enhanced gut microbial diversity and altered microbiota composition, increasing beneficial bacteria (Akkermansia_muciniphila, Dubosiella_sp910585105, and Ruminococcus_sp910584195) while reducing harmful bacteria (Treponema_D_sp910584475 and Duncaniella_sp910584825). Furthermore, significant changes in serum levels of metabolites including raffinose, ursolic acid, spermidine, inosine, hypoxanthine, thiamine, and pantothenic acid were observed post-treatment with curcumin or alendronate. Importantly, these beneficial metabolites and microorganisms were negatively correlated with inflammatory cytokines. In conclusion, curcumin holds promise for use against GIOP by modulating the gut microbiome and serum metabolome as well as reducing systemic inflammation.

Most cells are decorated with distinct sugar sequences that can be recognized by carbohydrate-binding proteins (CBPs), such as antibodies and lectins. While humans utilize ten monosaccharide building blocks, bacteria biosynthesize hundreds of activated sugars to assemble diverse glycans. Monosaccharides absent in mammals are termed "rare" and are enriched in deoxy l-sugars beyond the "common" sugar l-fucose (l-Fuc) found across species. While immune proteins recognize microbial surfaces, there are limited probes to identify CBPs for the many rare sugars that may mediate these interactions. Here, we devise chemoenzymatic strategies to defined glycoconjugates containing l-Fuc and its structural analog l-colitose (l-Col), a bacterial dideoxysugar believed to bind immune proteins. We report a concise synthesis of l-Col and semisynthetic routes to several activated l-sugars. Incorporation of these sugars into glycans is evaluated using bacterial and mammalian glycosyltransferases (GTs) annotated to transfer l-Col or l-Fuc, respectively. We find that each GT can transfer both l-sugars, along with the rare hexose l-galactose (l-Gal), onto various glycan acceptors. Incorporation of these l-sugars into the resulting glycoconjugates is confirmed using known CBPs. Finally, these glycan ligands are employed to detect rare sugar-binding proteins in human serum. Overall, this work reveals similarities between bacterial and mammalian GTs that may be exploited for in vitro glycoconjugate construction to unveil novel mediators of host-pathogen interactions.

Salmonella is a major foodborne pathogen that causes salmonellosis, which is characterized by symptoms such as diarrhea, fever, and abdominal cramps. Existing methods for detecting Salmonella, such as culture plating, ELISA, and PCR, are accurate but time-consuming and unsuitable for on-site applications. In this study, we developed a rapid and sensitive electrochemical sensor using a molecularly imprinted polymer (MIP) to detect Salmonella typhimurium (S. typhimurium) by targeting lipopolysaccharides (LPS). Polydopamine (PDA) was used as the polymer matrix because of its cost-efficiency and functional versatility. The sensor demonstrated high sensitivity and selectivity, with a detection limit of 10 CFU/mL and a linear response over the 10²-10⁸ CFU/mL range. The specificity of the sensor was validated against other gram-positive and gram-negative bacteria and showed no significant cross-reactivity. Furthermore, the sensor performed effectively in real food samples, including tap water, milk, and pork, without complex preprocessing. These results highlight the potential of the LPS-imprinted MIP sensor for practical on-site detection of S. typhimurium, improving food safety monitoring and preventing outbreaks in food-handling environments.

Diabetes significantly increases the risk of serious health issues, including prolonged skin inflammation and delayed wound healing, owing to inferior glucose control and suppression of the immune system. Although traditional hydrogen (H2) therapy is slightly effective, its ability to tailor the release of H2 on the skin is limited. Accordingly, this study proposed a novel strategy for electrocatalytic H2 release under neutral conditions to promote wound healing in diabetic mice and rabbit. Herein, a defect-engineered cobalt phosphide (CoP) catalyst was designed by introducing a neutral single-metal electrocatalytic Hydrogen valence state channel into CoP. By effectively regulating the formation and transfer of *H active species during the CoP catalytic process, a considerable enhancement in neutral electrocatalytic H2 evolution performance was achieved (-78.0 mV@-10.0 mA cm-2). Based on this superior catalytic performance, we developed a flexible electrode (namely, CoP/flexible gold electrode made by screen printing (FGSP) by combining a convenient electrolysis platform with continuous electrolyte supply and FGSP, enabling customized H2 release and accelerating wound healing in diabetic mice and rabbits. Notably, the designed flexible electrode features adjustable dimensions, interchangeable substrates, and material adaptability, meeting the diverse needs of clinical and basic research and demonstrating significant potential for applications in clinical medicine.

The intricate relationship between nutrition, gut microbiome, and mental health has gained increasing attention. We aimed to determine how vitamin C supplementation improves mental vitality through the gut microbiome and associated neurological and immunological changes. We used 16S rRNA sequencing to analyze gut microbiota profiles of participants from our previous trial, in which healthy young adults (20-39 years) with inadequate serum vitamin C levels (< 50 μM) received 500 mg vitamin C or a placebo twice daily for 4 weeks (vitamin C, n = 21; placebo, n = 19). We examined whether changes in gut microbiota correlated with previously determined mental vitality indices, including Stroop test performance, work engagement, and serum brain-derived neurotrophic factor (BDNF) levels. Serum concentrations of microbial-derived molecules, cytokines, and neurotransmitters were analyzed using enzyme-linked immunosorbent assay, electrochemiluminescence-based immunoassay, or ultra-high-performance liquid chromatography-mass spectrometry. Monocyte subpopulations in peripheral blood were quantified using fluorescence-activated cell sorting analysis. Vitamin C supplementation increased the relative abundance of Bacillaceae and Anaerotruncus, while decreasing Desulfovibrio, with the Desulfovibrio reduction correlating with Stroop test performance. Moreover, participants showing a substantial Desulfovibrio reduction ("responders") demonstrated greater BDNF increases and stronger correlations between serum L-DOPA levels and work engagement scores than did non-responders. In addition, vitamin C supplementation suppressed inflammatory responses with concurrent reduction in serum lipopolysaccharide levels, and responders showed greater decreases in IL-10 levels and classical monocyte frequencies than non-responders. In conclusion, vitamin C supplementation modulates gut microbiota composition, particularly by reducing Desulfovibrio abundance, with the extent of reduction correlating with mental vitality improvements and decreased inflammation. This study provides insights into vitamin C supplementation as a critical dietary intervention, as it may modulate mental health through its influence on the gut-brain-immune axis.