Qing-Hua Zou

Background/Objectives Coffee is widely consumed worldwide and is rich in bioactive compounds with potential metabolic benefits. Recently, lipid-enriched coffee formulations have gained popularity; however, their biological effects and underlying mechanisms remain poorly understood. Methods In this study, we employed an integrated multi-omics approach to investigate the impact of coffee and high-fat coffee on the plasma metabolome and gut microbiota of C57BL/6J mice. Eighteen male mice were randomly assigned to three groups (n = 6) and received water, coffee, or high-fat coffee by oral gavage daily for 14 days. The plasma metabolome was analyzed via UHPLC-MS/MS, and the gut microbiota was profiled u 16S rRNA gene sequencing. Results Metabolomic analysis revealed distinct clustering patterns among groups. A total of 200 metabolites were significantly altered in the coffee group compared with the water group, while 86 metabolites were altered in the high-fat coffee group compared with the coffee group, with 56 overlapping metabolites suggesting a core metabolic response. Microbiome analysis showed that coffee consumption increased the abundances of Akkermansia and Bifidobacterium and decreased the levels of Ligilactobacillus and Muribaculum. Coriobacteriaceae UCG-002 and Turicibacter were significantly enriched in the high-fat coffee group, whereas Lachnospiraceae NK4A136 group, Mucispirillum and unclassified Lactobacillaceae were reduced. Association analysis highlighted the top 20 metabolites with the highest degree of connection to gut microbial genera, two of which belong to the chlorogenic acid pathway. Conclusions Reduced levels of ferulic acid and 3-hydroxybenzoic acid, two metabolites potentially involved in antioxidant and anti-inflammatory activities, were observed in the high-fat coffee group, suggesting that dietary cream influences microbiota-associated chlorogenic acid metabolism.

Salmonella enterica subsp. enterica serovar typhimurium ( S. typhimurium ) is a facultative intracellular pathogen causing significant gastrointestinal infections in both humans and animals. The type VI secretion system (T6SS) plays a crucial role in its virulence, facilitating competition with host gut microbiota and promoting infection. While S. typhimurium possesses a single T6SS, it encodes three hcp genes, which are essential for its functionality. To investigate the non-redundant roles of each hcp gene, in this study, we used 16S rRNA sequencing to analyze gut microbiota in BALB/c mice after infection with wild-type (WT) S. typhimurium or three mutant strains (Δ hcp1 , Δ hcp2 , and Δ hcp3 ). Our findings revealed that S. typhimurium infection induced severe gut dysbiosis especially on the second day post-infection, which was characterized by a notable increase in Firmicutes and activation of the energy pathways. The deletion of each hcp gene led to distinct changes in bacterial taxa, underscoring the non-redundant function of each hcp. Lactobacillus showed significant effects in both strain main effects and strain-time interaction effects. Despite having only one T6SS, S. typhimurium achieves precise modulation of its functions through the divergent roles of its Hcp proteins, enabling efficient colonization and persistence in the host gut. These findings provide insights into the intricate mechanisms of bacterial adaptation and host-pathogen interactions, offering potential avenues for therapeutic interventions targeting T6SS-mediated dysbiosis. IMPORTANCE: Salmonella enterica subsp. enterica serovar typhimurium ( S. typhimurium ) is a facultative intracellular pathogen causing significant gastrointestinal infections in humans and animals. The type VI secretion system (T6SS) plays a crucial role in its virulence, facilitating competition with host gut microbiota and promoting infection. While S. typhimurium possesses a single T6SS, it encodes three hcp genes, which are crucial for its functionality and may exhibit non-redundant roles. In this study, we used 16S rRNA sequencing to analyze gut microbiota in BALB/c mice after infection with wild-type (WT) S. typhimurium or mutant strains (Δ hcp1 , Δ hcp2 , Δ hcp3 ). Our findings revealed that the deletion of individual hcp genes led to distinct bacterial taxa changes, underscoring the non-redundant functions of each hcp . Despite having only one T6SS, S. typhimurium achieves precise modulation of its functions through the divergent roles of its Hcp proteins, enabling efficient colonization and persistence in the host gut.

Cell-autonomous immunity represents evolutionarily conserved defense mechanisms present in both immune and non-immune cells. One of such mechanisms is mediated by cytoskeletal septins that entrap cytosolic bacterial pathogens within cage-like structures. To promote infection, Shigella flexneri delivers effector proteins directly into host cells via a type III secretion system. Here we demonstrate that OspC effectors enable Shigella flexneri to evade septin cage entrapment. Mechanistically, OspC catalyzes ADP-riboxanation of SEPT9 at Arg561, a site essential for stabilizing septin hetero-oligomers. Notably, Arg561 ADP-riboxanation impairs septin polymerization and hence assembly of higher-order structures, including filaments and cage-like structures. Furthermore, we provide evidence that OspC effectors act synergistically with OspG to antagonize septin cage entrapment via two distinct post-translational modifications, thereby facilitating cell-to-cell spread and intracellular replication. Overall, our work reveals the elegant strategies of bacterial pathogens to evade septin-mediated cell-autonomous immunity and offers avenues for therapeutic intervention.

Escherichia coli is a significant pathogen capable of inducing a variety of infections in both human and animal hosts. Despite its clinical significance, there is a lack of longitudinal research aimed at elucidating the genomic attributes that facilitate antimicrobial resistance and clonal dissemination in this bacterium. In this study, we investigated the genetic dynamics of antibiotic resistance and virulence factors within a collection of 137 E. coli isolates retrieved from a Chinese hospital over a 12-year period. Notably, a substantial increase in resistance to various antibiotics, including broad-spectrum β-lactams, aminoglycosides, and quinolones, was observed. Additionally, our study revealed the acquisition of diverse antibiotic resistance and virulence genes across different sequence types (STs). Among the STs, ST131 emerged as the most prevalent, exhibiting a high level of multidrug resistance. In contrast, ST73 and ST12 demonstrated a higher prevalence of virulence genes, suggestive of a potential trade-off between antibiotic resistance and virulence. What’s more, we identified significant intra-clonal diversification and convergence of antibiotic resistance and virulence traits within the dominant ST131 group. These findings underscore the importance of longitudinal studies in understanding the evolution of bacterial pathogens and the necessity for ongoing research to inform public health strategies.

The presence of sub-minimal inhibitory concentration (sub-MIC) antibiotics in our environment is widespread, and their ability to induce antibiotic resistance is inevitable. Acinetobacter baumannii, a pathogen known for its strong ability to acquire antibiotic resistance, has recently shown clinical resistance to the last-line antibiotic tigecycline. To unravel the complex mechanism of A. baumannii drug resistance, we subjected tigecycline-susceptible, -intermediate, and -mildly-resistant strains to successive increases in sub-MIC tigecycline and ultimately obtained tigecycline-resistant strains. The proteome of both key intermediate and final strains during the selection process was analyzed using nanoLC-MS/MS. Among the more than 2600 proteins detected in all strains, we found that RND efflux pump AdeABC was associated with the adaptability of A. baumannii to tigecycline under sub-MIC pressure. qRT-PCR analysis also revealed higher expression of AdeAB in strains that can quickly acquire tigecycline resistance compared with strains that displayed lower adaptability. To validate our findings, we added an efflux pump inhibitor, carbonyl cyanide m-chlorophenyl hydrazine (CCCP), to the medium and observed its ability to inhibit tigecycline resistance in A. baumannii strains with quick adaptability. This study contributes to a better understanding of the mechanisms underlying tigecycline resistance in A. baumannii under sub-MIC pressure.

The mechanism of tigecycline resistance in A. baumannii remains largely unclear. In this study, we selected a tigecycline-resistant and a tigecycline-susceptible strain from a tigecycline-susceptible and a resistant strain, respectively. Proteomic and genomic analyses were performed to elucidate the variations associated with tigecycline resistance. Our study showed proteins associated with efflux pump, biofilm formation, iron acquisition, stress response, and metabolic ability are upregulated in tigecycline resistant strains, and efflux pump should be the key mechanism for tigecycline resistance. By genomic analysis, we found several changes in the genome that can explain the increased level of efflux pump, including the loss of the global negative regulator hns in the plasmid and the disruption of the hns gene and acrR gene on the chromosome by the insertion of IS5. Collectively, we not only revealed the phenomenon that the efflux pump is mainly responsible for tigecycline resistance, but also highlighted the mechanism at the genomic level, which will help in understanding the resistance mechanism in detail and provide clues for the treatment of clinical multiple drug-resistant A. baumannii.

Acinetobacter baumannii is one of the main pathogens causing nosocomial and community-acquired infections. Tigecycline is an important antibiotic for the treatment of multidrug-resistant A. baumannii infections, but strains resistant to tigecycline have also emerged. There are still many unclear questions concerning the mechanism of tigecycline resistance in A. baumannii. In this study, tigecycline-susceptible and tigecycline-intermediate strains were gradually cultured with sub-minimum inhibitory concentrations of tigecycline to select for tigecycline-resistant mutants, and a tigecycline-resistant strain was cultured under 42 °C to select for tigecycline-susceptible mutants. We found that the acquisition of tigecycline resistance affected the susceptibility of the strains to other antibiotics. Resistance to ampicillin–sulbactam is negatively correlated with tigecycline resistance. The strains will experience fitness costs along with the acquisition of tigecycline resistance. Tigecycline resistance in the strains was not related to 16S rRNA target variation or outer membrane integrity alteration. By constructing a transposon mutation library, we found that transposon insertion of the adeL gene reduced the sensitivity of A. baumannii to tigecycline. This study provides important clues for understanding the mechanism of tigecycline resistance in A. baumannii.

Intestinal mycobiome dysbiosis plays an important role in the advancement of HIV- and HCV-infected patients. Co-infection with HCV is an important risk factor for exacerbating immune activation in HIV-infected patients, and gut fungal microbial dysbiosis plays an important role. However, no systematic study has been conducted on the intestinal fungal microbiome of HIV/HCV co-infected patients to date. Patients infected with HIV and HCV, either alone or in combination, and healthy volunteers were included. Stool samples were collected for fungal ITS sequencing and for further mycobiome statistical analysis. We found that the abundance of fungal species significantly decreased in the HIV/HCV co-infection group compared to in the healthy control group, while no significant differences were found in the mono-infection groups. Low-CD4 + T-cell patients in the HIV group and high-ALT-level patients in the HCV group were discovered to have a more chaotic fungal community. Furthermore, the opportunistic pathogenic fungal profiles and fungal inter-correlations in the co-infection group became less characteristic but more complicated than those in the mono-infection groups. Intestinal fungal dysregulation occurs in HIV- and HCV-infected patients, and this dysregulation is further complicated in HIV/HCV co-infected patients.

Salmonella enterica is one of the most important food-borne pathogens, whose motility and virulence are highly related to flagella. Flagella alternatively express two kinds of surface antigen flagellin, FliC and FljB, in a phenomenon known as flagellar phase variation. The molecular mechanisms by which the switching orientation of the Hin-composed DNA segment mediates the expression of the fljBA promoter have been thoroughly illustrated. However, the precise regulators that control DNA strand exchange are barely understood. In this study, we found that a putative response regulator, STM0347, contributed to the phase variation of flagellin in S. Typhimurium. With quantitative proteomics and secretome profiling, a lack of STM0347 was confirmed to induce the transformation of flagellin from FliC to FljB. Real-time PCR and in vitro incubation of SMT0347 with the hin DNA segment suggested that STM0347 disturbed Hin-catalyzed DNA reversion via hin degradation, and the overexpression of Hin was sufficient to elicit flagellin variation. Subsequently, the Δstm0347 strain was outcompeted by its parental strain in HeLa cell invasion. Collectively, our results reveal the crucial role of STM0347 in Salmonella virulence and flagellar phase variation and highlight the complexity of the regulatory network of Hin-modulated flagellum phase variation in Salmonella.