Project description:Flash proteotyping is a methodology for ultra-fast identification of microorganisns by tandem mass spectrometry. Here, we obtained results on five reference strains and ten new bacterial isolates. The methodology is based on direct sample infusion into the mass spectromete and an original, highly sensitive procedure for data processing and taxonomic identification.
Project description:Uterine diseases in dairy cattle are associated with reduced fertility, but the specific mechanisms of disease mediated subfertility are not fully elucidated. We hypothesized that an intrauterine infusion of Escherichia coli and Trueperella pyogenes would alter the endometrial transcriptome at a time coincident with conceptus elongation, and compromise oocyte developmental competence. Sixty-one non-lactating Holstein cows received an intrauterine infusion of either saline (n = 29) or pathogenic E. coli and T. pyogenes (n = 32) to induce acute endometrial inflammation. Endometrial samples were collected via cytobrush 41 d after infusion and analyzed by RNA sequencing. Bacteria infusion altered the expression of 203 genes in the endometrium 41 d after infusion. Top upregulated genes were GSTA3, PVALB, JAKMIP2, FOLH1B, TCF23, and top downregulated genes were MYMK, LOC104974498, CDRT1, KIAA0408, SLC45A2. The differentially expressed genes were functionally annotated to 54 canonical pathways with glutathione-mediated detoxification being the top predicted pathway altered following bacteria infusion. Collectively, this work demonstrates that bacterial infections can have lasting effects on the uterus which may provide a link to the long-term subfertility of cows after uterine disease.
Project description:To better understand the impact of infection on oocyte quality we employed global transcriptomics of oocytes collected from heifers after receiving intrauterine infusion of pathogenic Escherichia coli and Trueperella pyogenes. We hypothesized that oocyte transcriptome would be altered in response to intrauterine infection. A total of 452 differentially expressed genes were identified in oocytes collected from heifers 4 days after bacteria infusion compared to vehicle infusion, while 539 differentially expressed genes were identified in oocytes collected from heifers 60 days after bacteria infusion. Only 42 genes were differentially expressed in bacteria infused heifers at both day 4 and day 60. Interferon, HMGB1, ILK, IL-6 and TGF-beta signaling pathways were downregulated in oocytes collected at day 4 from bacteria infused heifers, while interferon, ILK and IL-6 signaling were upregulated in oocytes collected at day 60 from bacteria infused heifers. These data suggest that bacterial infusion alters the oocyte transcriptome differently at day 4 and day 60, suggesting different follicle stages are susceptible to damage. Characterizing the long-term impacts of uterine infection on oocyte transcriptome aids in our understanding of how infection causes infertility in dairy cattle.
Project description:To investigate changes in noradrenergic neurons of the LC following infusion of human Tyrosinase (hTyr) We performed gene expression profiling analysis using data obtained from RNA-seq of Pre-IP and IP samples following hTyr or EYFP infusion into the LC.
Project description:Previous studies have demonstrated that the iron content in marine heterotrophic bacteria is comparatively higher than that of phytoplankton. Therefore, they have been indicated to play a major role in the biogeochemical cycling of iron. In this study, we aimed to investigate the potential of viral lysis as a source of iron for marine heterotrophic bacteria. Viral lysates were derived from the marine heterotrophic bacterium, Vibrio natriegens PWH3a (A.K.A Vibrio alginolyticus). The bioavailability of Fe in the lysates was determined using a model heterotrophic bacterium, namely, Dokdonia sp. strain Dokd-P16, isolated from Fe-limited waters along Line P transect in the Northeastern Pacific Ocean. The bacteria were grown under Fe-deplete or Fe-replete conditions before being exposed to the viral lysate. Differential gene expression following exposure to the viral lysate was analyzed via RNA sequencing to identify differentially expressed genes under iron-replete and iron-deplete conditions. This study would provide novel insights into the role of viral lysis in heterotrophic bacteria in supplying bioavailable iron to other marine microorganisms under iron-limiting and non-limiting conditions. First, the marine heterotrophic bacterium genome, Dokdonia sp. strain Dokd-P16, was sequenced to provide a genomic context for the expression studies. Subsequently, the relative gene expression in Dokdonia sp. strain Dokd-P16 grown under Fe limiting and non-limiting conditions were analyzed. This transcriptomic approach would be utilized to elucidate genes regulated by Fe availability in Dokdonia sp. strain Dokd-P16, which indicate its Fe-related response viral lysate exposure. Taken together, in this study, the transcriptomic responses of Fe-limited and non-limited marine heterotrophic bacteria were analyzed, which provided novel insights into the biological availability of Fe from the viral lysates.
Project description:Here we tested the hypothesis that intrauterine infusion of pathogenic bacteria leads to changes in the transcriptome of the reproductive tract in dairy cattle three months later. We used virgin Holstein heifers to avoid the confounding effects of periparturient problems, metabolic stress and lactation. Animals were infused intrauterine with endometrial pathogenic bacteria Escherichia coli and Trueperella pyogenes to induce clinical endometritis (n = 4) and compared with control animals (n = 6). Three months after infusion, the caruncular and intercaruncular endometrium, isthmus and ampulla of the oviduct, and granulosa cells from dominant ovarian follicles were profiled by RNA-sequencing. Compared with control, bacterial infusion altered the transcriptome of all the tissues. Most differentially expressed genes were tissue-specific, with 109 differentially expressed genes unique to the caruncular endometrium, 57 in the intercaruncular endometrium, 65 in the isthmus, 298 in the ampulla, and 83 in granulosa cells. Surprisingly, despite infusing the bacteria into the uterus, the granulosa cells had more predicted upstream regulators of differentially expressed genes than all the other tissues combined. In conclusion, there was evidence of long-term changes to the transcriptome of the endometrium, oviduct and even the granulosa cells after intrauterine infusion of pathogenic bacteria, which implies that all these tissues contribute to the infertility that persists after endometritis.
2019-11-30 | GSE140469 | GEO
Project description:Bacteria extracted from bear saliva