Project description:This experiment set contains the raw data for 8 arrays that were used in the genomic typing of the pre- and post-mouse H. pylori strains SS1 and SS2000. 10700 is the pre-mouse clinical isolate of SS1 and PMSS2000 is the pre-mouse clinical isolate of SS2000. gDNA from these strains were labeled and hybridized to H. pylori microarrays as described in Salama et al. {Salama et al. 2000 PNAS 97:14668-73}. In each case the test gDNA sample was labeled with Cy5 (red) and this was hybridized with Cy3 (green) labeled reference DNA of equal amount. The reference DNA consisted of equal amounts of gDNA from the two H. pylori strains used to make the H. pylori microarray, 26695 and J99. This data was used for the manuscript: L. J. Thompson, S. J. Danon, J.E. Wilson, J. L. O'Rourke, N. R. Salama, S. Falkow, H. Mitchell, and A. Lee. (2004) Chronic Helicobacter pylori infection in C57BL/6 and BALB/c mice using SS1 and a newly identified mouse-adapted strain (SS2000). Infect. Immun. (in press).
Project description:The oral cavity is considered an extra-gastric reservoir for Helicobacter pylori (H. pylori) and oral H. pylori can contribute to the gastric eradication inability and recurrence. However, the oral environment is not ideal for H. pylori survival, and the factors promoting oral colonization and survival of H. pylori have not been elucidated. In this study, we explored the effects of extracellular polysaccharides (EPS), the fundamental building blocks of dental Streptococcus mutans (S. mutans) biofilm, on H. pylori colonization and drug resistance in the oral cavity, as well as stomach. In the co-culture system of H. pylori Sydney strain (SS1) and three S. mutans biofilms with different EPS contents (UA159 wild-type, UA159ΔgtfB, UA159ΔgtfBC), it was found that the adhesive force between SS1 and biofilms increased correspondingly with the increase in EPS content. Moreover, with the increase in EPS content of biofilms, the number of colonized SS1 increased. Proteome analysis revealed that SS1 co-cultured with UA159 biofilm exhibited 149 differentially expressed proteins compared to that co-cultured with UA159ΔgtfB biofilm, with significant enrichment in β-lactamase activity pathway. SS1 co-cultured with UA159 biofilm exhibited 154 differentially expressed proteins compared to that co-cultured with UA159ΔgtfBC biofilm, with significant enrichment in β-lactamase activity, aminoglycoside nucleotidyltransferase activity and antioxidant activity pathways. Both in vivo and in vitro, EPS synthesized by glucosyltransferases (Gtfs) surrounding SS1 was verified to protect SS1 against β-lactam and aminoglycoside antibiotics. These findings demonstrated that S. mutans biofilms mediate oral adhesion, colonization, and antibiotic resistance of H. pylori through a Gtfs-driven EPS biosynthesis mechanism.
Project description:The aim of this study is to identify alterations induced in gastric mucosa of mice exposed to Pteridium aquilinum and/or infected with Helicobacter pylori, in order to identify genes that are induced by bracken fern exerts exacerbating effects on gastric lesions associated to the infection. Six groups of C57Bl/6 mice were be used: 1) control, 2) infected Helicobacter pylori, 3) treated with Bracken fern extract orogastrically, 4) treated with Bracken fern extract in drinking water, 5) infected Helicobacter pylori + treated with Bracken fern extract orogastrically, 6) infected Helicobacter pylori + treated with Bracken fern extract in drinking water. The infection procedure was performed using an orogastric inoculation of H.pylori (strain SS1) twice in the first week. The RNA isolation was done in triplicate (3 mice per each condition). Further evaluation of morphological alterations on gastric mucosa, proliferative index and induction of DNA strand breaks will be performed in the mice stomach exposed to Pteridium aquilinum infected or not with Helicobacter pylori. Alterations of glycosylation in gastric tissues will also evaluated.
Project description:Helicobacter pylori (H. pylori) is a human pathogen that infects almost half of the world’s population. Infection with H. pylori is frequently associated with chronic gastritis and can even lead to gastric and duodenal ulcers and gastric cancer. Although the persistent colonization of H. pylori and the development of H. pylori-associated gastritis remain poorly understood, it is believed that, in gastric mucosa, the modulated gastric epithelial cells (GECs) by H. pylori are key contributors. We used microarrays to detail the global programme of gene expression in Helicobacter pylori infected-gastric epithelial cell line AGS cells and identified up-regulated genes induced by Helicobacter pylori infection.