Anti-stress proteins produced by Bacteroides thetaiotaomicron after nutrient starvation.

Bacteroides thetaiotaomicron maybe one of the most adaptable intestinal bacteria due to its complex genome. Known to be an opportunistic pathogenic anaerobe, B. thetaiotaomicron has recently been described as a symbiont with anti-inflammatory properties. In this study, peptide mass finger printing technique was used to identify the stress proteins (maybe anti-stress proteins for the host) extracted from B. thetaiotaomicron grown under nutrient starvation (without heme, blood or bile) prior to be placed in an aerobic solution containing a mild non-ionic detergent derived from cholic acid. We focus here on proteins related to stress, knowing that superoxide dismutase was previously identified in the extract. In parallel, the morphology of the bacterial cells was observed using electronic microscopy before and after the extraction process. The effective antioxidant effect of the extract was evaluated in vitro against hydrogen peroxide. This work highlights the B. thetaiotaomicron ability to produce a large amount of stress proteins and to remain viable during the extraction. Budding vesicles were observed on its cell wall. The extraction process did not exceed 20 h in order to preserve the bacterial viability that decreased significantly after 24 h in preliminary studies. In our experimental conditions, an inhibitory effect of the extract was found against hydrogen peroxide. Animal models of inflammation will later check in vivo if this extract of anti-stress proteins is able to counter the respiratory burst beginning an inflammation process.

Comparison of the in vitro effects of amoxicillin and ampicillin on the polymorphonuclear neutrophil respiratory burst.

OBJECTIVES: The aim was to compare the in vitro effects of amoxicillin and ampicillin on the oxidative metabolism of polymorphonuclear neutrophils (PMNs). METHODS: Superoxide radical anion production by PMNs, stimulated or not by various exogenous stimulants and in contact with increasing antibiotic concentrations, was measured using spectrophotometric methods. RESULTS: Whereas a pro-oxidative action of amoxicillin on PMNs was obtained without exogenous stimulation or with opsonized zymosan (OZ), the O(2)(-) production by PMNs incubated with ampicillin did not increase significantly. CONCLUSIONS: This amoxicillin pro-oxidative effect could be due to the activation of the PMN NADPH oxidase, to its induction by a membrane effect or via the OZ pathway. It probably reinforces amoxicillin intrinsic bactericidal action and might partly explain the severe rashes sometimes occurring with amoxicillin treatment.

Antioxidative effect of Bacteroides thetaiotaomicron extracts: superoxide dismutase identification.

Bacteroides thetaiotaomicron, a bowel anaerobic commensal, seems to release enzymes detoxifying reactive oxygen species according to our recent work. This opportunistic pathogen would be beneficial in the case of an inflammatory process. To explore its role after an oxidative or nutritive stress, six to seven separate experiments were performed. The bacteria were grown on media restricted in growth factors or supplemented with bile. Their viability was checked after surface protein extraction. The extracts underwent 2D electrophoresis. Gel images were statistically analysed to construct "master" gels. Proteins were identified (peptide-mass fingerprinting technique). The effect of each extract on superoxide anions was evaluated (spectrophotometric method). Superoxide dismutase was identified and a major superoxide anion inhibition was shown by extracts obtained after a nutritive and oxidative stress without significant bacterial death. So, a therapeutic antioxidant potential is firmly hoped for. [figure: see text]