Study of Linkage between Glutathione Pathway and the Antibiotic Resistance of Escherichia coli from Patients’ Swabs
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Date
2015-05-31
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Mark
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MDPI
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Abstract
V této práci jsme se zaměřili na rozdíly mezi bakteriálních kultur E. coli, získaných ze stěrů infekčních ran pacientů ve srovnání s laboratorním E. coli. Navíc blokování proteinu odpovědné za syntézu glutathion (-glutamyl-cystein synthasy GCL) za použití 10 mM buthionin sulfoximin byla zkoumána. Každý E. coli ukázala výrazné rozdíly v rezistenci vůči antibiotikům. Podle odhodlaný odolné, E. coli byly rozděleny do experimentálních skupin na základě statistického vyhodnocení jejich vlastností jako odolnější a citlivější. Tyto skupiny byly také použity pro nalezení rozdíly mezi nimi v závislosti na glutathion dráhy o rezistenci na antibiotika. Citlivější E. coli vykazovaly stejné kinetiky glutathionu syntézy zatímco blokuje GCL (km 0,1 um), ve srovnání s non-blokování. Kromě toho, nejčastější mutace v genech glutathion syntetázy, glutathion peroxidázy a glutathionreduktasy byly pozorovány v této skupině ve srovnání s laboratorní E. coli. Skupina "odolnější" E. coli vykazovaly rozdíly v km mezi 0,3 a 0,8 uM. Počet mutací ve srovnání s E. coli laboratoře byla podstatně nižší v porovnání s jinou skupinou.
In this work, we focused on the differences between bacterial cultures of E. coli obtained from swabs of infectious wounds of patients compared to laboratory E. coli. In addition, blocking of the protein responsible for the synthesis of glutathione (-glutamylcysteine synthase—GCL) using 10 mM buthionine sulfoximine was investigated. Each E. coli showed significant differences in resistance to antibiotics. According to the etermined resistance, E. coli were divided into experimental groups based on a statistical evaluation of their properties as more resistant and more sensitive. These groups were also used for finding the differences in a dependence of the glutathione pathway on resistance to antibiotics. More sensitive E. coli showed the same kinetics of glutathione synthesis while blocking GCL (Km 0.1 M), as compared to non-blocking. In addition, the most frequent mutations in genes of glutathione synthetase, glutathione peroxidase and glutathione reductase were observed in this group compared to laboratory E.coli. The group of “more resistant” E. coli exhibited differences in Km between 0.3 and 0.8 M. The number of mutations compared to the laboratory E. coli was substantially lower compared to the other group.
In this work, we focused on the differences between bacterial cultures of E. coli obtained from swabs of infectious wounds of patients compared to laboratory E. coli. In addition, blocking of the protein responsible for the synthesis of glutathione (-glutamylcysteine synthase—GCL) using 10 mM buthionine sulfoximine was investigated. Each E. coli showed significant differences in resistance to antibiotics. According to the etermined resistance, E. coli were divided into experimental groups based on a statistical evaluation of their properties as more resistant and more sensitive. These groups were also used for finding the differences in a dependence of the glutathione pathway on resistance to antibiotics. More sensitive E. coli showed the same kinetics of glutathione synthesis while blocking GCL (Km 0.1 M), as compared to non-blocking. In addition, the most frequent mutations in genes of glutathione synthetase, glutathione peroxidase and glutathione reductase were observed in this group compared to laboratory E.coli. The group of “more resistant” E. coli exhibited differences in Km between 0.3 and 0.8 M. The number of mutations compared to the laboratory E. coli was substantially lower compared to the other group.
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International Journal of Molecular Sciences. 2015, vol. 16, issue 4, p. 7210-7229.
http://www.mdpi.com/1422-0067/16/4/7210
http://www.mdpi.com/1422-0067/16/4/7210
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Peer-reviewed
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en