02012nas a2200181   4500008004100000022001400041245013800055210006900193260000800262300001300270490000700283520133400290100001701624700002001641700001901661700001901680856013101699       2019                        eng d  a1932-620300aUsing diphenyleneiodonium to induce a viable but non-culturable phenotype in Mycobacterium tuberculosis and its metabolomics analysis0 aUsing diphenyleneiodonium to induce a viable but nonculturable p  cAUG  ae02206280 v143 a<p>Depletion of oxygen levels is a well-accepted model for induction of non-replicating, persistent states in mycobacteria. Increasing the stress levels in mycobacterium bacilli facilitates their entry into a non-cultivable, dormant state. In this study, it was shown that diphenyleneiodonium, an inhibitor of NADH oxidase, induced a viable, but non-culturable state in mycobacteria, having similar features to dormant bacilli, like loss of acid-fastness, upregulation of stress-regulated genes and decreased superoxide levels as compared to actively growing bacilli. Comprehensive, untargeted metabolic profiling also confirmed a decrease in biogenesis of amino acids, NAD, unsaturated fatty acids and nucleotides. Additionally, an increase in the level of lactate, fumarate, succinate and pentose phosphate pathways along with increased mycothiol and sulfate metabolites, similar to dormant bacilli, was observed in the granuloma. These non-cultivable bacilli were resuscitated by supplementation of fetal bovine serum, regaining their culturability in liquid as well as on agar medium. This study focused on the effect of diphenyleneiodonium treatment in causing mycobacteria to rapidly transition from an active state into a viable, but non-cultivable state, and comparing their characteristics with dormant phenotypes.</p>
1 aYeware, Amar1 aGample, Suwarna1 aAgrawal, Sonia1 aSarkar, Dhiman  uhttps://library.ncl.res.in/content/using-diphenyleneiodonium-induce-viable-non-culturable-phenotype-mycobacterium-tuberculosis01959nas a2200181   4500008004100000022001400041245012100055210006900176260000900245300001000254490000700264520130300271100001901574700002001593700001701613700001901630856012801649       2021                        eng d  a2045-232200aNovel gene similar to nitrite reductase (NO forming) plays potentially important role in the latency of tuberculosis0 aNovel gene similar to nitrite reductase NO forming plays potenti  cOCT   a198130 v113 aThe development of the latent phenotype of Mycobacterium tuberculosis (Mtb) in the human lungs is the major hurdle to eradicate Tuberculosis. We recently reported that exposure to nitrite (10 mM) for six days under in vitro aerobic conditions completely transforms the bacilli into a viable but non-cultivable phenotype. Herein, we show that nitrite (beyond 5 mM) treated Mtb produces nitric oxide (NO) within the cell in a dose-dependent manner. Our search for the conserved sequence of NO synthesizing enzyme in the bacterial system identified MRA2164 and MRA0854 genes, of which the former was found to be significantly up regulated after nitrite exposure. In addition, the purified recombinant MRA2164 protein shows significant nitrite dependent NO synthesizing activity. The knockdown of the MRA2164 gene at mRNA level expression resulted in a significantly reduced NO level compared to the wild type bacilli with a simultaneous return of its replicative capability. Therefore, this study first time reports that nitrite induces dormancy in Mtb cells through induced expression of the MRA2164 gene and productions of NO as a mechanism for maintaining non-replicative stage in Mtb. This observation could help to control the Tuberculosis disease, especially the latent phenotype of the bacilli.1 aAgrawal, Sonia1 aGample, Suwarna1 aYeware, Amar1 aSarkar, Dhiman  uhttps://library.ncl.res.in/content/novel-gene-similar-nitrite-reductase-no-forming-plays-potentially-important-role-latency