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1.
Tuberculosis (Edinb) ; 103: 1-9, 2017 03.
Article in English | MEDLINE | ID: mdl-28237027

ABSTRACT

It is known that cholesterol plays a key role for Mycobacterium tuberculosis (Mtb) adaptation and survival within the host, thus contributing to the establishment of dormancy. It has been extensively demonstrated that fatty acids are the main energy source of Mtb during infection and dormancy, and it has been proposed that these molecules are implicated in reactivation of bacilli from a dormant state. We used in vitro models to analyze Mtb gene expression during dormancy and reactivation when fatty acids and cholesterol are the unique carbon source in the media. Our results suggest that cholesterol might function as a signal to trigger Mtb expression of some genes required for stress protection earlier than the one induced by fatty acids alone, indicating that cholesterol is very favorable for its development. This process is so conducive that cholesterol-adapted bacilli can reactivate their growth after NRP2 dormancy state even 10 min post ventilation. Thus, we hypothesize that cholesterol is not only involved in Mtb dormancy but that it also plays a critical role for favorable and almost immediate reactivation from an in vitro long-lasting dormant state induced by hypoxia.


Subject(s)
Cholesterol/metabolism , Latent Tuberculosis/metabolism , Mycobacterium tuberculosis/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Energy Metabolism , Fatty Acids/metabolism , Gene Expression Regulation, Bacterial , Latent Tuberculosis/microbiology , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/growth & development , Mycobacterium tuberculosis/pathogenicity , Oxygen/metabolism , Signal Transduction , Virulence
2.
Int Microbiol ; 13(1): 9-13, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20890835

ABSTRACT

Major differences regarding the pathology and host immune response of the Beijing and Canettii genotypes of Mycobacterium tuberculosis have been reported; however, studies on the genetic expression of these genotypes during in vitro dormancy are scarce. This study examined the expression of five cell-cycle-related genes and two dormancy-related genes in M. canettii, M. tuberculosis H37Rv, and M. tuberculosis Beijing during the Wayne model of dormancy. The results showed that under hypoxic conditions the three tuberculosis genotypes were able to transcribe genes involved in DNA replication and cellular division. In addition, dosR was found to be up-regulated in M. tuberculosis Beijing during the exponential growth phase but down-regulated under hypoxic conditions. In this genotype, the replication-related gene dnaA was also strongly down-regulated. These latter two findings suggest that, compared to M. tuberculosis H37Rv and M. canettii, the Beijing genotype has a lower capacity to synthesize dosR, hspX, and dnaA mRNAs during in vitro dormancy.


Subject(s)
Bacterial Proteins/biosynthesis , DNA-Binding Proteins/biosynthesis , Gene Expression Regulation, Bacterial , Mycobacterium tuberculosis/physiology , Oxygen/metabolism , Protein Kinases/biosynthesis , Antigens, Bacterial/biosynthesis , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , DNA-Binding Proteins/genetics , Gene Expression Profiling , Humans , Hypoxia , Mycobacterium tuberculosis/growth & development , Mycobacterium tuberculosis/metabolism , Protein Kinases/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/metabolism
3.
BMC Microbiol ; 10: 82, 2010 Mar 17.
Article in English | MEDLINE | ID: mdl-20236539

ABSTRACT

BACKGROUND: The prevalence of infections with Mycobacterium tuberculosis (MTb) and nontuberculous mycobacteria (NTM) species in HIV-infected patients in Mexico is unknown. The aims of this study were to determine the frequency of MTb and NTM species in HIV-infected patients from Mexico City, to evaluate the genotypic diversity of the Mycobacterium tuberculosis complex strains, to determine their drug resistance profiles by colorimetric microplate Alamar Blue assay (MABA), and finally, to detect mutations present in katG, rpoB and inhA genes, resulting in isoniazid (INH) and rifampin (RIF) resistance. RESULTS: Of the 67 mycobacterial strains isolated, 48 were identified as MTb, 9 as M. bovis, 9 as M. avium and 1 as M. intracellulare. IS6110-RFLP of 48 MTb strains showed 27 profiles. Spoligotyping of the 48 MTb strains yielded 21 patterns, and 9 M. bovis strains produced 7 patterns. Eleven new spoligotypes patterns were found. A total of 40 patterns were produced from the 48 MTb strains when MIRU-VNTR was performed. Nineteen (39.6%) MTb strains were resistant to one or more drugs. One (2.1%) multidrug-resistant (MDR) strain was identified. A novel mutation was identified in a RIF-resistant strain, GAG --> TCG (Glu --> Ser) at codon 469 of rpoB gene. CONCLUSIONS: This is the first molecular analysis of mycobacteria isolated from HIV-infected patients in Mexico, which describe the prevalence of different mycobacterial species in this population. A high genetic diversity of MTb strains was identified. New spoligotypes and MIRU-VNTR patterns as well as a novel mutation associated to RIF-resistance were found. This information will facilitate the tracking of different mycobacterial species in HIV-infected individuals, and monitoring the spread of these microorganisms, leading to more appropriate measures for tuberculosis control.


Subject(s)
AIDS-Related Opportunistic Infections/microbiology , HIV Infections/microbiology , Mutation , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/microbiology , Cluster Analysis , Drug Resistance, Multiple, Bacterial , Humans , Mexico , Mycobacterium tuberculosis/isolation & purification , Polymorphism, Restriction Fragment Length , Serotyping , Tuberculosis, Multidrug-Resistant/virology
4.
Int. microbiol ; 13(1): 9-13, mar. 2010. ilus
Article in English | IBECS | ID: ibc-87667

ABSTRACT

Major differences regarding the pathology and host immune response of the Beijing and Canettii genotypes of Mycobacterium tuberculosis have been reported; however, studies on the genetic expression of these genotypes during in vitro dormancy are scarce. This study examined the expression of five cell-cycle-related genes and two dormancy-related genes in M. canettii, M. tuberculosis H37Rv, and M. tuberculosis Beijing during the Wayne model of dormancy. The results showed that under hypoxic conditions the three tuberculosis genotypes were able to transcribe genes involved in DNA replication and cellular division. In addition, dosR was found to be up-regulated in M. tuberculosis Beijing during the exponential growth phase but down-regulated under hypoxic conditions. In this genotype, the replication-related gene dnaA was also strongly down-regulated. These latter two findings suggest that, compared to M. tuberculosis H37Rv and M. canettii, the Beijing genotype has a lower capacity to synthesize dosR, hspX, and dnaA mRNAs during in vitro dormancy (AU)


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Subject(s)
Humans , Protein Kinases/biosynthesis , Oxygen/metabolism , Mycobacterium tuberculosis/physiology , Gene Expression Regulation, Bacterial , Bacterial Proteins/biosynthesis , DNA-Binding Proteins/biosynthesis , RNA, Messenger/biosynthesis , RNA, Messenger/metabolism , Protein Kinases/genetics , Mycobacterium tuberculosis/growth & development , Mycobacterium tuberculosis/metabolism , Bacterial Proteins/genetics , DNA-Binding Proteins/genetics , Gene Expression Profiling , Hypoxia , Antigens, Bacterial/biosynthesis , Antigens, Bacterial/genetics
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