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Ilina E.N.,Karpov Institute of Physical Chemistry | Shitikov E.A.,Karpov Institute of Physical Chemistry | Ikryannikova L.N.,Karpov Institute of Physical Chemistry | Alekseev D.G.,Moscow Institute of Physics and Technology | And 14 more authors.
PLoS ONE | Year: 2013

Tuberculosis caused by multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis (MTB) strains is a growing problem in many countries. The availability of the complete nucleotide sequences of several MTB genomes allows to use the comparative genomics as a tool to study the relationships of strains and differences in their evolutionary history including acquisition of drug-resistance. In our work, we sequenced three genomes of Russian MTB strains of different phenotypes - drug susceptible, MDR and XDR. Of them, MDR and XDR strains were collected in Tomsk (Siberia, Russia) during the local TB outbreak in 1998-1999 and belonged to rare KQ and KY families in accordance with IS6110 typing, which are considered endemic for Russia. Based on phylogenetic analysis, our isolates belonged to different genetic families, Beijing, Ural and LAM, which made the direct comparison of their genomes impossible. For this reason we performed their comparison in the broader context of all M. tuberculosis genomes available in GenBank. The list of unique individual non-synonymous SNPs for each sequenced isolate was formed by comparison with all SNPs detected within the same phylogenetic group. For further functional analysis, all proteins with unique SNPs were ascribed to 20 different functional classes based on Clusters of Orthologous Groups (COG). We have confirmed drug resistant status of our isolates that harbored almost all known drug-resistance associated mutations. Unique SNPs of an XDR isolate CTRI-4XDR, belonging to a Beijing family were compared in more detail with SNPs of additional 14 Russian XDR strains of the same family. Only type specific mutations in genes of repair, replication and recombination system (COG category L) were found common within this group. Probably the other unique SNPs discovered in CTRI-4XDR may have an important role in adaptation of this microorganism to its surrounding and in escape from antituberculosis drugs treatment. © 2013 Ilina et al.

Shitikov E.,Karpov Institute of Physical Chemistry | Ilina E.,Karpov Institute of Physical Chemistry | Chernousova L.,Central TB Research Institute of RAMS | Borovskaya A.,Karpov Institute of Physical Chemistry | And 8 more authors.
Infection, Genetics and Evolution | Year: 2012

Identification and typing of mycobacteria is very important for epidemiology, susceptibility testing and diagnostic purposes. This paper describes the development and validation of the alternative methods for species identification and typing of mycobacteria based on a matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry (MALDI-ToF MS). Altogether there were 383 clinical isolates analyzed which include 348 strains of Mycobacterium tuberculosis complex (MTBC) (342 strains of M. tuberculosis and 6 strains of M. bovis) and 35 strains of nontuberculous mycobacteria (NTM) represented by 16 different species. Direct bacterial profiling (DBP) by means of MALDI-ToF MS was carried out. Cluster analysis of DBP mass spectra divided them into two large separate groups corresponding to MTBC and NTM, and also demonstrated the possibility of isolate identification at the species level. Spoligotyping protocol based on mass spectrometry was developed and validated, it matched completely to classical spoligotyping data. Our results suggest that MALDI-ToF MS has potential as a rapid and reproducible platform for the identification and typing of Mycobacterium species. © 2011 Elsevier B.V..

Makar'Yants N.N.,Central TB Research Institute of RAMS | Shmelev E.I.,Central TB Research Institute of RAMS
Vestnik Rossiiskoi Akademii Meditsinskikh Nauk | Year: 2012

In order to improve treatment of patients with exogenous allergic alveolitis morphologically different variants of the disease, i.e. acute, subacute and chronic were identified and confirmed. For each variant of exogenous allergic alveolitis new therapy schemes were proposed. The study included 74 patients who were divided into 5 groups. In the first group with acute exogenous allergic alveolitis inhalation glycocorticosteroids in high doses in combination with plasmapheresis were prescribed, in the second group standard therapy with systemic glycocorticosteroids was prescribed. The third and the fourth group consisted of patients with subacute exogenous allergic alveolitis. The protracted ambroxol inhalation using nebulizers and the reduced dose of systemic glycocorticosteroids were used in the third group; and the standard dose of systemic glycocorticosteroids was used in the fourth. The fifth group consisted of patients with chronic exogenous allergic alveolitis, who received the standard dose of glycocorticosteroids and cytostatic drugs. After one month of therapy, it was ascertained that the use of high doses of inhalation glycocorticosteroids in combination with plasmapheresis in patients with acute exogenous allergic alveolitis led to significant improvements in clinical and CTpresentation, physical activity tolerance, as well as the use of systemic glycocorticosteroids. The use of ambroxol inhalation in patients with subacute exogenous allergic alveolitis led to a significant improvement in clinical symptomatology, functional parameters and CTpresentation, thus enabling to reduce the dose of glycocorticosteroids used and to avoid unwanted side effects.

Kaminskaya G.O.,Central TB Research Institute of RAMS | Abdullaev R.Y.,Central TB Research Institute of RAMS | Komissarova O.G.,Central TB Research Institute of RAMS
Vestnik Rossiiskoi Akademii Meditsinskikh Nauk | Year: 2012

81 patients with active pulmonary tuberculosis were investigated. The morpho-functional status of vascular endothelium was evaluated by plasma levels of stable metabolites of nitric oxide, endothelin-1 and von Willebrand factor antigen. Typical increase of endothelin-1 in positive correlation with systemic inflammatory response syndrome (SIRS) expression was established. Nitric oxide level decreased in patients with chronic and severe course of disease. Decrease of nitric oxide level was not associated with SIRS but was consequence of specific intoxication, von Willebrand factor antigen decreased in patients with recent and limited spread of tissue damage but increased progressively with intensity of SIRS. This complex of changes (contrast shifts of nitric oxide and endothelin-1 and von Willebrand factor antigen increase) manifested in endothelium metabolic dysfunction syndrome and developed pre-conditions for microcirculation disturbances.

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