[Molecular and genetic characterization of rifampicin- and/or izoniazid-resistent strains of M. tuberculosis, isolated in Novosibirsk and Tomsk regions].

The purpose of the work was to study rifampicin- and izoniazid-resistent strains of M. tuberculosis, circulating in Western Siberia, by VNTR and IS6110 typing. The authors also studied genetic causes of resistance to these antibiotics and undertook a search of new VNTR loci, displaying polymorphism in genomes of closely related clonally-disseminated variants of Mycobacterium tuberculosis in W-Beijing family model analysis.

[The genetic diversity of Mycobacterium tuberculosis and an assessment of risk factors of tuberculosis spread in Russia's Siberian region by molecular epidemiological methods]. / Geneticheskoe raznoobrazie Mycobacterium tuberculosis i otsenka faktorov riska rasprostraneniia zabolevaniia tuberkulezom v Sibirskom regione Rossii metodami molekuliarnoi épidemiologii.

Molecular epidemiology approaches provided for a new interpretation of the TB infection transmission dynamics, contributed to changing the focuses of attention and updated the monitoring practice. On the basis of 101 cases of isolates of Mycobacterium tuberculosis (MBT) complex sampled from 84 patients with pulmonary tuberculosis in the Siberian region, we proved that the independent methods of IS6110 RFLP genetic typing and VNTR-typing by five accurate repeat tandems of ETR A, B, C, D, and E bring about similar results and can be used in studying the MTB clonal structure population in the Siberian region for the purpose of defining the TB infection transmission dynamics. The most widespread genetic types were detected, i.e. Beeijing family strains, the S42 spoligotype, and the 31323 VNTR type, which account for 52.3% of all samples. The general parameters describing the epidemic process intensity were evaluated, i.e. those characterizing the strains (91.6%) and the transmission activity factor (72%). Consequently, each three of the four analyzed TB cases resulted from a recent transmission. However, there is a trend, within the analyzed samples, towards a higher percentage of clusterization in the age group ranging from 40 to 60. Such trend is typical of a prevalence of TB reactivation cases caused by MBT complex strains spread intensively in the discussed territory. As for the clusterized isolates, which are endemic for the territory, such data should be interpreted as a recent transmission only cautiously. 28.5% of the studied isolates are resistant to anti-TB drugs used in medical practice; and 35.7% of them are resistant to izoniazide and rifampicin, therefore, according to the WHO classification they are considered to be poly-antibiotics-resistant (PAR). No strict associations were found between the spectrum of antibiotics-resistance and any of genotypes, however, 30% of PAR strains are 32525 and 42525 types VNTR (spoligotype S1 or Beejing type).

[Use of the major components methods in epidemiological studies]. / Primenenie metoda glavnykh komponent v épidemiologicheskikh issledovaniiakh.

When multifactorial processes are analyzed, it is most rational to use the methods of multidimensional statistical analysis, including the major components methods. This method is rather new in epidemiological surveys. Its advantage is that it may be used to get information that is difficult derived by other ways. Noteworthy is the versatility of the method, which lies in that the problems of analysis, prediction, and classification (stratification) of temporal series are solved by using actually the same techniques. How to apply the major components method are considered by using an example of investigating the general trends in the development of an epidemic tuberculous process in a specific area. Statistical data on tuberculosis morbidity in the areas of the Novosibirsk Region were employed as the initial material.

[Diagnostic value of the forced oscillation test in detection of bronchial obstruction]. / Diagnosticheskaia znachimost' metoda forsirovannykh ostsilliatsii v vyiavlenii bronkhial'noi obstruktsii.

Clinical testing of the forced oscillation test (FOT) yielded criteria for bronchial obstruction: an increase in viscous respiratory resistance (VRR) (Rfo, Rin, Rex); the frequency dependence of VRR; and a decrease in a phase angle. The study of VRR promoted detection of bronchial malpatency in additional 20.7% of the individuals who had no bronchial obstruction, as evidenced by spirography (SG) and forced expiration volume-current curve (FEV-CC) recording. To improve the diagnosis of bronchial patency impairments, it is expedient to use FOT of VRR along with the routine methods (SG and FEV-CC recording). A relationship has been established between the values of VRR and the clinical signs of chronic bronchitis and the degree of bronchial obstruction.

[Informative value of the major component technique in the assessment of the epidemic tuberculosis situation]. / Informativnost' metoda glavnykh komponent v otsenke épidemicheskoi situatsii po tuberkulezu.

The paper describes a method for comprehensive analysis of the epidemic situation which is based on the major component technique. The algorithm proposed for comprehensive assessment is highly effective in the retrospective analysis and in the analysis of the current material which includes a great deal of various characteristics that are at first sight occasionally unrelated. The use of the algorithm is shown by the example of analyzing 19 statistical variables that characterize the epidemic situation and different aspects of the work of a tuberculosis-controlling facility in 30 districts of the Novosibirsk Region in a period of 15 years (1985-1999). During processing, the variables fall into two groups: a group of variables that characterize the quality of antituberculosis work and a group of those reflecting the epidemic situation, which were used to divide the districts of the region into several groups.

[Specific features of the course of pulmonary tuberculosis concurrent with liver pathology]. / Osobennosti techeniia tuberkuleza legkikh u bol'nykh s patologiei pecheni.

The study was undertaken to reveal clinical, biochemical, morphological, and immunological features of pulmonary tuberculosis concurrent with liver pathology. Patients with pulmonary tuberculosis who suffered from liver diseases were found to have more marked clinical signs, such as subfebrile temperature, weight loss, elevated levels of aminotransferases and gamma-glutamyltransferase. Morphological studies of liver biopsy specimens indicated that the rates of necrosis and inflammatory infiltration were higher in patients with chronic hepatitis C (CHC) and chronic hepatitis B (CHB) and CHC and substantially lower in CHB and alcoholism. Impaired immunity involved primarily the phagocytic link.

[Molecular genetic characteristics of rifampicin-resistant Mycobacterium tuberculosis isolates in Novosibirsk]. / Molekuliarno-geneticheskaia kharakteristika ustoichivykh k rifampitsinu izoliatov Mycobacterium tuberculosis, vydelennykh v Novosibirske.

Forty rifampicin-resistant clinical isolates from patients living in Novosibirsk were studied. Six alleles earlier described in the literature were identified by the sequencing technique. The frequency of mutations in the studied samples slightly differs from that earlier reported for other geographic regions: 21 (52.5%) strains carried the mutated codon TTG in position 531 (Ser-->Leu), 7 (17.5%) had GTC in position 516 (Asp-->Val) and 2 (5%) had the GAC substitution in position 526 (His-->Asp), which is prevalent elsewhere. Sequence analysis revealed no mitations in 5 (12.5%) of the 40 isolates although this isolate was repeatedly resistant to rifampicin. VNTR-typing targeted to tandem repeats (ETR A, B, C, D, and E) was carried out to establish a genetic relationship for rifampicin-resistant isolates. Nine genetic types with VNTR-profiles termed as 12322, 32122, 32123, 32124, 32125, 32522, 23524, 12223, 22222, 33433 were revealed. There was no strict correlation between the type of mutation in the rpoB gene and the VNTR-type, which reflects different rates of evolution and the level of selective pressure on these genetic targets. The isolates of VNTR-types 32123 and 32125 with mutations in codon 531, and type 32122 in codons 531, 526, 516 showed a high clustering. This is likely to reflect the recent transmission and clonal dissemination of the epidemic strains of Mycobacterium tuberculosis. Thus, mutations in the rpoB gene did not reduce the virulence and transmissivity of these clones. Twenty-six of 27 clinical isolates selected by rifampicin-resistance were also resistant to isoniazid, which confirms the known fact that rifampicin-resistance may be used as a marker of isoniazid-resistance.

[Dynamics of major constituents of tuberculosis morbidity in the population of Novosibirsk region]. / Dinamika osnovnykh sostavliaiushchikh zabolevaemosti tuberkulezom naselenia Novosibirskoi oblasti.

After processing the dynamic series of tuberculosis morbidity in the population of the Novosibirsk region by the smooth component methods, two major constituents were defined in morbidity dynamics. In total, they make up about half the total variability of tuberculosis morbidity. The first constituent is due to socioeconomic causes that are common for the whole region. The second one shows rather evident 10-year fluctuations. The same 10-year fluctuations are observed in the dynamics of tuberculosis morbidity among the cattle of the Novosibirsk region by out-stripping that in the population in the northern and eastern areas by 1-3 years.

[Examining fibrotic process in lung during treatment of chronic murine tuberculosis with lysosomotropic drug isonaizid]. / Ussledovanie protsessov fibrozirovaniia v legkikh pri lechenii lizosomotropnym preparatom izoniazida khronicheskogo tuberkuleza u myshei.

Light and ultrastructural morphometries were used to evaluate the effects of dextran-conjugated isoniazid (MW 30-40 kDa) on the morphology of chronic pulmonary tuberculous inflammation induced by BCG vaccine in mice. The revealed antifibrotic effect of the conjugated drug is accompanied by much less damage to the alveolar macrophageal ultrastructures (the mitochondrial apparatus in particular) than that of free isoniazid. It is concluded that the lysosomotropic properties imparted to isoniazid basically change its effect on fibrogenesis and are essential in the prevention of antiinflammatory pneumoscleroses.

[Evolution of granulomas induced by BCG vaccine administered in the experiment]. / Evoliutsiia granulem, indutsirovannykh vvedeniem vaktsiny BTsZh v éksperimente.

The time course of hepatic granulomatosis was studied on a CBAxC57B1/6 murine model of a vaccinal process induced by BCG vaccine. It was found that over the natural course of inflammation, there were changes in the spectrum of granulomas from epithelioid cellular to macrophageal and fibrotic ones with stabilization of their sizes, averaged cellular composition and density in the organ. Long-term use of isoniazid caused a considerably (2-fold) reduction in the number of granulomas in the organ and thus a decrease in the level of its fibrotic process. Presumably, in the treated animals, the mechanism responsible for a reduction in the number of granulomas is the "dissociation" of macrophageal cells from them.

[Effectiveness of liposomal antibacterial drugs in the inhalation therapy of experimental tuberculosis]. / Effectivnost liposomalnoi lekarstvennoi formy antibakterialnykh preparatov v ingaliatsionnoi terapii éksperimentalnogo tuberkulëza.

The model of generalized tuberculosis in BALB/c mice was used to compare efficacy of daily and intermittent (2 times a week) regimens of ultrasound inhalation therapy. This employed water-soluble and liposomal dosages of antituberculous drugs. The highest effect was achieved in daily inhalations of liposomal drugs. In spite of the fact that intermittent regime of the liposomal drugs required a 2-fold lowered dosage, its efficacy was not inferior to daily water-soluble drug therapy and was only by 22% less than maximum. High response to liposomal form stems from prolonged persistence of high rifampicin concentrations in pulmonary tissue.

[Lymphogenic introduction into lesser circulation of orally administered rifampicin in standard and liposomal forms]. / Limfogennoe postuplenie v malyi krug krovoobrashcheniia peroral'no vvodimogo v obychnoi i liposomal'noi formakh rifampitsina.

Pharmacokinetics of water and liposomal suspensions of lipophilic antibiotic rifampicin introduced intragastrically through a tube was studied in experiments on Wistar rats. After administration of water suspension rifampicin concentration reached its peak in all the tissues in 3 hours, lung concentration being 1.6 and 1.9 times higher than blood one. After liposomal suspension administration lymphatic rifampicin concentrations were maximal in 3 hours, while in the lungs and blood they were the highest in 6 hours. It is concluded that oral liposoluble drug enter primarily chyle vessels and via thoracic lymphatic duct directly lesser circulation. Liposomal inclusion of the drug warrants its high concentration in lymphatic system and lung tissue.

[Ultrastructure of epithelioid cells in the dynamics of forming tubercular granules]. / Ultrastryktura épitelioidnykh kletok v dinamike formirovaniia tuberkuleznoi granulemy.

As shown by ultrastructural findings in epithelioid cells, their number and differentiation degree increase in granulomas of animals with chronic generalized tuberculosis. Isoniazid treatment stimulated differentiation of the monocytes into epithelioid cells and differentiation of the latter. A large number of epithelioid cells with adequate lysosomal apparatus may denote a risk of the destructive process in the affected organs. The efforts in the search for new potent anti-tuberculous drugs should be focused on their lysosomotropy, ability to stay long in the lysosomal apparatus being bactericidal and bacteriostatic, and to stimulate plastic processes in the macrophages.

[Changes in the granuloma in the experimental treatment of tuberculosis with a prolonged form of isoniazid]. / Izmeneniia granulemy pri lechenii tuberkuleza prolongirovannoi formoi izoniazida v éksperimente.

The main difficulties encountered in the treatment of the persistent forms of a chronic tuberculous process are associated with a difficult access to mycobacteria persisting in the vacuolar apparatus of macrophages that form granulomas as well as with a high level of toxic affections of the parenchymatous liver cells. The complex long-acting isoniazid preparation on the dextran basis and the scheme of its application prevented, to a considerable degree, the above complications under the experimental conditions and hence can be considered as promising for clinical use.

[Effectiveness of a liposomal form of rifampicin in the treatment of experimental tuberculosis in mice]. / Effektivnost' liposomal'noi formy rifampitsina pri lechenii éksperimental'nogo tuberkuleza myshei.

The therapeutic effectiveness of rifampicin as a free form and a component of multilamellar phosphatidylcholine cholesterol liposomes was studied on a model of generalized tuberculosis in BALB/c mice. Rifampicin as a liposomal form was found to have no advantages over its free form. Possible mechanisms of the phenomenon and prospects for using liposomes in the chemotherapy of tuberculosis are discussed.

[Effects of indomethacin and BCG vaccine on growth of transplanted tumors in mice]. / Vliianie indometatsina i vaktsiny BTsZh na rost perevivnykh opukholei u myshei.

Continuous administration of indomethacin (IM) in drinking water (20 micrograms/ml) causes moderate inhibition of the growth of transplantable tumors in control mice but significantly increases the antitumor effect of BCG vaccine inoculated in combination with transplanted tumor cells. Hepatoma 29, insensitive to the vaccine becomes highly sensitivity to it under IM administration.