[Molecular markers of M. tuberculosis virulence in lung tissue (experimental study)]. / Molekulyarnye markery virulentnosti M. tuberculosis v tkani legkikh (eksperimental'noe issledovanie).

More than a quarter of the world's population is infected with Mycobacterium tuberculosis. However, only about 10% of those infected develop active TB. This indicates a key role for innate immunity in limiting M. tuberculosis replication. Most often, bacteria can regulate the expression of host-specific molecules and weaken host immunity. OBJECTIVE: To use a biological model, in order to determine significant molecular immunohistochemical markers characterizing the virulence of the "Buryat" and "Omsk" subtypes of the M. tuberculosis Beijing genotype in lung tissue. MATERIAL AND METHODS: Lung samples of the C57BL/6 male mice were obtained during experimental infection with M. tuberculosis strains: the reference laboratory strain H37Rv, multidrug-resistant clinical strains 396 (highly lethal and hypervirulent «Buryat¼ genotype Beijing 14717-15) and 6691 (low-lethal and low-virulent "Omsk" genotype Beijing 1071-32) on days 14, 21, 60 and 120. They were studied by histological and immunohistochemical methods. The relative areas of expression of IL-6, IL-12A, iNOS, and TNF-α in the lung tissue of model animals were established. RESULTS: A study of strain 396 showed that both disease progression and damage to lung tissue are associated with a highly reactive immune response and increased synthesis of iNOS and strain characteristics that block the production of TNF-α. On the contrary, for strain 6691 a low reactivity of the immune response was revealed, with statistically significantly lower values of the relative area of expression of NOS and TNF-α during all observation periods (days 14-120). All animals that survived to day 120 showed a similar morphological picture with differences in cytokine levels, indicating a nonlinear relationship between proinflammatory factors and the damage substratum. CONCLUSION: The progression of the disease and damage of lung tissue were associated with a highly reactive immune response and increased synthesis of iNOS, strain properties that block the TNF-α production. Thus, iNOS and TNF-α can act as molecular markers characterizing the virulence of the "Buryat" and "Omsk" subtypes of M. tuberculosis in lung tissue.

[Preclinical and clinical trials of the new tuberculosis drug perchlozon].

The paper sets forth the stages of design and introduction of the new Russian tuberculosis (TB) drug perchlozon registered in the Russian Federation in 2012. Based on the results of Phases I-III clinical trials, the authors evaluate the efficacy and safety of the agent and consider the adverse effects of its treatment for respiratory TB. The use of perchlozon as a component of combination therapy versus standard chemotherapy regimens significantly reduces abacillation time in pulmonary TB caused by its drug-resistant pathogen. In terms of the higher prevalence of TB induced by its pathogen resistant to many drugs (with multiple and broad-spectrum drug resistance), perchlozon is an essential drug that has antituberculous activity mainly against multidrug-resistant Mycobacterium tuberculosis strains and gives patients with the severest and epidemiologically poor form of TB the chance to recover.

[The influence of immunotropic drugs on reparative processes in the lungs experimental chemotherapy drug resistant tuberculosis].

It is revealed that Roncoleukin (12.5 mg/kg, intraperitoneally, 5 injections a day), Betaleukin (0.1 mg/kg, intraperitoneally, 1 time in 3 days (5 weeks)), Bestim (0.1 mg/kg, intraperitoneally, 10 injections), cycloferon (3.6 mg/kg, intraperitoneally, 3 times a week for 6 weeks), Glutoxim (40 mg/kg subcutaneously (4 weeks)and the preparation of succinic acid remaxol (at a dose of 25 mg/kg intraperitoneally, daily 14 introduction), when you enter them in a comprehensive drug therapy pilot MDR tuberculosis in mice produce a positive effect on the regression of inflammation in the lung tissue, stimulate local immunity of the lungs, activate and digestive absorption capacity of peritoneal macrophages an average of 1.4 and 1.9, p < 0.05, inhibited the tuberculosis infection and chemotherapy.

[The hepatotropic activity of runihol and ademethionine against experimental liver damage caused by first-line antituberculosis drugs].

Experiments on 62 male albino outbred rats investigated the impact of 14-day use of the oral hepatotropic agents ademethionine and runihol in different doses in liver damage caused by a antituberculosis combination (HRZ). HRZ-induced liver damage was shown to be accompanied by the development of cytolysis and cholestasis. The test drugs in the doses under study had a mixed effect, by reducing the magnitude of biochemical manifestations of these syndromes. Both drugs favored the recovery of the structure of the liver and the reduction of the extent of carbohydrate, protein, and fat dystrophies of the organ. As in previous investigations, ademethionine was found to have enhanced activity of alterative processes in the liver.

[Reparative osteogenesis after incorporation of roncoleukin into therapy for experimental tuberculous gonitis].

The authors evaluated reparative osteogenesis after incorporation of roncoleukin into combination therapy for experimental drug-resistant tuberculous osteitis. Roncoleukin (12.5 pg/kg, 5 injections, once every 3 days) was used during antituberculosis therapy before and after combined plasty (autobone + OsteoSet-T) in repairing surgical defects. When used in the postoperative period of combined osteoplasty, roncoleukin was shown to contribute to a reduction in the spread of specific inflammatory foci in bone tissue, cessation of an alternative necrotic component, an increase in the rate of osteogenesis with newly formed bone trabeculae, and activation of hematopoietic processes in the bone marrow. In parallel with intensified bone reparative processes, there was immunomodifier-induced stimulation of the absorption and digestion of peritoneal macrophages suppressed in experimental tuberculous osteitis.

[The hepatoprotective activity of remaxol and S-adenosyl-L-methionine for liver damage caused by reserve-series antituberculosis drugs].

Experiments on male albino outbred rats investigated the hepatoprotective activity of remaxol and ademethionine in liver damage caused by reserve-series antituberculosis drugs. The test drugs had a mixed action, by reducing the degree of cytolytic and cholestatic syndromes. Remaxol and ademethionine assist in diminishing the degree of liver structural and functional impairments occurring with reserve-series antituberculosis drugs, by restoring the girder structure of lobes and decreasing the magnitude of dystrophic changes to stimulate reparative processes. Histological examination of liver tissue sections from rats receiving ademethionine revealed the enhanced activity of alterative processes in the liver.

[Immunotropic and antihypoxant therapy of experimental drug-sensitive and drug-resistant tuberculosis].

The results of pre-clinical research of cycloferon, remaxol and runihol on the model of experimental generalized tuberculosis, caused by the MBT with a different spectrum of drug sensitivity are presented. A considerable increase of the curative effect of the therapy with the used of cycloferon and remaxol. There was manifested the strengthening of lung clearance from the office, reducing the prevalence of specific inflammation in the lungs of the index of lung damage, stimulation of sorption and destructive ability of peritoneal macrophages, inhibited in the course of development of experimental tuberculosis infection. Runihol has no impact on the effectiveness of chemotherapy in the absence of a stimulating influence on the phagocytic function of the peritoneal macrophages.

[Pharmacological activity of runihol and S-adenosyl-L-methionine in rats with experimental liver damage by reserve antituberculosis drugs].

The hepatoprotective action of runihol and S-adenosyl-L-methionine (ademethionine) has been studied in a group of 47 white outbred male rats with model liver injury induced by reserve antituberculosis drugs (PAS, prothionamide, cycloserine). The ability of test drugs to correct structural and functional liver disorders is established. Both runihol and ademethionine favored decrease in the signs of structural and functional liver disorders induced by reserve antituberculosis drugs, Showing mixed type of action, the test drugs promoted recovery of the liver parenchyma and reduced manifestations of hepatocyte dystrophy to the same extent, without manifestations of necrobiotic processes and a mononuclear infiltration.

[Comparative efficacy of clinical use of reamberin, remaxol and ademethionine in patients with tuberculosis of the respiratory organs and liver drug-injury].

The efficacy ofreamberin, remaxol, S-adenosyl-L-methionine (ademethionine) and 5% glucose solution was estimated in the treatment of patients with tuberculosis of the respiratory organs and drug hepatotoxicity signs confirmed by higher activity of liver indicative enzymes and nitrogen oxide levels. Remaxol showed a pronounced positive effect on the cytolytic syndrome signs, evident from lower activity of alanine aminotransferase and aspartate aminotransferase. At the same time ademethionine was superior to remaxol in the effect on the cholestatic signs and inferior in the effect on the cytolytic signs. By the effect on the activity of alanine aminotransferase and aspartate aminotransferase, reamberin was inferior to remaxol and superior to ademe-thionine, its effect on the cholestasis markers level vs. the other drugs being superior only to that of 5% glucose solution. As compared to reamberin, ademethionine and 5% glucose solution, remaxol promoted higher integral indices of the host antioxidant protection (total antioxidant capacity and total antioxidant status), that partially explained the drug pronounced hepatoprotective effect.

[Hepatotropic therapy in treatment of liver injury].

At present, the conception of the use, efficacy and safety of hepatotropic agents in treatment of drug-induced liver injury, in particular due to antituberculosis drugs is not yet final, which is conditioned by extremely rare clinical trials on the subject adequate to the up-to-date principles of the conclusive medicine. The review presents data on the hepatotoxic effect of antituberculosis drugs, analysis and systematization of the data on the use of hepatotropic agents in liver injury induced by antituberculosis drugs, the principles and characteristics of their clinical use. The mechanism of action of remaxol, a new original hepatotropic agent and the indications of its use are discussed. The experimental findings on the remaxol ability to decrease the antituberculosis drug-induced liver injury through lowering the carbohydrate, albuminous and fatty degeneration and activating the organ reduction are presented. The clinical trials are evident of the most efficient action of remaxol on the signs of toxemia, as well as cytolysis and cholestasis, which along with its antiasthenic and antidepressant action allows to use remaxol as an universal hepatotropic agent in the treatment of diverse drug-induced liver injuries in both the therapeutic and prophylactic schemes.

[An experimental study of the efficiency of cycloferon in the complex chemotherapy of generalized drug-resistant tuberculosis].

The results of an experimental study of the efficiency of cycloferon included in a complex chemotherapy of generalized drug-resistant mycobacterium tuberculosis (MBT) are presented. It is established that cycloferon (3.6 mg/kg) produces a significant therapeutic effect, which is manifested by an increase in the lung clearance from MBT, a decrease in the spread of specific inflammation in the lungs, and the disappearance of MBT-induced alterations. In addition, activation of the signs of tension in the local immunity of lung tissues is observed as manifested by changes in the cellular composition of granulomas and more frequent detection of large lymphocytic and macrophage infiltration. The administration of cycloferon significantly increases the absorptive and digestive activity of phagocytosis by peritoneal macrophages, which has been inhibited during the development of experimental MBT infection.

[Effect of succinate- and methionine-containing drug runihol on reparative regeneration processes in liver under experimental hepatectomy conditions].

The effect of runihol and ademethionine on the processes of reparative regeneration in the liver has been experimentally studied in a group of 78 white male rats subjected to partial hepatectomy, in comparison to intact and drug-untreated operated control animals. The administration of ademethionine and runihol within the first four days after operation led to a decrease in the relative area of discomplexation in liver beams and the appearance of prerequisites of the activation of regeneration processes. A morphometric investigation showed that ademethionine stimulated intracellular regeneration reactions on all terms after the partial extirpation of liver, the most pronounced effect being observed on the 4th and 10th days, while the administration of runihol led to comparable effects only on the 14th day. The activation of regeneration processesin the liver parenchyma is retained after cancellation of the preparations.

[Investigation of remaxol efficacy in complex therapy of experimental generalized tuberculosis on mice].

Efficacy of remaxol in complex chemotherapy of generalized drug resistant tuberculosis was studied on mice. Mycobacterium tuberculosis 5419 SPBNIIF isolated from a patient with freshey diagnosticated pulmonary tuberculosis resistant to isoniazid (10 mcg/ml), rifampicin (40 mcg/ml), streptomycin (10 mcg/ml) and ethionamide (30 mcg/ml) was used in the experiments. The main polychemotherapy included 4 antituberculosis drugs in the highest therapeutic doses: isoniazid, amikacin, ethambutol and tavanic, the treatment course was 8 weeks. Remaxol was administered in a dose of 25 ml/kg intraperitoneally 5 times a week (14 injections). Significant activating effect of remaxol on the tension of the lung tissue local immunity was revealed by changes in the granuloma cell composition (from mainly epitheliod to mainly lymphoid) and by more frequent large lymphohistiocytic infiltrates. The use of remaxol also greatly increased the absorptive and digestive activity of the peritoneal macrophages phagocytosis, inhibited in the process of the experimental tuberculosis development.

[Prevention of tuberculosis: current approaches to development of vaccines].

This review is focused on recent advances in development of new vaccines for the prevention of tuberculosis. The main reasons for lack of BCG vaccine efficacy in different populations and geographic regions are presented. Design of new vaccines based on live modified strains of Mycobacterium bovis BCG, attenuated strains of Mycobacterium tuberculosis, recombinant proteins and viral vectors is considered in the specific examples. The usage of the heterologous "prime-boost" vaccination strategy against tuberculosis is discussed.

[The effect of runihol and exogenous S-adenosyl-L-methionine on the morphological pattern of the liver upon hepatotoxic exposure to reserve-series antituberculous drugs].

Experiments on 160 male albino outbred rats investigated the hepatoprotective activity of S-adenosyl-L-methionine (SAM) and runihol in liver damage caused by subtoxic doses of reserve-series antituberculous drugs (ATD) (PASA, cycloserine, prothionamide) and their combination. It was established that a combination of ATDs had the maximum hepatotoxic activity, cycloserine had the least. There was evidence that SAM versus runihol had a more pronounced ability to correct ATD-induced evolving cytolysis syndrome. Histological study indicated that SAM and runihol also showed a marked hepatoprotective effect. When added to PASA, prothionamide, or a combination of ATDs, both drugs promoted recovery of the hepatic architectonics and a reduction in the prevalence of albuminous dystrophy. The use of SAM additionally led to activation of alterative processes in the hepatic parenchyma.

[Comparative study of hepatoprotective action of remaxol, reamberin and ademethionine in liver injury induced by antituberculosis drugs (experimental study)].

The hepatoprotective activity of remaxol, reamberin and ademethionine was studied on a model of the liver injury induced by antituberculosis drugs. The study included 30 male uninbred albino rats. The following antituberculosis drugs were used: isoniazid (50 mg/kg) subcutaneously + rifampicin (250 mg/kg) intragastrically + pyrazinamide (45 mg/kg) intragastically (by the procedure of Yu. I. Slivka, 1989). Remaxol, reamberin and ademethionine were administered 1.5 hour prior to the antituberculosis drugs. The treatment course was 14 days. It was shown that remaxol, reamberin and ademethionin were able to correct the structural and functional disorders in the liver due to the use of the antituberculosis drugs. By the impact on the biochemical indices, evident of the liver function condition, remaxol showed the maximum effect. The effect of ream-berin was somewhat lower and the results of the ademethionine use were less significant. Remaxol had also a distinct effect as for lowering the level of the structural injuries in the liver, evident from recovery of the organ histoarchitectonics, less extended carbohydrate, albuminous and fatty degeneration, more active intracellular regeneration. It was noted that ademethionine had an insignificant effect on necrobiosis. Moreover, there was once detected a large necrosis focus, evident of possible stimulation of the liver tissue alteration by the drug.

[Comparative study of remaxol and ademethionine effects on reparative regeneration processes in the liver subjected to surgical intervention].

The influence of remaxol, reamberin and ademethionine on the process of reparative regeneration in the liver was experimentally studied in a group of 100 white male rats subjected to partial hepatectomy in comparison with intact and drug-untreated operated control animals. The administration of remaxol and ademethionine favored more intensive and early accumulation of the regenerated liver weight, restoration of the general structure of the liver, and appearance of the signs of reparative regeneration. Only rats treated with ademethionine exhibited clearly pronounced enhanced alterative processes in the liver. A morphometric investigation showed that remaxol and ademethionine improved the quality of a current compensatory process and stimulated intracellular regeneration reactions. Under the conditions of remaxol administration, the most pronounced effect of liver regeneration was observed on the 4th day after operation, while in the case of ademethionine the maximum effect was observed on the 10th day.

[Effectiveness of new diagnostic drug Diaskintest in children for tuberculosis diagnostic].

DST was ascertained to have a high sensitivity in virtually all patients with tuberculosis and a positive reaction was first noted in the infected. With stabilization and regression, the response to DST was much less pronounced than that in clinical and primary infection (that to the Mantoux test being more evident). DST showed its use as a marker of active tuberculosis not only in its local forms, but also in latent tuberculous infection. This makes it possible to apply DST when preventive treatment is performed. The agent may be used to monitor the progress of treatment. DST has a high specificity--healthy individuals had a negative response to DST while the Mantoux test was positive in many cases. The high specificity of DST was suggested by the fact that the persons vaccinated with (this caused BCG ostitis) had a negative reaction to DST while the Mantoux test was positive in all cases BCG-vaccinated BCG. The findings warrant the use of DST for the differential diagnosis of tuberculosis and BCG-associated complications and the possibility of differentiating postvaccinal and infection allergy in children and adolescents.

[The effectiveness of influenza vectors expressing the protective mycobacterial antigen ESAT-6 in the complex therapy of generalized tuberculosis in mice].

The use of Flu/ESAT-6 in chemotherapy for experimental tuberculosis has been found to cause a significant improvement of its results, which appears as the higher clearance of lung tissue from mycobacteria, the reduced extent of specific inflammation foci, and the altered cell composition of granulomas with mainly epithelioid to mainly lymphoid one. Flu/ESAT-6 of Tx-1 immunity (spontaneous and phytohemagglutinin-induced proliferation of splenocytes, spontaneous production of gamma-IFN) and ESAT-6-specific stimulation of a cell immune response (the proliferative activity of splenic lymphocytes and the production of IL-2 in response to recESAT-6) is shown to activate. The influenza vector A/PR8/NS1-125 that contains no ESAT-6 insert shows immunoadjuvant properties, which is likely to cause some reduction in the severity of tuberculosis in the control group of the vaccine.

[A role of the HLA-DRB1* genotype in the complex therapy of pulmonary tuberculosis].

It is as for now evident that in-depth researches of the genotype of a patient are needed to enhance the efficiency of therapeutic measures. The purpose of the present study was to enhance the efficiency of complex antituberculous therapy, by genetically identifying the allelic polymorphism of the HLA-DRB1* gene in a person who had ill with tuberculosis. The material of the study was the results of a followup and treatment of 100 patients with pulmonary tuberculosis. The subject of a special study was the molecular typing of the HLA genes of the DRB1* locus by polymerase chain reaction (PCR-SSP). The findings suggest that an individual approach to choosing treatment regimens (chemotherapy and pathogenetic therapy) for patients with pulmonary tuberculosis, by taking into account the HLA-DRB1* genotype enables one to enhance the efficiency of treatment in the major clinical and X-ray parameters.