Drug Resistance of Mouse Somatic Cells to Rifampicin in Experimental Tuberculosis.

We have demonstrated that long-term exposure of intact mice to rifampicin (6 months) induces resistance to this drug, which manifested in inability of rifampicin to suppress the growth of Mycobacterium tuberculosis in the lungs and spleen during subsequent infection. It the same time, isoniazid is still effective in these mice. In this case, the phenomenon of somatic resistance to rifampicin in mice was observed if the treatment was started in a short period (within 4 days) after infection with M. tuberculosis. If the interval between infection and rifampicin administration was longer (3 weeks), the resistance disappeared.

Plasticity of Human THP-1 Cell Phagocytic Activity during Macrophagic Differentiation.

Studies of the role of macrophages in phagocytosis are of great theoretical and practical importance for understanding how these cells are involved in the organism's defense response and in the development of various pathologies. Here we investigated phagocytic plasticity of THP-1 (acute monocytic human leukemia) cells at different stages (days 1, 3, and 7) of phorbol ester (PMA)-induced macrophage differentiation. Analysis of cytokine profiles showed that PMA at a concentration of 100 nM induced development of the proinflammatory macrophage population. The functional activity of macrophages was assessed on days 3 and 7 of differentiation using unlabeled latex beads and latex beads conjugated with ligands (gelatin, mannan, and IgG Fc fragment) that bind to the corresponding specific receptors. The general phagocytic activity increased significantly (1.5-2.0-fold) in the course of differentiation; phagocytosis occurred mostly through the Fc receptors, as shown previously for M1 macrophages. On day 7, the levels of phagocytosis of gelatin- and Fc-covered beads were high; however, the intensity of ingestion of mannan-conjugated beads via mannose receptors increased 2.5-3.0-fold as well, which indicated formation of cells with an alternative phenotype similar to that of M2 macrophages. Thus, the type and the plasticity of phagocytic activity at certain stages of macrophage differentiation can be associated with the formation of functionally mature morphological phenotype. This allows macrophages to exhibit their phagocytic potential in response to specific ligands. These data are of fundamental importance and can be used to develop therapeutic methods for correcting the M1/M2 macrophage ratio in an organism.

[Necrotizing sarcoid granulomatosis]. / Nekrotiziruyushchii sarkoidnyi granulematoz.

Necrotizing sarcoid granulomatosis (NSG) belongs to productive small-vessel vasculitis with the formation of sarcoid-like granulomas, which is accompanied by ischemic necrosis of varying degrees and duration. The disease involves the lung only. The clinical symptoms of the disease are nonspecific so the latter is detected rather rarely. The main diagnostic technique is morphological examination. Immune complex inflammation develops in the vessel walls with the formation of macrophage-histiocytic granulomas that do not contain epithelioid cells. The etiology and pathogenesis of NSG remain little studied. Its differential diagnosis is mainly presented with tuberculosis, sarcoidosis, and granulomatosis with polyangiitis.

Peculiarities of the Inflammatory Process in the Reproductive Organs of C57Bl/6 Female Mice with Experimental Tuberculosis.

Intravenous infection of C57Bl/6 female mice with M. tuberculosis H37Rv led to involvement of the lungs and dissemination of the tuberculous infection to the abdominal and pelvic organs. M. tuberculosis were detected in the lungs and spleen in 14, 35, and 90 days and in the uterine horns in 90 days after infection. Morphological analysis of organs showed successive development of exudative necrotic tuberculosis of the lungs, acute and chronic nonspecific inflammation in the reproductive organs (vagina, uterus, and uterine horns). The inflammatory process in the reproductive organs was associated with the development of anaerobic dysbiosis, that was most pronounced in 35 days after infection. Antituberculous therapy was followed by reduction of M. tuberculosis count in the lungs and spleen in 60 and 90 days after infection, eliminatation of M. tuberculosis in the uterine horns, arrest of nonspecific inflammation in female reproductive organs, recovery of the balance between aerobic and anaerobic microflora, and development of candidiasis of the urogenital mucosa.

[EFFECTIVENESS OF FULLERENE-(TRIS-AMINOCAPRONIC ACID) HYDRATE IN THE MODEL OF EXPERIMENTAL VIRAL-BACTERIAL PNEUMONIA OF MICE].

AIM: Study the effectiveness of the substance and various drug formulations of fullerene-(tris-aminocapronic acid) hydrate (FTAAH onwards) in the model of experimental viral-bacterial pneumonia of mice. MATERIALS AND METHODS: BALB/c mice were infected with influenza virus A/California/04/2009 and subsequently infected with Staphylococcus aureus. The animals were treated after viral infection with the substance and various drug forms of FTAAH, as well as comparative preparations--oseltamivir and arbidol. Therapy effectiveness was evaluated by clinical indicators (survival, lifespan, animal mass decrease reduction), virological (virus titer), microbiological (density of bacteria in lungs) parameters, confirmed by pathomorphological characteristics of lungs. RESULTS: FTAAH therapy in injectable form was effective in the model of a combined viral-bacterial pneumonia of mice by all the studied criteria: treatment increased mice survival, reduced the decrease of their body weight, resulted in a reduction of virus titers and density of bacteria in lungs, that correlated with the data from morphological study and signs of bronchopneumonia resolution in mice. FTAAH therapy in rectal form depended on animal infection schemes, as well as preparation dose, increasing with its increase. CONCLUSION: FTAAH substance is effective in the model of experimental viral-bacterial pneumonia of mice.

[Umifenovir (Arbidol) efficacy in experimental mixed viral and bacterial pneumonia of mice].

Pneumonia often occurs as a secondary infection after influenza and accounts for a large proportion of the morbidity and mortality associated with seasonal and pandemic influenza outbreaks. The efficacy of umifenovir (Arbidol) was investigated on a murine model of S. aureus pneumonia following A/California/04/2009 (H1N1) influenza virusinfection. Oral treatment with umifenovir (40 and 60 mg/kg/day) in all the contamination schemes increased the survival rate in the mice from 0% to 90% and lowered the animal weight loss. The umifenovir treatment also decreased the virus titer by ≥ 2 logs and the viable bacteria counts in the lungs of the mice. The lungs of the mice treated with umifenovir had less severe histopathologic lesions compared to the control group.

In vitro effects of pulmonary surfactant on macrophage morphology and function.

The effects of pulmonary surfactant on the morphology and functioning of young macrophages were studied on the model of monocyte/macrophage differentiation in vitro and on macrophages of the bronchial alveolar lavage fluid. Surfactant is not a differentiation inductor, but it stimulated the maturation and phagocytic activity of young macrophages. The stimulatory effect of surfactant on phagocytic activity of macrophages persisted even after its removal from the culture medium.

[Complex morphological diagnostics of tuberculosis and sarcoidosis of the lungs].

Differential diagnosis of disseminated lung diseases, particularly tuberculosis and sarcoidosis, presents certain difficulties for clinicians. In the verification of the diagnosis a decisive role belongs to the morphological study, for which the most commonly used material is transbronchial biopsy. Diagnostic signs of active disseminated pulmonary tuberculosis are presence of different-sized granulomas with signs merger, necrosis, infiltration of polymorphonuclear leukocytes, weakly expressed fibrillogenesis. Cytological markers of a specific process are young biosinteziruyuschie macrophages and acid-fast bacilli. The diagnostic features of pulmonary sarcoidosis are monomorphic granulomas with no tendency to merge and necrotisation expressed fibrosis and hyalinization processes. Cytological marker for the disease is the high content of epithelioid cells with signs of secretion.

[Optimisation of diagnostics and differential diagnostics disseminated pulmonary tuberculosis].

One of the reasons of dramatic situation with tuberculosis in Russia is untimely diagnostics of tuberculosis. The aim of the study was to identify the causes of diagnostic mistakes when we deal with disseminated pulmonary tuberculosis at current stage and to modernize the diagnostic process. The analysis of the diagnostic activity of the consultative diagnostic center of Central Tuberculosis Research Institute of Russian Academy Medical Sciences for 2011 was performed with special attention on the results of the survey of 505 patients with pulmonary dissemination. The frequency of discrepancies of disseminated pulmonary tuberculosis diagnostics was 96.1%. Based on the studies carried out the main causes diagnostic mistakes in patients with disseminated pulmonary tuberculosis were determined. New directions of improving of tuberculosis diagnostics were developed: overall high-technology examination of patient, adherence to the diagnostic procedure, developed by consultative diagnostic center of Central Tuberculosis Research Institute (CTRI), timely performing fiber-optic bronchoscopy with complex biopsy and diagnostic surgery procedures, further training of primary health care doctors. Implementation of proposed activities will significantly (by 3-5 times) reduce the time for diagnostics of respiratory system disease.

[Selective effects of pulmonary surfactant on various subpopulations of alveolar macrophages in the model of experimental tuberculosis].

Pulmonary surfactant is necessary component for maintenance of high level of phagocytic activity of alveolar macrophages. Tuberculosis inflammation reduces the production of surfactant by type II cells and phagocytic activity of alveolar macrophages. The effects of exogenous pulmonary surfactant on the ultrastructural changes of various subpopulations of alveolar macrophages were studied by TEM-method. For investigations the bronchial alveolar lavage fluid from guinea pigs infected of M. tuberculosis and treated by isoniatid or isoniazid + exogenous pulmonary surfactant were used. It was shown that isoniazid + exogenous pulmonary surfactant normalizes the heterogeneous population of alveolar macrophages providing stimulating effects on their maturation and phagocytic activity more effectively than isoniazid therapy.

[Cytologic features of bronchoalveolar lavage in evaluation of course of exogenous allergic alveolitis].

Application of complex of modern cytologic methods of research bronchoalveolar lavage allowed to allocate most characteristics of development of lymphocytic and macrophagic reaction of bronchial tree in different course of exogenous allergic alveolitis. The most indicative in assessment of origin of exogenous allergic alveolitis development is the characteristics of macrophagic population. In acute course of exogenous allergic alveolitis the considerable number of young activated and non-activated macrophages, increased number of mature phagocytes is observed. Even more significant increase of phagocytic macrophages is observed at dissemination which is primarily is connected with participation of these cells in lymphocytic apoptosis which takes place in high percentage of lymphocytes (up to 49%). Increased number of mature phagocytes is observed at chronic course of exogenous allergic alveolitis that is an important diagnostic pattern of this option of development of exogenous allergic alveolitis in association with the lowest T-helpers/T-supressors index.

[Up-to-date technologies in preoperative preparation and postoperative period in patients with progressive destructive pulmonary tuberculosis].

The impact of complex pathogenetic treatment involving the latest advances of science and technology in the correction of pulmonary surfactant system (PSS) disorders, extracorporeal therapy and immunomodulation on the enhancement of efficiency of surgical treatment in patients with progressive destructive tuberculosis of the lung. The results of examinations and treatment were studied in 199 patients with progressive and complicated pulmonary tuberculosis. According to the used treatment, the patients were divided into 2 groups: a study group (n = 102) and a control one (n = 97). In the pre- and postoperative periods, the study group patients received complex pathogenetic therapy by the developed algorithm: for pharmacological activation of PSS, nebulizer aerosol therapy with lasolvan (ambroxol) was first used in various modifications by the developed algorithms. The efficiency of preoperative preparation in the study group patients in accordance with the applied methods of pathogenetic therapy indicated that the best results were achieved in the context of stabilization of a specific process, compensation of complications and comorbidity in the patients who underwent modified drug-induced PSS activation with small-volume plasmapheresis and leukinferon (LI) in 93.3% and drug-induced PSS activation with small-volume medical plasmapheresis (MI) in 81.4%. Overall, comprehensive preoperative preparation involving correction of homeostatic disorders by the developed algorithm proved to be 30.9% more effective (81.4%) than the conventional preoperative preparation in the controls (50.5%). Analysis of the results of surgical treatment depending on the methods of pathogenetic therapy used in the pre- and postoperative period showed that the efficiency of surgical treatment was observed in 25 (80%) patients receiving nebulizer aerosol therapy and MP, in 20 (86.9%) who had nebulizer aerosol therapy + MP and in only 8 (65%) treated with nebulizer aerosol therapy.

[Pathomorphological changes in the lung in tuberculosis patients died from HIV infection at the stage of AIDS].

The authors revealed the typical morphological changes of lung tuberculous lesion in HIV infection at the stage of AIDS: these included alterative changes without typical tuberculosis granulomas; a well-defined exsudative inflammatory component with a predominance of leukocytic infiltration and a drastically decrease of and, occasionally, a complete disappearance of macrophages and lymphocytes; formation of pyonecrotic foci; the focal monomorphic pattern illustrating the loss of the signs of process indulation. These signs suggest the specific features of immunity and the course of specific inflammation as immediate hypersensitivity with the acutest progression of tuberculosis.

[Results of use of surfactant in complex therapy of patients with destructive pulmonary tuberculosis].

Many years' experience in studying the surfactant system in patients with pulmonary tuberculosis has allowed recommendation of using surfactant agents in the treatment of tuberculosis. The purpose of the study was to evaluate the efficacy of surfactant-BL (Russia) as a pathogenetic agent in chemotherapy in patients with destructive pulmonary tuberculosis. The results of treatment were compared in two groups of 70 persons in each, which were matched by gender, age, the extent of a tuberculous process, and the presence of drug resistance in the causative agent, including multidrug resistance. In the study group, the patients received surfactant inhalations (8 weeks) during chemotherapy while the control patients had only chemotherapy. Drugs were chosen on an individual basis, by taking into account the pathogen's drug sensitivity and a patient's tolerability of a drug. The cumulative dose of the surfactant was 700 mg. There were no adverse reactions to the surfactant in the study group of patients. After 2-5 surfactant inhalations, the amount of sputum increase, its discharge became easier, and cough diminished. Following 4 weeks, the level of bacterial isolation decreased in 49 (70.0%) patients from the study group and in 20 (28.6%) from the control one. Two months of treatment, bacterial isolation ceased in 82.9 and 64.3% in the study and control groups, respectively. In the study group, X-ray trend in infiltration resolution and cavernous closure was significantly better in the study group. By month 4, cavernous closure was achieved in 72.9% in the study group and in 41.4% in the control one. The changes in the cellular composition of the lung in surfactant-treated and untreated patients were analyzed by the data on broncho-alveolar lavage. The findings indicate that inhaled Surfactant-BL as a two-month therapy has a pathogenetic effect and during chemotherapy improves the efficiency of treatment reduces its time in pulmonary tuberculosis.

[Morphological signs of acute lung lesion in patients with caseous pneumonia].

The authors conducted light optical and electron microscopic studies of caseum-free resected lung parenchymal portions from 11 patients with caseous pneumonia. All cases were found to have severe vascular bed permeability impairments, developed extensive intraalveolar edema, destruction of both type 1 respiratory alveolocytes and surfactant-producing type 2 alveolocytes. Release of plasma proteins into the intraalveolar space and their interaction with extracellular phospholipoproteins gave rise to hyaline masses.

[On the morphological diagnosis of drug-resistant pulmonary tuberculosis].

The intraoperative samples taken from 15 patients with acutely progressive drug-resistant fibrocavernous pulmonary tuberculosis were studied. There were typical signs of granulomatosis inflammation, a predominance of an exudative tissue reaction, and an extensive vascular bed lesion. Two types of perifocal cellular infiltrates were identified. Mononuclear infiltrates with epithelioid cellular transformation along the periphery were defined as specified. Nonspecific infiltrates were composed of foam macrophages-lipophages and they reflected lipid metabolic disturbances. In addition, the severity of the process was determined by an extensive specific bronchial lesion of all generations. A morphological study of the samples could reveal the tissue and cellular features of respiratory organs in drug-resistant tuberculosis and identify the diagnostically significant signs of specific and nonspecific inflammation.

[Evaluation of the efficiency of activity of the liposomal form of isoniazid against different types of mycobacteria in vitro].

M. avium and three M. tuberculosis strains were used as an example to study the bacteriostatic activity of the liposomal form of isoniazid in the liquid nutrient medium by serial dilutions. Unlike isoniazid in solution, its liposomal form was found to suppress the multi plication of all study mycobacterial samples and diminishes their viability, but at different concentrations. The liposomes that did not contain the drug exerted an inhibitory effect only on the M. tuberculosis strains H37Rv and Erdman. It is suggested that the change in isoniazid sensitivity results from the effect of liposomal phospholipids on some mycobacterial enzymes. Electron microscopy indicated that the liposomal form of isoniazid is able to penetrate into the alveolar macrophageal cytoplasm of guinea pigs just 30 min after incubation.

[Impact of phthisiosurgery on the development of morphological studies of the lung].

The paper describes the most important developmental stages of phthisiomorphology in chronological order, by using as an example the work of the Pathomorphology Laboratory, Central Tuberculosis Research Institute, since its organization. It also shows the stages of phthisiosurgery with the inestimable scientific and practical contribution of the works by L. K. Bogush and his followers from the formation of lung surgery to today's achievements. The authors give examples of the long-term working partnership of surgeons and morphologists in the development of these two disciplines, which was fruitful in deciding many issues of phthisiology.

[Antimycobacterial activity of a dry birch bark extract on a model of experimental pulmonary tuberculosis].

The study deals with the evaluation of antimycobacterial activity of betulinol, a dry birch bark extract (BBE), on a model of infection caused by Mycobacterium tuberculosis (MBT). It has established the inhibitory action of betulinol on the in vitro and in vivo growth of MBT and its positive effect on reparative processes in the lung, liver, and spleen of tuberculosis-infected mice.