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Tuberculosis (TB) is a disease caused by Mycobacterium tuberculosis (Mtb) infection with the formation of a broad range of abnormal lung lesions within a single patient. Although host–pathogen interactions determine disease outcome, they are poorly understood within individual lesions at different stages of maturation. We compared Mtb load in a tuberculoma wall and the lung tissue distant from tuberculomas in TB patients. These data were combined with an analysis of activation and bactericidal statuses of alveolar macrophages and other cell subtypes examined both in ex vivo culture and on the histological sections obtained from the same lung lesions. The expression of pattern recognition receptors CD14, CD11b, and TLR-2, transcription factors HIF-1α, HIF-2α, and NF-κB p50 and p65, enzymes iNOS and COX-2, reactive oxygen species (ROS) biosynthesis, and lipid production were detected for various lung lesions, with individual Mtb loads in them. The walls of tuberculomas with insufficient inflammation and excessive fibrosis were identified as being the main niche for Mtb survival (single or as colonies) in non-foamy alveolar macrophages among various lung lesions examined. The identification of factors engaged in the control of Mtb infection and tissue pathology in local lung microenvironments, where host–pathogen relationships take place, is critical for the development of new therapeutic strategies.

Introduction: Immunological lab tests do not allow to reliably determine TB activity when the calcifications of the lung tissue and intrathoracic lymph nodes (Ca) are observed. Aim: To create a predictive model for determining the TB activity in children with Ca. Materials: A training set of patients (n=509), combined according to 6-12 months dynamics of the X-ray, clinical and epidemiological data, immunological tests with ESAT-6CFP-10 as well as Mtb detection, was created. We revealed a relationship between real and calculated levels of leukocytes (CLL), eosinophils (EO, %) and γ-globulins (γ-GF), with the activity of humoral immunity, provoking an active state of the infection. A level of α2-globulins (α2-GF) indicated a chronic inflammation activity. The model is based on a comparison of CLL in the patient’s blood within the acceptable upper (UB) and lower bounds (LB). The bounds were calculated according to the following data: Ca number (x1), age (x2), hemoglobin (x3), EO (x4), α2-GF (x5), γ-GF (x6) levels. UB-formula: UB1 = ((-3.944*x1)-1.01*x2-0.356*x3+1.664*x4-6.679*x5+0.574*x6+121.46); UB2 = (36.833*x1+3.681*x2+1.126*x3+0.916*x4-2.638*x5+1.119*x6-157.1)). LB-formula: LB1 = (0); LB2 = (-25.25*x1+0.653*x2+0.42*x3+2.877*x4+0.523*x5+0.54*x6-74.34)). Condition: if LB1 Results: In the training sample the accuracy of determination of inactive TB was 97.5%, while for active - 96%. Conclusion: The developed model did not require any special equipment and allowed to observe Ca-possessing patients timely.

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is a highly infectious and successful human pathogen. In human lungs, Mtb are recognized and engulfed by alveolar macrophages, leading to the release of many antimycobacterial factors, including reactive oxygen species (ROS) and nitric oxide produced by the enzyme inducible nitric oxide synthase (iNOS). However, Mtb can survive, replicate, and persist in human lungs for a long time. In this study with using immunofluorescence assay, we examined the number of Mtb-infected alveolar macrophages and the quantity of ROS- and iNOS-positive cells in the ex vivo cell cultures obtained from the resected lung tissues with different TB lesions: tuberculomas and lung parts distant from macroscopic TB lesions, - in the same patients with pulmonary MDR-TB as described in (Ufimtseva, E. et al. PLOS ONE 2018; 13:e0191918). We found more alveolar macrophages with single Mtb or Mtb in colonies, including those with cording morphology, in the wall of tuberculomas than in the distant lung tissues of the same TB patients. Conversely, many alveolar macrophages with the antimicrobial agents examined were detected in the distant lung tissues, but not in the wall of tuberculomas. Thus, TB patients’ alveolar macrophages were characterized by different microbicidal potential and contained a different number of Mtb in various TB legions of the same resected lungs. Finally, our findings provide a crucial insight into Mtb pathogenesis during human TB disease. Studies of the mechanisms of Mtb survival within alveolar macrophages during human TB disease are extremely important for the development of new methods for TB treatment.

2-Substituted 5-aryltetrazoles containing alkyl and aminoalkyl fragments were synthesized using alkylation and Mannich reactions. The tuberculostatic activity of the synthesized compounds against strain M. tuberculosis H37Rv was studied. Pronounced tuberculostatic activity (MIC ≤ 1.5 ig/mL) was found for tetrazoles containing N(2) dimethyl- and diethylaminoethyl fragments.

Studies of the antimycobacterial activity of 2-amino-6-chloropurine and its N-acyl derivatives against laboratory (Mycobacterium tuberculosis H37Rv, M. avium, M. terrae) and clinical Mycobacterium tuberculosis strains (MDR-TB) identified compounds with high tuberculostatic activity (MIC from 0.35 – 1.5 μg/ml). The cytotoxic activities of the compounds with antimycobacterial activity against human embryo fibroblasts were studied in the MTT test, which showed that the study compounds were essentially nontoxic (IC50 > 50 μM). Further chemical modification may yield compounds with potential for creating drugs for the treatment of tuberculosis. With the aim of identifying the possible mechanism of the tuberculostatic activity, the ability of the study compounds to inhibit mycobacterial serine-threonine protein kinases (STPK) was assessed. The tuberculostatic activity of 2-amino-6-chloropurine derivatives was not linked with their inhibition of STPK.

A facile two-step synthetic approach to fluorinated and non-fluorinated 5-aryl-4-(5-nitrofuran-2-yl)-pyrimidines from readily available 5-bromo-4-(furan-2-yl)pyrimidine has been developed. All synthesized compounds were screened in vitro for their antibacterial activities against twelve various bacterial strains. It is demonstrated that some of these compounds exhibited significant antibacterial activities against strains Neisseria gonorrhoeae and Staphylococcus aureus, comparable and even higher with that commercial drug Spectinomycin.

Tuberculosis (TB) is a significant global health threat, with one-third of the world’s population infected with causative agent M. tuberculosis (Mtb). With the increasing prevalence of multidrug and extensively drug resistant (MDR and XDR) TB, there is a need to study the mechanisms of Mtb survival in patients’ alveolar macrophages. The Mtb shape and size alterations during TB disease are expected to be as the markers of virulence, specific defense against host responses and the peculiarities of microbe’s pathogenesis in individual patients. We estimated Mtb shape and size in alveolar macrophages isolated from the resected lungs of patients with pulmonary MDR- and XDR-TB (Ufimtseva, E. et al. Tuberculosis 2019; 114:77-90) after ex vivo culture for 16-18 hours and found its significant variations in Mtb as single and in colonies, including with cord morphology. Mycobacterial shape within alveolar macrophages varied from shorter oval, approximately 0.5 to 1 µm in length, to the classical rods with a mean length in 2-4 µm, and long filamentous forms over 6-7 µm in length, while Mtb width did not change significantly. The Mtb ovoid and classical rods were observed simultaneously in the same ex vivo cell cultures of patients with MDR- and XDR-TB, but often in the different alveolar macrophages, probably reflected some environmental fluctuations in patients’ lungs. The presence of filamentous Mtb was associated with increased virulence of pathogen from the lungs of these patients in guinea pig TB model. Thus, the shape and size variations demonstrated pleomorphic phenomena in Mtb populations during persistence in TB patients’ lungs and the specific features of MDR- and XDR-TB disease.

Aim: to еvaluate the possibility of historical transthoracic needle under CT-navigation (THBL) and transbronchial biopsy (TBL) to verify the etiology of focal lesions of the lungs. Material and Methods: two randomized groups of patients (50) with limited focal lung lesions, the etiology of which could not be clarified by non-invasive diagnostic methods. THBL with the area of the biopsy under CT was performed on 20(group 1), TBL with the material sampling of the 2nd segment of the right lung was performed on 30(group 2). The biopsy material in the form of 1-2 columns of tissue 1-2 mm in diameter and a length of 1-2 mmby patients of the 1st group and 2-4 fragments diameter of 3 mm by patients of the 2nd group was subjected to a complex histological and molecular genetic (DNA search of M. tuberculosis). Results: Pathologically altered lung tissue was obtained in 85% in the 1st group and 56.3% in the 2nd group (p Conclusion: THBL under CT navigation is the method of choice for verification-focal lesions of the lungs.

A series of 6-fluoroaryl substituted [1,2,4]triazolo[1,5-A]pyrimidines have been synthesized by using the microwave-Assisted Suzuki cross-coupling reaction from readily available 6-bromo-[1,2,4]triazolo[1,5-A]pyrimidine. The antimicrobial activity of new compounds has been evaluated in vitro against Mycobacterium tuberculosis H37Rv and gram-negative (Neisseria gonorrhoeae ATCC 49226) bacteria. © ARKAT-USA, Inc.

A method for the preparation of 3-oxobutanoate-containing podands differing in the length of the polyether fragment by acetoacetylation of oligo(ethylene glycols) with 2,2,6-trimethyl-4H-1,3-dioxin-4-one upon heating in toluene was optimized. A multi-component Biginelli reaction was used for the formation of the dihydropyrimidine ring on the ether matrix obtained. Tuberculostatic activity of dihydropyrimidine podands in the in vitro experiments against a laboratory strain H37Rv was studied. A combination of dihydropyrimidine and podand fragments imparts tuberculostatic activity to the compounds obtained, which noticeably enhances with the increase in the oxyethylene spacer length.

Tuberculosis (TB) is a dangerous airborne disease caused by Mycobacterium tuberculosis (Mtb) and characterized by a tight interplay between pathogen and host cells, mainly alveolar macrophages. Studies of the mechanisms of Mtb survival within human cells during TB disease are extremely important for the development of new strategies and drugs for TB treatment. We have used the ex vivo cultures of alveolar macrophages and histological sections obtained from the resected lungs of patients with pulmonary TB to establish the unique features of Mtb lifestyle in host cells. Our data indicate that Mtb with different virulence, as single and in colonies, with or without cording morphology, are exclusively intravacuolar pathogens with intact phagosomal membranes in viable host cells of TB patients and Mtb-infected guinea pig. Mycobacteria were detected in the cytoplasm and/or damaged vacuoles only in alveolar macrophages with morphological signs of cell death after prolonged ex vivo culture, however Mtb were found inside phagosomes in viable alveolar macrophages or cells with apoptotic/necrotic morphology in the same ex vivo cell culture. The Mtb phagosomes interacted with human different endocytic pathways, but inhibited phagolysosomal biogenesis, while intracellular vesicles containing Mtb products were fused with lysosomes in the same host cells.

Background: Mechanisms of natural tuberculosis (TB) resistance are not still clear enough. It does not allow the creation if effective means of pathogenic therapy. It is important for drug-resistant TB. Objectives: Studying of influence of arginine on the course of TB. Methods: A model of experimental TB was done by laboratory animals, infected historic stain of TB (H37Rv) from Central Tuberculosis Research Institute, Moscow. Groups of animals: 3 groups of guinea pigs (healthy, infected of TB, infected of TB with a course of arginine) and 2 groups of rats (healthy, infected of TB). It was indicated the levels of nitric oxide (3/5), total lipid peroxides in plasma by ELISA (Biomedica, Gruppe-R&D Syst.). Results: The primary level of nitric oxide (3) of plasma was 56,3±5,4μmol/l in the control group whereas in study grope without the course of arginine - 88,8±3,2μmol/l and with the course of arginine - 250,6±57,9μmol/l. Wherein animals with the course of arginine had significantly heavier tuberculosis than other groups. Morphological examination of lesions of internal organs was showed 4 points in groups with arginine in comparison with 2 points - without arginine. It was showed significantly increased in the level of oxystate in guinea pigs but not in rats after infection of TB. Conclusions: Natural tuberculosis resistance of animals is multifactorial phenomenon. It can depend on the reactivity level of the active oxygen- containing and nitrogen-containing compounds after infection. Animals without natural tuberculosis resistance have high level of product of nitric oxide in plasma. It is negative factor producing progression of TB. With that phagocytic activity of peripheral blood leukocytes is depressed.

Introduction: Clinical situations and anatomic variants in tracheal stenosis (TS) are quite various so different approaches for artificial lung ventilation (ALV) during tracheal resection are required. The choice of the method of ALV depends on the extent of the lesion, the width of the lumen of the stenotic area, the presence of tracheostomy, and the stage of the operation. Aims: The aim of the study is to elaborate tactics of ALV which should provide good conditions for the surgeon as well as proper gas exchange at every stage of the operation. Methods: The tactics of ALV was developed during circular resection of the trachea in 52 patients with TS located in cervical part of the trachea. Orotracheal tube was inserted in any case. At the stage of trachea dissection and preparation for resection convective mandatory ventilation (CMV) was conducted when the orotracheal tube could be passed distal to the lesion. If the diameter of the stenotic area was too small the catheter was passed from the tube via the stenosis, and high frequency jet ventilation (HFJV) was conducted. The regimen of HFJV was selected to prevent barotrauma and hypoxemia. In the presence of tracheostoma CMV was conducted via the stoma. At the stage of anastomosis suturing the HFJV regimen was continued via the cateter. After the anastomosis completion the tube was passed to the distal trachea, and CMV was continued. Results: No complications occurred during the operation. A proper gas exchange was provided. Conclusions: The combination of CMV and HFJV provided proper conditions for surgical manipulation and adequate gas exchange at any stage of tracheal resection irrespectively of the duration of open airways period.

M. tuberculosis (Mtb), the causative agent of tuberculosis (TB), is spread through aerosols and taken up by alveolar macrophages, where pathogen is internalized in phagosomes for survival and persistence. It had been recently reported that Mtb vacuole-to-cytosol escape is required for Mtb replication and dissemination in TB disease progress. We isolated alveolar macrophages from the resected lungs of patients with pulmonary TB and found the large number of viable cells with Mtb as single and in colonies, including with cord morphology (Ufimtseva, E. et al. PLOS ONE 2018; 13:e0191918). For detecting the location of Mtb in TB patients’ alveolar macrophages we applied fluorescence microscopy assay of phagosomal integrity, based on the use of digitonin to label Mtb (primary antibodies to Mtb lipoarabinomannan) that are in cytosol or within disrupted phagosomes, and, then, microscopy assay of same macrophages as in fluorescent images re-stained for acid-fast Mtb by the Ziehl-Neelsen method. Our results document that Mtb with different virulence, as single and in colonies, including with cord morphology, are exclusively intravacuolar pathogens in healthy alveolar macrophages of studied TB patients (Fig. 1). Fig. 1. The lack of Mtb (white arrow) in left image and the presence of Mtb (black arrow) in the same host cell in right image indicate the phagosomal location of Mtb in alveolar macrophage of patient 6.

Tuberculosis (TB) is an airborne dangerous disease caused by M. tuberculosis (Mtb) and characterized by a tight interplay between Mtb and host cells. Instead of being eliminated, Mtb demonstrates the remarkable ability to survive, replicate, and persist in human lungs on a long time. Studies of the mechanisms of Mtb survival within alveolar macrophages during TB infection are extremely important for the development of new vaccines and methods for TB treatment. Previously, we established the intravacuolar location of Mtb with different virulence, as single and in colonies, in alveolar macrophages of patients with pulmonary TB. Here, we analyzed the relationships between Mtb and TB patients’ alveolar macrophages in phagosomal compartments. In confocal fluorescence microscopy assay with the use of antibodies and other reagents, we determined that Mtb phagosomes can have the sings both the early (the presence of CD14 receptor and the lack of reactive oxygen species) and the late (the presence of iNOS) phagosomes. The lack of colocalization of the acidophilic LysoTracker probe and Mtb phagosomes was observed. Also, no colocalization of Mtb phagosomes with filamentous actin was identified. However, we detected numerous host cell vesicles with Mtb secreted products fused with lysosomes and filamentous actin. Therefore, alveolar macrophages retained their function to utilize vesicles with Mtb lipoarabinomannan, glycoprotein Ag 38, and ESAT-6 protein, but Mtb avoided host killing in lysosomes. Our findings support that the live Mtb are responsible for the ability to avoid degradation and survive in macrophage phagosomes. The exact mechanisms of these processes are still not clear.

Sarcoidosis is a disease involving abnormal collections of inflammatory cells that form lumps known as granulomas. The disease can affect the lungs, skin, lymph nodes, eyes, liver, heart, and brain. As for the eyes it occupies 3-4 place among organs-target. Diagnosis of eye sarcoidosis is quite complicated and is usually found in ophthalmologic examination. Objective: Studying of diagnostic significance of additional methods of ophthalmology examination for sarcoidosis of lung and other localizations. Methods: It was a prospective investigation. The study group consisted of patients with the first identified sarcoidosis in different organs. The control group consisted of patients with tuberculosis (TB) with various localizations. It was interrogated patients by special questionnaire, and then it was done conventional ophthalmology examination and optical coherence tomography (iVue-100v3.2, Optovue, USA). Results: The study group. First identified sarcoidosis: interthoracic lyphnodes was 66, 8% of cases, interthoracic lyphnodes and lung - 22, 2%, lung - 5, 5% and first identified general sarcoidosis (3 and more localizations) - 5, 5%. Retinal lesions: macular degeneration was 33, 3% of cases, macular retinal edema – 11, 1%, retinal detachment – 17, 6%, epiretinal membrane – 5, 6%, healthy retina – 33, 3%. The control group. TB of lung was 89% of cases, TB of bones - 5, 5% and combined TB lesions - 5, 5%. It was not detected any morphological retinal lesions by optical coherence tomography. Conclusions: Degenerative-dystrophic eye processes for sarcoidosis can be diagnosed by optical coherence tomography. This makes it possible to identify specific eye lesions in the early stages.

Development of effective vaccine candidates against tuberculosis (TB) is currently the most important challenge in the prevention of this disease since the BCG vaccine fails to guarantee a lifelong protection, while any other approved vaccine with better efficiency is still absent. The protective effect of the recombinant fusion protein CFP10–ESAT6–dIFN produced in a prokaryotic expression system (Escherichia coli) has been assessed in a guinea pig model of acute TB. The tested antigen comprises the Mycobacterium tuberculosis (Mtb) proteins ESAT6 and CFP10 as well as modified human γ-interferon (dIFN) for boosting the immune response. Double intradermal immunization of guinea pigs with the tested fusion protein (2 × 0.5 µg) induces a protective effect against subsequent Mtb infection. The immunized guinea pigs do not develop the symptoms of acute TB and their body weight gain was five times more as compared with the non-immunized infected guinea pigs. The animal group immunized with this dose of antigen displays the minimum morphological changes in the internal organs and insignificant inflammatory lesions in the liver tissue, which complies with a decrease in the bacterial load in the spleen and average Mtb counts in macrophages.

Mycobacterium tuberculosis (Mtb) is an infectious agent that causes tuberculosis (TB) in humans. A study of the volume of Mtb population and the detection of Mtb virulence in the lungs of patients with pulmonary TB are of great importance for understanding the infectious process and the outcome of the disease. We analyzed the functional state of Mtb and their number in alveolar macrophages obtained from the resected lungs of patients with TB in ex vivo culture and determined that the number of Mtb, referred mainly to the Beijing genotype family (A0 and B0/W148 clusters), were significantly different in cells between different patients. Only single Mtb were found in alveolar macrophages of some patients, while Mtb were actively replicated in colonies in alveolar macrophages of other patients, including cord morphology of Mtb growth (the indicator of Mtb virulence). Our data demonstrated association between the formation of Mtb cording in alveolar macrophages of patients and increased virulence of Mtb from the lungs of these patients in guinea pig TB model. The find of cording formation by replicating Mtb in human alveolar macrophages may be used for preliminary quick estimation of increased Mtb virulence in individual patients with pulmonary TB.

Morphological heterogeneity and plurality of pulmonary lesions at tuberculosis defines need of improvement of methods of their visualization for the choice of tactics and volume of surgical treatment. Purpose: to estimate possibilities of magnetic resonance (MRI) and the x-ray computer tomography (XRCT) in identification of caseous-necrotic focal changes at patients from tuberculosis of the lungs. Material and Methods: 43 patients of the lung TB, subjected to a pulmonary resection within a lobe on the termination of a basic course of a chemotherapy are examined. Results of visualization of focal lesions in a zone of the planned surgical resection are verified by data of morphological and bacteriological researches of a resectat. Results: RCT with a standard width and level of a window taped at all patients focal dissemination without destruction, that assumed subsiding of activity with formation of productive type of an inflammation. However at the MRI in STIR and T2W TSE sequences at 25 of 43 patients (58%) regarding focuses hyper intensive includings, according to morphological comparison corresponding to zones of a caseous necrosis were visualized. Bacteriological researches of operational material of these patients taped significantly higher frequency of identification of viable MBT (against 22% other patients have 84%) and their medicinal fastness (MDR - 92%, XDR in 36,1%). Conclusion: MRI in STIR and T2W TSE sequences can be useful for the purpose of identification of the caseous focal lesions which are clinically associated with high activity of tuberculosis, viability and drug resistance of MBT.

Mycobacterium tuberculosis (Mtb) is an infectious agent that causes tuberculosis (TB) in humans. Studies of the mechanisms of Mtb survival and their replication in the hosts are extremely important for the development of new vaccines and methods for TB treatment because of the emergence and spread of high-virulence strains of Mtb. Previously, we have established the method for the production of ex vivo cultures of alveolar macrophages (Mph) from resected lungs of patients with pulmonary TB. We have observed the formation of irregular aggregates in colonies or, for the first time, cords (Fig. 1) by replicating Mtb in alveolar Mph of some TB patients. Cording is morphology of Mtb growth in colonies, in which Mtb are arranged in the form of plaits, bundles, or cords, while drawing up in a line along their long axes and in close parallel arrangement. As is known, the cord phenotype of Mtb growth on agar correlates with the virulence of Mtb. Here, our aim is to determine the correlation between Mtb cording in human alveolar Mph and virulence of Mtb in guinea pig TB model. In a result, we established the correlation between the formation of Mtb cords in human Mph and high Mtb virulence in guinea pigs. Thus, this marker may be used for quick estimation of Mtb virulence for TB patients. Fig. 1. Alveolar Mph from TB patient no. 6 with Mtb cording after Ziehl-Neelsen staining in ex vivo culture.

Background: Previously, the ex vivo cultures of alveolar macrophages were developed from the surgical samples of the lungs in patients with pulmonary tuberculosis (TB) to establish the unique features of Mycobacterium tuberculosis (Mtb) lifestyle in host cells, but the question has remained whether Mtb-infected cells are isolated from the human lungs or they may be the result of Mtb phagocytosis in ex vivo culture. The study was aimed to investigate Mtb uptake by TB patients' cells after ex vivo expansion. Methods: Alveolar macrophages were infected with the Mtb clinical isolates in ex vivo culture, and the acid-fast Mtb loads in the cells were analyzed. Immunofluorescent staining and the examination of cytological and histological preparations by confocal microscopy were applied to detect Mtb ligands and macrophage surface markers. Results: The studies shown the lack of Mtb uptake by patients' alveolar macrophages during experimental infection with highly virulent Mtb clinical isolates containing pathogen-associated molecular patterns lipoarabinomannan and Ag38 at all used multiplicity of infection including a very high dose of infection. This fact was probably determined by the absence of pattern recognition receptors CD14, TLR2, and CD11b on the plasma membrane of human cells, likely, as a result of cellular processing from the resected lung tissues of patients. Conclusion: The findings indicate that alveolar macrophages with single Mtb or Mtb in colonies, including those with cord-morphology, found in the ex vivo cell cultures of all TB patients examined, were isolated from the lungs, and they characterize the Mtb infection in patients at the time of surgery.

New podands containing pyrrole or pyrazoline fragments have been synthesized. A hypothesis was proposed that closure of the pyrrole ring in the presence of alumina nanoparticles may proceed through 1,4-addition of nitroethane to the chalcone podand to give the Michael adduct rather than through an iminium intermediate usually observed in the reactions of α,β-unsaturated ketones (chalcones) in the presence of Lewis acids. Moderate tuberculostatic activity was found for the starting chalcone podands. Much greater such activity was found upon subsequent modification of the chalcone fragment to give a five-membered heterocycle. Pyrrole derivatives displayed greatest activity among the hetaryl podands obtained.

The Petasis reaction of 6-alkoxy adducts of 1-alkyl-2,3-dicyano-5- arylpyrazinium salts with aromatic boronic acids, such as 2-thienylboronic, 2-furanylboronic and 3-thienylboronic acids, or their benzo analogs in dichloromethane proceeds smoothly at room temperature with the formation of the corresponding 5-aryl-6-hetaryl substituted 1,6-dihydropyrazine derivatives. All dihydropyrazines were separated as pure enantiomers by chiral HPLC, and their absolute configurations for each pair of enantiomers have been determined by X-ray analysis. Individual enantiomers were screened in vitro for their antimycobacterial activities against Mycobacterium tuberculosis H37Rv, avium, terrae and extensively drug-resistant and multi-drug-resistant strains isolated from tuberculosis patients in Ural region (Russia). It has been shown that several compounds exhibit a good level of antituberculosis activity compared to the reference drugs. © ARKAT-USA, Inc.

A series of novel 3,6-disubstituted 1,2,4,5-tetrazines and pyridazines bearing aminopyridyl, hydrazono, or aminoalkyl substituents were synthesized. Ten of these compounds showed marked tuberculostatic activity (MIC ≤ 12.5 mkg mL−1).

Aims: Disseminated pulmonary diseases to date remains difficult to diagnose the problem. Material and methods : This retrospective clinical study conducted on 258 patients (132 female) between 2006 and 2015. Median age was 43 years (range, 17-75 years). Pulmonary dissemination was accompanied intrachest lymphadenopathy (12), spontaneous pneumothorax (3), pleural effusion (7). Incisived lung biopsy was made at 77 patients, stapler biopsy – 179 and pleura biopsy - 72. The median operative time and intraoperative bleeding were 21,5 min (range 5-105) and 7,4 ml (2-70), respectively. Results: At complex inspection of patients following diagnoses have been established: sarcoidosis (124), disseminated pulmonary tuberculosis (33), pneumoconiosis (8), alveolitis (17), interstitialis pneumonitis (35), pulmonary canceromatosis (13), chronic obstructive diseases of a lung (7), visceral pulmonary defeat at system diseases (2), histiocytosis X (6), diseases of accumulation (2), leiomyomatosis (2), pulmonary candidiasis (1), sarcoidosis and tuberculosis (1), пpочие (7). Additional intervention (diagnostic thoracotomy) was required after videothoracoscopy with incisived lung biopsy which has changed the diagnosis. Thus diagnostic efficiency videothoracoscopy with incisived lung biopsy has made 98,7 %, but stapler biopsy – 99,4%. Complications it was observed on 6(2,3%) patients. Conclusions: 1. Surgical methods keep the value at the final stage of diagnostics disseminated pulmonary diseases. 2. Development endoscopic videoequipment and linear staplers allows to establish authentically the diagnosis at disseminated pulmonary diseases.

Background: The use of one-lung high-frequency jet ventilation (OHFJV) during minimally invasive surgery for limited forms of pulmonary tuberculosis has advantages over one-lung convective ventilation (OCMV): elimination of transmission of respiratory movements of the ventilated lung through the mediastinum to the operated lung and creating comfortable conditions for the surgeon in a limited rib cage space pleural cavity, absenceof depression of hemodynamics and adequate oxygenation. Nevertheless, several authors noted an increasing of PaCO 2 during application of OHFJV. Objective: Carry out a comparative study the influence of two OHFJV modes (with expiratory pause and without it) on gas exchange and central hemodynamic in patients with minimally invasive operations on the lungs. Methods: In two groups of 23 patients, conducted a study of gas exchange and central hemodynamic during a minimally invasive lung surgery at the limited forms of tuberculosis using two ventilation modes: I. OHFJV with expiratory pause; II OHFJV without expiratory pause. Results: During the II ventilation mode PaCO 2 marks an increasing of 22% in 30-35 minutes from the beginning of the application OHFJV with a tendency to develop respiratory acidosis. In parallel with this increase in cardiac index recorded by the rise in heart rate at 10%. Conclusions: Application of OHFJV with expiratory pause prevents hypercapnia and ensures adequate gas exchange, optimizing on this background the regulation of central hemodynamic. These results allow us to recommend this mode of OHFJV during minimally invasive surgery of the lung.

Various 5-(fluoroaryl)-4-(hetero)aryl substituted pyrimidines have been synthesized based on the Suzuki cross-coupling and nucleophilic aromatic substitution of hydrogen (SN(H)) reactions starting from commercially available 5-bromopyrimidine and their antitubercular activity against Mycobacterium tuberculosis H37Rv has been explored. The outcome of the study disclose that, some of the compounds have showed promising activity in micromolar concentration against Mycobacterium tuberculosis H37Rv, Mycobacterium avium, Mycobacterium terrae, and multidrug-resistant strains isolated from tuberculosis patients in Ural region (Russia). The data concerning the 'structure-activity' relationship for fluorinated compounds have been discussed.

Synthetic routes to novel N-(purin-6-yl)- and N-(2-aminopurin-6-yl) conjugates with amino acids and glycine-containing dipeptides were developed. In vitro testing of 42 new and known compounds made it possible to reveal a series of N-(purin-6-yl)- and N-(2-aminopurin-6-yl) conjugates exhibiting significant antimycobacterial activity against Mycobacterium tuberculosis H37Rv, Mycobacterium avium, Mycobacterium terrae, and multidrug-resistant M. tuberculosis strain isolated from tuberculosis patients in the Ural region (Russia). N-(2-Aminopurin-6-yl)- and N-(purin-6-yl)-glycyl-(S)-glutamic acids were the most active compounds.

The toxicity and tuberculostatic activity of new isoniazid derivatives containing sterically hindered phenols were investigated. The investigated isoniazid derivatives exhibited lower activity and toxicity than isoniazid.

Tuberculosis (TB), with the Mycobacterium tuberculosis (Mtb) as the causative agent, remains to be a serious world health problem. Traditional methods used for the study of Mtb in the lungs of TB patients do not provide information about the number and functional status of Mtb, especially if Mtb are located in alveolar macrophages. We have developed a technique to produce ex vivo cultures of cells from different parts of lung tissues surgically removed from patients with pulmonary TB and compared data on the number of cells with Mtb inferred by the proposed technique to the results of bacteriological and histological analyses used for examination of the resected lungs. The ex vivo cultures of cells obtained from the resected lungs of all patients were largely composed of CD14-positive alveolar macrophages, foamy or not, with or without Mtb. Lymphocytes, fibroblasts, neutrophils, and multinucleate Langhans giant cells were also observed. We found alveolar macrophages with Mtb in the ex vivo cultures of cells from the resected lungs of even those TB patients, whose sputum smears and lung tissues did not contain acid-fast Mtb or reveal growing Mtb colonies on dense medium. The detection of alveolar macrophages with Mtb in ex vivo culture as soon as 16-18 h after isolation of cells from the resected lungs of all TB patients suggests that the technique proposed for assessing the level of infection in alveolar macrophages of TB patients has higher sensitivity than do prolonged bacteriological or pathomorphological methods. The proposed technique allowed us to rapidly (in two days after surgery) determine the level of infection with Mtb in the cells of the resected lungs of TB patients and, by the presence or absence of Mtb colonies, including those with cording morphology, the functional status of the TB agent at the time of surgery.

Background: The percentage of minimally invasive operative interventions in total amount of surgeries for limited forms of lung tuberculosis is growing. Besides standard objectives (maintaining of adequate gaseous exchange and circulatory dynamics, healthy lung isolation in order to protect it from contamination, limitation of respiratory movements) the minimal invasive operative interventions require elimination of respiratory movements transition through interpleural space to the operated lung and providing the most comfortable conditions for the surgeon operating in the pleural space limited by ribs. In cases of thoracotomy all these objectives can be fulfilled by one-lung high frequency jet ventilation (OHFJV). Aims: To study OHFJV impact on gaseous exchange and circulatory dynamics of minimally invasive lung surgery patients in comparison with one-lung convectional mechanical ventilation (OCMV). Methods: Two groups (comparable by volume of surgery, pathology and age) were studied comparatively for gaseous exchange and circulatory dynamics maintained under use of OCMV and OHFJV. Results: Use of OHFJV there was an increase in CI and SI by 36% and 14% correspondingly in conditions of afterload reduction – PVR by 36%, and also PaO2 increase by 42% under normal pH and PaCO2 indications in both groups. Conclusions: Use of OHFJV in minimally invasive operative lung interventions, unlike OCMV, optimizes regulation of central circulatory dynamics indications under afterload reduction and oxygenation improvement at adequate gaseous exchange. The results received allow using OHFJV in minimally invasive operative lung interventions.

Background: MDR/XDR has become a crucial trouble in pulmonary tuberculosis. Persisting lung cavity prevents disease healing and promotes growth of drug resistance. Objectives: Collapse therapy (CT) improved prognosis in drug resistant CPTB when therapy alone fails. The aim of the study was to find out if combined application of artificial pneumothorax (AP), valve bronchial blockade (VBB) and pneumopertitoneum (PP) expanded the possibilities of CT. Materials and methods: CT was applied in 364 patients with CPTB. MDR was in 288 cases, XDR in 76 ones. AP was applied in 297 patients with CPTB located in upper and middle lobes. We used VBB in 67 patients with fibrotic caverns and lower lobe lesions inaccessible for AP. PP was added in 67 patients with AP or VBB where foci in lower lobes took place. Duration of AP was 3 - 6 months, VBB - 6 – 18 months. Individualized therapy regimens based on drug susceptibility test were applied in all cases. Results: No complication followed CT procedures. AP/PP resulted in sputum culture conversion in 254 patients (85,5%). Complete atelectasis of diseased segment after VBB was achieved in 38 patients with sputum culture conversion in 36 (94,7%). Incomplete atelectasis resulted in conversion in 17 of 29 patients (58,6%). Total effectiveness of VBB was thus 79,1%. Total effectiveness of CT was 84,3% (307 conversions of 364 cases). The results of treatment in patients with CPTB treated without CP were significantly lower: 45 of 109 ones (41,3%)(p Conclusion: VBB complements the capabilities of CT. Combined CT is promising in cases with multiple lung cavities included bilateral ones. Proper endobronchial placement of the valve remains a problem to be solved.

Background: Out of 12392 children born by HIV-infected mothers 779 (6,3%) were prenatally infected. We have studied therapeutic and clinical epidemiologic characteristics of 28 HIV-infected children with tuberculosis. Aim: to show high susceptibility of HIV-infected children of tender and preschool age to tuberculosis and fast development of HIV-infection in cases of prenatal contamination. Materials and methods: Diagnostics of 779 children prenatally infected with HIV was proved during first months of life of these children by findings in blood serum of antibodies to HIV (by ELISA) and virus RNA (by PCR) and by immune blotting. Tuberculosis in 28 children was proved by clinical, laboratory and roentenological (CT) technologies as well as by allergy skin tests with tuberculosis antigens. Results: Sourses of tuberculosis infection were defined in 67,9% of cases, notably in 80% of cases they appeared to be parents of the infected children. Tuberculosis of intrathoracic lymphonodus was diagnosed in 19 children (67,9%), primary tuberculosis complex – in 6 children (2,1%). Generalized tuberculosis was diagnosed in 1 child, generalized BCG-infection – in 1 child. In 60% of children clinical course of tuberculosis was complicated by: lung dissemination, bronchopulmonary lesion, destruction of lung tissue. All children were clinically cured from tuberculosis. Conclusion: HIV-infection in prenatally infected children without treatment reaches the AIDS stage within the 1 st year of life (CD4 reduction to 50 cells/mcl and less). However tuberculosis is responsive to treatment even in cases of pronounced immunodeficiency if antiviral and antituberculosis therapy is administered.

Aim: of this search is to determinate possibility of using the videothoracoscopy Material and methods: Above 4000 patients with exudative pleurisy (years 1986-2011) were examined In Chelyabinsk Region Clinical Tuberculosis Dispensary. Design of research: 434 patients documents were studied retrospectively. The patients had exudative pleurisy of unknown etiology, diagnostic surgery was carried out for patients between 2008 – 2010 years. There were 145 female patients and 289 male patients. Average age - 41 years(from 12 to 83 years). 24 patients have had bilateral pleurisy, 11 have had relapse, 8 - iatrogenic pneumothorax. 439 videothoracoscopy and 6 opened biopsy of the pleura were performed. Except common pleura biopsy forceps pleura biopsy was implemented in 15 cases. Pleurodesis with use of talcum was carried out for 27 patients. Results: tuberculous pleurisy was diagnosed in 235 (54%) patients; HIV – 29(7%); nonspecific pleurisy – 61(14%); parapneumonic pleurisy – 58(13%); tumoral pleurisy – 51(12%); other illnesses – 29(7%); verification in primary operation equals to 98%. Endoscopic view of pleura internal was not always pathognomonic for tuberculosis and tumoral etiology. There were postoperative complications in 7(1,6%) cases: 5 – slow straighten of lung and 5 – empyema. Conclusions: 1. Videothoracoscopy is the method of choice in identification of etiology and treatment of exudative pleurisy. 2. Verification in primary operation equals to 98% and becomes higher in earliest application. 3. Videothoracoscopy allows to carry out treatment manipulations (defragmentation of pleurisy,sanitation and aeration of pleural cavity, drainage ets.), reduces time of treatment and rehabilitation of patients.