Актуальность. Учитывая важность пуриновой регуляции воспалительного процесса, раскрытие её участия в патогенезе хронической обструктивной болезни лёгких (ХОБЛ) может дать дополнительную информацию о патофизиологических механизмах воспаления и компенсации, приводящих к развитию ХОБЛ на фоне хронического воспаления, поддерживаемого туберкулёзной инфекцией. Цель настоящего исследования - выявить связь параметров аденозинового метаболизма с характеристиками окислительного взрыва, генерацией оксида азота и функциональными показателями внешнего дыхания у больных туберкулёзом лёгких в сочетании с ХОБЛ. Материалы и методы. В исследование включены мужчины активные курильщики с верифицированным диагнозом туберкулёз лёгких (ТЛ) и ТЛ в сочетании с ХОБЛ (ТЛ+ХОБЛ). Пуриновый метаболизм оценивали по активности аденозиндезаминазы (АДА-1 и АДА-2) в сыворотке крови (экто-АДА), мононуклеарах (мн) и нейтрофилах (нф), уровню экто-5’-нуклеотидазы (экто-5’-НК) в сыворотке крови, CD26 (дипептидилпептидазы-4, ДПП-4) в сыворотке и мононуклеарах. Окислительный взрыв фагоцитов оценивали по тесту восстановления нитросинего тетразолия (НСТ-тесту). Генерацию оксида азота - по концентрации метаболитов NO в сыворотке крови, мононуклеарах и нейтрофилах. Результаты. У больных ТЛ и ТЛ+ХОБЛ получены разнонаправленные изменения концентрации внеклеточного аденозина (рост активности экто-АДА-2, уровня экто-5’-НТ, снижение активности экто-АДА-1), в то время как внутриклеточные концентрации этого медиатора могут быть повышены (снижение активности АДА-1 и CD26 (ДПП-4) мононуклеаров). У больных ТЛ+ХОБЛ выявлено усиление респираторного взрыва мононуклеаров и нейтрофилов. У больных ТЛ регистрировали усиление продукции реактивных радикалов кислорода только в индуцированном НСТ-тесте нейтрофилов. В обеих группах отмечено значимое снижение продукции нитритов и нитратов как в моноцитах, так и нейтрофилах. У больных ТЛ параметры функции внешнего дыхания (ФВД) были связаны с активностью экто-5’-НТ и неспецифической пептидазы CD26 (ДПП-4), с продукцией нитритов нейтрофилами и моноцитами. В то время как у больных ТЛ+ХОБЛ показатели ФВД были ассоциированы с активностью экто-АДА-1 и АДА-1 в моноцитах, с сывороточной продукцией нитратов и продукцией нитритов нейтрофилами. Выводы. У больных ТЛ нарушение ФВД связано с избыточным образованием аденозина при чрезмерной активации формирующих его ферментов, а также с продукцией нитритов нейтрофилами и моноцитами, активных участников бактерицидных реакций, направленных против микобактерии туберкулеза (МБТ). Тогда как при ТЛ+ХОБЛ ведущим является нарушение деградации аденозина при снижении активности АДА-1, развитие эндотелиальной дисфункции и изменение функционального состояния нейтрофилов. Таким образом, разработка целенаправленных воздействий, приводящие к нормализации пуринергического обмена у больных ТЛ, в частности, к увеличению активности изоформы АДА-1, сможет обеспечить либо превенцию с ХОБЛ, либо, у лиц с уже развывшейся патологией, привести к стабилизации процесса. Background: Purine regulation plays an important role in inflammation. Therefore, disclosing the role of purine regulation in the pathogenesis of chronic obstructive pulmonary disease (COPD) may provide additional information about inflammation pathophysiology and compensation, which lead to COPD in chronic inflammation supported by tuberculosis infection. The aim of this study was to identify the relationship of adenosine metabolic indexes with characteristics of an oxidative burst, nitric oxide generation and functional parameters of external respiration (ER) in patients with pulmonary tuberculosis in combination with COPD. Materials and methods. The study included male active smokers with a verified diagnosis of pulmonary tuberculosis (PT) and PT in combination with COPD (PT+COPD). Purine metabolism was evaluated by adenosine deaminase (ADA-1 and ADA-2) activity in serum (ecto-ADA), mononuclear cells (mnc), and neutrophils (nph); serum concentration of ecto-5’-nucleotidase (ecto-5’-NT); and serum and mnc concentrations of CD26 (dipeptidyl peptidase-4, DPP-4). Oxidative burst in phagocytes was evaluated by the nitroblue tetrazolium conversion test (NBT-test). Nitric oxide generation - by concentrations of NO metabolites in blood serum, mnc, and nph. Results. Patients with PT and PT+COPD had multidirectional changes in extracellular adenosine concentration (increased activity of ecto-ADA-2, level of ecto-5’-NT, decreased activity of ecto-ADA-1). At the same time, intracellular adenosine concentrations could be increased (decreased mnc activities of ADA-1 and CD26 (DPP-4)). In patients with PT+COPD, the respiratory burst was observed only in mnc and nph. In patients with PT, increased production of reactive oxygen species was observed only in nph in the inductive NBT-test. In both groups, the nitrite and nitrate production significantly decreased both in monocytes and nph. In patients with PT, parameters of external respiration (ER) were linked to ecto-5’-NT and nonspecific peptidase CD26 (DPP-4) activities and to nitrite production by nph and monocytes. At the same time, in patients with PT+COPD, ER indexes were linked to monocyte activities of ecto-ADA-1 and ADA-1 and to serum nitrate and nph nitrite. Conclusion. In PT patients, impaired ER was associated with excessive formation of adenosine and excessive activation of adenosine-forming enzymes, as well as with nitrite production by nph and monocytes, which are active participants in bactericidal reactions directed against mycobacterium tuberculosis (MBT). However, in PT+COPD, the leading factor is impaired adenosine degradation with decreased ADA-1 activity, development of endothelial dysfunction, and changed nph functionality. Therefore, development of targeted means for normalizing the purinergic metabolism in patients with PT, specifically, by increasing the ADA-1 isoform activity, may provide either prevention in COPD or stabilization of the process in patients with already developed pathology.

Bactericidal activity of leukocytes was investigated in 63 patients with new infiltrate pulmonary tuberculosis and 28 patients with fibrous cavernous pulmonary tuberculosis. The diverse oxygen-dependent bactericidal activity of phagocytes is typical of the patients suffering from both clinical forms of tuberculosis: reduction of nitrosative stress rates, which is more frequent in those suffering from fibrous cavernous tuberculosis, and increase of oxidative stress rates, which is more intensive in case of fibrous cavernous tuberculosis. The associated functions were detected among immune-competent cells, involved in the bactericidal function: in case of fibrous cavernous tuberculosis – between oxidative stress rates, and in case of infiltrate pulmonary tuberculosis – between nitrosative stress rates; the synergistic effect of respiratory burst was observed. In case of chronic pulmonary tuberculosis, both types of cells were equally responsible for bactericidal functions, while neutrophils were dominating in new infiltrate pulmonary tuberculosis, without prior treatment, being the cells of the first line of defense. The obtained results allowed concluding that in case of new pulmonary tuberculosis without prior treatment, nitrosative stress played the important role in the killing of tuberculous mycobacteria, while in case of chronic tuberculosis – it was oxidative stress.

Host-pathogen relations were analyzed on basis of the results of examinations of 42 patients with newly diagnosed, previously untreated infiltrative pulmonary tuberculosis (IPT) with regard for the biological characteristics of Mycobacterium tuberculosis (MBT) and the biomarkers of the inflammatory reaction — the acute phase proteins (APPs). The genotypes of MBT (Beijing and the others) and the efficacy of threeonth antituberculosis therapy were used as the grouping factors. Genotype Beijing MBT patients were significantly often characterised by multiple drug resistant, had widespread pulmonary damage and association of MBT cytoxicity with the effect of therapy. The patients with the best postreatment effect had initial (pretreatment) APPs levels in the range of the referent ones. These conclusions were confirmed by analysis of the correlation pleads of the characteristics of the “host-pathogen” system and their discriminant analysis with regard for MBT genotypes and results of antituberculosis three-month therapy. It was shown that the constellation of four host’s APP characteristics (haptoglobin, ceruloplasmin, elastase and adenoaminase activities) has above 90% prognostic efficacy of treatment in spite of MBT genotype. It is suggested that in IPT patients the first component of the “host-pathogen” system reflecting the reaction of the patient’s organism to the MBT induced inflammatory process is more prognostically important.

The comprehensive prospective examination of 66 patients with first established non-treated infiltrative tuberculosis of lungs was used to analyze the possibility of optimization of assessment of course of specific inflammatory process on the basis of levels of proteins-reactants of acute phase. The characteristics of dynamics of clinical roentgenologic data and terms of coming of abacillarization as a result of three months anti-tuberculosis therapy has been used as grouping factors. It is established that the constellation of 3 out of 12 basic (before treatment) analyzed level indicators are the most prognostic informative--haptoglobin, ceruloplasmin and blood albumin. Their combined application according the proposed decisive rule provides 90.6% of effectiveness of prognosis of the results of treatment in the discussed category of patients.

The comprehensive prospective examination of 66 patients with first established non-treated infiltrative tuberculosis of lungs was used to analyze the possibility of optimization of assessment of course of specific inflammatory process on the basis of levels of proteins-reactants of acute phase. The characteristics of dynamics of clinical roentgenologic data and terms of coming of abacillarization as a result of three months anti-tuberculosis therapy has been used as grouping factors. It is established that the constellation of 3 out of 12 basic (before treatment) analyzed level indicators are the most prognostic informative haptoglobin, ceruloplasmin and blood albumin. Their combined application according the proposed decisive rule provides 90.6% of effectiveness of prognosis of the results of treatment in the discussed category of patients.

The authors have compared the informative value of the tests determining the activity of adenosine deaminase (ADA) and the levels of interferon-gamma (INF-gamma) and neopterin in the diagnosis of pleural effusions of tuberculous (n = 67) and nontuberculous (n = 30) origin. The equally high diagnostic value has been found in the study of the activities of ADA (94.8%) and INF-gamma, which are markers of lymphocytic cellular immunity. There are great differences in the sensitivity of the neopterin test depending whether tuberculosis is isolated or complicates the course of tuberculosis of the lung or intrathoracic lymph nodes, which may be regarded as a reflection of the different status of macrophageal cellular immunity.

The paper analyzes whether it is possible to predict a risk for postoperative empyema, by studying a totality of characteristics of a patient, a pathological process, and the properties of MBT in 46 patients with progressive fibrocavernous tuberculosis. It also shows it actual to accomplish this task with a prediction accuracy of 89.5-100%, by simultaneously taking into account both the bacteriological properties of MBT (the magnitude of Mycobacterium tuberculosis excretion and viability) and different combinations of serum biological parameters that reflect the activity of an inflammatory process.

The paper compares the characteristics of the informative value of tests determining the activity of adenosine deaminase (ADA) activity and the level of gamma-interferon (gamma-IFN) for the differential diagnosis of tuberculous pleurisy (n = 35) and non-tuberculous pleural effusion (n = 53). Both tests were ascertained to have the similar differentially diagnostic capacities with their threshold values of 35 U/l and 180 pg/ ml, respectively. At the same time, the sensitivity and specificity of the ADA test were 98.4% and these of the gamma-IFN test were 94.3 and 96.2%, respectively, with the positive and negative predictive value being 91.3 and 98.3% and 94.3 and 96.2%, respectively, and with the diagnostic value of 94.5% for the ADA test and 96.4% for the gamma-IFN test. There was a high unidirectionality of changes in both values: it was higher than the cut-off points in 94.1% of the patients with tuberculous pleurisy and below them in 88.7% of those with other pleural effusions.

The results of examination of 56 patients with destructive pulmonary tuberculosis, which involved the study of biochemical parameters characterizing different stages of an inflammatory process, as well as clinical and X-ray findings were analyzed, by applying various multivariate statistical methods. This approach was shown to be promising for estimating the severity of the process and for specifying the stage of development of an inflammatory process in some patients.

A hundred and three patients with destructive pulmonary tuberculosis were examined. These patients had heterodirectional changes in the serum levels of alpha2-macroglobulin (MG) and its forms. The lowering of the level of the F-form of the inhibitor outstripped the decrease in the activity of total alpha2-MG and it was detectable in the lobar processes with moderate symptoms of intoxication and with the inoculation of adjacent segments. The preservation of the low total activity of the inhibitor and its forms during antituberculous therapy characterize a diminished process. The high content of the S form of alpha2-MG is a feature in patients showing positive changes before and 2 months after treatment whereas the changes in total alpha2-MG and its F form may vary. Whether it is expedient to study the fractional composition of alpha2-MG is determined by the heterodirectionality of changes in its S and F forms depending on the pattern of a process.

Sixty-three patients with fibrocavernous pulmonary tuberculosis were examined before and 1 month after preoperative therapy with antituberculosis agents. The time course of changes in the clinical and X-ray characteristics of the severity of the disease were compared with the alterations of different biochemical indices, including acute-phase proteins, proteolysis inhibitors, intoxication severity. It was demonstrated that improvement of clinical and X-ray characteristics of the severity of a process after preoperative preparation could be predicted with an accuracy of 83% from the baseline serum concentrations of albumin and ceruloplasmin. The accuracy of prediction does not increase when the levels of protease inhibitors, C-reactive protein, adenosine deaminase, medium-weight molecules are simultaneously studied. The likelihood of the clinical and X-ray characteristics becoming better is higher with enhanced ceruloplasmin activity and decreased albumin levels.

All the steps of osteoarticular tuberculosis evolution were accompanied by an increase in proteolytic activity in rabbit blood serum, which was compensated by activation of alpha 1-inhibitor of proteinases at the initial steps of osteitis. The system "proteinases-inhibitors" was imbalanced during the severe phases of arthritis independently on the rate of its development as follows: predominance of proteolytic potential over inhibitory activity, dissimilar activity of alpha 1-inhibitor of proteinases simultaneously with equal rate of alpha 2-macroglobulin content decrease. Dissimilar alterations in the system studied enabled to suggest that differential application of proteinases is suitable for treatment of inflammation with respect to the step of osteoarticular tuberculosis development.

Changes in lipid peroxidation parameters (LP), antioxidant system and proteinase inhibitors were studied at different evolutionary stages of experimental osteoarticular tuberculosis in 30 rabbits. All stages of the process were characterized by LP activation which in animals of all groups (except rabbits having the acute stage of arthritis) is accompanied by compensatory increase of superoxide dismutase and ceruloplasmin activity. Deficiency of superoxide dismutase found in the acute stage of arthritis with a growth of LP activity shows its inadequate production at a given stage of articular inflammation. Decrease in alpha 1-proteinase inhibitor activity at the stage of osteitis with reactive synovitis can be caused by the influence of LP products. The importance of this mechanism in the inhibition of alpha 1-proteinase inhibitor, together with reduction in its production at the acute stage of arthritis, cannot be ignored. The results obtained make it possible to recommend further study of the perspectives of antioxidants use in osteoarticular tuberculosis.

A correlation between the informativeness of the proteinogram, blood ceruloplasmin and haptoglobin levels and connective tissue metabolism indices used to characterize the inflammatory process activity was performed in 55 patients with different stages of tuberculosis of bones and joints. Among the acute phase proteins examined, the most sensitive indices include ceruloplasmin and haptoglobin found in their combination. A higher urinary oxyproline and glycosaminoglycan excretion in different forms of osteoarticular tuberculosis (both of an inflammatory and a degenerative type) should not be interpreted as an indication of activity of the specific process.

Role of nitric oxide in the pathogenesis of pulmonary tuberculosis has been studied in 77 patients with new infiltrate pulmonary tuberculosis and 34 patients with fibrous cavernous pulmonary tuberculosis. The level of cumulative and endogenous nitrite depended on the clinical form of tuberculosis: in infiltrate pulmonary tuberculosis patients it was within the limits of reference ranges, and in fibrous cavernous pulmonary tuberculosis patients it was significantly lower. Results of statistical analysis point out at the significant homogeneity (monofunctionality) of the set of rates, defining the level of NO metabolites in blood serum in infiltrate pulmonary tuberculosis, namely: impact of adenosine deaminase, levels of α1-protease inhibitor, ceruleoplasmin and age. On the contrary in case of fibrous cavernous pulmonary tuberculosis the diverse (multi-functional) complexes were detected including clinical blood rates providing influence on the reduction of NO level in blood. Nitric oxide in the patients with both clinical forms of tuberculosis correlated with classical markers of system inflammatory response, thus nitric oxide can be regarded as an integral component of inflammatory response with potential evaluation of prognosis of specific lesions during follow-up of changes.