[Change in the expression of gene transcription negative regulator SOCS1 in the patients with bronchial asthma and metabolic disorders].

AIM: Comprehensive study of the negative regulation components of cell signaling in the bronchial asthma (BA) patients with metabolic disorders. MATERIALS AND METHODS: 113 people were examined: 63 patients with allergic BA (ABA), 50 patients with a non-allergic variant of the disease (NABA). SOCS1 mRNA expression was evaluated by reverse transcription PCR (RT-PCR). SOCS1 protein expression was investigated by immunoblotting. The determination of cytokine levels was carried out according to the standard protocol on a Bio-Plex flow fluorimeter. RESULTS: A significant and multidirectional change in the expression of SOCS1 mRNA was found at a body mass index 25 (greater than normal) in ABA and NABA. The positive correlations between SOCS1 mRNA expression and body mass index indicate the regulatory role of SOCS1 in leptin signaling. The spectra of correlations in ABA and NABA are different, it indicates the probable existence of specificity in the pathogenesis of these variants of the diseases. CONCLUSION: The obtained data allow us to consider the complexity of regulation disorders occurring at different levels of cell signaling. The multifunctionality of the SOCS1 regulator provides complex control of cytokine signaling simultaneously in different signaling pathways in the BA with metabolic disorders.

[Characteristics of the expression of the transcription factor pSTAT3 in asthma]. / Kharakteristika ekspressii transkriptsionnogo faktora pSTAT3 pri bronkhial'noi astme.

AIM: To investigate the transcription factor pSTAT3 in overweight asthmatics on a model of peripheral blood mononuclear cells at baseline and during recombinant leptin modulation. MATERIAL AND METHODS: A flow fluorometric assay was used according to the standard Bio-Plex protocol on a Bio-Plex immunoanalyzer (a flow fluorimeter) (Bio-Rad, USA) using xMAP technology at baseline and during modulation with recombinant leptin (Leptin, human, recombinant, expressed in a E. coli, Sigma, USA). RESULTS: There was an obvious reduction in the level of the transcription factor pSTAT3 in patients with non-allergic asthma and an increase in that in patients with allergic asthma (AA). Recombinant leptin modulation of pSTAT3 levels caused their paradoxical decrease in both overweight women younger than 45 years of age with AA and in those with non-allergic asthma. CONCLUSION: The elevated level of the transcription factor pSTAT3 in AA is probably due to the overexpression of pSTAT3 in this group of patients. The paradoxical decrease in pSTAT3 levels in overweight women under 45 years of age with AA, which is similar in the non-allergic asthma group, can be explained by the enhanced expression of negative SOCS3 regulators and by leptin resistance.

[Role of negative regulators of SOCS1, SOCS3, and SOCS5 gene transcription in the negative cell signaling regulation system in asthma]. / Rol' negativnykh regulyatorov transkriptsii genov SOCS1, SOCS3 i SOCS5 v sisteme negativnoi regulyatsii kletochnoi signalizatsii pri bronkhial'noi astme.

AIM: To conduct a comprehensive study of the components of negative cell signaling regulation in different types of asthma. SUBJECTS AND METHODS: A total of 171 people, including 80 patients with allergic asthma (AA), 60 patients with non-allergic asthma (NAA), and 31 apparently healthy individuals, were examined. SOCS5 mRNA expression was assessed by reverse-transcription polymerase chain reaction. The expression of SOCS1 and SOCS3 proteins was investigated by immunoblotting. The concentration of total serum IgE was determined by enzyme immunoassay; the level of cytokines was measured according to the standard protocol using a Bio-Plex fluorometer. RESULTS: The findings show that the patients with AA generally display more marked changes in the expression of all three investigated SOCSes (SOCS1, SOCS3, and SOCS5) at baseline and when interleukin 4 (IL-4) acts. In NAA, there are pronounced changes in the expression of SOCS3 only and, to a lesser extent, SOCS5. The results of investigating the concentrations of IL-4 in the examined groups demonstrate its significant decrease in the AA group, whereas in the NAA group, it is similar to those in healthy individuals. On the contrary, IL-10 concentrations in AA tend towards those in the control group, but much exceed in NAA. CONCLUSION: The findings allow one to consider the complexity of regulatory disorders arising at various levels of cell signaling in the context of the multifunctional nature of the molecules from the family of negative regulators of transcription of the SOCS1, SOCS3, and SOCS5 genes, which provide the comprehensive control of cytokine signaling simultaneously in different signal pathways.

[Bronchial asthma and obesity: common mechanisms].

The prevalence of bronchial asthma and obesity has grown in the recent decades. The mechanisms of these pathologies remain unclear despite a number of publications on the relationship between the two diseases. Analysis of the association of BA with obesity should take into account that both diseases develop in young children undergoing effects of breast milk and specific nutrients, intestinal colonization pattern (neonatal and early childhood), body mass at birth and it growth rate, sedentary lifestyle, and adipokine level in early ontogenesis. The available data suggest that the phenotype of BA associated with obesity is characterized by a number of clear-cut peculiarities (more severe clinical manifestations with frequent exacerbaions and impaired control of the disease). Moreover; such patients show no signs of cellular inflammation. There are several distinct mechanisms characterizing obesity-associated BA, viz. co-morbid conditions such as gastroduodenal reflux disease, sleep apnea, obesity-associated systemic inflammation (elevated cytokine (IL-6, TNF-alpha) levels), oxidative stress, production of obesity hormones (leptin. adiponektin, resistin). Thus, BA and obesity have some common potential mechanisms, including genetic factors, systemic inflammation, mechanical factors, and concomitant diseases. Understanding the common mechanisms of these diseases will promote the development of new therapeutic strategies.

[Evaluating the activity of the apoptosis-regulating genes from Bcl-2, Bax expression, and caspase-3 activity in bronchial epithelial cells in patients with asthma].

The specific features of bronchial epithelial cell apoptosis in asthma were studied, by evaluating the activity of the apoptosis-regulating genes Bcl-2, Bax, and caspase-3. Fibrobronchoscopy with target biopsy of the bronchial mucosa was carried out in 21 asthmatic patients (the latter had given informed consent). The expression of Bcl-2, Bax, and CPP32 (caspase-3 activity) was determined in the bronchial epithelial cells by the immunohistochemical technique using the DAKO kits. Allergic asthma is characterized by higher Bcl-2 and lower Bax expression than non-allergic asthma and in patients receiving systemic glucocorticosteroids. The similar nature of changes was found in the analysis of the Bcl2/Bax ratio. Regardless of the type of asthma, the expression of caspase-3 was rather high. The specific features of impaired bronchial epithelial cell apoptosis in different types of asthma can determine the pathogenetic value of apoptotic disorders in the persistence of allergic inflammation.

[Plasma leptin level in patients with bronchial asthma].

The aim of this study was to measure plasma leptin level in patients with bronchial asthma (BA) and compare it with clinical features of the disease. It included 21 healthy subjects, 9 patients with extrapulmonary allergy, and 18 with partially controllable BA. Plasma leptin was measured by ELISA. Patients with BA showed a much higher leptin level and its significant difference depending on body mass index. Correlation analysis revealed significant correlation between leptin level and degree of eosinophilia in peripheral blood and phlegm, epidermal and iatrogenic sensitization, per os glucocorticoid therapy, concomitant pathology, such as type 2 diabetes, thyroid diseases, cardiological problems (CHD, hypertensive disease), and their complications (acute myocardial infarction, stroke). The reported phenomenon of elevated plasma leptin level in BA patients and its correlation with a number of clinical features open up a possibility for further investigation into the role of obesity in BA pathogenesis and its progress, for the search of new ways to manage the disease.

[Gustatory sensitivity to sodium chloride and potassium chloride and certain parameters of sodium metabolism in patients with bronchial asthma].

The aim of the study was to determine threshold gustatory sensitivity (TGS) to sodium chloride and potassium chloride in patients with bronchial asthma (BA) taking into account a number of factors responsible for the enhanced risk of development and progress of this disease. Forty five practically healthy subjects were compared with 139 asthmatic patients; a separate group comprised patients treated permanently with oral glucocorticoids and those having non-pulmonary allergy. Chemically pure solutions of sodium and potassium chlorides were used in the dripping test to assess TGS. Asthmatic patients had enhanced TGS to sodium chloride compared with healthy subjects. TGS values were especially high in non-allergic bronchial asthma and minimal in allergic asthma when they were unrelated to the phase of the disease. TGS to potassium chloride in asthmatic patients also differed from that in healthy subjects; more patients were bitter-insensitive or sensitive only to high concentrations of potassium chloride. Patients treated with systemic glucocorticoids had the highest TGS to sodium chloride. Patients with non-pulmonary allergy and allergic BA had very similar TGS to potassium chloride. TGS to sodium chloride in the former was lower than in healthy subjects and patients with allergic BA. Asthmatic patients had a higher sodium ion concentration in erythrocytes than normal subjects; it reached maximum values in persons used to add salt to fully cooked food.

[Disturbance of apoptosis of peripheral blood lymphocytes in different variants of bronchial asthma].

AIM: To reveal disturbances of peripheral blood lymphocytes apoptosis in different variants of bronchial asthma (BA). MATERIAL AND METHODS: Apoptosis was studied in 20 healthy subjects, 70 BA patients: 32 patients with allergic BA (ABA) and 26 patients with non-allergic (NABA), 12 patients with BA treated with oral glucocorticosteroids. The following parameters of apoptosis were assessed: cell readiness for apoptosis (expression of CD95/Fas/APO-1 receptors), an early reversible stage of apoptosis (expression of phosphatidylserine on outer leaflet of lymphocyte membrane) and a late stage of apoptosis (the key effector caspase 3 activity). RESULTS: ABA is characterized by increased peripheral blood lymphocytes resistance to apoptosis: a decrease in expression of CD95/Fas/APO-1 receptors and phosphatidylserine on lymphocyte surface and caspase 3 activity. NABA is characterized by peripheral blood lymphocyte apoptosis activation at all stages: high expression of CD95/Fas/APO-1 receptors, expression of phosphatidylserine and caspase 3 activity. BA patients on systemic glucocorticosteroids are characterized by maximal expression of CD95/Fas/APO-1 receptors and phosphatidylserine on lymphocyte surface and the highest caspase 3 activity. CONCLUSION: The findings allow more accurate definition of the features of peripheral blood lymphocytes apoptosis in different variants of bronchial asthma and characteristics of relations between disturbed apoptosis and inflammation. Disturbances of apoptosis may be of pathogenetic importance in persistence ofallergic inflammation.

[Surface architectonics and cytoskeleton of erythrocytes and their modulation by adrenergic agents in patients with bronchial asthma]. / Poverkhnostnaia arkhitektonika i tsitoskelet éritrotsitov i ikh moduliatsiia adrenergicheskimi agentami pri bronkhial'noi astme.

AIM: To evaluate erythrocyte surface architectonics and cytoskeleton in bronchial asthma (BA) under modulation of adrenergic agents. MATERIAL AND METHODS: 24 healthy persons, 61 patients with bronchial asthma (BA) were examined. Of them, 28 patients had allergic BA (ABA) and 33 ones had nonallergic BA (NABA). Erythrocyte surface architectonics was studied by phase-contrast microscopy. Fixation in 0.5% solution of glutar aldehyde was used, preparation "squashed drop" was prepared. In all preparations adrenalin hydrochloride and obzidan were used in equimolar final concentrations of 4.10(-5) M. The mean morphologic index of transformation that reflects the shift to stomatocytosis or to echinocytosis was estimated to characterize integrally erythrocyte surface architectonics. The cytoskeleton was studied by the modified method of Chentosov's et al. The integral optic density was evaluated on imaging analyzer of Ista-Videotest company (St-Petersburg). RESULTS: In healthy persons there was a correlation between erythrocyte surface architectonics and cytoskeleton under modulation of adrenergic system. Adrenalin-induced decrease of cytoskeletal proteins correlated with the shift to stomatocytosis. In NABA erythrocyte surface architectonics was characterized by a pronounced shift to stomatocytosis. This shift was accompanied with the most pronounced decrease of cytoskeletal proteins and it reduced in remission. In ABA manifestation of the shift was minimal and did not depend on the phase of the disease. In ABA no changes in erythrocyte surface architectonics in the presence of adrenalin were revealed. In the same conditions a decrease in cytoskeletal proteins was found. CONCLUSION: A correlation was found between erythrocyte surface architectonics and cytoskeleton in healthy persons. In ABA, under modulation of adrenergic system by adrenalin dissociation was revealed between receptor and cytoskeletal mechanisms of cell form induction. This dissociation does not depend on the phase of the disease and is thought to be one of the important postreceptor disorders typical for this variant of the disease.

[Bronchoobstructive syndrome in bronchoectasias]. / O bronkhoobstruktivnom sindrome pri bronkhéktaziiakh.

AIM: To specify mechanisms of bronchial obstruction in bronchoectasia (BE). MATERIAL AND METHODS: Clinical, x-ray, bronchological and spirographic examinations with broncholytic test were made in 26 BE patients aged 16-66 years (14 males, 12 females). The membrane-receptor complex of erythrocytes was based on the effect of adrenoantagonist propranolol on osmotic erythrocytic resistance (obsidan test) and effect of adrenoagonist adrenalin on dynamics of ESR. RESULTS: Allergological anamnesis was detected in 69% of BE patients, hereditary allergological anamnesis--in 22%, membrane-receptor changes characteristic for atopic condition by obsidan test--in 46%, fractional erythrocytic sedimentation with adrenalin--in 42%. Bronchoobstruction syndrome occurred in 37% cases with mean values FEV1 46 +/- 6.2%, MSV50 17.4 +/- 3%, MSV75 14.4 +/- 0.9%. The broncholytic test detected a mean increment of FEV1 by 12.2 +/- 3.1%. Latent bronchospasm was revealed in 33% patients with BE. CONCLUSION: The correlation analysis suggests that among mechanisms of bronchial obstruction in bronchoectasia, bronchospasm is of great importance especially in allergological anamnesis pathogenesis of which involves disorders of membrane-receptor complex.

[Phenomenon of hyperexpression of ribosomic cistrons in bone marrow hemopoietic elements in bronchial asthma]. / Fenomen povyshennoi ékspressii ribosomnykh tsistronov v krovetbornykh élementakh kostnogo mozga bol'nykh bronkhial'noi astmoi.

AIM: To study expression of ribosomic cistrons (RC) of bone marrow hemopoietic elements (BMHE) in bronchial asthma (BA). MATERIAL AND METHODS: A total of 37 bronchial asthma (BA) patients were examined. Of them, 20 ones had allergic BA (ABA) and 10--nonallergic BA (NABA). Seven patients received systemic glucocorticoid hormones (SGH). Control included cells of bone marrow from 7 healthy donors. Transcription and maturation of pre-rRNA in BMHE were analysed using bone marrow samples obtained during the sternal puncture (a modified method of Howell and Black). RESULTS: RC activity was high. It depended on BA variant and the disease phase. In ABA high RC activity persisted in remission. GCH suppress RC activity rise. The maximal effect of GCH suppressive action was seen in myeloid lineage, minimal--in erythroid one. The effect is absent in more mature erythroid cells and bone marrow lymphocytes. CONCLUSION: The phenomenon of RC activity rise in myeloid, erythroid and lymphoid lineages of the bone marrow of BA patients may reflect contribution of the bone marrow to formation of BA pathogenesis.

[Effect of adrenergic and histaminergic disorders on the subclinical course of atopic bronchial asthma]. / Vyiavlenie narushenii v adrenergicheskoi i gistaminergicheskoi sistemakh na doklinicheskom étape pri atopicheskoi bronkhial'noi astme.

To characterize membrane-receptor peculiarities of the adrenergic and histaminergic systems under the model of peroxide and hyperosmolar effect in atopic bronchial asthma (ABA) and preclinical stage of BA, we have examined 25 patients with ABA, 19 patients with nonpulmonary allergy, 28 healthy men and 21 healthy blood relatives of ABA patients. The phenomenon of inversion of the effect of beta-adrenoblocker (obzidan) and of the combined effect of histamine and H1-antagonist (dimedrol) was revealed in ABA patients. The same phenomenon was registered at the preclinical stage of BA. In ABA and in preclinical BA the shift of H1/H2-histaminergic balance to increased H1-activity exists. Peroxide and hyperosmolar effects modeled on red cell membranes allowed us to characterize reactivity of adrenergic and histaminergic systems not only in ABA but also in preclinical BA.

[Erythrocyte insulin-reactive system and carbohydrate metabolism in bronchial asthma]. / Osobennosti insulinreaktivnoi sistemy éritrotsitov i nekotorye pokazateli uglevodnogo obmena pri bronkhial'noi astme.

AIM: Analysis of insulin-reactive system of erythrocytes and carbohydrate metabolism in bronchial asthma (BA) at cell and other levels. MATERIAL AND METHODS: 106 patients with BA, 26 healthy subjects and 10 healthy subjects with familial history of atopy participated in the study. Glucose tolerance and colorimetric tests were made to measure glucose, glycosylated hemoglobin, respectively. Erythrocytic insulin reactivity was assessed using an original method. Bronchial resistance was determined and specific bronchial conduction was calculated. Acetylcholine test was conducted before and after a single inhalation of 5% isoosmolar glucose solution. RESULTS: Atopic BA was characterized by a trend to hypoglycemia, low glycosylation of red cell hemoglobin, impaired membrane transport of glucose in red cells, a hypoglycemic type of glucose tolerance test curve, a paradoxical reaction of erythrocytes to insulin and adrenaline--release of glucose. CONCLUSION: The detected changes in cell insulin reactivity in atopic BA and positive effects of 5% glucose solution inhalations on bronchial hyperreactivity in BA patients provide grounds for development of new therapeutic approaches in BA.

[Membrane-receptor disorders in adrenergic and histaminergic systems under peroxide exposure in bronchial asthma patients]. / Membranno-retseptornye narusheniia v adrenergicheskoi i gistaminergicheskoi sistemakh pri perikisnom vozdeistvii u bol'nykh bronkhial'noi astmoi.

AIM: To characterize membrane-receptor peculiarities in adrenergic and histaminergic systems under model peroxide effect in bronchial asthma (BA). MATERIALS AND METHODS: 172 patients with BA, 17 patients with nonpulmonary allergy, 38 healthy men and 19 healthy blood relatives of BA patients were examined for peroxide resistance of erythrocytic membrane to modulation with adrenergic and histaminergic agonists and antagonists. The modified method by A. A. Abramov and A. A. Pokrovsky was used. RESULTS: In atopic BA the phenomenon of inversion of the effect of beta-adrenoblocker (obzidan) and of combined effect of histamine and H1-antagonist (dimedrol) was revealed. The same phenomenon was registered at the stage of preasthma and at the preclinical stage of BA. In infection-dependent BA this phenomenon occurred less frequently being rare in remission. In both BA variants the shift of H1/H2-histaminergic balance to increased H1-activity exists which is more significant in atopic BA, preasthma and preclinical BA. CONCLUSION: Peroxide effects modelled on red cell membranes enabled to characterize reactivity of adrenergic and histaminergic systems not only in BA but also in preasthma and preclinical BA.

[Role of biological defects in formation and development of bronchial asthma and potentialities of premorbid diagnosis]. / Rol' biologicheskikh defektov v formirovanii i razvitii bronkhial'noi atmy i vozmozhnosti premorbidnoi diagnostiki.

The paper considers bronchial asthma as a polygenic disease. It presents some data available in the literature on the role of genetic disorders in the development of atopic states and the authors' own findings of various biological defects detected in apparently healthy blood relatives of patients with bronchial asthma. It is concluded that premorbid diagnosis is of importance in high risk persons.

[Membrane receptor disorders in bronchial asthma]. / Membranoretseptornye narusheniia pri bronkhial'noi astme.

AIM: To give impedance characteristics of erythrocytes in bronchial asthma (BA) and to determine the relation between the impedance parameters and specific features of the membrane-receptor complex reflecting the condition of cell membrane lipid matrix. MATERIALS AND METHODS: 164 patients with BA. 19 patients with preasthma, 21 normal blood relatives of BA patients and 36 healthy controls were examined. Parameters of electrical impedance (capacity and temperature-dependent) were measured by modified method of S. Takashima et al. (1988). Peculiarities of low-temperature hyperosmolarity hemolysis were studied before and after modulation with adrenergic and lipotropic agents. RESULTS: Atopic BA was characterized by increased erythrocyte electrical capacity and inversion of adrenaline and obzidan effects on electrical impedance, initially high erythrocyte resistance to low-temperature hyperosmolarity effects. Bacterial asthma demonstrated the opposite trend in both impedance and hemolysis parameters. CONCLUSION: There is low erythrocyte membrane microviscosity in atopic BA, but high microviscosity in bacterial asthma. Basing on these findings the authors propose to use essential phospholipids in the treatment of bacterial BA.

The specific binding of beta-adrenergic ligands and morphofunctional properties of erythrocytes in bronchial asthma.

Peripheral blood erythrocytes from patients with bronchial asthma (BA) and preasthma (PA) were investigated using the following methods: 1) the radiochemical method for determining the specific binding of beta-adrenergic ligands; 2) direct erythrocytometry for erythrocyte diameter; and 3) the cytochemical method [paraldehyde fuchsin (PAF) reaction] for membrane glycoproteins. The specific binding of beta-adrenergic ligands in erythrocytes was decreased in BA and PA patients. The number of erythrocytes of small diameter (ESD) showed a tendency to increase in patients with atopic BA and extrapulmonary allergies, as well as in healthy subjects with a family history of atopy. The percent of PAF-negative erythrocytes was found to be considerably increased in atopic BA. It is concluded that general disorders of the membrane-receptor system are a fundamental cause of reactivity changes in allergy.

[Acupuncture therapy in the treatment of patients with bronchial asthma]. / Iulorefleksoterapiia bol'nykh bronkhial'noi astmoi.

Erythron membrane impairment was investigated using membrane -active drugs (obsidan, delagil, morphium, ethanol) to specify indications to acupuncture in bronchial asthma (BA). The patients were studied during the treatment and followed up for a year. It was found that both immediate and long-term results of acupuncture were related to patterns of the membrane impairment. BA patients with a sharp, greater than 1.5 fold diminution of microcyte count upon delagil test should be assigned to repeated courses of acupuncture during a year. To monitor the treatment effect, it is recommended to define osmotic resistance of the red blood cells in obsidan test in addition to clinicofunctional indices.

[Membranoreceptor disorders in the pathogenesis of bronchial asthma]. / Membranoretseptornye narusheniia v patogeneze bronkhial'noi astmy.

The purpose of the work was to study some characteristic features of the membranoreceptor complex in patients suffering from atopic bronchial asthma (BA) and infection-dependent BA. That was done with the aid of the erythrocytic model (phytohemagglutinin /PHA/ reception, electrophysical analysis of the membranous properties). Electrophoretic mobility (EPM) of red blood cells and red blood cell sedimentation under the action of PHA were determined. The data obtained indicate that in patients with infection-dependent BA, the membranoreceptor complex is characterized by a decrease of EPM of red blood cells, inhibition of red blood cell sedimentation in the presence of PHA. The results of studying the membranoreceptor complex in BA patients are summarized as are the marker characteristics of atopic and infection-dependent BA, reflecting pathology of reception, glycocalyx, and lipid matrix.