Destruction of the bronchial epithelium in patients with severe asthma according to different patterns of inflammation and cold airway hyperresponsiveness.

AIM: The aim of the research was to study the state of the bronchial mucosa epi-thelium in relation to the severity of clinical manifestations in severe uncon-trolled asthma depending on the pattern of inflammation and the presence of cold airway hyperresponsiveness. MATERIALS AND METHODS: In 48 patients with severe uncontrolled asthma, there were assessed asthma symptoms, clinical signs of cold airway hyperre-sponsiveness, and lung function; the samples of slides were analyzed in the cytological examination of the sputum; the degree of damage to epithelial cells and granulocytes was estimated using the total cell destruction index (CDI). RESULTS: According to the analysis of sputum cytograms, the patients were divided into two groups: group I (22 patients) included persons with eosin-ophilic inflammation pattern (31.0±3.1% of eosinophils and 22.0±2.2% of neutrophils), group II (26 patients) was with mixed inflammation pattern (7.2±1.4 and 71.8±4.2%, respectively). The patients of group II had lower disease control according to Asthma Control Test (ACT; 12.1±0.7 and 17.8±0.2 points, respectively; р<0.05), a greater frequency of exacerbations (4.1±0.3 and 3.2±0.2 per year, respectively; р<0.05), greater incidence of clinical signs of cold airway hyperresponsiveness (79 and 19%, respectively; χ2=14.18; р<0.001); lower lung function (midexpiratory flow rate MEF25-75 was 14.6±1.6 and 20.7±1.9%, respectively; р<0.05); they received a higher dose of the combined medications of inhaled glucocorticosteroid in controller anti-inflammatory therapy (salmeterol/fluticasone at a dose of 705.3±19.7 and 650.7±14.8 µg/day for fluticasone propionate; р<0.05) In patients of group II the correlations of epithelial CDI with neutrophil CDI (r=0.61; p<0.01) and eosinophil CDI (r=0.48; p<0.05), as well as correlation of ACT with neutrophil CDI (r=-0.71; p<0.01) and eosinophil CDI (r=-0.53; p<0.05) were found. CONCLUSION: The degree of destruction of the epithelium and granulocytes in the inflammatory patterns has diagnostic relevance for the assessment of the severity of the disease, clinical manifestations of the airway response to the cold trigger, and the inertia of achieving control in patients with severe un-controlled asthma.

[The problem of obstructive sleep apnea syndrome in asthmatic patients]. / Problema sindroma obstruktivnogo apnoé vo sne u patsientov s bronkhial'noi astmoi.

Asthma and obstructive sleep apnea syndrome (OSAS) are one of the most common chronic respiratory diseases. These have common risk factors that include obesity, gastroesophageal reflux disease (GERD) and impaired nasal breathing (allergic rhinitis, sinusitis). At the same time, experimental evidence demonstrates common pathophysiological mechanisms of these diseases, such as involvement in the process of the respiratory tract, a systemic inflammatory response, and implementation of neuromechanical reflexes. Thus, there is an obvious synergism between these conditions, which affects symptoms, response to therapy, and prognosis. The available data allow discussion on whether there is a need to identify and treat OSAS in asthmatic patients. By keeping in mind the high incidence of OSAS in patients with severe asthma, it may be suggested that treatment for OSAS can reduce the number of exacerbations, improve the quality of life, and decline the number of obstinate asthma cases. It is very important for general practitioners to assess risk factors, such as body weight, nasal stuffiness, and GERD, and to utilize screening tools for more efficient use of healthcare resources. Considering the known positive effects of CPAP therapy in short-term studies, future investigations should focus on the impact of CPAP therapy on asthma symptoms in the long-term, as well as on the effects of asthma drugs on the course of OSAS.

Inflammatory Pattern of the Bronchial Mucosa in Patients with Asthma with Airway Hyperresponsiveness to Hypoosmotic Stimulus.

Positive reaction of the bronchi to distilled water inhalation in asthmatics is associated with significant stimulation of the respiratory epithelium desquamation against the background of increased content of eosinophilic and neutrophilic leukocytes in induced sputum, predomination of eosinophil and neutrophil cytolysis, and lower activity of myeloperoxidase in leukocyte granules (in comparison with the parameter in patients with a negative response to bronchostimulation). Enhanced cytolysis and destruction of leukocytes and high myeloperoxidase concentration in the extracellular space are essential for the development of bronchial hyperresponsiveness to hypoosmotic stimulus in asthma.

Influence of TRPV4 gene polymorphisms on the development of osmotic airway hyperresponsiveness in patients with bronchial asthma.

The effect of single nucleotide polymorphisms (SNP) of TRPV4 gene on the development of airway hyperresponsiveness (39.7% of cases) in response to the decrease in osmolarity under inspiration of distilled water aerosol was studies in 189 patients with uncontrolled bronchial asthma. rs6606743 SNP was found to significantly contribute to the development of osmotic airway hyperresponsiveness. Analysis of the dominant genetic model revealed substantial prevalence of AG + GG genotype frequency in the group of patients with asthma with osmotic hyperresponsiveness in comparison with the patients who had negative response to bronchoprovocation. In addition, carriers of GG or AG genotypes had significantly more profound decrease of lung function parameters in relation to A homozygous patients.

[Pneumonia: Regional experience with prevention programs]. / Pnevmoniya: regional'nyi opyt organizatsii profilakticheskikh programm.

AIM: to generalize the regional experience in implementing a package of organizational and methodical and antiepidemic measures for preventing pneumococcal infections. MATERIAL AND METHODS: How the prevention programs were implemented using the materials and methods of the epidemiological and statistical monitoring of the incidence of pneumonia in the Amur Region was analyzed. Pneumococcal conjugate vaccine (Prevenar-13) and influenza vaccines were used for immunoprophylaxis against acute respiratory viral and pneumococcal infections. Information on the incidence of acute respiratory viral infections and pneumonia over time in the period 2010 to 2015 must be taken into account. Indicators and special criteria are used to evaluate the efficiency of vaccination. RESULTS: The comparative statistical analysis revealed the high efficiency of regional programs using the methods for immunoprophylaxis against pneumococcal infections: the vaccination prophylactic efficiency index in terms of the incidence of pneumonia might be as high as 75-100%. Pneumonia morbidity rates became 2.3 times lower in the vaccinated population of the region. CONCLUSION: The results of the investigation suggest that the Program for the clinical and epidemiological monitoring and prevention of community-acquired pneumonias, by using the vaccine against pneumococcal infection in the Amur Region, has a high medical and socioeconomic efficiency.

Cigarette smoke induces mucin hypersecretion and inflammatory response through the p66shc adaptor protein-mediated mechanism in human bronchial epithelial cells.

The p66Shc adaptor protein is a newly recognized mediator of mitochondrial dysfunction and might play a role in cigarette smoke (CS)-induced airway epithelial cell injury. CS can induce an excessive amount of reactive oxygen species (ROS) generation, which can cause mitochondrial depolarization and injury through the oxidative stress-mediated Serine36 phosphorylation of p66Shc. The excessive production of ROS can trigger an inflammatory response and mucin hypersecretion by enhancing the transcriptional activity of pro-inflammatory cytokines and mucin genes. Therefore, we speculate that p66Shc plays an essential role in airway epithelial cell injury and the process of mucin generation in CS-induced chronic inflammatory airway diseases. Our present study focuses on the role of p66Shc in ROS generation, and on the resulting mitochondrial dysfunction, inflammatory response and mucus hypersecretion in CS-stimulated human bronchial epithelial cells (16HBE). We found that CS disturbed the mitochondrial function by increasing the level of phosphorylated p66Shc in these cells and that the effects were significantly reduced by silencing p66Shc. Conversely, the ectopic overexpression of wild-type p66Shc enhanced these effects. We also found that high levels of ROS inhibited FOXO3a transcriptional activity, which led to NF-κB activation. Subsequently, activated NF-κB promoted pro-inflammatory cytokine production and mucin hypersecretion. Thus, manipulating p66Shc might offer a new therapeutic modality with which to treat chronic inflammatory airway diseases.

[CLINICAL AND EPIDEMIOLOGICAL ASSESSMENT OF THE EFFECTIVENESS OF VAC- CINATION OF CHILDREN AGAINST PNEUMOCOCCAL INFECTION IN THE AMUR REGION].

AIM: To study the effectiveness of anti-pneumococcal vaccination of children in the organiza- tion of anti-epidemic measures in the areas of the flood in the Amur region. MATERIAL AND METHODS: The monitoring program included 4988 children aged 2 to 5 years who have risk factors for pneu- mococcal infection. Pneumococcal conjugate vaccine Prevenar- 13 was used for immunization. Data on the incidence of child with acute respiratory infection, acute otitis media, pneumonia, meningitis during the post-vaccination period were taken into account. To evaluate the effective- ness of vaccination we used indicators and specific criteria (coefficient prophylactic vaccination and infection index). RESULTS: The level of total morbidity of children in post-immunization pe- riod decreased by 13.6%; the number of cases ofpneumonia in the population of observed children decreased by 2.3 times; the total duration of the illness in children decreased by 14.6%, the number of.courses of antibiotic therapy was reduced by 21.3%, the number of hospital admissions of children - 38.4%, the number of days of temporary disability ofparents - 11.1%. Direct dependence of the degree of effectiveness of vaccination against pneumococcal disease by the age of children is determined. CONCLUSION: The findings suggest that implementation of the program of clinical and epidemiological monitoring and prevention of community-acquired pneumonia with use of a vaccine against pneumococcal infections in the territory of the Amur Region has a high level of medical and socio-economic efficiency.

Multi-walled carbon nanotubes increase anxiety levels in rats and reduce exploratory activity in the open field test.

The results of the first study on the effects of multi-walled carbon nanotubes (MWNTs) on the exploratory activity and the emotional state in laboratory rats assessed by the open field test are reported. During three or ten days, rats received 8-10 nm MWNTs added to their food at a dose of 500 mg/kg. It was demonstrated that, in the group of rats which were fed with MWNTs, the integrated anxiety level index began to increase as early as the third day of the experiment; on the tenth day, it appeared to be twice increased. It was also demonstrated that MWNTs decreased the integrated exploratory activity index nearly twofold on the third day and nearly fourfold on the tenth day.

[POSSIBILITIES FOR THE USE OF THE ANTI-LEUKOTRIENE DRUG MONTELUKAST IN COMBINED THERAPY OF PATIENTS WITH BRONCHIAL ASTHMA AND COLD-INDUCED BRONCHIAL HYPERACTIVITY].

AIM: To study effectiveness of the use of the anti-leukotriene drug montelukast in combination with inhalation glucocorticoid and long-acting beta-agonist in patients with bronchial asthma (BA) and cold-induced bronchial hyperactivity (CBHA) with a view to optimizing control of the disease. MATERIALS AND METHODS: We carried out an open comparative prospective study of patients with persistent BA in a cold season under conditions of real clinical practice. The patients were divided into 2 groups. Group 1 included patients with CBHA, group 2 consisted of subjects with constant bronchial reactivity in response to cold in the standard provocative test. The patients were followed up for 24 weeks. During the first 12 weeks of the treatment, the patients of group I were given sodium montelukast with budesonide/formoterol. In the next 12 weeks they received only budesonide/formoterol at the same dose. Patients of group 2 were treated with budesonide/formoterol during the entire study period. Efficacy of therapy was assessed by asthma control test (ACT). RESULTS: Control of BA in group 1 (20-25 scores in A CT) was achieved in 83% of the patients within the first 12 week period The result was comparable (87%) with that in group 2. Impairment of control (to 52%) was documented in group I during the last 12 weeks although it was preserved (20-25 scores) in group 2 (81%). CONCLUSION: The use of sodium montelukast in combination with budesonide/formoterol for the treatment of BA with CBHA in winter season ensured control of the disease in most patients during 12 weeks. Withdrawal of montelukast in the subsequent period leads to the loss of control and a rise in the frequency of exacerbation.

[Possibility of achieving and maintaining asthma control in patients with bronchial cold hyperreactivity].

AIM: To evaluate the clinical efficiency of tactics to widen the scope of monotherapy with inhaled glucocorticosteroids (IGCS) in asthmatic patients with bronchial cold hyperreactivity (BCHR) during winter to achieve control of the disease in real clinical practice. SUBJECTS AND METHODS: An open-label longitudinal study was conducted in a cold period in 106 asthmatics divided into 2 groups: 1) those with BCHR and 2) those with unchanged bronchial reactivity to a cold stimulus. The study involved monitoring the symptoms by the asthma control test, peak expiratory flow rate (PEFR), and spirometry results before and after cold bronchoprovocation testing; assessment of the pattern of bronchial inflammation from the ratios of induced sputum (IS) cell populations; and estimation of the number of asthma exacerbations and emergency care recourses. Group 1 used a stepwise increase of the scope of basic therapy with beclomethasone dipropionate 1000 microg/day until asthma control was achieved, which was followed by the therapy with the stable dose. Group 2 received monotherapy with beclomethasone dipropionate as the stable dosage of < or = 500 microg/day. RESULTS: After the first 12 weeks of a follow-up, Group 1 showed the most marked positive changes in the intensity of clinical symptoms, forced expiratory volume in one second, and PEFR that remained within the following 12 weeks during the continued therapy with the stable dose of the drug. A preponderance of the eosinophilic and neutrophilic pattern of inflammation was seen in the patients of this group. By the end of the study, there was a decline in the number of IS inflammatory cells. A discriminant model was developed as a tool to predict asthma control achievement in patients with BCHR. CONCLUSION: A stepwise increase in the scope of IGCS monotherapy in asthmatic patients with BCHR during winter can yield the results of disease control and the incidence of exacerbations, which are similar to those seen in asthmatics with no signs of BCHR (53 and 49%, respectively).

[Correlation of brain electrical activity and motivation in healthy people].

Motivation dominates in the structure of the personality and is one of the basic notions which explains the dynamics of the behavior. The literature has little data about neurophysiology of motivation. The aim of the research was to study the correlation between the motivational sphere and electrical activity of the brain at the influence of different provocations. 24 healthy people at the age of 26-36 years were examined. The results of motivation tests turned out to be uniform (the motivation to success was of a moderate or high level, there were mean values of readiness to risk and low motivation to achievement and approval). Multiple correlations between different types of motivation and electrical activity of the brain at rest, at hyperventilation with room temperature air and at isocapnic cold air hyperventilation were revealed.

[Electrical activity of the brain in different ventilation mode and its relationship with pattern of quite breathing and forced expiratory indices].

The influence of hyperventilation on electric activity of the brain has traditionally been the subject of study of physiologists. The role of the brain in the processes of adaptation to cold air inhalation is still studied not enough. The aim of the work is to define the peculiarities of brain reaction to isocapnic hyperventilation with cold air and identify correlation between its electric activity against different modes of ventilation and the pattern of breathing and parameters of the forced expiration. 21 healthy volunteers went through complex functional examination. Qualitative differences of relative power of theta rhythm at isocapnic hyperventilation with cold air were found out. The peculiarities of correlation between brain electric activity during cold provocation and breathing pattern and bronchomotor tone were revealed.

Study on TRPV1-mediated mechanism for the hypersecretion of mucus in respiratory inflammation.

Transient receptor potential vanilloid 1 (TRPV1) is sensitized by the high affinity TrkA receptor, which promotes pro-inflammatory cytokine production and mediates mucus hypersecretion in bronchial epithelial cells. The purpose of this study was to investigate the mechanism of TRPV1-mediated mucus hypersecretion and respiratory inflammation. Firstly, using Western blot analysis we found that epidermal growth factor receptor (EGFR) and TRPV1 were highly co-expressed in human bronchial epithelial cells (HBE16) with HNE and capsaicin co-treated, the levels of pro-inflammatory cytokines and MUC5AC were also highly co-expressed; however, TRPV1 receptor expression was low in these cells with only HNE stimulation, which demonstrated that sensitization of TRPV1 was not increased in HBE16 cells treated with HNE alone. Secondly, the EGF receptor antagonist (AG1478) and the TrkA receptor inhibitor (K252a) significantly inhibited TRPV1 sensitivity and the expression of MUC5AC and pro-inflammatory cytokines. Furthermore, the PI3K inhibitor LY294002, the PKC inhibitor bisindoylmaleimide (BIM) and the TRPV1 antagonist capsazepine completely abrogated the EGF sensitizing effect. Furthermore, the hypoxia-inducible factor (HIF-1α) inhibitor 2-methoxyestradiol (2-ME2) decreased the activity of PKC by a specific pathway. These findings strongly suggest that TRPV1 sensitization influences the hypersecretion of mucus and inflammatory cytokines, is associated with the PI3K and PKC signaling pathways and is involved HIF-1α activity.

Role of ß2 adrenoreceptor gene polymorphism in the formation of cold hyperreactivity of the airways in asthmatics.

The relationship between ß(2)-adrenoreceptor gene Arg16Gly polymorphism and bronchial cold reactivity has been studied. Genotype Arg16Arg and allele Arg16 carriership is associated with the development of cold bronchospasm in asthmatics. In addition, a significant reduction of 3,5'-cyclic adenosine monophosphate (cAMP) level has been recorded in Arg16Arg homozygotes on minute 30 after cold provocation in comparison with Gly16Gly genotype carriers. These data indicate the influence of primary dysfunction of ß(2)-adrenoreceptors for the formation of bronchial cold hyperreactivity in the patients. Reduced cAMP synthesis by cells in Arg16Arg carriers indicates congenital liability of their ß(2)-adrenoreceptors to desensitization during cold air isocapnic hyperventilation test.

Naringenin attenuates mucous hypersecretion by modulating reactive oxygen species production and inhibiting NF-κB activity via EGFR-PI3K-Akt/ERK MAPKinase signaling in human airway epithelial cells.

Naringenin (Nar) is a flavonoid derived from plant foods. It has been shown to have anti-inflammatory properties. Many studies have shown that overexpression of reactive oxygen species (ROS) and nuclear factor-κB (NF-κB) leads to increased mucin (MUC) 5AC expression in chronic inflammation of the airway. In addition, some studies have reported that naringenin inhibits NF-κB activity in a murine model of asthma. We speculated that naringenin might be associated with mucous hypersecretion, but the molecular mechanisms remain to be defined. Our study has also investigated whether naringenin could inhibit production of ROS and the activity of NF-κB on the inflammatory pulmonary diseases induced by human neutrophil elastase (HNE) and reduce the mRNA and protein levels of MUC5AC as shown by reverse transcriptase-polymerase chain reaction and real-time PCR (RT-PCR). Serum total MUC5AC protein was detected by enzyme-linked immunosorbent assay (ELISA), the protein morphological changes of MUC5AC were also observed by immunofluorescence and confocal laser technology. Hyperactivation of epidermal growth factor receptor (EGFR) signaling is commonly involved in the mucous hypersecretion process and initiates both the activation of extracellular signal-related kinases 1/2 (ERK1/2) and of phosphatidylinositol 3-kinase (PI3K) and Akt kinase. NF-κB is a key factor downstream of PI3K/Akt signaling, which induces overexpression of the MUC5AC gene. Our data revealed that naringenin inhibited the activation of EGFR resulting in the downregulation of the enzyme activities. Naringenin also reduced the protein expressions of p-EGFR, PI3K, p-Akt, p-ERK1/2, and NF-κB as shown by western blotting. Furthermore, naringenin significantly inhibited PI3K/Akt and ERK MAPKinase signaling with a concurrent reduction in production of ROS and NF-κB activities. These results suggest that naringenin may play a protective role by minimizing mucous production during airway inflammation by down-regulating ROS production and inhibiting the NF-κB activity via EGFR-PI3K-Akt/ERK MAPKinase signaling pathway.

[The psychotherapy of astheno-hypochondriacal disorders in patients with somatic pathology at a clinic for borderline states]. / Psikhoterapiia astenoipokhondricheskikh rasstroistv u bol'nykh s somaticheskoi patologiei v klinike pogranichnykh sostoianii.

Overall 90 patients aged 35 to 55 years with somatic pathology treated at the clinic for borderline conditions were examined. Astheno-hypochondriac disorders appeared dominant in 41 patients while in 22 patients the disorders were of more partial character, the hypochondriac symptoms, even pronounced to the degree of hyperquantivalent ideas, were relatively unstable, bearing the traces of exaggeration, "defensiveness", affective disorders were labile. Nineteen patients showed subdepressive disorders with a tinge of "endogenicity", masked by astheno-hypochondriac and somatic complaints. In the first group, the psychotherapeutic policy was based on the unraveling and destruction of the "conventionally profitable" mechanisms of neurotic responding and on the formation of the emotionally saturated dominant in the solution of real life problems. Cognitive psychotherapy was predominant in the second group receiving small doses of psychotropic agents (antidepressants and tranquilizers). Psychotherapy made it possible not only to reduce the main astheno-hypochondriac disorders but also to attain steady compensation for somatic disorders.