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Introduction Tuberculous spondylitis is the most common extrapulmonary tuberculosis. The thoracolumbar lesion due to tuberculous spondylitis is one of the most difficult locations for surgical treatment. Analysis of the recent literature shows a limited amount of data on the results of various current surgical reconstruction techniques. Purpose To review the literature on surgical treatment of thoracolumbar tuberculous spondylitis published during the last five years and judge upon an optimal method. Materials and methods A systematic literature review was performed of the sources from eLibrary, PubMed, Cochrane Library databases. Inclusion criteria: etiologically verified tuberculous spondylitis of thoracolumbar location, follow-up ≥ 1 year, patients older than 18 years. Twenty-one studies that summarize 1,209 cases were selected. Patients were divided into three groups depending on the method of spinal reconstruction (group 1 – ventral approach, group 2 – combined approach, group 3 – dorsal approach). Surgical indicators, correction of kyphotic deformity and its dynamics in the long-term period, rates of complications and the length of hospital stay were analyzed. Results and discussion Blood loss and duration of the intervention were significantly lower in the reconstruction of the thoracolumbar spine from the dorsal approach (599.6 ± 195.1 ml and 196.3 ± 35.6 min). Correction of kyphotic deformity from posterior and combined approaches was higher than in the reconstruction from the ventral approach (64 and 69 %, respectively). At the same time, an inverse proportional dependence of the degree of correction loss in the long-term period was revealed, which was higher with anterior fusion (7.3° ± 1.7° according to Cobb). The duration of hospital stay was shorter in patients with reconstructions from the dorsal approach (13.7 ± 8.2 days). The rate of complications in group 3 was significantly lower (p < 0.0001), while the assessment of their structure indicates prevalence of neurological deficits in dorsal reconstructions, while in ventral and combined reconstructions, infectious complications, pneumothorax, and chronic pain syndrome in the area of autologous costal graft harvesting. Conclusion The optimal method of surgical treatment of thoracolumbar tuberculous spondylitis is a three-column reconstruction from the dorsal approach. The advantages of the method are a decrease in the rate of postoperative complications, a reduction in the duration of inpatient treatment, surgical blood loss and duration of surgical intervention.

the dorsal part of the vertebrae and normal growth of the ventral part. The most common nosological variant of the defect is lordoscoliotic deformity of the thoracic spine, which often occurs due to syndromic defects (spondylocostal dysostosis) and systemic diseases of the musculoskeletal system (arthrogryposis). Along with the vertebral syndrome, the pathological complex in congenital lordosis of the thoracic or thoracolumbar spine includes thoracic insufficiency syndrome, which in the overwhelming cases leads to the development of respiratory failure caused by bronchial obstruction. The purpose of the study is to summarize information and analyze literature data on a rare pathology of the spine, which is clinically manifested by the formation of thoracic lordosis due to congenital anomaly in the development of the vertebrae. Materials and methods An analysis of the literature on the diagnosis and treatment of congenital lordosis and lordoscoliosis in children was carried out. The depth of the search was more than 100 years. Inclusion criteria: presence of lordotic deformity of the thoracic, cervicothoracic or thoracolumbar spine. Results and discussion Respiratory failure in this group of patients develops due to reduction in the anteroposterior size of the chest (an increase in the chest penetration index), a decrease in the height of the thoracic spine, changes in the mechanics of rib movements, bronchial obstruction, worsening ventilation and perfusion on the convex side of the deformity. The leading component of disability and impairment of the quality of life of patients is respiratory failure syndrome. A decrease in lung function, primarily due to reduction in VC, is directly proportional to the magnitude of the deformation of the spine and chest. Surgical correction of spinal and chest deformity is the main component of treatment for this group of patients. Surgical treatment consists in the mandatory multilevel mobilization of all three columns of the spine (preferably combined access) and bilateral resection of the ribs to reduce chest rigidity. Postoperative respiratory support is critical in respiratory failure. Conclusion Congenital spinal deformities in the form of thoracic lordosis and lordoscoliosis are extremely rare. The key aspects of treatment are the fight against respiratory failure and the selective choice of surgical technologies for deformity correction with a trend towards simultaneity of interventions. The literature review confirmed the insufficient knowledge on the issues of this pathology.

The results of recent studies indicate the potential role of gamma-aminobutyric acid (GABA), as an inhibitory mediator of the central nervous system, in the pathogenesis of obstructive sleep apnea syndrome (OSAS) ‒ a common disorder that often accompanies asthma. The aim of the study was to investigate the possible role of some GABAergic system genetic polymorphisms in the formation of OSAS in asthma patients. Overnight cardiorespiratory monitoring was performed to diagnose OSAS and spirometry was conducted to evaluate the airway reactivity to the bronchodilator fenoterol in 184 asthma patients. Polymorphisms of GAD1, GAD2, GABBR1 and GABBR2 genes (15 polymorphisms in total) were genotyped by LATE-PCR method. Significant results were obtained for rs3749034 polymorphism of GAD1 gene and rs35400353 of GABBR2 in association analysis with the presence of OSAS. rs3749034 significantly influenced the presence of OSAS in males, which was accompanied by the predominance of the CC genotype among patients with OSAS, while CT+TT genotypes were more common in patients without OSAS (OR 3.9 95%CI [1.36–11.67], p=0.01). In total sample GAD1 rs3749034 polymorphism was an independent factor increasing the likelihood of having OSAS after adjustment for significant confounders (OR 1.9 95%CI [1.23–3.15], p=0.005). rs35400353 polymorphism was also associated with OSAS after adjustment for confounders, although its relationship was less significant (OR 1.5 95%CI [1.1–2.3], p=0.04). There was a tendency for interrelation with airway hyperresponsiveness to bronchodilator for both polymorphisms: rs3749034 ‒ in case of CT+TT genotypes, rs35400353 ‒ in case of DD genotype. rs3749034 polymorphism also significantly influenced lung function parameters. After additional verification of the results, the identified genetic polymorphisms may be used to individually predict the risk of OSAS as well as for the development of personalized approaches in asthma treatment using GABA.

Obstructive sleep apnea syndrome (OSAS) is a pathogenetically heterogeneous condition that aggravates the course of asthma. For this reason, timely diagnosis and treatment of OSAS may improve asthma control, reduce the number of exacerbations and the frequency of short-acting bronchodilators use. The aim of the study was to describe the clinical and functional features of patients with asthma and OSAS, and to identify risk factors that allow suspecting the comorbidity. A total of 194 patients with asthma were examined. The research methods included assessment of symptoms using the ACT questionnaire, spirometry with bronchoprovocational tests, body plethysmography and determination of lung diffusion capacity, analysis of the cellular composition of induced sputum, echocardiography, overnight cardiorespiratory monitoring. According to the results of cardio-respiratory monitoring OSAS of varying severity was detected in 33% of patients (n=65). Epworth sleepiness scale did not allow specific discrimination of OSAS among the examined patients. In general, patients with the comorbid pathologies were more often men of older age groups, they had a long history of disease and smoking. In patients with OSAS diabetes and cardiovascular diseases were more common in the structure of concomitant pathology. The presence of OSAS did not significantly affect spirometry but was associated with a pronounced airway hyperresponsiveness regardless of the provocative stimulus. In addition, a higher airway resistance and an increase in lung diffusion capacity were noted in OSAS patients. Neutrophilic phenotype of inflammation was three times more common in case of concomitant OSAS, and signs of cardiac remodeling and an increase in pulmonary artery pressure were also observed in these patients. Thus, knowledge of the above features of asthma-OSAS phenotype will facilitate timely diagnosis and optimal therapeutic care for patients with a combination of pathologies.

The aim of the study was to analyze the peculiarities of TRPM8 receptor expression at protein level in induced sputum and nasal epithelium of asthma patients, and to evaluate their relationship to maintenance therapy and cold airway hyperresponsiveness. The study enrolled 43 patients, including those with persistent mild-to-moderate asthma and those with chronic non-obstructive bronchitis (control group). Analysis of TRPM8 expression was performed by indirect flow cytometry. In addition, patients underwent spirometry and a bronchoprovocation test with 3-minute cold air hyperventilation. As a result, we found that TRPM8 was expressed on macrophages of induced spleen and nasal epithelium. Its expression in patients with asthma who did not receive maintenance therapy was more pronounced as compared to the treated patients or the control group. Moreover, the relationship between increased expression of TRPM8 on macrophages and cold airway hyperresponsiveness was established. Further studies should reveal the possibility of TRPM8 usage as a prognostic biomarker of asthma at various stages of the disease development.

This work is devoted to the review of genetic researches in the field of etiology and development of depressive disorders. The interest to depression is caused by high prevalence and increasing frequency of this disease in the world and its negative influence on the ability to work and social activity of a man. Depression is a heterogenic disorder, in the pathogenesis of which both genetic factors and the conditions of micro-and macroenvironment play an important role. The study of polymorphisms of gene-candidates as well as consequences from the totaling of these effects can help to define the degree of disposition to depressive disorders. According to modern theories, a special role in pathogenesis and dynamics of depression belong to the pathology of molecular components of neurochemical structures of central nervous system and genes that code them. The most known genes, for which the correlation with anxiety, depression or their separate symptoms was found, are the genes of 5-serotonin transporters HIT, noradrenalin NET, dopamines DAT1 and D4DR, neurotropic brain factor BDHF, serotonin receptors of HTR1A, HTR2A, genes of enzymes of tryptophan hydroxylase TPH, catechol-O-methyl transferase COMT, methylene tetra hydro folate reductase MTHFR, genes of anti-inflammatory mediators IL-6, IL-1beta, TNF-a, CRP, genes participating in the immune response PSMB4 and TBX21, rare variants of gene NKPD1 and others. The study of the genetic background in relation with affective manifestations seems to be important and promising. In-time and accurate diagnosis of the disposition to depressive disorders can be the most important one in successful prophylactics and effective treatment of patients.

In the present article there are considered approaches to problem solving of selection of suitable vacancies for young experts on the basis of correlation of a competency based matrix of the graduate with employer’s requirements. The offered technique involves five stages: correlation of disciplines and competences, prioritization of employer’s requirements, initial selection of compared competences from the competency based matrix of a graduate, correlation of requirements and competences, assessment of the graduate’s competence relevance in relation to the vacancy. Technique automation using a self-learning neural network will provide flexibility and constant relevance of the obtained results.

To date, the role of dysfunction of the bronchial ciliated epithelium (BCE) in the formation of mucociliary disorders during cold exposure has not been fully established. The aim of the study was to investigate the nature and severity of changes in the ciliary motility of the BCE under the influence of a cold stimulus in vitro. Ten volunteers with asthma underwent a bronchoscopy with biopsy of the lobar bronchus mucosa. The biopsy specimens were placed in a Hank’s balanced salt solution (HBSS) on a slide, located on the thermal stage, used to simulate the effect of various temperature on the BCE. Ciliary beat frequency (CBF, Hz) was recorded using a microscope, high-sensitivity digital camera and computer with specially developed software. The initial recording of CBF was performed at 24ºC, after which the temperature of HBSS was gradually increased to physiological level and repeated recording was made at the control points (28ºC and 36ºC). Then, the temperature of the solution was lowered and CBF was registered again at 28ºC, 21ºC and 17ºC. The initial CBF of the BCE varied from 6.74 to 3.77 Hz with average of 5.25±1.48 Hz (M±m). There was a statistically significant increase in CBF when the solution was heated: at 28ºC it was 6.33±1.36 Hz (p=0.004) and remained at this level up to 36ºC – 7.14±1.33 Hz (p=0.002). A gradual decrease in the solution temperature produced a reduction in CBF in comparison with the physiological conditions: 28ºC – 6.12±1.31 Hz (p=0.121), 21ºC – 5.27±1.32 Hz (p=0.001) and 17ºC – 3.95±1.18 Hz (p=0.0001). By the end of the experiment, CBF of the BCE decreased almost 2-fold in comparison with the physiological conditions. A mathematical model that characterizes the behavior of BCE cilia under cold stress was developed. Thus, the results of the study demonstrate a marked decrease in the motor activity of the BCE cilia under cold stress, which underlines a significant contribution of the functional disturbance of BCE to the pathophysiological mechanisms of hypersecretory disorders induced by inhalation of cold air.

Asthma is a widespread chronic disease of the bronchopulmonary system with a heterogeneous course due to the complex etiopathogenesis. Natural-climatic and anthropogenic factors play an important role in the development and progression of this pathology. The reception of physical and chemical environmental stimuli and the regulation of body temperature are mediated by thermosensory channels, members of a subfamily of transient receptor potential (TRP) ion channels. It has been found that genes encoding vanilloid, ankyrin, and melastatin TRP channels are involved in the development of some asthma phenotypes and in the formation of exacerbations of this pathology. The review summarizes modern views on the role of high and low temperatures in airway inflammation in asthma. The participation of thermosensory TRP channels (vanilloid, ankyrin, and melastatin TRP channels) in the reaction to high and low temperatures and air humidity as well as in the formation of bronchial hyperreactivity and respiratory symptoms accompanying asthma is described. The genetic aspects of the functioning of thermosensory TRP channels are discussed. It is shown that new methods of treatment of asthma exacerbations caused by the influence of temperature and humidity should be based on the regulation of channel activity.

The aim of the study was to determine the role of rs11562975 polymorphism of TRPM8 gene and tobacco smoking in the formation of severe bronchial obstruction in asthma patients. According to this aim, 416 patients with varying severity of asthma were examined. The study design included standard spirometry, genotyping of rs11562975 TRPM8 gene polymorphism and evaluation of the status and duration of smoking. The study revealed the detrimental effect of smoking on the respiratory function in patients with a pack/year index ≥10. rs11562975 polymorphism had a modulating effect on the smoking in patients with asthma. The influence of smoking ≥10 packs-years on the formation of severe bronchial obstruction was more pronounced among carriers of GC genotype, compared to patients with GG genotype. In case of pack-year history ≥20, the GC genotype significantly increased the risk of severe bronchial obstruction in the examined patients

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.Бронхиальная астма (БА) и синдром обструктивного апноэ во сне (СОАС) - одни из самых распространенных хронических заболеваний дыхательной системы. Они имеют общие факторы риска, к которым относятся ожирение, гастроэзофагеальная рефлюксная болезнь (ГЭРБ) и нарушение носового дыхания (аллергические риниты, синуситы). При этом экспериментальные данные показывают общие патофизиологические звенья этих заболеваний: вовлечение в процесс дыхательных путей, системный воспалительный ответ, а также реализация нейромеханических рефлексов. Таким образом, очевиден синергизм этих заболеваний, который влияет на симптомы у пациентов, ответ на терапию и прогноз. Имеющиеся данные позволяют обсуждать необходимость выявления и лечения СОАС у пациентов с БА. Учитывая высокую распространенность СОАС у пациентов с тяжелой БА, можно предположить, что лечение СОАС может снизить количество обострений, улучшить качество жизни и уменьшить число случаев трудно контролируемой БА. Для врачей практического здравоохранения очень важно оценивать факторы риска, такие как масса тела, заложенность носа, наличие ГЭРБ, а также применять инструменты скрининга для более эффективного использования ресурсов здравоохранения. Учитывая известные положительные эффекты от CPAP-терапии в непродолжительных исследованиях, будущие работы должны быть направлены на изучение воздействия CPAP-терапии на симптомы БА в долгосрочной перспективе, а также активнее исследовать влияние препаратов для лечения БА на течение СОАС.

The aim of our study was to evaluate the role of airway cellular inflammation and the lipid peroxidation level in the development of airway hyperresponsiveness (AHR) to inhalation of hypertonic saline (IHS). Methods and Results: The study included the estimation of inflammatory-cellular composition, intracellular concentration of myeloperoxidase (MPO) in induced sputum (IS), serum levels of lipid hydroperoxides (LHP), ceruloplasmin, and vitamin E in 29 patients with asthma and 12 healthy persons. AHR to IHS was assessed by spirometry after 3-min IHS via ultrasonic nebulizer. Patients with asthma had higher indices of leukocytes destruction and cytolysis intensity with the increased leukocyte count in IS. Maximum values of neutrophils cytolysis intensity and leukocytic MPO were found in IS of the patients with AHR to IHS. After the bronchial provocation, serum concentration of LHP was higher in these patients in comparison with the patients without the AHR and control groups. In addition, patients with asthma had lower level of antioxidants than healthy subjects. Conclusion: Marked inflammation involving MPO-activated leukocytes and intensive lipid peroxidation underlie the excessive airway response to IHS.

Background: Role of interferon-gamma (IFNγ) in asthma remains controversial. While certain studies found association of this cytokine with severe asthma and bronchial hyperresponsiveness, the other ones established its rather beneficial effect on mucin secretion and asthma development. Objectives: The aim of the study was to investigate IFNγ levels in exhaled breath condensate (EBC) of asthma patients. Methods: The study included 46 patients with asthma (mostly newly diagnosed - 52% and moderate - 30%) and 23 patients with chronic bronchitis. Mean age of patients was 38.1±1.60 years. Among asthma patients, 78% had uncontrolled disease according to ACQ5. Lung function was tested by spirometry. IFNγ was measured in EBC by BD cytometric bead array method with enhanced sensitivity. Results: Asthma patients had higher level of IFNγ than patients of control group (2654 (1328.9; 3123.9) fg/ml vs. 1872 (1707.8; 2506.3) fg/ml) but the difference was not significant. IFNγ was increased in moderate asthma (2199 (1764.8; 2519) fg/ml) compared to newly diagnosed asthma (1666 (928.8; 2048.5), p=0.01). Moreover, IFNγ was positively correlated with ACQ5 score (R=0.39, p=0.01). In uncontrolled asthma concentration of the cytokine was more than two times higher compared to patients with partially or fully controlled disease (2807 (1512.9; 3123.9) fg/ml vs. 1158 (995.5; 1589.6) fg/ml, p=0.02). 10-days treatment with oral glucocorticoids decreased IFNγ from the initial (2654 (1328.9; 3123.9) fg/ml) to “normal” values (1790 (1124.4; 3070.1) fg/ml, p=0.08). Conclusions: Our observations suggest detrimental effect of IFNγ in asthma pathogenesis. The study was supported by RFBR (#18-54-53019).

Background: Airway hyperresponsiveness (AHR) to osmotic stimuli is common and clinically relevant in asthma. AQP5 is a membrane water channel that may be functionally implicated in airway inflammation and hypersecretion. Objectives: The aim was to assess the effect of AQP5 SNPs on osmotic AHR in asthma patients. Methods: The study enrolled 274 patients with mild-to-moderate asthma, mean age 36.2±0.67 years. The patients underwent bronchoprovocation with 3-min inhalation of distilled water (hypoosmotic stimulus) and hypertonic saline aerosol (hyperosmotic stimulus). Lung function was measured by spirometry before and after each challenge. AQP5 SNPs (rs3759129, rs3736309, rs296756, rs1964676) were genotyped by LATE-PCR. Results: Association with hypoosmotic AHR was found for rs3759129 and rs296756. C allele of rs3759129 was protective against hypoosmotic AHR as AC heterozygotes had less profound response to the challenge compared to AA carriers (ΔFEV1 -1.4 (-6.3; 2.33)% vs. -5.4 (-12.0; -1.0)%, p=0.001; ΔFEF25-75 -1.0 (-18.0; 14.0)% vs. -9.1 (-21.1; 1.2)%, p=0.009). G allele of rs296756 in homozygous state also had protective effect. AA and AG (but not GG) carriers showed marked reduction in airway patency (ΔFEV1 -5,5 (-12,0; -0,8)% vs. -0,2 (-4,5; 3,4)%, p=0.004; ΔFEF25-75 -9,6 (-22,0; 2,3)% vs. 2,0 (-8,9; 9,0)%, p=0.008, for AA and GG genotypes, respectively). rs3759129 also influenced hyperosmotic AHR: all patients with the AHR had AA genotype while 27% of patients without the AHR had AC genotype (p=0.006). Conclusions:AC genotype of rs3759129 and GG genotype of rs296756 exert a protective effect on hypoosmotic AHR. AC genotype of rs3759129 also protects asthma patients from hyperosmotic AHR.

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.

Background: Cold airway hyperresponsiveness (CAH) is common in asthma. However, it is not clear, if any changes in the expression of particular mucins are induced by cold air inhalation in vivo. Objectives: The aim of the study was to investigate MUC5AC and MUC5B expression in nasal epithelium of asthma patients under cold temperature. Methods: The study enrolled 33 patients with mild-to-moderate asthma and mean age 37.9±1.89. Isocapnic (5% CO2) cold air (-20°C) hyperventilation was performed through the nose and the mouth during 3-min. Nasal epithelium was obtained by superficial scrape biopsy before and after the hyperventilation. Expression of mucins was evaluated by qRT-PCR and 2-ΔΔCT method. Results: Basal expression of MUC5AC in nasal epithelium was 3-times higher in patients with CAH when compared to patients without the hyperresponsiveness. After the cold air hyperventilation MUC5AC expression in CAH patients remained increased up to 2.3-fold. Generally, cold air produced 1.6-fold increase in MUC5AC expression with slightly higher response in patients without CAH (1.7-fold) and lower – in patients with CAH (1.4-fold). At the same time, CAH was associated with down-regulation of MUC5B before (0.8-fold) and after (0.05-fold) the challenge. Interestingly, cold air inhalation resulted in decrease of MUC5B expression (0.2-fold) in patients with CAH, while in patients without CAH it increased (3.4-fold). Conclusions: The obtained results suggest that cold air may produce differential response in terms of mucins expression. Cold airway hyperresponsiveness is associated with increased MUC5AC and decreased MUC5B expression, what is typical for asthma. This study was supported by RFBR (project 17-54-53162).

Background and objective Cold-induced airway hyperresponsiveness (CAH) is common in bronchial asthma (BA) patients and represents a problem for those living in cold climate. Transient receptor potential melastatin 8 (TRPM8) channel is the main cold temperature sensor in humans that could mediate cold response in asthmatics with CAH. No associations between TRPM8 gene polymorphisms and CAH have been reported. Methods The present study involved 123 BA patients. CAH was assessed by 3-min isocapnic (5% CO2) cold air (−20°C) hyperventilation challenge. The c.750G > C (rs11562975), c.1256G > A (rs7593557), c.3048C > T (rs11563208) and c.3174C > G (rs11563071) polymorphisms of TRPM8 gene were genotyped by allele-specific polymerase chain reaction (PCR) and PCR with subsequent restriction fragment length polymorphism analysis. Results: GC genotype and C allele carriers of the c.750G > C (rs11562975) polymorphism were more frequently observed to exhibit CAH. The estimated odds ratio for the GC genotype was 3.73 95%CI (1.48; 9.37), P = 0.005. Furthermore, GC heterozygotes had a prominent decrease in forced expiratory volume in 1 s after the challenge as compared to GG homozygotes (−12% (−16; −8.1) vs −6.45% (−11; −2.1), P C (rs11562975) polymorphism is associated with CAH in patients with BA, which suggests a potential role of TRPM8 in CAH development.

Background: Some of TRPV channels subfamily are known osmoreceptors for low and high osmolality and also have been shown to contribute to inflammatory responses. Objective: The aim of the study was to evaluate mRNA expression of TRPV1 , TRPV2 and TRPV4 genes in upper and lower airway epithelium of asthma patients with different airway responses to hypo- and hyperosmotic stimuli. Methods: The study included 33 patients with mild-to-moderate asthma. Osmotic airway reactivity was assessed by bronchoprovocation challenges with distilled water or hypertonic saline aerosol. Epithelium was obtained during nasal and bronchial brush biopsy. mRNA expression was evaluated by RT-qPCR. Results: Airway hyperresponsiveness to hypo- and hyperosmotic stimuli was observed in 30% and 41% of patients, respectively. Patients with lower baseline FEV1 had higher expression of TRPV1 (3.2-fold, p=0.05) and TRPV2 (6.2-fold, p=0.013) in bronchial epithelium. TRPV1 was also overexpressed in the nasal epithelium of these patients (3.4-fold, p=0.04). Hypoosmotic airway hyperresponsiveness was associated with up-regulation of TRPV1 (5.96-fold, p=0.014) and TRPV2 (10.8-fold, p=0.015) genes in bronchial epithelium. Airway hyperresponsiveness to hypertonic stimulus was also marked by higher expression of TRPV1 (4.56-fold, p=0.032) and TRPV2 (9.54-fold, p=0.05). No difference in TRPV4 expression levels was found. Conclusions: TRPV1 and TRPV2 are up-regulated at mRNA level in bronchial epithelium of asthma patients with hypo- and hyperosmotic airway hyperresponsiveness what may indicate their role in the development of osmotic airway responses and asthma pathogenesis. The study was supported by RSF (#14-25-00019).

Osmolarity of the airways being influenced by air humidity serves as an important factor affecting lung function. It has been shown that high as well as low humidity may provoke bronchoconstriction in patients with asthma. TRPV1 , TRPV2 and TRPV4 cation channels are expressed in the respiratory tract and were shown to mediate responses to hyper- and hypoosmotic stimuli. The aim of the study was to investigate the effect of Thr469Ile and Ile585Val SNPs of TRPV1 gene and c.-323A>G SNP of TRPV2 gene on hypo- and hyperosmotic airway hyperresponsiveness (AH) in asthma. Methods: The study involved 116 patients diagnosed with mild or moderate persistent uncontrolled asthma. All subjects underwent spirometry before and after the challenge tests with inhalation of nebulized distilled water (DW), hypertonic saline (HS) and test with physical exercise (PE). FEV1 fall by 10% or more was considered as a positive response. SNPs were genotyped by PCR with RFLP analysis. Results: AA genotype and A allele of c.-323A>G SNP were significantly associated with hypoosmotic AH. Carriers of AA genotype prevailed among the patients with the AH (69% vs 48.6%, p=0.02 for additive model). Frequency of A allele was also higher in these patients (84.5% vs 68.6%, p=0.02; OR 2.5 95%CI (1.13-5.53)). Thr469Ile SNP was found to influence AH to PE. Ile469Ile variant comprised 31% of patients with positive response and only 7% in the group without the AH (p=0.04). No SNPs were associated with hyperosmotic AH to HS. Conclusion: TRPV1 and TRPV2 SNPs may serve as factors predisposing to the development of AH to DW and PE in patients with asthma. The study was supported by RSF (14-25-00019).

Background: TRPM8 is a member of TRP channels family known for its sensitivity to cool temperature, increased osmotic pressure, particulate matter, cigarette smoke and products of oxidative stress. This cation channel is widely expressed in the respiratory tract including airway epithelium and vagal nerve endings where it can mediate inflammatory and secretory responses. Previous studies have reported the increased expression of TRPM8 in the respiratory tract of COPD patients. Objective: To investigate TRPM8 expression in nasal epithelium and induced sputum of asthma patients and to estimate its relation to airway hyperresponsiveness induced by cold air. Methods: The study enrolled 43 subjects with mean age of (39.8±1.76) years including 35 patients with mild-to-moderate asthma and 8 patients with chronic bronchitis. Among the asthma patients 43% were steroid-naïve. Lung function was measured by standard spirometry before and after bronchoprovocation challenge with 3 min isocapnic cold air (-20 °C) hyperventilation. Cold air hyperresponsiveness was diagnosed in case of FEV1 falling by 10% from baseline. TRPM8 expression was measured by indirect flow cytometry in nasal brushings and induced sputum. Nasal epithelium cells were gated after exclusion of dead cells and staining with anti- cytokeratin 19 antibodies. Induced sputum was processed with dithiothreitol, filtered and stained for CD45 and cell viability. Extracellular expression of TRPM8 was detected by staining with primary unconjugated anti-TRPM8 antibodies (Alomone Labs) and secondary antibodies labeled with Alexa Fluor 488 (Abcam). Expression level was calculated as normalized median fluorescence intensity (nMFI) and percent of positively stained cells (%Pos). The data are presented as median, lower and upper quartiles (Me (Q1; Q3)). Results: TRPM8 protein was detected on the epithelial cells and sputum macrophages. The expression levels of TRPM8 were significantly correlated in these cells (nMFI R=0.41, P=0.04; %Pos R=0.53, P=0.008). Expression of TRPM8 was increased both in nasal epithelium and macrophages of steroid-naïve asthma patients as compared to the patients receiving maintenance therapy or patients with chronic bronchitis. nMFI values for macrophages were 2.1 (1.95; 2.79), 1.3 (1.10; 1.91) and 1.2 (1.06; 1.35), respectively (P=0.001). %Pos values for macrophages were 38.5 (13.1; 51.1)%, 11.6 (2.55; 20.1)% and 8.0 (3.20; 9.20)%, respectively (P=0.01). nMFI and %Pos values for the epithelium kept the same trend but the differences were mostly insignificant. In addition, we found a correlation between FEV1 in response to cold air hyperventilation challenge and TRPM8 expression (macrophages %Pos R=-0.43, P=0.02; epithelium %Pos R=- 0.43, P=0.08). Cold airway hyperresponsiveness was accompanied by higher TRPM8 expression on macrophages (21.6 (17.1; 40.7)% vs. 11.6 (7.45; 16.55)%, P=0.03) and nasal epithelium (4.2 (3.30; 5.40)% vs. 2.8 (1.10; 4.30)%, P=0.2). Cold airway hyperresponsiveness was not affected by maintenance therapy in our patients. Conclusions: TRPM8 is overexpressed in steroid-naïve asthma patients as well as in asthma patients with cold airway hyperresponsiveness and may be implicated in asthma pathogenesis. The utility of TRPM8 as a diagnostic biomarker or therapeutic target in chronic obstructive airway diseases is of great interest.

Background: Air humidity is considered to be one of the main climatic triggers of asthma symptoms. Bronchospastic responses to humid air are common in asthma and may complicate the course of the disease in damp climate. Osmotic-induced reactions are likely to be mediated by hypoosmolality-sensitive ion channels TRPV4 which are widely expressed in the respiratory tract. Objective: The aim of the study was to evaluate the effect of rs6606743 and rs7971845 SNPs of TRPV4 gene on osmotic airway hyperresponsiveness (OAH) in patients with asthma. Methods: The study included 189 subjects diagnosed with mild or moderate uncontrolled asthma (mean age 36±0.78 years). All of them underwent spirometry before and after the bronchoprovocation challenge with 3-minute inhalation of ultrasonically nebulized distilled water (UNDW). The SNPs were genotyped by PCR-RFLP analysis. Results: Significant association with OAH was observed for rs6606743 SNP. G was defined as a risk allele with larger prevalence in the OAH group (OR 1.58 95%CI (1.04-2.40), p=0.03). A/G+G/G genotypes in dominant model were linked with positive reaction to UNDW inhalation (p=0.03). A/G and G/G carriers had a significant decrease in FEV1 after the challenge in comparison with A/A subjects (-7.3 (-12; -3.1)% and -8.6 (-16; -4.2)% vs. -2.25 (-10.3; 2.1)%, respectively, p Conclusion: TRPV4 variants may serve as factors predisposing to the development of OAH in patients diagnosed with asthma.

As it has been proved by recent studies, asthma may lead to the development of obstructive sleep apnea (OSA). However, there are little data available on interrelation between OSA and airway hyperresponsiveness (AHR) as one of the main asthma hallmarks. Objective: The aim of the study was to evaluate mutual influence of AHR and OSA in patient with asthma. Methods: 122 patients diagnosed with asthma (mean age 42.5±1.07) underwent spirometry before and after the challenges with 3-minute isocapnic cold air hyperventilation, 3-minute ventilation with ultrasonically nebulized distilled water, and tests for reversibility of airflow limitation with salbutamol and ipratropium. Results: OSA was diagnosed in 31.9% of all subjects. Patients with OSA had more excessive responses (ΔFEV1) to inhalation of cold air (-13 (-17; -5) vs. -3.8 (-7.8; -0.01), p=0.001), distilled water (-9.7 (-12.5; -5.7) vs. -4 (-7.1; -1.5), p Conclusion: The obtained data suggest mutual burdening of OSA and AHR that appears as an increase of AHI in patients with AHR and higher grade of AHR in OSA.