Evgeny Kulikov

Morphological and Molecular Characteristics of “Difficult” Asthma

Background. There are several clinical variants of severe difficult-to-treat asthma: asthma with persistent airflow limitation, brittle asthma, and fatal asthma; but the differences between the pathogenetic mechanisms underlying the disease heterogeneity are unknown. Objectives. The aim was to evaluate the morphological and molecular characteristics of brittle asthma type I and asthma with persistent airflow limitation compared to mild-to-moderate asthma, by the analysis of the cellular structure and gene expression in the bronchial mucosa. Methods. Bronchoscopic evaluation was performed in 42 asthmatic patients: 10 with brittle asthma, 10 with severe asthma with persistent airflow limitation, and 22 with mild-to-moderate asthma. Morphometric and cytological analyses of the bronchial mucosa were performed. The mRNA levels for the ADRB2, HRH1, and CHRM3 genes in the bronchial mucosa were measured by quantitative real-time polymerase chain reaction (PCR). Results. A predominance of eosinophils (29.48/mm2, 95% confidence interval [CI] 25.24–33.72) and neutrophils (40.13/mm2, 95% CI 32.77–47.49) was observed in patients with mild-to-moderate asthma; however, histiocytes-macrophages (65.80/mm2, 95% CI 56.95–74.65) and lymphocytes (52.94/mm2, 95% CI 42.83–63.06) were more common in patients with brittle asthma, and neutrophil counts (81.11/mm2, 95% CI 58.33–103.89) were significantly increased in subjects with persistent airflow limitation. An increase in the expression of the M3-cholinoreceptor and the β2-adrenoreceptor genes was demonstrated in severe asthmatics compared to mild-to-moderate asthma patients. Significantly higher levels of CHRM3 (57.17%, 95% CI 55.04–59.29) and HRH1 (38.82%, 95% CI 35.84–41.81) mRNAs were observed in patients with brittle asthma. The level of ADRB2 gene expression (71.41%, 95% CI 63.54–85.09) was maximal in patients with asthma with persistent airflow limitation. Conclusions. There is evidence of significantly different morphological characteristics and molecular mechanisms of inflammation and bronchoconstriction underlying the clinical heterogeneity of severe asthma.

Risks and benefits of trial participation: A qualitative study of participants’ perspectives in Russia

Background: The Russian Federation is one of the emerging clinical trial regions where the numbers of international clinical trials have been significantly rising over the course of recent years. Purpose: Our aims were to describe and explain risk–benefit calculus by clinical trial participants in Russia and to analyse the significance of the results for the ethical regulation of globalizing clinical trials. Methods: In-depth semi-structured interviews were conducted with 21 individuals participating in trials for cardiovascular disease. Analysis was based on the inductive constant comparative method. Results: Interviewed participants perceived multiple benefits in trial enrolment including regular check-ups, provision and explanation of individual test results, the opportunity to ask investigators for advice and the provision of treatment recommendations for those with limited access to a physician outside of the trial. Participants tried to manage risks of trial enrolment by paying attention to how they felt and reporting changes to investigators. Regular monitoring, the opportunity to drop out of the trial and health insurance provision in case of adverse events were viewed as further minimizing individual risks. Importantly, interviewed trial participants did not assess the risks and benefits of a single trial independently of wider social situation or particularities of their own health condition. Value of trial enrolment benefits for participants was enhanced by the healthcare system that was viewed as being unresponsive to the needs of people with cardiovascular disease. Therefore, in their risk–benefit assessment, participants weighed enrolment risks against the risks of dealing with their fragile health without continuous contact with a medical professional. Limitations: A relatively small number of interviews was conducted, only participants of cardiovascular disease trials were interviewed and the extent to which the described perspectives are generalizable is not established. Conclusion: The risk–benefit assessment as performed by most interviewed trial participants involved multiple components, including the ones unrelated to the trial itself, and was largely context-dependent. Perspectives of research participants can enrich frameworks for the evaluation of trial risks and benefits.

Expression of adenosine receptors in monocytes from patients with bronchial asthma

Adenosine is generated from adenosine triphosphate, which is released by stressed and damaged cells. Adenosine levels are significantly increased in patients with bronchial asthma (BA) and mediate mast cell degranulation and bronchoconstriction. Over the last decade, increasing evidence has shown that adenosine can modulate the innate immune response during monocytes differentiation towards mature myeloid cells. These adenosine-differentiated myeloid cells, characterized by co-expression of monocytes/macrophages and dendritic cell markers such as CD14 and CD209, produce high levels of pro-inflammatory cytokines, thus contributing to the pathogenesis of BA and chronic obstructive pulmonary disease. We found that expression of ADORA2A and ADORA2B are increased in monocytes obtained from patients with BA, and are associated with the generation of CD14(pos)CD209(pos) pro-inflammatory cells. A positive correlation between expression of ADORA2B and IL-6 was identified in human monocytes and may explain the increased expression of IL-6 mRNA in asthmatics. Taken together, our results suggest that monocyte-specific expression of A2 adenosine receptors plays an important role in pro-inflammatory activation of human monocytes, thus contributing to the progression of asthma.

Роль компонентов микробиоты в модификации иммунного ответа при отдельных вариантах течения хронической обструктивной болезни легких

Background. According to the World Health Organization, chronic obstructive pulmonary disease (COPD) is one of the leading causes of morbidity and mortality in the world. COPD with frequent exacerbations is a most challenging variant of the disease. Currently it is not clear how respiratory microbiota can modify the immune response in this disease. Aim. To establish the role of bacterial oligonucleotides in modification of the immune response in patients with COPD. Materials and м ethods. In accordance with the protocol of the study, 10 patients with stable COPD with frequent exacerbations and 10 patients without frequent exacerbations were included. Immature dendritic cells were obtained by culturing the monocyte fraction of the peripheral blood of patients with COPD. The cells were stimulated by addition of bacterial lipopolysaccharide and small oligodeoxynucleotides (CpG-ODN) of A or B classes. Then the immunophenotypical profile of the obtained cells was determined by flow-cytometry with the use of monoclonal antibodies to antigens CD40, CD83, CD86. To determine the antigen-presenting properties, these dendritic cells were cultivated with CD4+, and then the phenotypic profile of the obtained T-lymphocytes was evaluated by using antibodies to CD4, CD25, CD127, and CD45RO. Results. Cultivation of stimulated dendritic cells by СpG-ODN of A class with T-cells in COPD patients without exacerbations leads to an increase of the amount of lymphocytes of CD25+CD45RO phenotype (15% increase after stimulation), in contrast to the group of patients with frequent exacerbations of COPD (p = 0,018). It may indicate inadequate control of persistent inflammation, mediated by CD25+CD45RO pool of cells in the group of COPD patients with frequent exacerbations. Conclusion. This study demonstrated the presence of discoordination of the immune response of a bidirectional nature in patients with COPD with frequent and infrequent exacerbations.

Clinical characteristics of newborn with different birth weight (results of a multicenter cohort study)

This article is dedicated to analysis of clinical characteristics of children with different birth weight included in the multicenter cohort study of newborns. The study analyzed data 572 children who were stratified according to birth weight (normal, low, very low and extremely low body weight). As part of the protocol incorporating the patient’s visit was provided at the time of birth (visit 0), a retrospective collection of anamnestic data about the mother (Visit -1) and visit prospective study of children aged 12 months (Visit 1). As a result of the analysis, it was found that the increase in frequency of operative delivery, and reduction of Apgar score and the presence of the studied pathological conditions, to the greatest extent, common in children who had birth weight of less than 1500 in this case. The probability of a combination of 3 or more conditions (described in this paper), in the case of a child with an extremely low birth weight was higher in the 44-fold (OR = 44.52; CI95% 15,5-127,5), while when the newborn had very low birth weight - a chance of development was significantly higher only in the 7-fold (OR = 7.12; CI95% 2,4-20,9) compared with children who had a low birth weight. In this regard, there is a need not only to the use of modern technologies nursing of low birth weight infants, but also provide preventive care in a group of women with a high degree of perinatal risk and risk of preterm delivery, allowing prolong pregnancy up to 32 weeks and/or weight of the fruit body of more than 1000 g, which can improve the survival rates of patients and the prevention of premature severe disabling conditions in the future.