The aim is to assess the levels of certain cytokines IL-6, TNF-α, IL-1β и IL-10, and BDNF in the blood plasma of the elderly, depending on degree of cognitive impairment. 65 elderly with (vascular dementia) or without cognitive impairment were enrolled in the study. The level of cytokines and BDNF were measured in plasma by ELISA. It was found that, regardless of the degree of cognitive impairment, the condition of systemic chronic low-grade inflammation is characteristic of the elderly. Against this background, plasma levels of BDNF were increased in elderly with vascular dementia, reaching statistical significance compared with healthy individuals. Such changes in the level of BDNF may reflect a compensatory repair mechanism in neurodegeneration or be associated with a defective axonal transport or utilization of BDNF in the central nervous system paralleled by increased serum concentrations.

The study of the small intestine microbiota in humans is difficult due to the low availability of biomaterial. Non-invasive methods of metabolomics and bioinformatic data analysis can expand our understanding of the structure and role the small intestine microbiota in maintaining homeostasis of the body. The article presents the trajectory of age-related changes in the microbial community of the small intestine in healthy individuals in the context of interaction with the cytokine and neuroendocrine systems within the metaorganism, using the methods of gas chromatography - mass spectrometry of microbial markers (GCMS MM) and optimal scaling. 110 practically healthy individuals: children, adults and elderly, were included into the study. The number of the main types of small intestine microbiota (Bacteroidetes, Firmicutes, Actinobacteria, Proteobacteria, and Fusobacteria) was determined in peripheral blood by the GCMS MM method. To construct the trajectories of changes in the small intestine microbiota and indicators of the cytokine and neuroendocrine systems with age, the optimal scaling technique based on the multivariate Gifi transformation (CATPCA method) was used. It was found, that the bacterial community of the small intestine of both children and the elderly and seniors has a significantly low total number of microorganisms, due to the low content of bacteria of the types Firmicutes and Actinobacteria against the background of a high number of representatives of the types Proteobacteria and Fusobacteria, in comparison with similar indicators in adults. Assessment of the trajectory of age-associated changes in the microbiota of the small intestine showed: 1) children have strong dynamic fluctuations in the number and connections within the community of microorganisms against the background of the formation of connections between the main regulatory systems of the metaorganism – immune and neuroendocrine; 2) adults present the plasticity and consistency of the functioning of the immune and nervous systems, what determine the state of dynamic balance of the small intestine microbiota; 3) healthy aging characterize by hight degree of cooperation between the main members of the bacterial community, which ensures the stability of the system at a new level, as one of the mechanisms of adaptation of the organism. Thus, the using the methods of GCMS MM and optimal scaling, allows us to expand our understanding of the age-associated trajectory of changes in the small intestine microbiota and its cooperation with the immune and neuroendocrine systems within the metaorganism, which can be used in the development of new methods of therapy of an infectious and non-infectious diseases.

Резюме. В последние годы слюна все чаще используется в качестве диагностической жидкости для оценки различных биологических параметров, а именно уровней активности информационных сигнальных молекул метаорганизма -иммуннонейроэндокринной природы, но реже -метаболитов микробного сообщества и структуры бактериального социума. В работе проведена оценка микробного социума ротовой полости (слюна/мазок с поверхностей проживания микробиоты) здоровых детей с целью создания микробных образов «здоровья» -контроля, который может использоваться в изучении микробного сообщества при развитии локальных и/или системных патофизиологических процессов, в том числе инфекционной природы, в организме ребенка. С помощью метода газовой хромaто-масс-спектрометрии микробных маркеров определены специфические химические маркеры 38 таксонов микроорганизмов в полости рта здоровых детей от 1,5 до 14 лет. Для выявления распределения различных представителей микробных социумов между экологическими нишами (слюна/мазок) в ротовой полости и оценки влияния на них возраста детей был использован многомерный статистический метод -канонический анализ соответствий. Обнаружено высокое сходство структуры микробиоты слюны и мазка с поверхностей проживания микробиоты у здоровых детей, что, возможно, свидетельствует о перекрестных путях движения бактериальных представителей различных видов и родов микробного сообщества или об их функциональной пластичности. Наибольший интерес представляют данные о количестве бактерий рода Alcaligenes spp. в мазке с поверхностей проживания микробиоты, которое в два раза превышает аналогичный показатель в слюне. Alcaligenes продуцирует антибиотики и оригинальные антибактериальные компоненты, дезорганизующие рост широкого круга бактерий, а также инициирует В-лимфоциты лимфоидных фолликулов к продукции Alcaligenes-специфичных антител, для создания из них собственного «плащевого» покрытия, облегчающего ее поступление в Пейеровы бляшки через М-клетки. Можно предположить, что уровень Alcaligenes spp. в слюне в какой-то степени отражает миграцию представителей данного рода как из небных, так и из назофарингеальных миндалин. Определены возрастные особенности микробиоты экологической ниши -ротовой полости: с возрастом у детей повышается число представителей рода Clostridium spp. и снижается количество бифидобактерий. Полученные нами результаты могут использоваться в качестве контроля при системных патофизиологических процессах, в том числе инфекционной этиологии, а также в ходе терапии.

A large and diverse repertoire of antibodies encodes the history of past immunological experience, creating a global network of the body’s regulation system. In this article, we propose to use a peptide microarray (“immunosignature”) for evaluating global individual antibody patterns and bioinformatic data analysis for differential diagnosis of autism spectrum disorders. The peptide microarray consists of 124 000 antigen mimetics with random sequences covalently bound to the surface of the glass slide. A drop of plasma is tested for the presence of antibodies of distinct specificity, by measuring their binding to each antigen mimetic in the microarray detectable by fluorescent staining with secondary IgG antibodies, and this reaction is registered by laser activation assay. For bioinformatic analysis, we used the files of digitalized fluorescence intensity data, which presented the reactivity of plasma antibodies bound to antigen mimetics. Data processing was carried out by packages of the Bioconductor project for the R software environment to perform statistical evaluation. At the stage of primary data processing, the quantile normalization was used in order to equalize the distributions of antibodies’ reactivity. The sample data and other necessary information were combined into the discrete ExpessionSet container files. To compare the control and experimental groups, the Welch’s one-way ANOVA (for unequal variances) was used. The obtained estimates of the mean value differences and statistical significance of P levels were used further for constructing a volcano diagram, in order of ranking and selecting the most promising antibody reactivity parameters. For differential diagnosis of autism, and evaluation of diagnostic significance of the immunosignature method, a heatmap was constructed. The standardized values of the logarithms of antibody reactivity, and the results of the hierarchical cluster analysis performed by the Ward method using Pearson correlation, as a measure of similarity were used in constructing the heatmap. As a result of the bioinformatiс analysis of the data, 73 antibodies were selected whose reactivity had statistically significant differences in groups of children with autism and normally developing children. These antibodies were used for differential diagnosis, the value of which was determined in the heatmap construction. It was found that the group of children with autism spectrum disorders by the antibody reactivity exhibits marked heterogeneity, and consists of at least two subgroups. In addition, 60 antibodies in children with autism showed predominantly medium and low reactivity, i.e. these antibodies had a weak binding power with antigenic mimetics, and only 13 antibodies showed high reactivity. In general, diagnostic specificity of the autism spectrum disorders using immunosignature approach was 96.0% (95% CI 82.8 to 99.6%), sensitivity was 78.3% (95% CI 64.9 to 88.2%), and diagnostic efficiency was 82.7%. Our pilot study allows us to propose a method of immunosignature for differential diagnosis of autism and, possibly, to expand our understanding of autism spectrum disorders.