Особенности цитокинового профиля у детей с расстройством аутистического спектра

The aim of the study: to reveal the particularities of the concentration of cytokines IL4, IL6, IL10, IL17, IFNγ in blood serum in children with autism spectrum disorder (ASD).Materials and methods. The blood samples obtained from children of two study groups: children with autism spectrum disorder (n = 93) and clinically healthy children (n = 30), served as the material for the  study. Cytokine concentrations were determined in blood serum using the Bender Medsystems (Austria) kits  for IL17A and Vector-Best (Russia) kits for IL4, IL6, IL10, IFNγ. Serum cytokine concentrations were determined by enzyme immunoassay using kits for IL17A (Bender Medsystems, Austria), IL4, IL6, IL10, IFNγ (Vector-Best, Russia). Assessment of cognitive and psychophysiological indicators in children was performed using the  Autism Treatment Evaluation Checklist (ATEC).Results. The concentrations of IL17A (U = 54; p = 0,015) and IFNγ (U = 4.64; p = 0,006) were increased and the concentrations of IL6 (U = 327; p = 0.001) and IL4 (U = 177; p = 0.001) were decreased in children with ASD. The concentration of IL6 correlates with the concentration of IL4 (r = 0.68; p < 0.05). The concentration of IL17A correlates with the concentration of IFNγ (r = 0.41; p < 0.05), IL6 (r = 0.87; p < 0.05) and ATEC score (r = 0.24; p < 0.05) in the group of children with ASD.Conclusion. The cytokine disbalance in children with ADS, which was observed in our study, confirms the  hypothesis of their participation in the development of the disease and clearly shows the Th17  immunoregulation pathway in the pathogenesis of the autism spectrum disorder.Цель работы: выявить уровень концентрации цитокинов IL-4, I-L6, IL-10, IL-17, IFNγ в сыворотке крови у детей с расстройством аутистического спектра (РАС).Материалы и методы. Материалом исследования служили образцы крови, полученные от детей двух групп исследования: детей с расстройством аутистического спектра (n = 93) и клинически здоровых детей (n = 30). Средний возраст в обеих группах составил (7 ± 2) лет. В сыворотке крови методом иммуноферментного анализа определяли концентрацию цитокинов IL-17А (с применением набора Bender Medsystems, Австрия) и IL-4, IL-6, IL-10, IFNγ (Вектор-Бест, Россия). Оценку когнитивных и  психофизиологических показателей проводили с помощью анкеты Autism TreatmentEvaluation Checklist (АТЕС).Результаты. У детей с РАС повышены значения концентрации IL-17A (U = 54; p = 0,015) и IFNγ (U = 4,64; p = 0,006) и снижены – IL-6 (U = 327; p = 0,001) и IL-4 (U = 177; p = 0,001) по сравнению с этими показателями у детей в контрольной группе. Установлены корреляции между концентрацией IL-6 и IL-4 (r = 0,68; p < 0,05); между IL-17A и IFNγ (r = 0,41; p < 0,05), IL-6 (r = 0,87; p < 0,05), количеством баллов АТЕС (r = 0,24; p < 0,05) у детей с РАС.Заключение. Установленный нами дисбаланс цитокинов у детей с РАС подтверждает гипотезу его участия в развитии РАС и свидетельствует об Th17-направлении  иммунорегуляции в патогенезе расстройств аутистического спектра

Features of Food Hypersensitivity in Children with Autism Spectrum Disorder

Currently, the incidence of autism spectrum disorders (ASD) continues to increase in the children’s population. However, the pathogenesis of autism remains controversial.The purpose of this work is to evaluate the performance of the elimination nutritional approaches to the dynamics of indicators of food hypersensitivity, the balance of interleukins and psychophysiological parameters in children with ASD.Methods. A prospective cohort study of children aged 7 ± 2 years, healthy and diagnosed with ASD was performed. Determination of specific immunoglobulin concentrations to 111 food antigens and interleukins was measured by ELISA. Psychophysiological indicators of communicative competence in children with ASD were evaluated in the ATEC-test.Results. It was established that in children with ASD (n = 69), the frequency of occurrence of food hypersensitivity and indicators of specific sIgG concentrations to food antigens of dairy and grain products are higher (р < 0.05). A direct correlation was found between the concentration of INFγ, IgG concentrations to antigens of dairy, grain products and C. albicans. High ratios of the INFγ/IL4 and INFγ/IL10 ratios (p < 0.05) were revealed. As a result of compliance with the elimination diet was noted significant decrease in the concentration of sIgG to food antigens, sIgG to C. albicans, changes indicators following the results of ATEC testing.The data obtained revealed the unexplored effects the influence of specific food hypersensitivity on the process of initiation and maintenance of immune inflammation, as well as on the communicative competence in pathological conditions. A personalized elimination diet can improve the immunorehabilitation and the quality of life of children with autism

ELECTRONIC PROCESS OF REGISTRATION AND CERTIFICATION FOR TEST EQUIPMENT

The aim of our work is to optimize some forms of input and output parameters of the module for metrological support of equipment on the Ural Federal University science service and to describe the electronic processes of registration and certification for test equipment base on the quality management

PROCEDURE FOR INTERNAL CERTIFICATION OF TEST EQUIPMENT

Documentation support of the procedure for internal certification of test equipment is aimed at improving the functioning of the University’s quality management system. In our work describes the process for conducting internal certification of test equipment in the UrFU

Clinical-immunological phenotypes of the autistic spectrum disorders

Autistic spectrum disorders (ASD) affect about one in every 59 children. It is noteworthy that patients with ASD are more likely to have other comorbidities than the general population. Undoubtedly, they may aggravate clinical course of the underlying disease or affect the diagnostics. The aim of this work was to identify clinical and immunological phenotypes of the ASD clinical course. Patients and methods. The study included children classified in 2 groups: pediatric patients with ASD (n = 100), and clinically healthy children (n = 30). Based on the presence of comorbidities, the children were divided into 3 types of clinical patterns: convulsive, infectious, dermato-respiratory and gastrointestinal phenotypes. Cytokine concentrations in blood serum were determined by ELISA using Bender Medsystems (Austria) for IL-17А and Vector-Best (Russia) for IL-4, IL-6, IL-10, IFNγ. The concentration of spIgG to 111 nutritional antigens (IgG) was determined by a modified ELISA method using the Immunohealth™ technique. Assessment of cognitive and psychophysiological indices in children was carried out using the ATEC questionnaire. As a result of the study, clinical and immunological phenotypes were identified among the ASD patients, being associated with certain types of food tolerance, cytokine profile, clinical severity of psycho-physiological disorders and concomitant comorbid diseases. In all four phenotypes, were have revealed an increased synthesis of specific antibodies associated with humoral immunity for the studied food antigens, increased concentration of total spIgG to food antigens, concentration of spIgG to legumes and casein, and C-reactive protein levels.Moreover, in convulsive phenotype (concomitant epilepsy and convulsions), the maximal concentrations of spIgG are shown for Solanaceae products, the concentration of IL-10 is increased, IL-4 amounts are reduced, and the content of serum iron and ferritin is also lowered. In the infectious phenotype (frequently ill children) the spIg’s to grain and fermented products are detected, IL-10 and IFNγ concentrations are increased and IL-4 contents is reduced, along with increased absolute and relative number of lymphocytes and fibrinogen. In the dermato-respiratory phenotype (skin rashes) – to dairy products, the concentrations of IL-4 and IL-17A are increased. In the gastrointestinal phenotype, the highest number of elevated IgG responses to the largest range of food antigens was found in presence of changing cytokine profile , i.e., an increase in IFNγ in IFNγ/IL-4 and IFNγ/IL-10 ratios. Thus, the identified phenotypes of the ASD course are associated with the influence of food antigens and reflect a special variant of the immunological inflammatory pathogenesis, which makes it possible to personalize elimination diets, propose measures for correction and individual prevention, and, probably, to predict clinical course of the disease

Composition of EVs markers under normoxic and hypoxic conditions depends on the expression level of adaptor protein Ruk/CIN85 in mouse renal carcinoma Renca cells

Abstract Aim: To isolate and characterize extracellular vesicles (EVs) produced by mouse renal carcinoma Renca cells with different expression levels of the adaptor protein Ruk/CIN85 under normoxia and hypoxia conditions. Methods: The density gradient centrifugation was used to isolate EVs from the conditioned medium of Renca cells cultured under normoxia and hypoxia conditions. Further characterization of EVs was performed by using nanoparticle tracking analysis (NTA), electron microscopy and Western Blot analysis. Results: Significant differences in average particle size between EVs produced by sublines studied under experimental conditions were not found. At the same time, concentration of particles produced by Ruk/CIN85 overexpressing cells turned out to be an order of magnitude higher in hypoxia in comparison to normoxia conditions. It was shown that under normoxia conditions the content of both Ruk/CIN85 and EVs’ markers Alix and CD81 was increased in vesicles produced by Renca cells with Ruk/CIN85 overexpression in comparison with those from control mock-transfected cells. Under hypoxia conditions, the content of studied proteins decreased by more than two orders of magnitude in EVs secreted by Renca cells with up-regulation of adaptor protein whereas the content of Ruk/CIN85 and CD81 increased in EVs from mock-transfected cells. Conclusions: It has been demonstrated that the adaptor protein Ruk/CIN85 is a novel component of EVs produced by tumor cells that may play a role in the control of EV composition under normoxia and hypoxia