Studies of interrelations between neurotrophic antibodies and individual neurophysiological indices in patients with professional chronic mercury intoxication at the post-exposure period

Disturbances in the nervous system during chronic mercury intoxication (CМI) are accompanied by regular changes in the immune system. Diagnostic use of neurospecific protein markers of various pathological changes in the central nervous system is one of the promising areas of modern neuroimmunology. Prolonged exposure to the mercury vapors may cause disturbances of both CNS neurons and peripheral nerves. At the same time, the neuroimmune relationships underlying the evolving disease are not studied to sufficient grade, especially in the later post-contact period of chronic mercury intoxication. The aim of this work was to identify the relationship between the contents of antibodies to neurotrophic proteins of the nervous tissue, and changes of the neurophysiological indices characterizing the state of central and peripheral pathways in the nervous systems of the patients after chronic mercury intoxication (CMI). Our study immunological study included 30 males in the post-contact period of the CМI, who previously worked in contact with metallic mercury vapors. The clinical patterns of patients were dominated by encephalopathy, the main manifestation of which are mental disorders (more often in the form of organic asthenic disorder, or organic personality disorder with cognitive and emotional-volitional disorders of varying severity). The criteria for inclusion in the study were: verified diagnosis in all patients, written informed consent to participate in the study, a history of the harmful effects of metallic mercury vapor under production conditions. Statistical evaluation was carried out using Statistica for Windows 6.0 software. The levels of neurotrophic antibodies (AT), somatosensory evoked potentials (SSEP), electroneuromyography (ENMG) data were studied in terms of assessing the relationship between changes in specialized structures of the nervous tissue, and the state of the central and peripheral neural pathways in the patients after chronic mercury exposure. The following correlations were found: those between the concentrations of antibodies to endorphin p ф-END) and changes in central conducting structures, as well as between antibodies to proteins S-100, MBP, GFAP, NF-200, VG calcium channel, MAG, p-END, AH-R, Ser-R, M-OR and the speed of axonal impulse conduction within various peripheral neural structures. The interrelations revealed in this study indicate to pathological changes in specialized structures of the nervous tissue, which may be used as diagnostic indicators for clinical course of CМI in the post-contact period

OPPORTUNITIES OF USE OF THE BIOFEEDBACK METHOD IN REHABILITATION OF PATIENTS WITH OCCUPATIONAL DISEASES CAUSED BY CHEMICAL AND PHYSICAL FACTORS

The article shows the opportunity of use of biomonitoring in patients with exogenous toxic encephalopathy in the remote period of chronic mercury intoxication. (CMI) and with, vibration disease caused by local vibration. The efficiency of the biofeedback method was assessed on the basis of the data of electroencephalography, auditory, visual and somatosensoric generated potentials, electroneuromyography, thermometry, dosed cold test and. rheography. Biofeedback training in people with. CMI caused decrease of total brain changes in EEG, improvement of amplitude indices at of the brain generated potentials, in people with vibration disease it caused decrease of manifestation of angiodistonic syndrome and. restoration of neuromuscular conductivity

Experimental Peroxidase Conjugate for Detection of Specific Antibodies to Anthrax Agent in Enzyme Immunoassay

Anthrax poses a pressing issue for veterinary medicine and public health in many countries, including the Russian Federation, which necessitates the improvement and development of new, sensitive and specific diagnostic tools.The aim of the work was to create an experimental peroxidase conjugate for the detection of specific antibodies to the anthrax pathogen and to optimize the conditions for performing enzyme immunoassay (ELISA).Materials and methods. The peroxidase conjugate was constructed using horseradish peroxidase and Staphylococcus aureus protein A (Sigma-Aldrich, USA). Bacterial antigens isolated from strains of Bacillus anthracis 55ΔTPA-1Spo, B. anthracis Sterne 34 F2 were used as sensitizing agents. The developed experimental batches of the conjugate were tested in ELISA for the ability to bind antibodies in the blood sera of anthrax patients and vaccinated individuals. The sensitivity, specificity, and accuracy of the method were calculated using the built-in functions of the ROCR software package.Results and discussion. The peroxidase conjugate to detect specific antibodies to the anthrax pathogen in the study of clinical material has been developed; conditions for the ELISA performance have been optimized. To interpret the results of the study, a threshold value of the positivity coefficient was used, below which the result was considered negative, and at an equal or higher value, positive. The test demonstrated significant differences in the “positivity coefficient” indicator for the “Healthy”/“Sick” and “Healthy”/“Vaccinated” groups, while the differences between the “Sick”/“Vaccinated” groups were statistically insignificant. The maximum accuracy of the method was observed at blood serum dilutions of 1:250 and 1:500. 100 % intra-run, run-to-run and series-to-series reproducibility has been established for all positive samples. The sensitivity and specificity of the experimental peroxidase conjugates were 100 and 95.8 %, respectively, and the accuracy was 97.6 %

Development of a protective lyophilisation medium and conditions to stabilise the erythrocyte diagnostic preparation of tularaemia immunoglobulin

Liquid erythrocyte diagnostic preparations have a practical disadvantage; i.e., long-distance transportation involving possible non-compliance with cold-chain requirements may result in a complete loss of biological activity. A lyophilisation technology is necessary to ensure that the preparations retain their original properties for a long time. The aim of the work was to develop a protective medium and conditions for lyophilisation to stabilise the erythrocyte diagnostic preparation of tularaemia immunoglobulin. Materials and methods: Gelatin, thiourea, trehalose, sucrose, dextran, and Tween 80 were used as excipients for protective media. The authors used nine strains of homologous and heterologous microorganisms of different genera and species to control the lyophilised diagnostic preparation sensitivity and specificity. Evaluation of the main stability-related quality attributes (appearance of the dried preparation, loss on drying, solubility, appearance after reconstitution, appearance after settling, sensitivity, specificity) considered the temperatures specific to the climatic zones where the in vitro diagnostics is intended to be marketed and used. Results: The authors developed protective stabilising media with different compositions, used them in freeze-drying of the preparation and carried out control testing. The most promising was the lyophilisation medium containing a smaller amount of ingredients —6% of dextran, 0.06% of Tween 80 and up to 0.01% of sodium azide—as it was the simplest one to prepare and ensured complete preservation of the quality attributes. The authors carried out practical evaluation of lyophilisation procedures, and the 12–14-hour procedure proved to be the most cost-effective. Conclusions: The results of long-term, or real time, and accelerated stability testing of the lyophilised diagnostic preparation demonstrated the possibility of two-year storage at a labelled temperature of 2–8 °C, as well as at elevated and low temperatures of 30±2 °С and –18 °С, respectively. The tests showed no negative effects of the temperatures on the controlled quality attributes

STUDYING DEVELOPMENT OF POST-VACCINAL CELLULAR IMMUNITY AGAINST BRUCELLOSIS BY MEANS OF LYMPHOCYTE <i>IN VITRO</i> TESTS USING AN EXPERIMENTAL ANTIGENIC COMPLEX

Regulatory framework and methodological approaches to evaluation of immunological effects of vaccination against brucellosis are not established, and the degree of immunological post-vaccinal rearrangement is not yet developed. Due to leading role of cellular immunity in formation of immune protection against brucellosis, evaluation the cellular response in response to antigenic stimulation may be considered the most informative and objective approach to analysis of immune changes in the body during vaccination. In order to develop the most diagnostically informative methods for design of antigen-stimulation cell tests in vitro, a careful selection of a stimulating agent (antigen) is required, which should have a sufficient activating potential, thus providing specificity of reaction under in vitro conditions. The aim of the present study is to study the in vitro specific activity of a protein-polysaccharide antigenic complex from the Brucella abortus 19 BA strain (BrAg), and an opportunity of its application in order to assess the formation of post-vaccinal cellular immunity against brucellosis.The study was performed with white laboratory mice (n = 50) immunized with the Brucella abortus 19 BA strain. The control group (n = 50) consisted of laboratory mice that received a sterile saline solution in a volume of 0.5 ml. Blood samples were taken from immunized and control animals before vaccination, and 7, 14, 21, and 30 days after immunization. By means of flow cytometry, the activation molecules CD25, CD69, MHC II and CD95, expressed on T lymphocytes (CD3+CD69+, CD3+CD25+, CD3+CD95+, CD3+MHC+) were determined. To observe the development of immunity, the intensity of expression of T lymphocyte activation markers was calculated using the stimulation quotient. BrAg was used for specific in vitro stimulation of T lymphocytes. The liquid brucellosis allergen (brucellin) was used as an antigen for comparison, when studying opportunity of BrAg usage for assessing the postvaccinal immunity development.The following results were obtained: BrAg has pronounced specific activity, it did not cause non-specific in vitro reactions (activation) of T lymphocytes, thus enabling its application as a test antigen when evaluating development of adaptive vaccine immunity against brucella.Experimental testing of brucellosis antigen for carrying out the in vitro antigen-stimulated cellular reactions, aiming for evaluation of post-vaccinal immunity development against brucellosis, showed that the usage of BrAg promotes increase in diagnostic sensitivity of cellular reactions under in vitro experimental conditions. The applied experimental antigen is a quite promising tool for development of laboratory algorithms for brucellosis diagnostics, and assessment of actual vaccination efficiency in cohorts previously vaccinated against brucellosis

RESULTS AND PERSPECTIVES OF STUDYING THE OCCUPATIONAL DISEASES IN WORKERS OF AIRCRAFT INDUSTRY IN EAST SIBERIA

The materials of the long-studies among the employees working in the aircraft industry, of both the patients with occupational diseases and the practically healthy persons are discussed in the paper. Using the method of the biological feedback for the vibration-induced disease prevention has been grounded. The preliminary results of the experimental studies using the animals are presented in this paper

A critical period of translational control during brain development at codon resolution

Translation modulates the timing and amplification of gene expression after transcription. Brain development requires uniquely complex gene expression patterns, but large-scale measurements of translation directly in the prenatal brain are lacking. We measure the reactants, synthesis and products of mRNA translation spanning mouse neocortex neurogenesis, and discover a transient window of dynamic regulation at mid-gestation. Timed translation upregulation of chromatin-binding proteins like Satb2, which is essential for neuronal subtype differentiation, restricts protein expression in neuronal lineages despite broad transcriptional priming in progenitors. In contrast, translation downregulation of ribosomal proteins sharply decreases ribosome biogenesis, coinciding with a major shift in protein synthesis dynamics at mid-gestation. Changing activity of eIF4EBP1, a direct inhibitor of ribosome biogenesis, is concurrent with ribosome downregulation and affects neurogenesis of the Satb2 lineage. Thus, the molecular logic of brain development includes the refinement of transcriptional programs by translation. Modeling of the developmental neocortex translatome is provided as an open-source searchable resource at https://shiny.mdc-berlin.de/cortexomics

Analysis of Epizootiological-Epidemiological Situation on Brucellosis in the Russian Federation in 2018 and Forecast for 2019

Presented is the analysis of brucellosis incidence among humans and animals in the Russian Federation in 2018. Epizootiological situation in the regions of developed animal husbandry remains reasonably tense. In 2018, as in previous years, the foci of bovine cattle and small ruminant brucellosis were registered in the North Caucasian, Southern Federal Districts, Volga and Siberian Federal Districts, the share of which made up to more than 90% of all registered in Russia potentially hazardous as regards brucellosis areas and cases of the disease in animals. Against the background of long-term unfavorable epizootic condition, the incidence of brucellosis over the past three years was, on average, 14 % lower than the average long-term indicators. The greatest number of cases (94.1 % of the overall Russian incidence) is registered in the administrative subjects of the North Caucasus Federal District, Southern Federal District and Siberian Federal District, which have the maximum levels of brucellosis incidence in cattle (88.9 %) and small ruminants (95 %). In 2019, persistence of epidemiological problems in regard to brucellosis in the subjects of the North Caucasus Federal District (primarily the Republic of Dagestan, Stavropol Territory), the Southern Federal District (the Republic of Kalmykia, Volgograd and Astrakhan Regions), and the Siberian Federal District (the Tuva Republic, the Omsk and Tyumen Regions) is predicted. The number of human cases of brucellosis may be within the range of 290–310 cases (intensive incidence rate per 100 thousand population – 0.21)

ОПЫТ ИСПОЛЬЗОВАНИЯ ТЕХНОЛОГИИ БИОУПРАВЛЕНИЯ В КЛИНИКЕ ПРОФЕССИОНАЛЬНЫХ ЗАБОЛЕВАНИЙ

Biofeedback in toxic encephalopathy in the late period chronic intoxication by mercury and complex toxic substances led to decrease in the EEG changes, performance improvement of the amplitude and latency evoked potentials, in vibration disease – to decrease manifestations of angiodystonia syndrome, recovery of neuromuscular conductivity. The effectiveness of biofeedback is confirmed by changes in subjective measures of the patients.Биоуправление при токсической энцефалопатии в отдаленном периоде хронической интоксикации ртутью и комплексом токсических веществ привело к уменьшению общемозговых изменений по ЭЭГ, улучшению показателей амплитуды и латентности вызванных потенциалов мозга, при вибрационной болезни – к снижению проявлений ангиодистонического синдрома, восстановлению нервно-мышечной проводимости. Эффективность биоуправления подтверждена изменениями субъективных показателей состояния пациентов

Разработка защитной среды высушивания и режима лиофилизации для стабилизации диагностикума эритроцитарного туляремийного иммуноглобулинового

Liquid erythrocyte diagnostic preparations have a practical disadvantage; i.e., long-distance transportation involving possible non-compliance with cold-chain requirements may result in a complete loss of biological activity. A lyophilisation technology is necessary to ensure that the preparations retain their original properties for a long time. The aim of the work was to develop a protective medium and conditions for lyophilisation to stabilise the erythrocyte diagnostic preparation of tularaemia immunoglobulin. Materials and methods: Gelatin, thiourea, trehalose, sucrose, dextran, and Tween 80 were used as excipients for protective media. The authors used nine strains of homologous and heterologous microorganisms of different genera and species to control the lyophilised diagnostic preparation sensitivity and specificity. Evaluation of the main stability-related quality attributes (appearance of the dried preparation, loss on drying, solubility, appearance after reconstitution, appearance after settling, sensitivity, specificity) considered the temperatures specific to the climatic zones where the in vitro diagnostics is intended to be marketed and used. Results: The authors developed protective stabilising media with different compositions, used them in freeze-drying of the preparation and carried out control testing. The most promising was the lyophilisation medium containing a smaller amount of ingredients —6% of dextran, 0.06% of Tween 80 and up to 0.01% of sodium azide—as it was the simplest one to prepare and ensured complete preservation of the quality attributes. The authors carried out practical evaluation of lyophilisation procedures, and the 12–14-hour procedure proved to be the most cost-effective. Conclusions: The results of long-term, or real time, and accelerated stability testing of the lyophilised diagnostic preparation demonstrated the possibility of two-year storage at a labelled temperature of 2–8 °C, as well as at elevated and low temperatures of 30±2 °С and –18 °С, respectively. The tests showed no negative effects of the temperatures on the controlled quality attributes.Практическое применение эритроцитарных диагностических препаратов выявило недостатки, связанные с их транспортировкой на значительные расстояния с возможным несоблюдением режимов холодовой цепи, что может привести к полной потере их биологической активности. Для стабилизации основных свойств жидких форм эритроцитарных диагностических препаратов в настоящее время необходима разработка технологии получения лиофилизированных форм диагностикумов, которая позволит сохранить первоначальные свойства препарата в течение длительного времени. Цель работы: разработка защитной среды высушивания и режима лиофилизации для стабилизации диагностикума эритроцитарного туляремийного иммуноглобулинового. Материалы и методы: были использованы вспомогательные вещества для подготовки защитных сред (желатин, тиомочевина, трегалоза, сахароза, декстран, твин 80). Для контроля чувствительности и специфичности лиофилизированных диагностикумов использовали 9 штаммов гомологичных и гетерологичных микроорганизмов разных родов и видов. При изучении стабильности основных показателей качества препаратов для диагностики in vitro (внешний вид высушенного препарата; потеря в массе при высушивании; растворимость, внешний вид восстановленного препарата; внешний вид препарата после отстаивания; чувствительность; специфичность) учитывали температуры различных климатических зон, в которых предполагается их реализация и использование. Результаты: разработаны и использованы стабилизирующие защитные среды с различным составом, с последующей сублимационной сушкой препарата и постановкой контрольных исследований. Наиболее перспективной признана среда высушивания для эритроцитарных диагностикумов, содержащая в своем составе меньшее количество ингредиентов — 6% декстрана, 0,06% твин 80 и азид натрия до 0,01%, как наиболее простая в исполнении и обеспечивающая полное сохранение качественных показателей препарата. Отработан рентабельный 12–14-часовой режим лиофилизации. Выводы: по совокупности полученных результатов изучения стабильности в реальном времени (долговременная стабильность) и ускоренном исследовании стабильности лиофилизированных форм диагностикума показана возможность хранения в течение двух лет при регламентированной температуре от 2 до 8 °С, а также в условиях повышенных и пониженных температур при 30±2 °С и минус 18 °С соответственно. Отрицательного влияния указанных температур на результаты контролируемых показателей не выявлено