Immunogenicity of recombinant fragment of orthopoxvirus p35 protein in mice

Despite the elimination of smallpox, orthopoxviruses continue to be a source of biological danger for humans, as cowpox and monkey pox viruses circulate in nature and the last virus can cause both sporadic cases of human diseases and outbreaks of smallpox-like infection. In addition, periodic vaccination is necessary for representatives of some professions (scientists studying pathogenic orthopoxviruses, medical personnel, etc.). Vaccination against smallpox virus with live vaccinia virus, which was widely used during the elimination of smallpox, induces the formation of long-term immunity in vaccinated people. However, providing a high level of protection, the vaccination is often accompanied by serious post-vaccination complications, the probability of which is particularly great for individuals with compromised immunity. In this regard, the development of preparations for the prevention and treatment of infections caused by orthopoxviruses remains important today. The aim of this study was to assess the immunogenicity in the mouse model of recombinant protein р35Δ12, designed previously on the base of the cowpox virus protein p35. It was previously shown that the protein р35Δ12 was recognized by fully human neutralizing anti-orthopoxviral antibody with high affinity. In this work, recombinant protein р35Δ12 produced in E. coli cells XL1-blue and purified by chromatography was used for two-time immunization of mice. Two weeks after the second immunization, blood samples were taken from mice and serum antibodies were analyzed. It was shown by ELISA and Western-blot analysis that immunized mice sera contained IgG antibodies specific to recombinant protein р35Δ12. Confocal microscopy showed that antibodies induced by the р35Δ12 protein were able to recognize Vero E6 cells infected with the LIVP-GFP vaccinia virus. In addition, the antibodies in the serum of immunized mice were able to neutralize the infectivity of the vaccinia virus LIVP-GFP in the plaque reduction neutralization test in vitro. These experiments have demonstrated promising properties of the р35Δ12 protein if it were used as a component of vaccine for prophylaxis of orthopoxvirus infections

Development of a stable eukaryotic strain producing fully human monoclonal antibody on the basis of the human antibody against ectromelia virus

Fully­human antibodies have a great therapeutic importance; however, the development of stable strains providing a high level of production of full­size antibodies is a challenging task, as antibody molecules contain two types of polypeptide chains. To develop the producing strain, random integration of the plasmid containing the gene encoding the target protein into the genome of the host cells is commonly used. The aim of this study was the development of an original expression system, using gene targeting to integrate the gene encoding the fully­human antibody into the transcriptionally active region of the genome of eukaryotic suspension cells CHO­S. To develop a stable strain, the cassette vector plasmid pCDNA5/FRTDHFR­CH­CL containing the site of homologous recombination and the genes encoding heavy and light chains of the fully human antibody of the IgG1/kappa class was constructed at the first step. Notably, DNA of the plasmid pCDNA5/FRT­DHFR­CH­CL was organized in such a way that the restriction sites for rapid cloning of DNA fragments encoding the variable domains of heavy and light chains were inserted upstream of the sequences encoding constant domains of the heavy and light chains of the antibody. Secondly, DNA fragments encoding the variable domains of the heavy and light chains of antibody against orthopoxvirus protein p35 were inserted into the pCDNA5/FRT­DHFRCH­CL cassette plasmid. Then, CHO­S/FRT cells, which contain the FRT­site for homologous recombination and are able to produce green fluorescence protein GFP, were transfected with the constructed plasmid. After the insertion of the target genes into the FRT­site, GFP production was supposed to stop. Using this selection system, a stable clone producing target antibody fh8E was selected with the level of production of about 100 μg/ml. The binding affinity of purified antibody fh8E with the targeted protein, measured by surface plasmon resonance, was 12 nM. In addition, antibody fh8E demonstrated anti­vaccinia virus activity in the plaque reduction neutralization test in vitro

АВАРИЯ НА АЭС «ФУКУСИМА-1»: ПЕРВЫЕ ИТОГИ АВАРИЙНОГО РЕАГИРОВАНИЯ. СООБЩЕНИЕ 2: ДЕЙСТВИЯ ОРГАНОВ РОСПОТРЕБНАДЗОРА ПО РАДИАЦИОННОЙ ЗАЩИТЕ НАСЕЛЕНИЯ РОССИЙСКОЙ ФЕДЕРАЦИИ НА РАННЕЙ СТАДИИ АВАРИИ

Basic measures of the Rospotrebnadzor emergency response during the early stage of the «Fukushima-I» NPP radiation accident are being analyzed in the article. Radiation monitoring of the environmental objects of the territories of the Russian Federation Far East Federal District, radiation control of arriving from Japan vehicles, freights and passengers as well as imported from Japan food products were promptly organized. This allowed to get reliable evaluations of the levels of radioactive contamination at the Russian Federation territory and population exposure doses due to the «Fukushima-I» NPP accident, timely exclude the possibility of import to the Russian territory for the freights, vehicles, food products having contamination exceeding established in the Russian Federation standards.В статье проанализированы основные мероприятия по аварийному реагированию Роспотребнадзора в ранней стадии развития радиационной аварии на АЭС «Фукусима-1». Оперативно организован ради- ационный мониторинг объектов окружающей среды на территории субъектов Российской Федерации Дальневосточного Федерального округа, входной радиационный контроль транспортных средств, грузов и пассажиров, прибывающих из Японии, а также импортируемых из Японии пищевых продуктов. Это позволило получить достоверные оценки уровней радиоактивного загрязнения территории России и доз облучения населения вследствие аварии на АЭС «Фукусима-1», своевременно исключить возможность ввоза на территорию России грузов, транспортных средств, пищевых продуктов, имеющих радиоак- тивное загрязнение свыше установленных в Российской Федерации нормативов

Session 17 Ecophysiology

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Sanatorium-resort complex as a healthcare integral part in a period of socio-economic instability

The relevance of the research topic is due to the ambiguity of the influence of interrelated factors and conditions of the historical and cultural level of development of the sanatorium and resort complex of a large industrial region of Russia under the influence of changes in the socio-political and economic situation. The article summarizes and systematizes information and analytical materials that reflect the stages of formation and development of sanatorium organizations in the Sverdlovsk region. The author presents an attempt to identify mutually conditioning relationships and contradictions in the development of organizations of the sanatorium complex. The purpose of the study is to identify the influence of socio-cultural factors in the development of sanatorium organizations on increasing the level of competitiveness of tourist destinations in the industrial region of the Urals. Based on the processing, analysis and comparison of information-analytical and regulatory material of official sources of foreign and domestic literature, presented in the article, the author's conclusions are formulated about the role and contribution of the functioning of sanatorium-resort organizations in the socio-cultural and economic development of settlements, small towns and tourist destinations in the Sverdlovsk region

Pathogenetic mechanisms of atherosclerosis development in athletes

Aim of the study was to reveal the relationship between the performance of high-intensity physical activity and the development of endothelial dysfunction in people involved in professional sports. The development of endothelial dysfunction as one of the main markers of atherosclerotic damage to the vascular wall in athletes is an urgent problem in modern medicine in connection with the asymptomatic course and unpredictability of the development of cardiovascular complications. Material and methods. The publications on the topic of the scientific review from 2010 to 2020 were analyzed. The electronic databases of the Google Academy (https://scholar.google.ru/), UpToDate (www.uptodate.com), Oxford Medicine Online (https://oxfordmedicine.com/), PubMed (https://pubmed.ncbi.nlm.nih.gov/), scientific electronic library «Cyberleninka» and foreign journals Springer (https://www.springer.com/gp), Journal of the American College of Cardiology (https://imaging.onlinejacc.org/). Results. Recent foreign and domestic studies show a relatively high relationship between the level of physical activity and the development of coronary atherosclerosis in professional athletes. Conclusion. When performing prolonged and excessive physical exertion, trained athletes often experience oxidative stress, the presence of which causes the development of endothelial dysfunction, which from modern positions is a key link in the pathogenesis of atherosclerosis. Further study of the mechanism of atherogenesis will contribute to the use of new diagnostic methods in predicting the disease at an early stage and treating it, thus preserving the health of an athlete

Rapid test for the qualitative simultaneous determination of cardiac fatty acid-binding protein and cardiac troponin I in the diagnosis of acute coronary syndrome

Aim. To study the diagnostic characteristics of the rapid test for qualitative simultaneous determination of cardiac fatty acid-binding protein (FABPs) and cardiac troponin I (cTnI) CARD-INFO 1+1 in patients with acute coronary syndrome (ACS).Material and methods. The study included 168 patients undergoing inpatient treatment after ACS, with typical anginal pain lasting at least 20 minutes occurred in the previous 1-24 hours. In addition to routine diagnostic procedures, on admission, we determined FABPs and cTnI concentrations using the high-quality immunochromatographic rapid test CARD-INFO 1 1 (OOO CARDIO-Plus, Russia).Results. The sensitivity of the CARD-INFO 1+1 rapid test was 88,1%, specificity — 89,8%, diagnostic accuracy — 88,7%. The indicators of the diagnostic effectiveness of CARD-INFO 1+1 test in patients with STE-ACS and NSTE-ACS did not significantly differ (p>0,05). The sensitivity of the rapid test reached a maximum in the period from 3 to 6 hours from the onset of pain. Compared with the determination of cTnI performed on admission to the hospital, a higher sensitivity of the CARD-INFO 1+1 test was revealed in patients with STE-ACS (87,7% vs 75,3%; p=0,044), in the first 1-3 hours after the beginning of clinical manifestations (86,8% vs 60,5%; p=0,041) and in the entire sample as a whole (88,1% vs 77,1%; p=0,033), with comparable specificity (89,8% and 93,2%, respectively; p=0,741).Conclusion. Qualitative immunochromatographic CARD-INFO 1+1 rapid test for the simultaneous determination of the content of FABPs and cTnI I is highly effective in the diagnosis of various forms of ACS. The highest diagnostic characteristics of the test were observed in patients in the early stages of the disease (the first 1-6 hours after the onset of pain). Carrying out the CARD-INFO 1+1 test revealed 12 MI cases more (11%) than the first determination of cTnI. Further studies will clarify the place of this technique in the modern algorithm for the management of patients with ACS and evaluate the possibility of using the rapid test in predicting the course of the disease