ОСТРАЯ, ХРОНИЧЕСКАЯ ТОКСИЧНОСТЬ И РАЗДРАЖАЮЩЕЕ ДЕЙСТВИЕ СМ-КОМПЛЕКСА

The researchers elaborated pharmacological complex of SM at the Chair of Morphology, Physiology and Pharmacology atSouth-UralStateAgrarianUniversity. The complex contains butafosfan, vitamins, vitamin-like substances, selected on the basis of synergistic effect on the body. The parameters of acute toxicity of the SM-complex were explored in the experiment on clinically healthy adult white mice of both genders by a single injection of the solution in the maximum permissible dose according to GOST 31926-2013. Intolerance on the skin of animals was detected in line with GOST R ISO 10993.10-2009. GOST R ISO 10993.10-99 became a guideline for evaluating SM-complex intolerance on the eye conjunctiva. Chronic toxicity was investigated on non-linear rats; the rates were divided into 4 groups of 6. The first group became a control group, the second group received the SM-complex in a therapeutic dose (255mg/kg), the third group - in a 5 -fold therapeutic dose (1275mg/kg), the fourth group - in a 10-fold dose (2550mg/kg). Pharmacological substance was applied through the probe in the form of an aqueous solution for 30 days. The researchers found out that at single oral introduction of SM-complex in the maximum possible doses it does not affect mice organism and it is referred to the 4th class of danger according to GOST 12.1.007-76. Local application in the form of applications on the skin and mucous membranes of rabbits, the complex does not have a local irritant effect. Long-term application of pharmacological composition (30 days) in high doses causes functional and morphological changes of the liver in the form of gray foci and flabbiness, as well as it increases the volume of the organ. Due to the fact that the therapeutic doses are 5 and 10 times lower than the toxic ones and the period of application does not exceed 7-14 days, the authors make a conclusion that SM-complex is safe and secure and can be used in the recommended doses.В условиях кафедры морфологии, физиологии и фармакологии Южно-Уральского государственного аграрного университета был разработан фармакологический комплекс СМ, который в своем составе содержит бутафосфан, витамины, витаминоподобные вещества, подобранные по принципу синергического действия на организм. Параметры острой токсичности СМ-комплекса изучены в опыте на клинически здоровых половозрелых белых мышах обоего пола путем однократного введения через зонд раствора в максимально возможной дозе согласно ГОСТ 31926–2013. Определение раздражающего действия на кожу животных проводилось в соответствии с ГОСТ Р ИСО 10993.10–2009. Для оценки раздражающего действия СМ-комплекса на конъюнктиву глаза руководствовались ГОСТ Р ИСО 10993.10–99. Хроническая токсичность была изучена на нелинейных крысах, разделенных на 4 группы по 6 животных. Первая группа служила контролем, 2-я получала СМ-комплекс в терапевтической дозе (255 мг/кг), 3-я – в 5-кратной терапевтической дозе (1275 мг/кг), 4-я – в 10-кратной (2550 мг/кг). Фармакологическое средство задавали через зонд в виде водного раствора на протяжении 30 суток. В результате исследований определено, что при однократном пероральном введении СМ- комплекса в максимально возможных дозах он не оказывает летального действия на организм мышей и относится к 4-му классу опасности по ГОСТ 12.1.007–76. При местном применении в виде аппликаций на кожу и слизистые оболочки кроликов комплекс не оказывает местного раздражающего действия. Длительное использование фармакологической композиции (в течение 30 дней) в высоких дозах вызывает функциональные и морфологические изменения печени в виде серых очагов и дряблости, а также увеличение органа в объеме. Но в связи с тем, что терапевтические дозы ниже токсических в 5 и 10 раз и период использования не превышает 7–14 суток, можно сделать заключение, что СМ-комплекс относительно безопасен и может быть использован без ограничения в рекомендуемых дозах

Mathematical Model of Kinetics of O-Antigen Accumulation in the Process of Periodic Submerged Cultivation of <I>Vibrio cholerae</I> M-41 Ogava with Limitation on Carbon Substrate

Developed is the system of differential equations that characterize kinetics of biomass growth, glucose utilization and O-antigen accumulation in the process of submerged cultivation of V. cholerae strain M-41 Ogawa. The parameters of the mathematic model are identified. Using the developed software in Mathcad 15.0 determined are kinetic constants and coefficients. The mathematical model is demonstrated to describe adequately O-antigen biosynthesis process. The received data can be used in large-scale technology of V. cholerae strain M-41 submerged cultivation

Mathematical Model of Kinetics of Antigens Accumulation in the Process of Periodical Submerged Cultivation of <I>Vibrio cholerae </I>569В Inaba with Limitation as Regards Carbonic Substrate

Presented is mathematical model of kinetics of the process of O-antigen and cholera toxin synthesis during periodical submerged cultivation of V. cholerae 569В Inaba with limitation as regards carbonic substrate. The proposed model is based upon analysis of experimental data on V. cholerae 569В Inaba biomass and antigens accumulation, rate of growth and antigens release, and glucose utilization. Using Mathcad 15.0 software calculated are coefficients of differential equations entering into the mathematical model. Comparison of predicted and experimental data demonstrates that relative error of determination of concentrations of the synthesized substances, glucose and cholera vibrio is between 5 and 20 %. The proposed model permits to determine maximum output of final products and specify the parameters of cultivation process performance at different initial conditions

Experimental Evaluation of Application of Cross-Flow Ultrafiltarion Method for O Antigen Concentrating in Cholera Chemical Bivalent Vaccine Production

Demonstrated is possibility to apply cross-flow ultrafiltration method for O antigen of Vibrio cholerae M-41 Ogawa concentrating from germ-free centrifugate. Technological process of concentrating was optimized. Worked out were the regimes of conservation and cleaning of the ultrafiltration device. The prospects of cross-flow ultrafiltration method introduction in technology of cholera chemical bivalent vaccine production were determined

Deployment of Ultrafiltration for Concentrating and Purification of Antigens

Represented is domestic and foreign literature review dedicated to usage of ultrafiltration for concentrating and purification of antigens. Discussed are the issues of deployment of various ultrafiltration techniques. It is determined that filtering in the tangential mode by means of modules with flat-frame filtering elements is among the prospective ones. Demonstrated is the impact of such technological specifications as concentration rate, pressure, temperature, and membrane nominal cut-off on molecular mass on the quality of target products, the time elapsed, and preparation losses decrease (increase). Literature data analysis proves to be useful for the selection of the proper procedure for concentrating and purification of protective antigens of bacterial and viral origins. In addition, it allows for taking into account the parameters under discussion when developing specific manufacturing technologies for diagnostic and preventive medical immunobiological preparation production

Examination of Processes of Cultivation of <i>V. cholerae</i> strains - Producers of Protective Antigens of Cholera Bivalent Chemical Vaccine in Tablets - in the Engineered Bioreactor

Studied is the kinetics of major antigen accumulation at growing of Vibrio cholerae M-41 Ogawa and 569 B Inaba strains in industrial and engineered reactors. Demonstrated is the possibility to obtain conditional native protective antigens in the engineered bioreactor. Shown is the identity of physiological and morphological properties of industrial Vibrio cholerae strains during their submerged cultivation in industrial and engineered bioreactors. Cholera bivalent chemical vaccine obtained using engineered bioreactor possesses quality indices meeting the requirements of normative documents and equal to those of preparation received by traditional approach

Features of the frequency of occurrence of T-330G <i>IL2</i> gene polymorphism in patients with COVID-19

SARS-CoV-2 infection is the etiopathogenetic factor of the new coronavirus infection. Susceptibility to the virus and, accordingly, the incidence differs in children and adults. On the one hand, this reflects the age-related features of the immune response. On the other hand, it is realized through the production of a number of cytokines, including IL-2, and reflects the genetically determined features of cytokine production. The aim of the study was to analyze the frequency of occurrence of T-330G polymorphic variants of the IL2 gene in patients with a new coronavirus infection. A total of 145 patients were examined, including 31.0% of children (n = 45) and 69.0% of adults (n = 100). The diagnosis of a new coronavirus infection was verified by RT-PCR confirming the presence of the SARS-CoV-2 virus and identifying clinical symptoms of an upper respiratory tract infection. The control group consisted of 50 healthy volunteer donors. Allele-specific PCR with electrophoretic detection in 3% agarose gel (Litech, Russia) was used to analyze the T-330G polymorphism of the IL2 gene. To compare the frequencies of allele combinations, the χ2 test and the odds ratio OR and (95% CI) were used.The dominant genotype in patients with COVID-19 was the heterozygous GT genotype of the T-330G polymorphism of the IL2 gene. In the group of children at risk of developing a new coronavirus infection, the GG genotype of the T-330G polymorphism of the IL2 gene was associated (31.1% in children and 18.0% in the control group, p &lt; 0.05, OR = 2.047). While the homozygous TT genotype of the T-330G polymorphism of the IL2 gene was a protective genotype (its occurrence rate was 26.7% in patients, 54.0% in the control group, p &lt; 0.05, OR = 0.315). In adults, the heterozygous GT genotype of the T-330G polymorphism of the IL2 gene was associated with the risk of developing a new coronavirus infection (in the group of patients – 44.0% versus control – 28.0%, p = 0.028, OR = 2.020). A low risk of developing the disease was associated with the homozygous TT variant of the T-330G polymorphism of the IL2 gene (in the group of patients 37.0% versus control – 54.0%, p = 0.024, OR = 0.500).The T-330G polymorphism of the promoter zone of the IL2 gene differently affects its production. The direction of the immune response and its effectiveness depend on the level of IL-2. Understanding the individual factors that determine the features of the immune response can help in understanding the mechanisms of development of COVID-19-associated diseases and the selection of approaches to personalized methods of their treatment

Experimental Technology for O-Antigens Production of Non-Toxigenic Strains of <I>Vibrio cholerae</I>

Determined are the key bio-kinetic indexes of submerged cultivation of Vibrio cholerae non-toxigenic strains, producers of O-antigens. Evaluated is the technology of O-antigen concentration using tangential ultrafiltration technique. The results suggest the principal possibility of using these strains for biologically safe production of chemical cholera vaccines

Methods and Technologies of Cholera Vibrio Cultivation (Scientific Review)

Displayed is the review of domestic and foreign literature sources devoted to matters of cholera vibrio cultivation. Discussed is the information concerning the following methods utilized in the technological process of vibrio growth stimulation: batch cultivation, fed-batch, fermentation and dialysis, periodic and continuous cultivation. Analyzed is the impact of such parameters as dissolved oxygen concentration, count of carbon nutrition source, medium pH, temperature rate , concentration and physiological condition of inoculate, duration of technological process, affecting the growth of this microorganism and synthesis of its antigens. Consequently, literature data analysis has contributed to the selection of proper method for cholera vibrio cultivation and consideration of the factors mentioned above for the development of manufacture technology applied to the production of medical immunoglobulin preparations for diagnostics and prophylaxis of cholera

Improvement of Technology of Cholera Toxin B-Subunit Production

Consideration is given to implementation of state-of-the-art filtration technologies for up-scaled manufacturing of cholera toxin B-subunit, produced by recombinant Vibrio cholerae non O1 KM93 strain. Selected are micro- and ultra-filtration membranes to be incorporated into manufacturing method. Investigated are the properties of cholera toxin B-subunit, obtained applying the pilot technology. The engineered method for up-scaled manufacturing of cholera toxin B-subunit makes the procedure easier-to-maintain due to tangential micro- and ultra-filtration, performed at the stage of purification and concentration. It excludes labor-consuming chromatographic purification, while retaining B-subunit properties. The studies undertaken make it possible to manufacture cholera toxin B-subunit with the same characteristics as in the case of the pilot technology, but under production conditions, and use it as a component for chemical cholera vaccine