B1 plus exam : a key to higher education in Russia

The use of open electronic courses is a relatively new tendency in education and it is not yet fully discovered by researchers. This article examines the specifics of the introduction of online courses, including the requirements to them, as well as factors that slow down their active development. The following methods of investigation were used: analysis, synthesis, deduction and induction. As an example the Kozma Minin Nizhny Novgorod State Pedagogical University - Minin University was used. On the basis of the study of the "Regulations on the use of open courses in the educational activities of Minin University", it was noted, in addition to the functions of the coordination group on open education, that for the trainee at the end of the course, in the case of unsatisfactory result, the possibility of the recalculation is established. This possibility is used not by every university. In the article we make a conclusion that open courses are a new qualitative step in the development of world education, but online courses would be much more effective as narrow-minded, adult-oriented, and deeply motivated for learning. Taken as the basis of research, Minin University showed that the university is ready for the implementation of online courses. It is one of the few universities that actively use open courses in Russia.peer-reviewe

L-NAME-induced preeclampsia: correction of functional disorders of the hemostasis system with resveratrol and nicorandil

Preeclampsia is a formidable disease of the second half of pregnancy, leading to severe complications, including disability and even death. Many authors have recognized the correlation between the severity of preeclampsia and the degree of disturbances in the hemostasis system. In this regard, the objective of this study was to assess inhibition of platelet aggregation and the possibility of its correction with resverаtrol and nicorand

The influence of imidazoline receptor agonists on the ophthalmoscopic and morphometric parameters in retina in simulated retinal ischemia-reperfusion

An important task of experimental and clinical pharmacology and ophtalmology is to find highly effective drugs for correcting retinal ischemi

Search and evaluation of pharmacodynamic and pharmacokinetic parameters of selective blocker of TRPA₁ ion channels from the group of substituted pyrazinopyrimidinones

Search considerations evaluation of pharmacodynamic and pharmacokinetic parameters of selective blocker of TRPA₁ ion channels from the group of substituted pyrazinopyrimidinone

Review of a new concept of glaucoma pathogenesis based on the glymphatic theory of cerebrospinal fluid circulation

Clinical evidence for the glymphatic system hypothesis: There is some clinical, including case-based, and epidemiological evidence for similarities between glaucomatous optic nerve/retinal injuries and AD, since both occur in the form of improper secretion of neurotoxic metabolites, and both are often diagnosed togethe

ROLE OF MESENCHYMAL MULTIPOTENT STROMAL CELLS IN REMODELING OF BONE DEFECTS

Ability of mesenchymal multipotent stromal cells (MSCs) to differentiate into several types of mesenchymal tissues allows to consider these cells the main candidates for creating tissue engineering constructions for regenerative medicine. MSCs promote integration of bio-implants into the native bone and stimulate osteogenesis. MSCs are characterized by immunomodulatory properties, due to inflammation control and modification of immune cells. MSCs affect not only the in vivo immune response by preventing immunological rejection of implanted tissue engineering designs, but it can also influence the bone tissue immunity. MSCs play an important role in bone regeneration, by regulating the osteoblastic generation, and suppressing activity of inflammation effectors and osteoclastogenesis. Some pre-clinical and first clinical trials of bone bio-implants colonized with MSC, demonstrate promising outlooks for this strategy in order to obtain tissue engineering constructions for bone regeneration

Production and characterisation of a SARS-CoV-2 S-protein RBD homodimer with increased avidity for specific antibodies

Monitoring of the proportion of immune individuals and the effectiveness of vaccination in a population involves evaluation of several important parameters, including the level of virus-neutralising antibodies. In order to combat the COVID-19 pandemic, it is essential to develop approaches to detecting SARS-CoV-2 neutralising antibodies by safe, simple and rapid methods that do not require live viruses. To develop a test system for enzyme-linked immunosorbent assay (ELISA) that detects potential neutralising antibodies, it is necessary to obtain a highly purified recombinant receptor-binding domain (RBD) of the spike (S) protein with high avidity for specific antibodies.The aim of the study was to obtain and characterise a SARS-CoV-2 S-protein RBD homodimer and a recombinant RBD-expressing cell line, as well as to create an ELISA system for detecting potential neutralising antibodies.Materials and methods: the genetic construct was designed in silico. To generate a stable producer cell line, the authors transfected CHO-S cells, subjected them to antibiotic pressure, and selected the optimal clone. To isolate monomeric and homodimeric RBD forms, the authors purified the recombinant RBD by chromatographic methods. Further, they analysed the activity of the RBD forms by Western blotting, bio-layer interferometry, and indirect ELISA. The analysis involved mono clonal antibodies GamXRH19, GamP2C5, and h6g3, as well as serum samples from volunteers vaccinated with Gam-COVID-Vac (Sputnik V) and unvaccinated ones.Results: the authors produced the CHO-S cell line for stable expression of the recombinant SARS-CoV-2 S-protein RBD. The study demonstrated the recombinant RBD’s ability to homodimerise after fed-batch cultivation of the cell line for more than 7 days due to the presence of unpaired cysteines. The purified recombinant RBD yield from culture broth was 30–50 mg/L. Monomeric and homodimeric RBD forms were separated using gel-filtration chromatography and characterised by their ability to interact with specific monoclonal antibodies, as well as with serum samples from vaccinated volunteers. The homodimeric recombinant RBD showed increased avidity for both monoclonal and immune sera antibodies.Conclusions: the homodimeric recombinant RBD may be more preferable for the analysis of levels of antibodies to the receptor-binding domain of the SARS-CoV-2 S protein

К вопросу о критическом артериальном стенозе

Cardiovascular disease (CVD) is the leading cause of death worldwide. The main contribution to the structure of mortality from CVD is made by atherosclerosis. Indications for surgical treatment of patients with diseases caused by atherosclerotic lesions of the arteries are set taking into account the degree of their stenosis. Angiography has been considered the gold standard for screening patients with CVD for many years. Numerous studies carried out over the past several decades have revealed the weak side of this method in assessing the significance of borderline stenosis. Therefore, to analyze such changes, functional tests were introduced to clarify the indications for surgical intervention. Currently, criteria for the significance of stenosis of the iliac, renal and coronary arteries have been determined. The significance of stenosis of the arteries supplying the brain and intestines is still a matter of debate and requires further study.Сердечно-сосудистые заболевания (ССЗ) занимают 1-е место среди причин смерти во всем мире. Основной вклад в структуру смертности от ССЗ вносит атеросклероз. Показания к хирургическому лечению пациентов с заболеваниями, вызванными атеросклеротическим сужением артерий, выставляются с учетом степени их стеноза. На протяжении многих лет «золотым стандартом» обследования пациентов с ССЗ считается ангиография. Множественные исследования, проведенные за последние несколько десятков лет, выявили слабую сторону этого метода в оценке значимости пограничных стенозов. Поэтому для анализа таких изменений были введены функциональные тесты, уточняющие показания к хирургическому вмешательству. В настоящее время определены критерии значимости сужения подвздошных, почечных и коронарных артерий. Значимость стенозов артерий, кровоснабжающих головной мозг и кишечник, до сих пор остается предметом дискуссий и требует дальнейшего изучения

Получение и характеристика гомодимерной формы RBD S-белка SARS-CoV-2, обладающей повышенной авидностью к специфическим антителам

Monitoring of the proportion of immune individuals and the effectiveness of vaccination in a population involves evaluation of several important parameters, including the level of virus-neutralising antibodies. In order to combat the COVID-19 pandemic, it is essential to develop approaches to detecting SARS-CoV-2 neutralising antibodies by safe, simple and rapid methods that do not require live viruses. To develop a test system for enzyme-linked immunosorbent assay (ELISA) that detects potential neutralising antibodies, it is necessary to obtain a highly purified recombinant receptor-binding domain (RBD) of the spike (S) protein with high avidity for specific antibodies. The aim of the study w as t o obtain and characterise a SARSCoV-2 S-protein RBD homodimer and a recombinant RBD-expressing cell line, as well as to create an ELISA system for detecting potential neutralising antibodies. Materials and methods: the genetic construct was designed in silico. To generate a stable producer cell line, the authors transfected CHO-S cells, subjected them to antibiotic pressure, and selected the optimal clone. To isolate monomeric and homodimeric RBD forms, the authors purified the recombinant RBD by chromatographic methods. Further, they analysed the activity of the RBD forms by Western blotting, bio-layer interferometry, and indirect ELISA. The analysis involved monoclonal antibodies GamXRH19, GamP2C5, and h6g3, as well as serum samples from volunteers vaccinated with Gam-COVID-Vac (Sputnik V) and unvaccinated ones. Results: the authors produced the CHO-S cell line for stable expression of the recombinant SARS-CoV-2 S-protein RBD. The study demonstrated the recombinant RBD’s ability to homodimerise after fed-batch cultivation of the cell line for more than 7 days due to the presence of unpaired cysteines. The purified recombinant RBD yield from culture broth was 30–50 mg/L. Monomeric and homodimeric RBD forms were separated using gel-filtration chromatography and characterised by their ability to interact with specific monoclonal antibodies, as well as with serum samples from vaccinated volunteers. The homodimeric recombinant RBD showed increased avidity for both monoclonal and immune sera antibodies. Conclusions: the homodimeric recombinant RBD may be more preferable for the analysis of levels of antibodies to the receptor-binding domain of the SARS-CoV-2 S protein.Важным параметром, оцениваемым при мониторинге иммунной прослойки у населения и эффективности вакцинации населения, является уровень вируснейтрализующих антител. Разработка подхода к выявлению вируснейтрализующих антител к вирусу SARS-CoV-2 с помощью безопасного, простого и быстрого метода, не требующего использования живых вирусов, имеет большое значение для борьбы с пандемией COVID-19. Для разработки тест-систем для проведения иммуноферментного анализа (ИФА), детектирующих потенциально вируснейтрализующие антитела, необходимо получение высокоочищенного рекомбинантного рецептор-связывающего домена (RBD) S-белка, обладающего высокой авидностью к специфическим антителам. Цель работы: получение и характеристика гомодимерной формы RBD S-белка вируса SARS-CoV-2, а также клеточной линии, продуцирующей рекомбинантный RBD, для создания ИФА тест-системы для выявления потенциально вируснейтрализующих антител. Материалы и методы: дизайн генетической конструкции проводили in silico. Стабильную клеточную линию получали при помощи трансфекции клеток CHO-S, селекции на антибиотике и отбора оптимального клона. Рекомбинантный RBD очищали с использованием хроматографических методов, получали мономерную и гомодимерную формы RBD. Активность полученных форм анализировали с использованием методов Вестерн-блот, биослойной интерферометрии и непрямго ИФА. Для анализа использовали моноклональные антитела GamXRH19, GamP2C5 и h6g3, а также образцы сывороток крови добровольцев, вакцинированных препаратом Гам-КОВИД-Вак, и невакцинированных добровольцев. Результаты: получена клеточная линия CHO-S, стабильно продуцирующая рекомбинантный RBD S-белка вируса SARS-CoV-2. Показано, что при культивировании данной клеточной линии в режиме fed-batch более 7 суток рекомбинантный RBD способен образовывать гомодимеры за счет наличия неспаренных цистеинов. Количественный выход очищенного рекомбинантного RBD из культуральной жидкости составил 30–50 мг/л. Мономерная и гомодимерная формы RBD были разделены при помощи гель-фильтрации и охарактеризованы по способности взаимодействовать со специфическими моноклональными антителами, а также сыворотками крови от вакцинированных добровольцев. Продемонстрировано, что именно гомодимерная форма рекомбинантного RBD обладает повышенной авидностью к моноклональным антителам и антителам в сыворотке крови вакцинированных. Выводы: гомодимерная форма рекомбинантного RBD может являться более предпочтительной для анализа уровня антител к рецептор-связывающему домену S-белка вируса SARS-CoV-2

rAAV expressing recombinant antibody for emergency prevention and long-term prophylaxis of COVID-19

IntroductionNumerous agents for prophylaxis of SARS-CoV-2-induced diseases are currently registered for the clinical use. Formation of the immunity happens within several weeks following vaccine administration which is their key disadvantage. In contrast, drugs based on monoclonal antibodies, enable rapid passive immunization and therefore can be used for emergency pre- and post-exposure prophylaxis of COVID-19. However rapid elimination of antibody-based drugs from the circulation limits their usage for prolonged pre-exposure prophylaxis.MethodsIn current work we developed a recombinant adeno-associated viral vector (rAAV), expressing a SARS-CoV-2 spike receptor-binding domain (RBD)-specific antibody P2C5 fused with a human IgG1 Fc fragment (P2C5-Fc) using methods of molecular biotechnology and bioprocessing.Results and discussionsA P2C5-Fc antibody expressed by a proposed rAAV (rAAV-P2C5-Fc) was shown to circulate within more than 300 days in blood of transduced mice and protect animals from lethal SARS-CoV-2 virus (B.1.1.1 and Omicron BA.5 variants) lethal dose of 105 TCID50. In addition, rAAV-P2C5-Fc demonstrated 100% protective activity as emergency prevention and long-term prophylaxis, respectively. It was also demonstrated that high titers of neutralizing antibodies to the SARS-CoV-2 virus were detected in the blood serum of animals that received rAAV-P2C5-Fc for more than 10 months from the moment of administration.Our data therefore indicate applicability of an rAAV for passive immunization and induction of a rapid long-term protection against various SARS-CoV-2 variants