Interaction of non-profit organizations and state structures of the region in the field of adaptive physical culture

The features of the provision of services by state organizations, non-profit organizations within the framework of project activities and grant support in the field of adaptive physical culture and sports are revealed. Based on the results of the analysis of the involvement of non-profit organizations (NPOs) and socially oriented non-profit organizations (SO NPOs) in the scheme of interaction with government agencies, their effectiveness in providing services (works) to the population in the field of adaptive physical culture and sports, a scheme of interaction was developed and proposed to solve the identified problem

Развитие инновационной деятельности в Российской Федерации: проблемы и перспективы

The purpose of the article is to identify key trends in the development of scientific and innovative activities in the Russian Federation. The methodology consists of comparative and structural analyses, analysis and synthesis, and the method of logical generalization. The study established the main trends in the process of development, implementation, and diffusion of innovations in scientific and industrial activities in the Russian Federation. In recent years, the innovation ranking of the countries of the world has not highly assessed the state of the innovation system of the Russian Federation. The country occupies a stable 45–47th place among 130 surveyed countries of the world with weaknesses including institutions, infrastructure, energy efficiency, and environmental friendliness of businesses, as well as the amount of investment, including foreign ones. Strong but improperly implemented aspects include the high quality of human potential, the results of creative details, and the scale of the market. The factors hindering the implementation of the innovation process have been analyzed, and despite decreasing over the past twenty years, the lack of own funds, high innovation costs, and lack of state financial support are still significant obstacles to enterprises. The process of implementing innovative activities by enterprises and their sustainable development directly depend on the state of science in the country. The analysis has shown that decisions made in the process of implementing state policy do not have a significant positive impact on its development; moreover, a number of indicators such as the number of personnel involved in research and development generally decrease annually. Thus, there is clearly a need to revise the quality, quantity, and essence of public administration measures so that the innovative and scientific potential of the Russian Federation is realized, ensuring the country’s economy to switch to an innovative development path.Цель статьи заключается в выявлении ключевых тенденций развития инновационной деятельности в Российской Федерации. Методологию составляет сравнительный и структурный анализ, анализ и синтез, метод логического обобщения. В результате исследования были проанализированы основные тенденции разработки, реализации и диффузии инноваций в научной и производственной деятельности в Российской Федерации. Инновационный рейтинг стран мира последние годы невысоко оценивает состояние инновационной системы Российской Федерации –  страна занимает устойчивое 45–47 место из 130 обследованных стран. Анализу подвергнуты факторы, препятствующие осуществлению инновационного процесса. Установлено, что за последние двадцать лет в целом воздействие таких барьеров снижается, однако недостаток собственных денежных средств, высокая стоимость нововведений и недостаток финансовой поддержки со стороны государства по-прежнему выделяются предприятиями как самые значимые из всей совокупности факторов. Процесс осуществления инновационной деятельности у предприятий, их устойчивое развитие напрямую зависят от состояния науки в стране. Анализ позволил установить, что принимаемые в процессе осуществления государственной политики решения не оказывают значимого положительного воздействия на ее развитие, более того, ряд показателей, таких как число персонала, занятого научными исследованиями и разработками, вообще ежегодно снижается. Таким образом, явно назрела необходимость пересмотра качества, количества и сути мероприятий государственного управления и администрирования с тем, чтобы инновационный и научный потенциал Российской Федерации был реализован, давая возможность перейти экономике страны на инновационный путь развития

Интернализация рекомбинантной имиглюцеразы в перитонеальные макрофаги мыши и фибробласты мыши линии L929

Enzyme replacement therapy (ERT) is one of the most efficient treatments for lysosomal storage diseases. Type 1 Gaucher disease is caused by β-glucocerebrosidase enzyme deficiency, which may be compensated for by intravenous infusions of imiglucerase—a recombinant enzyme. Imiglucerase targets macrophages and enters these cells via interaction with mannose receptors on the cell membrane. Characterisation of internalization of enzymes by target cells is important in the context of the development of new medicines and production of existing ERT medicines. The peritoneal and alveolar macrophages, as well as macrophages of the spleen of small laboratory animals (rats and mice) are widely used in such studies. However, isolation of cells from animal sources raises ethical issues, and therefore continuous mammalian cell lines may offer an attractive alternative. The aim of the study: to conduct comparative studies on the internalization of recombinant imiglucerase into mouse peritoneal macrophages and L929 mouse fibroblasts. Materials and methods: CerezymeR batches 7HV0913, C6214H05, 7HV0888 (Genzyme Ltd., UK); Glurazim batches 020416, 011117, 021117 (LLC “IBC “Generium”, Russia). We used peritoneal macrophages obtained from BALB/c mice and L929 mouse fibroblasts. The cells were cultured in DMEM/F12 complete growth medium with 10% fetal bovine serum. The activity of imiglucerase internalized into the cells was evaluated spectrophotometrically by hydrolysis of the artificial substrate—4-methylumbelliferyl-β-Dglucopyranoside. Results: the study compared internalization of recombinant imiglucerase (the active ingredient of CerezymeR and Glurazim) by mouse peritoneal macrophages and L929 mouse fibroblasts. It was demonstrated that the medicines activity in the lysates of peritoneal macrophages is comparable with that in the lysates of L929 mouse fibroblasts. Regardless of the model system, the activity of Glurazim stayed within the acceptable range (80–125%) established for biosimilar products. Conclusions: the experiments proved that L929 mouse fibroblasts could be recommended for assessment of internalization of recombinant imiglucerase.Фермент-заместительная терапия (ФЗТ) является одной из самых действенных при лечении болезней лизосомального накопления. Болезнь Гоше первого типа характеризуется недостатком нативного фермента β-глюкоцереброзидазы, который возмещают внутривенными инфузиями рекомбинантного фермента (имиглюцераза). Клетками-мишенями имиглюцеразы являются макрофаги, в которые фермент проникает посредством взаимодействия с рецепторами маннозы на клеточной мембране. Оценка интернализации ферментов клетками-мишенями представляет интерес при разработке новых и воспроизведении существующих препаратов для ФЗТ. Для этих исследований широко применяются перитонеальные и альвеолярные макрофаги, макрофаги селезенки мелких лабораторных животных (крыс и мышей). Однако получение таких клеток затрагивает этические вопросы использования лабораторных животных. Альтернативой являются перевиваемые клеточные линии млекопитающих. Цель работы: провести сравнительные исследования интернализации рекомбинантной имиглюцеразы в перитонеальные макрофаги мыши и фибробласты мыши линии L929. Материалы и методы: Церезим®, серии 7HV0913, C6214H05, 7HV0888 (Джензайм Лтд., Великобритания); Глуразим, серии 020416, 011117, 021117 (ООО «МБЦ «Генериум», Россия). В работе использовали перитонеальные макрофаги, полученные от мышей линии BALB/c, и фибробласты мыши линии L929. Клетки культивировали в полной ростовой среде ДМЕМ/Ф12 c добавлением 10% сыворотки плода крупного рогатого скота. Активность имиглюцеразы, проникшей в клетки, оценивали спектрофотометрически по гидролизу искусственного субстрата 4-метилумбеллиферил-β-D-глюкопиранозида. Результаты: представлены данные сравнительной оценки интернализации рекомбинантной имиглюцеразы, действующего вещества препаратов Церезим® и Глуразим, перитонеальными макрофагами мыши и клетками фибробластов мыши линии L929. Показано, что активность препаратов в лизатах перитонеальных макрофагов сопоставима с их активностью в лизатах клеток фибробластов мыши линии L929, при этом активность разработанного препарата Глуразим независимо от типа клеток, была в границах допустимого диапазона (80–125%), установленного для биоподобных препаратов. Выводы: экспериментально доказано, что фибробласты мыши линии L929 могут быть рекомендованы для оценки интернализации рекомбинантной имиглюцеразы

Assessment of T-cell immunity to SARS-CoV-2 in COVID-19 convalescents and vaccinated subjects, using TigraTest^® SARS-CoV-2 ELISPOT kit

With the onset of the COVID-19 pandemic, a number of molecular-based tests have been developed to diagnose SARS-CoV-2 infection. However, numerous available serological tests lack sufficient sensitivity or specificity. They do not detect specific antibodies in a significant proportion of patients with PCR-confirmed COVID-19. There is evidence that some convalescents have a relatively short-lived humoral immunity. In contrast, a number of publications have shown that T-cell response to human coronaviruses, including SARS-CoV-1, MERS, and SARS-CoV-2, can be strong and long-term. Assessment of T-cell immunity to SARS-CoV-2 is important not only for stratification of risks and identification of potentially protected populations with immunity acquired as a result of previous infection, but also for determining immunogenicity and potential efficacy of vaccines under development. The existing methods of quantitative or semi-quantitative assessment of specific T-cell response are mainly used in scientific research and are not standardised. The aim of the study was to develop and verify experimentally a test kit to be used in a standardised procedure for in vitro determination of T-cells specific to SARS-CoV-2 antigens, in human peripheral blood. Materials and methods: the TigraTest® SARS-CoV-2 kit developed by GENERIUM, which determines the number of T-cells secreting interferon gamma in vitro, was tested in the study. Samples of venous blood of volunteers from three different groups were analysed in the study: presumably healthy volunteers; COVID-19 convalescents; individuals vaccinated against SARS-CoV-2. Results: the authors developed the TigraTest® SARS-CoV-2 kit for in vitro determination of T-cells specific to SARS-CoV-2 antigens in human peripheral blood, demonstrated its specificity and performed preliminary assessment of its sensitivity. The study analysed the range and magnitude of the T-cell response in convalescent and vaccinated individuals. A pronounced T-cell response was also shown in some individuals with no symptoms or with unconfirmed diagnosis. It was discovered that the mean T-cell response to peptides of the spike protein (S-protein) was higher in the vaccinated individuals than in the convalescent patients. A correlation was determined between the severity of the disease and the level of T-cell response. Specific contributions of various groups of antigens to the T-cell response after COVID-19 infection were also determined. Conclusions: the TigraTest® SARS-CoV-2 kit is a specific and sensitive tool for the assessment of T-cell immunity to the SARS-CoV-2 virus, which can also be used for vaccinated individuals. The kit may be used in clinical practice for comprehensive assessment of immunity to SARS-CoV-2

Internalization of Recombinant Imiglucerase into Mouse Peritoneal Macrophages and L929 Mouse Fibroblasts

Enzyme replacement therapy (ERT) is one of the most efficient treatments for lysosomal storage diseases. Type 1 Gaucher disease is caused by β-glucocerebrosidase enzyme deficiency, which may be compensated for by intravenous infusions of imiglucerase—a recombinant enzyme. Imiglucerase targets macrophages and enters these cells via interaction with mannose receptors on the cell membrane. Characterisation of internalization of enzymes by target cells is important in the context of the development of new medicines and production of existing ERT medicines. The peritoneal and alveolar macrophages, as well as macrophages of the spleen of small laboratory animals (rats and mice) are widely used in such studies. However, isolation of cells from animal sources raises ethical issues, and therefore continuous mammalian cell lines may offer an attractive alternative. The aim of the study: to conduct comparative studies on the internalization of recombinant imiglucerase into mouse peritoneal macrophages and L929 mouse fibroblasts. Materials and methods: CerezymeR batches 7HV0913, C6214H05, 7HV0888 (Genzyme Ltd., UK); Glurazim batches 020416, 011117, 021117 (LLC “IBC “Generium”, Russia). We used peritoneal macrophages obtained from BALB/c mice and L929 mouse fibroblasts. The cells were cultured in DMEM/F12 complete growth medium with 10% fetal bovine serum. The activity of imiglucerase internalized into the cells was evaluated spectrophotometrically by hydrolysis of the artificial substrate—4-methylumbelliferyl-β-Dglucopyranoside. Results: the study compared internalization of recombinant imiglucerase (the active ingredient of CerezymeR and Glurazim) by mouse peritoneal macrophages and L929 mouse fibroblasts. It was demonstrated that the medicines activity in the lysates of peritoneal macrophages is comparable with that in the lysates of L929 mouse fibroblasts. Regardless of the model system, the activity of Glurazim stayed within the acceptable range (80–125%) established for biosimilar products. Conclusions: the experiments proved that L929 mouse fibroblasts could be recommended for assessment of internalization of recombinant imiglucerase