Design and assembly of plant-based COVID-19 candidate vaccines: reсent development and future prospects

An outbreak of a new variant of the coronavirus infection, known as COVID-19, occurred at the end of 2019 in China, in the city of Wuhan. It was caused by the SARS-CoV-2 virus. This variant of the virus is characterized by a high degree of variability and, as the current situation with its spread across different regions of the globe shows, it can lead to a progressive spread of infection among the human population and become the cause of a pandemic. The world scientific community is making tremendous efforts to develop means of protection, prevention and treatment of this disease based on modern advances in molecular biology, immunology and vaccinology. This review provides information on the current state of research in the field of vaccine development against COVID-19 with an emphasis on the role of plants in solving this complex problem. Although plants have long been used by mankind as sources of various medicinal substances, in a pandemic, plant expression systems become attractive as biofactories or bioreactors for the production of artificially created protein molecules that include protective antigens against viral infection. The design and creation of such artificial molecules underlies the development of recombinant subunit vaccines aimed at a rapid response against the spread of infections with a high degree of variability. The review presents the state of research covering a period of just over two years, i. e. since the emergence of the new outbreak of coronavirus infection. The authors tried to emphasize the importance of rapid response of research groups from various scientific fields towards the use of existing developments to create means of protection against various pathogens. With two plant expression systems – stable and transient – as examples, the development of work on the creation of recombinant subunit vaccines against COVID-19 in various laboratories and commercial companies is shown. The authors emphasize that plant expression systems have promise for the development of not only protective means under conditions of rapid response (subunit vaccines), but also therapeutic agents in the form of monoclonal antibodies against COVID-19 synthesized in plant cells

Recommendations for the Programme of Clinical Trials of Medicinal Products for the Treatment of Influenza

The development of new medicinal products to treat influenza is motivated by the limitations of existing treatment options, the emergence of drug resistance, and the health consequences of influenza epidemics associated with the highly contagious nature of the virus. Proper planning and implementation of clinical programmes providing reliable data on the efficacy and safety of medicinal products under development requires adherence to recommendations of the regulatory authorities. At the moment, the Russian Federation, the Eurasian Economic Union, and the European Union lack documented recommendations on conducting clinical trials of anti-influenza medicines. There is a need in national guidelines that will reflect the procedure for conducting clinical trials and establish the required amount of data to be submitted with marketing applications for new anti-influenza products. The aim of this study was to analyse possible regulatory approaches to planning clinical development programmes for anti-influenza medicinal products. The article pays particular attention to phase III studies, as the main studies confirming efficacy and safety. The authors described a clinical development strategy and the requirements for the volume and quality of efficacy and safety data. This article is based on the current Russian recommendations for the design and development of medicinal products and guidelines on their evaluation, as well as the recommendations by the U.S. Food and Drug Administration (FDA). The analysis results demonstrate the necessity for elaborating Russian recommendations for clinical studies of medicinal products for the treatment of influenza that will take into account the national legislation and clinical development practices. Such recommendations will streamline the implementation of new effective anti-influenza medicinal products

Features of humoral immunity after COVID-19

At the present time, studying humoral immunity to the new coronavirus infection is among the most important tasks. The COVID-19 infection induces a protective pool of specific antibodies determining severity and duration of such immune protection after convalescence. The antibody testing is also necessary for assessing efficiency of anti-COVID vaccines in order to defeat the SARS-CoV-2 pandemic. Despite enormous interest of scientific community in this problem seen in the literature, there is still a lack for longitudinal observations of immunological status (more than 6 months) in the patients who have undergone COVID-19. The aim of this study is a long-term monitoring (9-14 months) of development and extinction of immune response to SARS-CoV-2 infection using quantitative assessment of IgA and IgG levels in peripheral blood of the patients who had COVID-19 in anamnesis. Monitoring of anti-SARS-CoV-2 levels over time has demonstrated significant individual variability, and made it possible to divide the study participants into three groups, according to characteristic features of humoral immunity after documented COVID-19. The study describes characteristic features of humoral immune response for each of these groups. The first group (30% of the study group) exhibited classical pattern of antibody response to viral infection. The second group (40% of study participants) presented with high plasma IgA levels, and their significant excess (about 2 times) over IgG levels throughout the observation period. The third group (30% of study participants), apparently comprised the subjects with increased humoral immunity to SARS-CoV-2 infection. Their plasma antibodies remain at high levels for at least 9-10 months after the onset of infection. The data obtained confirm the pattern of plasma IgA which is not quite typical to viral infections in dynamics after a sufficiently long time period after the disease in most study participants (2nd and 3rd groups; 70% of all volunteers who have recovered from COVID-19) and suggests an important role of this immunoglobulin against SARS-CoV-2 infection. The specific responses of anti-SARS-CoV-2 IgG are very similar to behavior of such antibodies in other viral infections including contacts with coronaviruses from earlier generations. Humoral immunity against SARS-CoV-2 may persist for more than 6 months, thus supporting an assumption that the naturally infected patients are able to resist re-infection for a long time

Решение проблемы повышенной чувствительности дентина: механизмы реминерализации при курсовом использовании зубной пасты с фторидом олова

The purpose of the study was clinical and laboratory analysis of the remineralizing properties and clinical efficacy of the course application of the Sensodyne Rapid Relief therapeutic and prophylactic toothpaste for sensitive teeth with tin fluoride. The study involved 50 overwise healthy patients aged from 18 to 25 years diagnosed with a localized form of hyperesthesia of hard dental tissues and no signs of acute inflammation of periodontal tissues. After 7 days of clinical observation and daily use of the toothpaste the hygienic condition of the oral cavity in all patients improved: oral hygiene index was 1.02±0.2 units; gum inflammation index (PMA) - 8.0±1.5%; the absence of hyperesthesia was diagnosed (1 point, p≤0.05). Examination of patients after 14 and 28 days confirmed the absence of pain in 100%, the organoleptic properties of the toothpaste under investigation were also highly appreciated by the participants. A laboratory study showed that patients have an increase in the pH value of mixed saliva and a decrease in its buffer capacity by 13.4±2.25%, which contributes to a more effective remineralizing effect of calcium ions by calcium-binding proteins of mixed saliva. The concentration of fluoride ions in the saliva of patients of both groups increased by average rate of 34±0.5% per week. The combination of tin and fluoride is more effective than fluoride alone. The presence of tin ions causes a higher ability of fluorides to bind with calcium ions on the surface of the dentin and dentinal tubules, inhibits the process of demineralization, enhances the additive interaction of the ions forming the acid-resistant layer on the surface of the dentin and dentinal tubules, leading to a decrease in hyperesthesia of the teeth.Цель исследования - клинико-лабораторный анализ реминерализующих свойств и клинической эффективности курсового применения лечебно-профилактической зубной пасты Sensodyne Мгновенный Эффект для чувствительных зубов с фторидом олова. Обследованы 50 практически здоровых пациентов в возрасте от 18 до 25 лет, у которых диагностированы локализованная форма гиперестезии твердых тканей зубов и отсутствие признаков острого воспаления тканей пародонта. По истечении 7 дней клинического наблюдения и ежедневного использования данной зубной пасты гигиеническое состояние полости рта у всех пациентов улучшилось - индекс гигиены составил 1,02±0,2 единицы; индекс воспаления десны (РМА) - 8,0±1,5%; диагностировано отсутствие гиперестезии (1 балл, р≤0,05). Обследование пациентов через 14 и 28 дней подтвердило отсутствие боли в 100% случаев, органолептические свойства исследуемой зубной пасты также высоко оценены участниками. Проведенное лабораторное исследование показало, что у пациентов наблюдается увеличение значения рН смешанной слюны и происходит снижение ее буферной емкости на 13,4±2,25%, что способствует более эффективной реминерализирующей отдаче ионов кальция кальцийсвязывающими белками смешанной слюны. Концентрация ионов фтора в слюне пациентов обеих групп за 1 нед увеличивается в среднем на 34±0,5%. Сочетание олова и фторида более эффективно, чем применение только фторида. Присутствие ионов олова обусловливает более высокую способность фторидов к связыванию с ионами кальция на поверхности дентина и дентинных канальцев, ингибирует процесс деминерализации, усиливает аддитивное взаимодействие ионов, формирующих кислотоустойчивый слой на поверхности дентина и дентинных канальцев, приводящее к снижению гиперестезии зубов

Information system on microbial collections as a part of bioresource collections portal for Russia’s FASO organizations: a working protocol

Nowadays, many scientific organizations of Russia own collections of microorganisms on which large volumes of information have been generated. These data represent the descriptions of objects of diverse nature (bacteria, archaea, fungi, protists) and their properties, which have been carefully collected and cataloged by generations of researchers. Not every organization that has such collections has an open access electronic catalog, which not only complicates work with these unique materials, but also even hides the fact of the existence of such collections. This state of affairs requires the development of electronic resources for presenting these materials to the scientific community. To put together the information on microorganism collections, we have developed an internet portal (http://www.biores.cytogen.ru/microbes/) of microbial bioresource collections of FASO organizations in the Russian Federation. The portal was created under the project developing the information system for bioresource collections of FASO institutes. It is a platform where collection organizations can place information about the storage units of their collections, as well as other information on collections, including links to their own catalogs. In this paper, we describe the principles of working with the portal. The portal’s graphical interface allows users, both registered and unregistered, to receive the following information about collections of microorganisms: a list of collections represented in the database, contact details of the organization and information about the curator of the collection, summary statistics for each collection, as well as information on storage units. Registered users – owners of collections – have the opportunity to create and modify records about the storage units of their collections, and to update their description. To automate work with the portal, software access to the database through the REST API has been implemented (http://api.biores.cytogen.ru/ microbes/). At present, the portal is still being filled, but it already contains a description of more than 13,000 items of storage (of which 3500 are in the microorganisms’ part) of 65 bioresource collections in Russia’s FASO organizations. Of these collections, 12 with microorganisms have a total diversity of funds of about 50,000 strains)

Collection of microorganisms of ICG SB RAS as a genetic resource for biotechnology

Genetic knowledge of microorganisms plays a critical role in the creation of new biotechnologies, since the effectiveness of any biotechnology is determined by the particular qualities of the structurally functional organization of molecular-genetic systems and their components used for the production of targeted products. Collections of microbial cultures play a decisive role in mobilizing biological resources and make it possible to form a solid base for genetic, molecular biological and biotechnological research. The aim of this work was to assess the key molecular-genetic and phenotypic characteristics of strains of the collection of microorganisms created in the “FRC Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences” as a genetic resource for biotechnology. Thirty strains of microorganisms of the collection were isolated by employees of the FRC ICG SB RAS from extreme natural ecosystems, the key molecular-genetic and phenotypic characteristics were described using modern methods of molecular biology and mass-spectrometry. DNA isolation and the sequencing of 16S rRNA gene sequences were performed. The strains of the collection were characterized by morphological, physiological, moleculargenetic and mass-spectrometric characteristics. The particular qualities of growing of strains on different substrates have been established, the study of cell morphology has been carried out. The physiological characteristics of the strains of the collection have been established: the attitude to oxygen, the type of nutrition, the range of temperature and pH, the attitude to NaCl and others. Different resistance of strains to antibiotics has been established. The creation of personal mass spectra of protein profiles of the studied strains of the collection was carried out. The resulting DNA sequences of the strains are deposited in the GenBank. The chemotaxonomic characteristics of strains have been determined. The biotechnological properties of the strains were assessed, the amount of metabolites (ethanol, lactic and acetic acids) in the culture liquid was determined. The value of the collection of microorganisms of the FRC ICG SB RAS as a genetic resource for biotechnology and bioengineering is determined not only by the species diversity of its strains, but also by a wide range of their area isolation and by the depth of their characterization using the widest arsenal of both classical and modern methods (including methods of genomics, proteomics, transcriptomics and bioinformatics)

An integrated method for taxonomic identif ication of microorganisms

For accurate species-level identification of microorganisms, researchers today increasingly use a combination of standard microbiological cultivation and visual observation methods with molecular biological and genetic techniques that help distinguish between species and strains of microorganisms at the level of DNA or RNA molecules. The aim of this work was to identify microorganisms from the ICG SB RAS Collection using an integrated approach that involves a combination of various phenotypic and genotypic characteristics. Key molecular-genetic and phenotypic characteristics were determined for 93 microbial strains from the ICG SB RAS Collection. The strains were characterized by means of morphological, physiological, moleculargenetic, and mass-spectrometric parameters. Specific features of the growth of the strains on different media were determined, and cell morphology was evaluated. The strains were tested for the ability to utilize various substrates. The strains studied were found to significantly differ in their biochemical characteristics. Physiological characteristics of the strains from the collection were identified too, e. g., the relationship with oxygen, type of nutrition, suitable temperature and pH ranges, and NaCl tolerance. In this work, the microorganisms analyzed were combined into separate groups based on the similarities of their phenotypic characteristics. This categorization, after further refinement and expansion of the spectrum of taxa and their metabolic maps, may serve as the basis for the creation of an “artificial” classification that can be used as a key for simplified and quicker identification and recognition of microorganisms within both the ICG SB RAS Collection and other collections

Рекомендации по программе клинических исследований лекарственных препаратов для лечения гриппа

The development of new medicinal products to treat influenza is motivated by the limitations of existing treatment options, the emergence of drug resistance, and the health consequences of influenza epidemics associated with the highly contagious nature of the virus. Proper planning and implementation of clinical programmes providing reliable data on the efficacy and safety of medicinal products under development requires adherence to recommendations of the regulatory authorities. At the moment, the Russian Federation, the Eurasian Economic Union, and the European Union lack documented recommendations on conducting clinical trials of anti-influenza medicines. There is a need in national guidelines that will reflect the procedure for conducting clinical trials and establish the required amount of data to be submitted with marketing applications for new anti-influenza products. The aim of this study was to analyse possible regulatory approaches to planning clinical development programmes for anti-influenza medicinal products. The article pays particular attention to phase III studies, as the main studies confirming efficacy and safety. The authors described a clinical development strategy and the requirements for the volume and quality of efficacy and safety data. This article is based on the current Russian recommendations for the design and development of medicinal products and guidelines on their evaluation, as well as the recommendations by the U.S. Food and Drug Administration (FDA). The analysis results demonstrate the necessity for elaborating Russian recommendations for clinical studies of medicinal products for the treatment of influenza that will take into account the national legislation and clinical development practices. Such recommendations will streamline the implementation of new effective anti-influenza medicinal products.Последствия вспышек эпидемий гриппа, связанных с высококонтагиозным характером инфекции, а также ограничения существующих методов лечения и возникновение лекарственной резистентности обуславливают актуальность разработки новых лекарственных препаратов для лечения этого заболевания. Надлежащее планирование программы и проведение клинических исследований, гарантирующих получение корректных данных об эффективности и безопасности разрабатываемых препаратов, обеспечивается соблюдением рекомендаций регуляторных органов. На данный момент в Российской Федерации и Евразийском экономическом союзе отсутствуют документы, содержащие рекомендации по проведению клинических исследований противогриппозных препаратов. Существует потребность в разработке отечественного руководства, отражающего порядок проведения клинических исследований, а также регламентирующего необходимый объем данных, которые должны быть представлены при регистрации новых противогриппозных препаратов. Цель работы — изучение основных возможных регуляторных подходов к планированию программ клинических исследований лекарственных препаратов для лечения гриппа. Особое внимание уделено клиническим исследованиям III фазы как основным исследованиям, подтверждающим эффективность и безопасность препарата. Описана стратегия клинических исследований препарата, требования к объему и качеству данных по эффективности и безопасности. Материал подготовлен на основе действующих рекомендаций к планированию и разработке лекарственных средств в Российской Федерации, руководства по экспертизе лекарственных средств и рекомендаций Управления по контролю за качеством продуктов питания и лекарственных средств США. В результате проведенного анализа показана необходимость разработки отечественных рекомендаций по проведению клинических исследований препаратов для лечения гриппа с учетом специфики юридических норм и традиций российской клинической практики. Разработка таких рекомендаций приведет к ускорению ввода в практику новых эффективных препаратов против гриппа

Hyperexpression of TLR2 and TLR4 in patients with ischemic stroke in acute period of the disease

Pathogenesis of ischemic stroke  is actively  involved  in the  system  of innate immunity. Under conditions of cerebral  ischemia, a number of biologically  active  substances are  released  that  interact with innate immunity receptors, in particular TLR2  and  TLR4, which  exacerbate inflammation in brain  tissue. Identification of predictor markers  at the level of the innate immunity system may foresee the clinical course of ischemic stroke and ensure timely treatment. Our objective was to study expression of TLR2 and TLR4 receptors in peripheral blood leukocytes  in patients with ischemic stroke in the dynamics of the disease. 27 people  were included in the study. The main  group consisted of patients with ischemic stroke of varying severity (n = 19). Patients of the main  group were divided into two subgroups:  with an NIHSS index value of < 10 (n = 10) and > 10 (n = 9). The control group included healthy  donors  with no history  of acute  and chronic inflammatory diseases (n = 8). Peripheral blood  leukocytes  were used as the  test material. To determine expression  of the TLR2  and TLR4  genes, RT-PCR in real time was used. Surface  expression  of TLRs was determined by flow cytometry. A study of the TLR2 and TLR4 gene expression showed that on the 1st, 3rd  and 7th  day post-stroke, the TLR4 gene expression  in patients was significantly  increased, when compared to the control group (p < 0.01), whereas TLR2 gene expression on the 3rd  day of the disease was not statistically different from the control group. A study of surface expression  of receptors showed that the average TLR2 fluorescence intensity on the patients’ peripheral blood monocytes was significantly  increased on the 1st  and 3rd  day of disease when compared to the control group.  The  surface  expression  of TLR4  on monocytes has a statistically significant  increase  only on day 7. Assessment  of surface expression  of TLRs in subgroups  with different  severity values by NIHSS showed that  patients with a NIHSS index > 10 had a significantly  higher  level of surface of TLR2  expression  over the observation period, while the largest difference in TLR4  expression  in the subgroups  was observed  on the 1st day of the disease (p < 0.05). Patients with ischemic stroke showed an increase  in TLR2 and TLR4 expression at the gene and protein level, compared to healthy  donors. These indices can be considered possible predictors for clinical  prognosis  of ischemic stroke

Menstrual function and mental health of medical students during the COVID-19 pandemic: a continuous cross-sectional study

Aim. To assess the impact of new coronavirus infection (NCI) and COVID-19 vaccination on menstrual function in comparison with the frequency of depressive disorders during the COVID-19 pandemic among female students of a medical university. Materials and methods. Data for a continuous transverse (cross-sectional) study were obtained using an online survey. The questions included demographics, characteristics of menstrual function, the history of COVID-19 and vaccinations against it, and the standard CES-D (Center for Epidemiologic Studies Depression Scale) questionnaire. The survey included 1.879 female medical students. The exclusion criteria were age under 18 and over 25 years, hormonal contraceptives, pregnancy, childbirth during the last year, and vaccination after COVID-19. After applying the exclusion criteria, three study groups were formed. Group 1 included female students with a history of NCI (n=140), group 2 included students with no history of NCI, who were initially vaccinated against COVID-19 (n=647), group 3 (control group) included unvaccinated students with no history of NCI (n=55). Results. There were no differences in the prevalence of changes in menstrual function in female students after the NCI (49.3%) and after COVID-19 vaccination (39.6%) compared to the control group (43.6%); p=0.477. The rate of depression in the overall study cohort was 43.3%, without any significant differences between the study groups. Significant predictors of changes in menstrual function during the pandemic were marriage (odds ratio OR 2.33 [1.513.61]), depression (OR 1.72 [1.282.3]), a history of menstrual dysfunction (OR 1.5 [0.121.99]), and later menarche (OR 1.76 [1.023.04]). Multivariate analysis did not show the significance of the history of NCI and vaccination as factors of menstrual dysfunction (OR 1.61 [0.892.90] and OR 0.91 [0.591.41], respectively). Conclusion. During the COVID-19 pandemic, female medical students reported frequent changes in menstrual function and depressive disorders. During the pandemic, the most significant predictors of menstrual disorders in female students were depression, a history of menstrual dysfunction, and marriage. A multicenter prospective study is necessary to clarify the mechanisms of the pandemic's impact on menstrual function