Making complex things simpler: modern tools to edit the plant genome

There are several technologies for plant genome editing, of which the most simple and universal is CRISPR/Cas. Currently, this technology is widely used for gene knockout, deleting genome fragments and inserting exogenous sequences in the plant genome. For each of these applications, many different types of genetic tools have been developed that are used by various research groups to solve specific problems. The CRISPR/Cas technology for plant genome editing is at an early stage of optimization, which is reflected by the ongoing search for the most effective, simple and flexible techniques. As a result, experimental work has to be preceded by a rather long and laborious process of selecting a genetic tool that will be optimal for a specific experimental task. In our review we describe the main variants of the CRISPR/Cas technology used to edit a plant genome. We classify them in terms of experimental tasks solved, major components and technology performance. In the first half of the review a detailed description of two major components of CRISPR/Cas technology – nuclease and guide RNA – is given, the effect of structural features of these elements on editing efficiency is analyzed. Experimental data on the relationship between editing efficiency and nucleotide sequence of guide RNA are generalized. We also give the characteristic for different variants of nucleases used for plant genome editing and discuss their benefits for different experimental purposes. In the second half of the review various strategies for expression of CRISPR/Cas elements in plant cells, in particular, advantages and disadvantages of stable transformation and transient expression, are discussed. The effect of various regulatory elements of genes encoding nuclease and guide RNA on editing efficiency is described. Special emphasis is placed on the techniques of increasing targeted gene replacement efficiency

Влияние развития интернет-торговли на инфляцию в России

Over the past few years in Russia and the world there has been a rapid development of goods and services e-commerce. Therefore, an issue of inflationary consequences of the process of e-commerce growth is topical. The subject of this research is the impact of the development of e-commerce on inflation. The purpose of the paper to identify key theoretical mechanisms of e-commerce development influence on inflation, in addition to empirically verify the presence of the de-inflation effect on Russian data. The paper bases on methods of systematization, descriptive analysis, panel data econometric analysis. Also systematize basic mechanisms of e-commerce influence and provide empirical evidence for the disinflationary effect of ecommerce in Russian regions as a result of the study. The study concludes that e-commerce causes predominantly downward pressure on prices and inflation. Econometric analysis confirms this effect on Russian regions data in 2014–2020. The scientific novelty of the research lies in systematization of channels of e-commerce influence on various spheres of economy and confirmation of disinflationary effect of e-commerce using Russian data. The study’s recommendation is to consider the aspect of price reduction pressure in Russia’s monetary policy during the further spread of e-commerce.В течение последних лет в России и в мире наблюдается скачок развития сегмента интернет-торговли, что актуализирует оценку дезинфляционного эффекта этого процесса. Предметом исследования является влияние развития интернет-торговли на инфляцию. Цель работы заключается в выявлении механизмов влияния развития интернетторговли на инфляцию и в эмпирической проверке наличия дезинфляционного эффекта на российских данных. Исследование проведено с  опорой на метод систематизации, дескриптивный анализ, эконометрический анализ на панельных данных. По результатам исследования представлено резюме странового опыта выявления влияния интернет-торговли на инфляцию, а также проведена эмпирическая проверка наличия дезинфляционного эффекта интернет-торговли в регионах России. Авторы приходят к выводу о доминировании понижательного давления развития интернет-торговли на цены. Эконометрический анализ подтвердил это на российских региональных данных в 2014–2020 гг. Научная новизна работы заключается в систематизации каналов влияния интернет-торговли на макроэкономические показатели и подтверждении наличия дезинфляционного эффекта распространения интернетторговли на российских данных. Рекомендация исследования состоит в учете фактора понижательного давления на цены в практике денежно-кредитной политики России по мере дальнейшего развития интернет-торговли

Changes in the oxidative-antioxidant status of blood in freestyle wrestlers under the influence of physical exertion

Purpose of the study: to evaluate the “lipid peroxidation-antioxidant protection” system in freestyle wrestlers under the influence of intense physical activity in order to identify the nature of oxidative processes and the antioxidant status of the athletes’ body.Materials and methods: the study involved athletes — freestyle wrestlers, boys 18–22 years old, with sports qualifications: candidate master of sports or master of sports, regularly involved in freestyle wrestling. The control group consisted of students who did not have a sports category and were not involved in sports activities, comparable in gender, age, weight category with the comparison group.  The material for the study was plasma and red blood cells. Blood sampling from athletes was carried out twice, at the end of the preparatory and competitive periods, in the morning, on an empty stomach from the ulnar vein, in accordance with generally accepted requirements. Components of lipid peroxidation and antioxidant defense were determined using spectrophotometric and fluorometric methods.Results: it was found that in athletes after intense physical activity (competitive period), there is a significantly significant decrease in substrates with double bonds by 61 %, primary products by 59 % and intermediate products by 44 %, an increase in the final products of lipid peroxidation by 25 % against the background of inhibition activity of antioxidant protection components: retinol by 40 %, superoxide dismutase by 7 % and reduced glutathione by 11 %.Conclusion: The level of free radical processes increases during physical activity due to the accumulation of oxygen in organs and tissues, and depends on the intensity and duration of the exercise. In the course of the study, it was established that the preparatory and competitive periods, in comparison with the control group, among freestyle wrestlers, depending on the previous level of physical activity, are characterized by activation of lipid peroxidation processes and inhibition of enzymatic and non-enzymatic components of antioxidant protection. The established changes are recommended to be taken into account when conducting comprehensive rehabilitation of athletes, when developing recommendations for improving the training process, as well as when using antioxidant complexes for athletes aimed at restoring various parts of the body’s antioxidant defense

Синтез 5-оксиметил-1,2,4-триазол-3-карбоксамидов

Objectives. A key step in the synthesis of natural nucleoside analogs is the formation of a glycosidic bond between the carbohydrate fragment and the heterocyclic base. Glycosylation methods differ in terms of regio- and stereoselectivity. A promising method for the highly specific synthesis of new pharmacologically active compounds involves an enzymatic reaction catalyzed by genetically engineered nucleoside phosphorylases. This study is devoted to the synthesis of a library of analogs of nucleoside heterocyclic bases—5-oxymethyl-1,2,4-triazole- 3-carboxamides—in order to investigate the substrate specificity of genetically engineered nucleoside phosphorylases.Methods. A method of cyclization of acylamidrazones obtained from the single synthetic precursor β-N-tert-butyloxycarbonyl-oxalamidrazone was used to parallel-synthesize new 5-alkoxy/ aryloxymethyl-1,2,4-triazole-3-carboxamides. Silica gel column chromatography was used to isolate and purify the synthesized compounds. A complex of physicochemical analysis methods (nuclear magnetic resonance spectroscopy, chromatography, and mass spectrometry) confirmed the structure of the compounds obtained in the work.Results. 5-alkoxy/aryloxymethyl-1,2,4-triazole-3-carboxamides were obtained to study the substrate specificity of genetically engineered nucleoside phosphorylases. The possibility of obtaining new nucleoside analogs by the chemico-enzymatic method was demonstrated on the basis of preliminary assessment results.Conclusions. The physicochemical characteristics of a series of novel 5-alkoxy/aryloxymethyl- 1,2,4-triazole-3-carboxamides were studied along with their potential to act as substrates for the transglycosylation reaction catalyzed by nucleoside phosphorylases.Цели. Ключевая стадия синтеза аналогов природных нуклеозидов – образование гликозидной связи между углеводным фрагментом и гетероциклическим основанием. Методы гликозилирования различаются по регио- и стереоселективности. Ферментативная реакция, катализируемая генно-инженерными нуклеозидфосфорилазами – перспективный метод высокоспецифичного синтеза новых фармакологически активных соединений. Данное исследование посвящено синтезу библиотеки аналогов гетероциклических оснований нуклеозидов – 5-оксиметил-1,2,4-триазол-3-карбоксамидов для изучения субстратной специфичности генно-инженерных нуклеозидфосфорилаз.Методы. Для параллельного синтеза новых 5-алкокси/арилоксиметил 1,2,4-триазол3-карбоксамидов применен метод циклизации ациламидразонов, получаемых из единого синтетического предшественника – β-N-третбутилоксикарбонил-оксаламидразона. Для выделения и очистки синтезированных соединений использована колоночная хроматография на силикагеле. Структура полученных в работе соединений подтверждена комплексом методов физико-химического анализа: спектроскопией ядерного магнитного резонанса и хромато-масс-спектрометрометрией.Результаты. Получены 5-алкокси/арилоксиметил-1,2,4-триазол-3-карбоксамиды для изучения субстратной специфичности генно-инженерных нуклеозидфосфорилаз. По результатам предварительной оценки показана возможность получения из них новых аналогов нуклеозидов химико-ферментативным методом.Выводы. Для серии новых 5-алкокси/арилоксиметил-1,2,4-триазол-3-карбоксамидов изучены физико-химические характеристики, а также их способность выступать в роли субстратов реакции трансгликозилирования, катализируемой нуклеозидфосфорилазами

Ethnic peculiarities of some indicators of metabolic and clinical status in infertile men

The article presents the results of the research of lipid peroxidation (LPO) and activity of components of antioxidant protection (AOP) in men of reproductive age with primary and secondary infertility, living on the territory of Buryatia and have abnormalities in spermogramme. Aim: to study the frequency structure of male infertility, the features of the system lipid peroxidation-antioxidant protection in men with normospermic and pathospermia Buryat and Russian nationality. A retrospective analysis was performed in 562 men of Russian and Buryat nationality, residing in the territory of the Republic of Buryatia. Study groups were as follows: 204 - Buryat, 358 - Russian. In this study we used clinical and laboratory methods and the statistical analysis of the obtained data. Study the contents of products of lipid peroxidation (LPO) and activity of components of the antioxidant protection (AOP) was studied in 113 infertile men (71 Buryat, 42 Russian). Group of infertile men were divided into a group with normospermia and pathospermia. High rate of male factor infertility in men of Russian and Buryat populations was found. No difference was found for frequency and structure of reproductive disorders in men depending on ethnicity. The frequency of male factor in infertile marriages and structure of the causes of male infertility and made comparisons based on ethnicity. Was found the presence of oxidative stress in patients with pathospermia. The findings suggest about the unfavorable pro-and antioxidant status of men with pathospermia, both Russian and Buryat populations. The differences indicate a different degree of activity of metabolic processes in infertile men of different ethnic groups

THE RELATIONSHIPS BETWEEN SERUM ANTIOXIDATIVE PARAMETERSAND HORMONAL LEVELS IN MEN WITH INFERTILITY

The following research analyses relationships between antioxidative parameters and hormonal levels in men with infertility. The getting data could be used for differentiated approach to diagnostics and prophylactics of men infertility

LIPID PEROXIDATION SYSTEM IN TYPE 1 DIABETES: ETHNIC ASPECTS

Oxidative stress is an acknowledged pathogenic mechanism in Type 1 diabetes mellitus. Acute increases in plasma glucose concentration may increase free radical production by the following mechanisms: labile glycation; auto-oxidation of glucose, and intracellular activation of the polyol pathway; which produces an imbalance in the NADh/NAD+ ratio and favours the production of free radicals. Incidence, severity, and rate of complications of Type 1 diabetes mellitus are associated with many factors including geographic location and ethnicity. The main features of lipid peroxidation in 30 women with type 1 diabetes in Buryat and Russian ethnic groups were examined. The materials for biochemical studies were blood serum and red cells. The content of the substrates and products of lipid peroxidation (diene conjugates, ketodienes and coupled trienes, malondialdehyde), total antioxidant activity of serum and its components (superoxide dismutase, α-tocopherol, retinol, reduced and oxidized glutathione) were evaluated by a spectrofluorophotometer «SHIMADZU-1501» (Japan). Statistical analysis was performed by non-parametric tests. It is shown that the intensity of lipid peroxidation processes in patients Buryat ethnic group is reduced in comparison with Russian ethnic group, that confirmed by the values of the coefficient of oxidative stress. The study was supported by grants of the President of the Council of the Russian Federation (Scientific School 494.2012.7)

FEATURES OF ANTIOXIDANT SYSTEM STATE IN ADOLESCENTS OF EASTERN SIBERIA IN DEPENDS ON THEIR GENDER AND ETHNIC ORIGIN

This research is devoted to studying of features of antioxidant protection system in adolescent boys and girls of various ethnic groups of Eastern Siberia. Study of the peculiarities of processes of antioxidant protection of populations of people of different ethnicity, living in the same geographical conditions, is of undoubted scientific interest. It is especially important to assess metabolic status of a young, growing body to ensure in the future the preservation of reproductive ability. We biochemically evaluated 58 boys of 14-17 years old. Out of them 19 boys were Russian, 29 boys were Buryat population and 10 boys were metises. Also we biochemically evaluated 57 girls. Out of them 21 were Russian, 27 girls were Buryat population and 9 girls were metises. The materials for biochemical studies were blood serum and red cells. The total antioxidant activity of serum and the content of its components (superoxide dismutase, α-tocopherol, retinol, blood-reduced glutathione) were evaluatedbya spectrofluomphotometer «SHIMADZU-1501» (Japan). Statistical analysis was performed by parametric tests. The study was supported by grants of the President ofthe Council of the Russian Federation (Scientific School 494.2012.7). The detected features of antioxidant protection processes in observed adolescents had gender differences, also may be explained by their ethnicity and characterized by different degree of activity of metabolic processes in adolescent boys and girls of different ethnic groups of Eastern Siberia. Study the state of the antioxidant system of the organism can be used as an additional criterion for complex examination of practically healthy adolescents, which significantly enhance the representation about the adaptation possibilities of the organism to external conditions and can be a basis for effective monitoring of reproductive health in the future

Markers of oxidative damage lipids and DNA in men with type 1 diabetes mellitus and different levels of albuminuria

BACKGROUND: Diabetic nephropathy is a specific kidney damage that affects up to 40% of type 1 diabetes mellitus patients. There is still insufficient knowledge about oxidative stress at the different levels of albuminuria.AIM: To assess the indicators of oxidative damage to lipids, DNA and antioxidant defense in men with type 1 diabetes mellitus and albuminuria different levels.MATERIALS AND METHODS: The study was conducted during 2018–2019. The main group included 56 men of reproductive age with type 1 diabetes mellitus (T1DM) divided into 2 groups: 24 patients with albuminuria level A1 (group A1) and 32 patients with albuminuria level A2 (group A2). The control group consisted of 28 healthy men. The oxidative stress indicators content, as well as the activity of antioxidant defense system various links was evaluated. Spectrophotometric, fluorimetric and enzyme immunoassay methods were used.RESULTS: In patients of the A1 group there were higher values of the median of primary — conjugated dienes, secondary — ketodienes and conjugated trienes, final — Schiff bases products and 8-hydroxy-2’-deoxyguanosine in comparison with the control. Similar changes in patients of the A2 group were found. Intergroup differences related to increased median values of TBARs and 8-hydroxy-2’-deoxyguanosine in patients of the A2 group compared with A1 group. The activity of the antioxidant defense system in A1 group relative to the control by increased values of the glutathione peroxidase, reduced glutathione, oxidized glutathione median, and retinol was characterized. A2 group had higher values of glutathione peroxidase, reduced glutathione and oxidized glutathione medians in comparison with controls. Correlation analysis in A1 group showed the relationships between the duration of the disease and the products of lipid peroxidation, glycated hemoglobin with 8-hydroxy-2’-deoxyguanosine. In A2 group, there was a relationship between the albumin/creatinine ratio and the duration of the disease, glomerular filtration rate and creatinine level.CONCLUSION: In men with T1DM, regardless of the albuminuria level, there are higher values of the oxidative damage lipids and DNA parameters, as well as the presence of close relationships between these parameters and the duration of the disease, which can be used to develop potential strategies for the prevention and early therapy of diabetic nephropathy

The influence of the heliogeophysical factors on the clotting of blood of pregnant women with placental insufficiency

Geomagnetic storms are one of the natural abiotic risk factors for human health. The response of the organism to their impact depends on the individual adaptive abilities, which were formed during the evolution, but can change in the conditions of pathological processes. Evaluation of the effect of a weak magnetic storm (G4v (22 < Ar < 39 nTl) on hemorheological parameters of blood of pregnant women with placental insufficiency compared to healthy pregnant women showed an increase in platelet count (p = 0.05), fibrinogen content (p = 0.02), thrombocyte (p = 0.03), erythrocyte sedimentation rate (p = 0.004). At the same time, a decrease in the number of red blood cells (p = 0.04), hemoglobin concentration (p = 0.001), hematocrit value (p = 0.0009) was registered. We found a strong direct correlation between subpopulations of platelets, which are characterized by different functional activity. Heterogeneity of platelets, their multidirectional correlations with indicators of blood coagulation system of healthy pregnant women can be considered as a response to the change in the disturbance of the earth's magnetic field. For pregnant women with placental disorders, the magnetic storm was associated with increased activity of the blood coagulation system and a significant increase in its components in the blood of patients. Despite the fact that changes in the blood coagulation system occur from the beginning of pregnancy and are often adaptive, in pregnant women with placental disorders, the additional impact of a weak magnetic storm can be a risk of hemostasis disorders and the development of platelet thrombosis, timely diagnosis and therapy of which is important