Analysis of the steel industry in Russia

This article presents an analysis of the steel industry in Russia. The external factors of the steel market have been studied. The results of the activities of  steel enterprises for 2019 with the subsequent calculation of the market share of each participant, – have been investigated. The calculation of the main indicators of the market structure in the form of the Herfindahl-Hirschman formulas and the concentration coefficient for the three largest sellers has been carried out. The dynamics of revenue and steel production in Russia by the main producers in the industry market have been estimated, as well as changes in the  level of use of average annual production capacity for a number of products  have been evaluated. The structure of the country’s exports and the influence of the products of the steel industry in it, as well as the factors of its internal and external demand, – have been analysed. Factors and trends of the industry  development have been considered

Clinical and biochemical changes and their correction in patients with metabolic phenotype of osteoarthritis and insomnia

BACKGROUND: Currently, OA is considered a polyetiological disease, where several phenotypes are distinguished based on the leading role of a specific risk factor. It is assumed that each clinical phenotype corresponds to its own fundamental changes in various organs and systems. Modern experimental evidence of chondro-osteogenic activity allows us to consider melatonin as a potential drug for various types of osteoarticular pathology.AIM: To study clinical and biochemical changes and effects of melatonin in the metabolic phenotype of OA and insomnia.MATERIALS AND METHODS: The study involved patients with a metabolic phenotype of OA and healthy volunteers. The subjects were collected complaints and anamnesis, as well as general clinical and orthopedic examination. In the blood serum, markers of bone and cartilage metabolism were determined. Patients were asked to answer the questions of clinical scales to assess the quality of sleep, the functional state of the joints and quality of life.RESULTS: The study involved 36 patients. Participants were divided into 3 groups: group health patients — patients without articular pathology, sleep quality disorders and normal body mass index; control group- patients with metabolic phenotype of OA, insomnia and basic treatment for 30 days; experimental group — patients with a metabolic phenotype of OA and insomnia, whose basic treatment included melatonin (Melaxen®) at a dosage of 3 mg for 30 days. Statistically significant differences were observed between the initial levels of acid phosphatase (AP), bone isoenzyme alkaline phosphatase (ALP) and calcium (Ca) in the control and experimental groups. According to the results of screening for the detection of insomnia among patients with the metabolic phenotype of, there were statistically significant insomnological disorders compared to the group health patients, as well as significant differences in terms of pain, symptoms, activity and quality of life in general according to the KOOS and SF-36 scales. Correlation analysis showed moderate correlations with biochemical parameters in patients with OA metabolic phenotype groups. After the treatment there was a certain increase in the level of Ca and a decrease in the activity of ALP and АP in patients of experimental group in comparison with patients of control groups. There was a positive trend in the range of criteria of the KOOS and SF-36 scale in experimental groups, pronounced decrease in pain syndrome (P), symptoms (S) and an increase in the levels of daily activity (A) and the total indicator (Sum) in comparison with patients of control groups. Also as improved sleep quality on all scales in comparison with the control group. In control group, problems with the quality and quantity of sleep remained at the same level.CONCLUSION: In patients with the most pronounced indicators of clinical manifestations of OA, more active processes of bone remodeling. The higher the level of bone resorption markers, the greater the severity of the clinical course in patients with osteoarthritis, and the worse the quality of sleep in general. The addition of melatonin to the treatment regimen was associated with a decrease in serum activity of the bone isoenzyme of alkaline phosphatase, acid phosphatase, an increase in calcium levels, as well as with an improvement in sleep and clinical symptoms ОА

Спектроскопическое исследование фотофизических свойств метиленового синего в биологических средах

A spectroscopic study of the photophysical properties of methylene blue (MB) in aqueous solutions was carried out. Absorption and fluorescence spectra as well as fluorescence lifetime were recorded. The concentration dependence of the intensity and shape of the spectra allowed establishing the ranges of MB concentrations for in vitro and in vivo studies at which aggregation is not observed (up to 0.01 mM, which corresponds to 3.2 mg/kg). Studies of photodegradation in biological media showed that photobleaching of more than 80% in plasma and culture media is observed already at a dose of 5 J/cm2 , while in water at this concentration and dose photobleaching is not yet observed, and at a dose of 50 J/cm2  photobleaching of MB is about 30%. It was found that in media containing proteins and having an alkaline pH, photobleaching occurs significantly faster than in neutral aqueous media. The ionic strength of the solution has no effect on the photobleaching rate. Such photobleaching is caused by the photodegradation of MB rather than the transition to the leucoform.The efficiency of singlet oxygen generation and photodynamic activity were evaluated in vitro. In the investigated range of MB concentrations, the efficiency of singlet oxygen generation is rather low, because positively charged MB binds to negatively charged cell membranes, which leads to a change in the type of photodynamic reaction. The emergence of other reactive oxygen species (ROS), different from singlet oxygen, in cells has been demonstrated. The generation of ROS and the low quantum yield of singlet oxygen generation indicate the tendency of MB to provide the type I photosensitization mechanism (electron transfer with the formation of semi-reduced and semi-oxidized MB+ radicals) rather than to the type II mechanism (energy transfer to oxygen with the formation of singlet oxygen) in biological media and in vivo.Проведено спектроскопическое исследование фотофизических свойств метиленового синего (МС) в водных растворах и биологических жидкостях. Зарегистрированы спектры поглощения и флуоресценции, а также времена жизни флуоресценции. По зависимости интенсивности и формы спектров от концентрации удалось установить диапазоны концентраций МС для исследований in vitro и in vivo при которых не наблюдается агрегация (до 0,01 мМ, что соответствует 3,2 мг/кг). Исследовано фотообесцвечивание МС под действием лазерного излучения. Исследования фотодеградации в биологических средах показали, что фотообесцвечивание более чем на 80% в плазме и культуральной среде наблюдается уже при дозе 5 Дж/см2 , в то время как в воде при такой концентрации при дозе 5 Дж/см2  фотообесцвечивания еще не наблюдается, а при дозе 50 Дж/см2  фотообесцвечивание МС составляет порядка 30%. Установлено, что в средах, содержащих белки и обладающих щелочным рН, фотообесцвечивание происходит существенно быстрее, чем в нейтральных водных средах. Ионная сила раствора не оказывает влияния на скорость фотообесцвечивания. Такое фотообесцвечивание вызвано фотодеградацией МС, а не переходом в лейкоформу. Проведена оценка эффективности генерации синглетного кислорода и фотодинамической активности in vitro. В исследуемом диапазоне концентраций МС эффективность генерации синглетного кислорода достаточно низкая, так как положительно заряженный МС связывается с негативно заряженными мембранами клеток, что приводит к изменению типа фотодинамической реакции. Продемонстрировано возникновение в клетках других активных форм кислорода (АФК), отличных от синглетного кислорода. Генерация АФК и невысокий квантовом выход генерации синглетного кислорода свидетельствуют о склонности МС к механизму фотосенсибилизации I типа (перенос электрона с образованием полувосстановленных и полуокисленных радикалов MB+), а не к механизму II типа (перенос энергии к кислороду с образованием синглетного кислорода) в биологических средах и in vivo

Спектроскопическое исследование метиленового синего in vivo: влияние на оксигенацию тканей и опухолевый метаболизм

Methylene blue (MB) is a promising photosensitizer (PS) for the treatment of pathological neoplasms, since it has both photodynamic activity (under laser irradiation) and redox and catalytic properties (in the absence of light). In the framework of this work, using spectroscopic methods, the effect of intravenous administration of MB on tissue oxygenation of hemoglobin in small animals in vivo in tumor and normal tissues was analyzed. The influence of MB on cell metabolism was analyzed. It has been shown that the use of MB promotes an increase in oxygen consumption by the tumor, and also leads to a shift in metabolism towards oxidative phosphorylation. It was shown that the use of MB contributes to an increase in oxygen consumption by the tumor, and also leads to a shift in metabolism towards oxidative phosphorylation.Метиленовый синий (МС) является перспективным фотосенсибилизатором для терапии патологических новообразований, поскольку обладает как фотодинамической активностью (при лазерном облучении), так и окислительно-восстановительными и каталитическими свойствами (в отсутствии света). В рамках данной работы при помощи спектроскопических методов было проанализировано влияние внутривенного введения МС на тканевую оксигенацию гемоглобина на малых животных in vivo в опухоли и нормальных тканях. Проведен анализ влияния МС на клеточный метаболизм. Показано, что применение МС способствует увеличению потребления кислорода опухолью, а также приводит к сдвигу метаболизма в сторону окислительного фосфорилирования

Concurrent Thermochemoradiotherapy in Glioblastoma Treatment: Preliminary Results

Glioblastoma is the most frequent and aggressive primary brain tumor. The patient can be alive with this pathology using the modern standard of intensive combined treatment less than 2 years. Between December 2013 and August 2017, 30 patients with newly diagnosed supratentorial glioblastoma had received concomitant chemoradiotherapy with transcranial radiofrequency hyperthermia. The gross total or the subtotal resection of the tumor was made previously in all cases. The median follow-up time after operation achieved 12 months (95% confidence interval (CI): 8.5–23 months) in this study. The median disease-free survival time was 9.6 months (95% CI: 7.2–19.0 months). The median overall survival time of patients included in the study was 23.4 months. No increase in the systemic side effects of chemotherapy was found compared with the frequency described in the population. Preliminary results had shown that the usage of concomitant thermochemoradiotherapy with transcranial radiofrequency hyperthermia improves progression-free survival rates. Overall survival rates also tended to increase. Given the absence of severe complications, it is necessary to continue research to achieve statistically significant results

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

Comparison of different technologies for producing recombinant adeno-associated virus on a laboratory scale

Adeno-associated virus vectors are among the most promising ones for the delivery of transgenes to various organs and tissues. Recombinant adeno-associated virus (rAAV) is able to transduce both dividing and non-dividing cells, has low immunogenicity, and is able to provide long-term expression of transgenes. Modern technologies make it possible to obtain rAAV for in vivo use, but they are not without drawbacks associated with laboriousness, scalability difficulties, and high cost, therefore, improvement of technological schemes for obtaining rAAV is an urgent issue. The aim of the study was to compare different technological approaches to rAAV production based on different conditions of the transfected HEK293 cell line cultivation on a laboratory scale. Materials and methods: HEK293 cell culture, AAV-DJ Packaging System, PlasmidSelect Xtra Starter Kit were used in the study. The technologies were compared using a model rAAV vector with a single-domain antibody transgene fused to the Fc-fragment of IgG1 specific to botulinum toxin. HEK293 cells were transfected with supercoiled plasmid DNA isolated by three-step chromatographic purification. The identity of the rAAV preparation was determined by electrophoresis, immunoblotting, and real-time polymerase chain reaction. Results: the study demonstrated the efficiency of the chromatographic method for obtaining a supercoiled form of plasmid DNA that can be used for efficient transfection of cell culture in order to produce rAAV. The study compared the following processes of rAAV production: using transient transfection and cultivation of the transfected HEK293 cell suspension in Erlenmeyer flasks, adherent culture in T-flasks, and adherent culture in a BioBLU 5p bioreactor on a matrix of Fibra-Cel disks. Conclusions: the data obtained showed the possibility of using the described approaches to purification of plasmid DNA, cell transfection, and cultivation of the transfected cells under various conditions to obtain rAAV samples that expresses the antibody gene. The BioBLU 5p reactor with Fibra-Cel discs was used for the first time to produce preparative quantities of rAAV on a laboratory scale, which increased the adherent surface area during cell culture and transfection, and, as a result, increased the yield of the target product

Dissemination of Invasive Mosquito Species, Aedes (Stegomyia) aegypti (L., 1762) and Aedes (Stegomyia) albopictus (Skuse, 1895) in the South of Krasnodar Region, Russia

Objective of the study was to investigate the current borders of the invasive species Ae. aegypti and Ae. albopictus distribution in the south of the Krasnodar Region and climatic factors limiting their expansion. Material and methods. Mosquito larvae, pupa and imago were collected in 21 inhabited localities in August–September, 2017, using conventional entomological methods, and identified through standard morphological keys. Climatic data found on the website pogoda. ru were analyzed using SPSS program. Results and conclusions. Ae. albopictus is widely spread along the Black Sea coast, from Adler to Novorossiysk (Ozereyevka), as well as on the north slopes of Caucasus up to Maikop. Additional investigations are required to specify the northern border of species distribution and to evidence the existence of established populations here. Ae. aegypti were not found in the south of the Krasnodar Region. The obtained data suggest a possibility of much wider spreading of dengue, Chikungunya and Zika fevers in case of pathogen importation to the south of Krasnodar Region, than was assumed earlier

Finding of the Low Molecular Weight Inhibitors of Resuscitation Promoting Factor Enzymatic and Resuscitation Activity

Background: Resuscitation promoting factors (RPF) are secreted proteins involved in reactivation of dormant actinobacteria, including Mycobacterium tuberculosis. They have been considered as prospective targets for the development of new antituberculosis drugs preventing reactivation of dormant tubercle bacilli, generally associated with latent tuberculosis. However, no inhibitors of Rpf activity have been reported so far. The goal of this study was to find low molecular weight compounds inhibiting the enzymatic and biological activities of Rpfs. Methodology/Principal Findings: Here we describe a novel class of 2-nitrophenylthiocyanates (NPT) compounds that inhibit muralytic activity of Rpfs with IC50 1–7 mg/ml. Fluorescence studies revealed interaction of active NPTs with the internal regions of the Rpf molecule. Candidate inhibitors of Rpf enzymatic activity showed a bacteriostatic effect on growth of Micrococcus luteus (in which Rpf is essential for growth protein) at concentrations close to IC50. The candidate compounds suppressed resuscitation of dormant (‘‘non-culturable’’) cells of M. smegmatis at 1 mg/ml or delayed resuscitation of dormant M. tuberculosis obtained in laboratory conditions at 10 mg/ml. However, they did not inhibit growth of active mycobacteria under these concentrations. Conclusions/Significance: NPT are the first example of low molecular weight compounds that inhibit the enzymatic an

Effect of Laser Optoperforation of the Zona Pellucida on Mouse Embryo Development in vitro

Laser operations on cells and embryos are an important field of current photobiology and biophotonics. The high power density of tightly focused laser irradiation provides an efficient impact on matter of cells or embryos. Precise focusing of the laser spot allows strictly controlled perforation of the membrane. The present work was devoted to studying the influence of optoperforation of mammalian embryonic zona pellucida with a tightly focused laser beam with 1.48-µm wavelength on further development of the embryo. Such a laser operation was proposed for application in in vitro fertilization (IVF) practice and intracytoplasmic sperm injection into the oocyte (ICSI). For cultured in vitro oocytes and embryos, the process of natural exiting from the zona pellucida ("hatching") is often impaired, which decreases probability of implantation and pregnancy The goals of the present work were to determine the influence of different manipulations on development of embryos in vitro until blastocyst formation and on the ISSN 0006-2979, Biochemistry (Moscow), 2015, Vol. 80, No. 6, pp. 769-775. © Pleiades Publishing, Ltd., 2015. Original Russian Text © E. O. Zakharchenko, A. D. Zalessky, A. A. Osychenko, A. S. Krivokharchenko, A. K. Shakhbazyan, A. V. Ryabova, V. A. Nadtochenko, 2015, published in Biokhimiya, 2015 769 * To whom correspondence should be addressed. Abstract-The effect of laser optical perforation of the zona pellucida on the viability and development of mouse embryos has been studied. Operations of zona pellucida thinning and single or double perforation were carried out on 2-cell embryo, morula, and blastocyst stages with a laser pulse (wavelength 1.48 µm, pulse duration 2 ms). Embryo development up to the blastocyst stage and hatching efficiency were statistically analyzed. It was found that 2-cell or morula stage embryo zona pellucida thinning or single perforation did not affect development to the blastocyst stage and number of hatched embryos, but it accelerated embryo hatching compared to control groups one day earlier in vitro. Double optoperforation on 2-cell embryo or morula stage did not significantly affect development to the blastocyst stage, but it strongly decreased the number of hatched embryos. Also, zona pellucida perforation at the blastocyst stage had a negative effect: hatching did not occur after this manipulation. Blastocyst cell number calculation after single zona pellucida perforation at 2-cell and morula stages showed that cell number of hatching or hatched blastocysts did not differ from the same control groups. This fact points out that the laser single optoperforation method is a useful and safe experimental tool that allows further manipulations within the zona pellucida. Effect of Laser Optoperforation of the Zon