Search results for: Eduard A. Shuralev

Authors: Albina V. Moskvicheva, Gevorg G. Kazarian, Anna R. Valeeva, Marina A. Efimova, Malik N. Mukminov, Eduard A. Shuralev, Rustam Kh. Ravilov, Kamil S. Khaertynov

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Improving the early diagnosis of tuberculosis and solving a number of problems associated with the differential diagnosis of Mycobacterium bovis infection, nonspecific tuberculin reactions caused by sensitization of the body by non-tuberculosis mycobacteria, is urgent. The filtrates and extracts of M. bovis cell surface components are promising antigens with diagnostic potential. The purpose of this study was to isolate and characterize antigenic proteins and determine the dominant M. bovis antigens recognized by the humoral immune system. The mycobacterial cells were homogenized on FastPrep-24. Gel-filtration chromatography was used to fractionate the lysates of cell surface component extracts and proteins isolated from M. bovis culture supernatant. The separated fractions were analyzed using two-dimensional gel electrophoresis followed by determination of antigen serological activity using immunoblot with specific hyperimmune rabbit blood serum. As a result of electrophoretic separation of components by molecular weight, 23 antigen fractions were obtained. Analysis of densitograms showed that the fractions contained two zones of antigens with pronounced serological activity, corresponding to molecular weights of 28 and 21 kDa. The high serological activity of the 28 kDa antigen was established by immunoblot using hyperimmune blood sera. Separated and characterized by M. bovis specific antigen with a molecular weight of 28 kDa was added to the collection of specific marker antigens for M. bovis.

Keywords: antigen, gel-filtration chromatography, immunoblot, Mycobacterium bovis

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Authors: Bezuglov Eduard, Ryland Morgans, Talibov Oleg, Kalinin Evgeny, Butovsky Mikhail, Savin Evgeny, Tzgoev Eduard, Artemii Lazarev, Bekzhan Pirmakhanov, Anthony C. Hackney

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Currently, there is conflicting evidence regarding the efficacy of citrulline for physical performance and post-exercise recovery. Moreover, the vast majority of studies conducted used physically active volunteers from the general population and heterogeneous exercise protocols that are not specific to most sports. A single use of citrulline, regardless of the dose, will not have a significant effect on physical performance and post-exercise recovery in highly trained soccer players performing sport-specific exercises at maximum intensity. To evaluate the effectiveness of a single administration of citrulline at various doses in adult male professional soccer players performing sport-specific exercise at maximum intensity. A randomized, double-blind, placebo-controlled study analyzing eighteen soccer players from the top divisions of several European countries. The participants were randomized into three groups of six and performed a field-based soccer-specific test at 115% VO2max for 18-minutes. Comparative analysis of the cardiovascular system, physical activity, subjective perceived fatigue and post-exercise recovery was conducted. There were no statistically significant differences in more than one analyzed parameter. A single application of 3 to 6 grams of citrulline does not affect physical performance, subjective feeling of fatigue and post-exercise recovery in adult professional soccer players who have performed a sport-specific test. Currently, citrulline cannot be recommended for use as a supplement in adult professional soccer players

Keywords: citrulline, performance, recovery, soccer players

Procedia PDF Downloads 99

Authors: Ana Nemsadze

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Contemporary era and society are called postindustrial era and postindustrial society and/or informational era and informational society. Today science intends to define concept of information and comprehend informations role and function in contemporary society. Organization of social environment and governance of public processes on the base of information and tools of communication are main characteristics of informational era. This was defined by technological changes which were accomplished in culture in the second half of twentieth century. Today Georgia as an independent state needs to create an informational discourse of the country and therefore it is very important to study political and social cases which accomplished in the country after collapse of the Soviet Union because they start to define the present and the future of the country. The purpose of this study is to analyze political cases of the latest history of Georgia in terms of culture and information, concretely to elucidate which political cases transformed social life of post Soviet Georgia most of all who accomplished these political cases which visual and verbal messages was each political case spread with. The research is conducted on the base of interview. Participants of the interview are people of various specializations. Their professional activity is related to reflections on culture and theme of visual communication. They are philosophers sociologists a journalist media researcher a politologist a painter. The participants of the interview enumerated political cases and characterized them separately. Every expert thinks that declaration of independence of Georgia is the most important fact among all facts which were implemented in Georgia after collapse of the Soviet Union. The research revealed important social and political cases. Most of the cases are related to independence and territorial integrity of the state. Presidents of Georgia Zviad Gamsakhurdia Eduard Shevardnadze Mikheil Saakashvili Catholocos-Patriarch of All Georgia, the Archbishop of Mtskheta Tbilisi and Metropolitan bishop of Bichvinta and Tskhum Abkhazia Ilia II, businessman Bidzina Ivanishvili assumed dominating roles in cases. Verbal narrative of the cases accomplished during Zviad Gamsakhurdia presidential term expresses national freedom and visual part of cases of the same period expresses ruin of social-political structure. Verbal narrative of the cases accomplished during Eduard Sevardnadze presidential term expresses Free State and stability and reestablishment of Georgias political function in international relations and visual part of cases of the same period describes the most important moment of his presidential term and Eduard Shevardnadzes face appears too. Verbal narrative of the cases accomplished during Mikheil Saakashvilis presidential term expresses social renewal and visual part of cases of the same period describes August war and Mikheil Saakashvilis face appears too. The results of the study also reveal other details of visual verbal narrative of political and social cases of post Soviet Georgia. This gives a chance to start further reflection.

Keywords: culture, narrative, post soviet, visual communication

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Authors: Eduard Fosch-Villaronga

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This paper pioneers Care Robot Impact Assessment (CRIA), a methodology used to identify, analyze, mitigate and eliminate the risks posed by the insertion of non-medical personal care robots (PCR) in medical care facilities. Its precedent instruments (Privacy and Surveillance Impact Assessment (PIA and SIA)) fall behind in coping with robots. Indeed, personal care robots change dramatically how care is delivered. The paper presents a specific risk-sector methodology, identifies which robots are under its scope and presents some of the challenges introduced by these robots.

Keywords: ethics, impact assessment, law, personal care robots

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Authors: Irina Maria Artinescu, Costin Radu Boldea, Eduard-Ionut Matei

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The paper is a comparative study of two classical variants of parallel projection methods for solving the convex feasibility problem with their equivalents that involve variable weights in the construction of the solutions. We used a graphical representation of these methods for inpainting a convex area of an image in order to investigate their effectiveness in image reconstruction applications. We also presented a numerical analysis of the convergence of these four algorithms in terms of the average number of steps and execution time in classical CPU and, alternatively, in parallel GPU implementation.

Keywords: convex feasibility problem, convergence analysis, inpainting, parallel projection methods

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Authors: Elizabeta Eduard Baltadzhyan

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This work presents a stage of an investigation about the metaphorical conceptualization of translation in Bulgarian language. The material is a linguistic corpus consisting of 38 interviews with several generations Bulgarian translators and interpreters. The aim of this presentation is to inform about the results of the organization of the source concepts in scenarios that dominate the discursive manifestations of the source domains. The data show that, on the one hand, translators from different generations share some basic assignments of source and target domains, e. g. translation is a journey or translation is an artistic presentation. On the other hand, there are some specific scenarios motivated by significant changes in the socio-economic structure of the country and the valuation of the translator´s mission and work, e. g., the scenario of pleasure and addictive activity marks the generation that enjoy great support and stimulation from the socialist government, whereas the war scenario marks the generation during the Perestroika time.

Keywords: Bulgarian language, metaphor, scenario, translation

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Authors: Spartak Makovskyi, Volodymir Klochykhin, Valery Zakharchenko, Konstantyn Balushok, Eduard Tsyvirko, Anatoly Shalomeyev

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A highly efficient, cost-effective grain refinement technique for ML5 magnesium alloy with a commercially pure carbon nanopowder has been proposed. An experimental casting of testing specimens with incremental additions of a carbon nanopowder (0.001 - 0.1 wt.% ) was performed. It has been found that the carbon nanoparticle inoculation of the alloy structure is efficient in a narrow concentration range. The additions of 0.005-0.01 wt. % the grain refiner in the alloy resulted in a maximum increase of ductility properties (appr. Twofold) and improved tensile strength. However, further expansion of the grain refiner content led to the deterioration of the alloy's mechanical properties. In particular, the introduction of 0.1 wt.% of the nanocarbon and more caused internal defects in the metal. The carbon nanoparticle inoculation is a promising way of improving the properties of the Mg-Al-Zn alloys for critical lightweight aerospace applications on an industrial scale.

Keywords: carbon nanopowder, inoculation, melt, tensile strength

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Authors: Hossein Alimohammadi, Kristina Vassiljeva, Hassan HosseinNia, Eduard Petlenkov

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This research article discusses the mechanisms for bandgap tuning of beam-type resonators to achieve ‎broadband vibration suppression through adjustable stiffness. The method involves changing the center of ‎mass of the cantilever-type resonator to achieve piezo-free tuning of stiffness. The study investigates the ‎effect of the center of masses variation (δ) of attached masses on the bandgap and vibration suppression ‎performance of a non-uniform beam-type resonator within a phononic structure. The results suggest that the ‎cantilever-type resonator beam can be used to achieve tunability and real-time control and indicate that ‎varying δ significantly impacts the bandgap and transmittance response. Additionally, the research explores ‎the use of the first and second modes of resonators for tunability and real-time control. These findings examine ‎the feasibility of this approach, demonstrate the potential for improving resonator performance, and provide ‎insights into the design and optimization of metamaterial beams for vibration suppression applications.

Keywords: bandgap, adjustable stiffness, spatial variation, tunability

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Authors: Farshad Zeinali, Sajedeh Norouzi, Nader Mokari, Eduard Jorswieck

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The capacity of fifth-generation (5G) vehicle-to-everything (V2X) networks poses significant challenges. To ad- dress this challenge, this paper utilizes New Radio (NR) and New Radio Unlicensed (NR-U) networks to develop a heterogeneous vehicular network (HetNet). We propose a new framework, named joint BS assignment and resource allocation (JBSRA) for mobile V2X users and also consider coexistence schemes based on flexible duty cycle (DC) mechanism for unlicensed bands. Our objective is to maximize the average throughput of vehicles while guaranteeing the WiFi users' throughput. In simulations based on deep reinforcement learning (DRL) algorithms such as deep deterministic policy gradient (DDPG) and deep Q network (DQN), our proposed framework outperforms existing solutions that rely on fixed DC or schemes without consideration of unlicensed bands.

Keywords: vehicle-to-everything (V2X), resource allocation, BS assignment, new radio (NR), new radio unlicensed (NR-U), coexistence NR-U and WiFi, deep deterministic policy gradient (DDPG), deep Q-network (DQN), joint BS assignment and resource allocation (JBSRA), duty cycle mechanism

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Authors: Daniela Istrati, Alina Morosan, Maria Stanca, Bogdan Purcareanu, Adrian Fudulu, Laura Olariu, Alice Buteica, Ion Mindrila, Rodica Cristescu, Dan Eduard Mihaiescu

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Our present work is related to the synthesis, characterization and applications of new nanocomposite materials based on silica mesoporous nanocompozites systems. The nanocomposite support was obtained by using a specific step–by–step multilayer structure buildup synthetic route, characterized by XRD (X-Ray Difraction), TEM (Transmission Electron Microscopy), FT-IR (Fourier Transform-Infra Red Spectrometry), BET (Brunauer–Emmett–Teller method) and loaded with Salvia officinalis plant extract obtained by a hydro-alcoholic extraction route. The sustained release of the target compounds was studied by a modified LC method, proving low release profiles, as expected for the high specific surface area support. The obtained results were further correlated with the in vitro / in vivo behavior of the nanocomposite material and recommending the silica mesoporous nanocomposites as good candidates for biomedical applications. Acknowledgements: This study has been funded by the Research Project PN-III-P2-2.1-PTE-2016-0160, 49-PTE / 2016 (PROZECHIMED) and Project Number PN-III-P4-ID-PCE-2016-0884 / 2017.

Keywords: biomedical, mesoporous, nanocomposites, natural products, sustained release

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Authors: Givi Gavardashvili, Eduard Kukhalashvili, Tamriko Supatashvili, Giorgi Natroshvili, Konstantine Bziava, Irma Qufarashvili

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In the article to research water levels in the Zhinvali water reservoirs by field and theoretical research and using GPS and GIS technologies has been established dynamic of water reservoirs changes in the suitable coordinates and has been made water reservoir maps and is lined in the 3D format. By using of GPS coordinates and digital maps has been established water horizons of Zhinvali water reservoir in the absolute marks and has been calculated water levels volume. To forecast the filling of the Zhinvali water reservoir by solid sediment in 2018 conducted field experimental researches in the catchment basin of river Tetri (White) Aragvi. It has been established main hydrological and hydraulic parameters of the active erosion-debris flow tributaries of river Tetri Aragvi. It has been calculated erosion coefficient considering the degradation of the slope. By calculation is determined, that in the river Tetri Aragvi catchment basin the value of 1% maximum discharge changes Q1% = 70,0 – 550,0 m3/sec, and erosion coefficient - E = 0,73 - 1,62, with suitable fifth class of erosion and intensity 50-100 tone/hectare in the year.

Keywords: Zhinvali soil dam, water reservoirs, water levels, erosion, debris flow

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Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

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The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability

Procedia PDF Downloads 331

Authors: Bezuglov Eduard, Achkasov Evgeniy, Emanov Anton, Shagiakhmetova Larisa, Pirmakhanov Bekzhan, Morgans Ryland

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Background: Previous studies have found that there is a wide range of the relative age effect (RAE) in young athletes, which is dependent on age and gender. However, there is currently scant data comparing the prevalence of the RAE in successful athletes across different age groups from the same sport during the same time period. We aimed to compare the prevalence of the RAE in different age groups of successful athletes. Materials and methods: The date of birth of all youth (under 18 years old) and senior (20 years and above) male and female track and field athletes were analyzed. All athletes had entered the World Top 20 rankings in disciplines where performance rules were the same at youth and adult levels. Data were collected from the website www. tilostopaja.eu between 1999 and 2006. Results: A significant prevalence of RAE in successful youth track and field athletes were reported. Early-born (61,1%) and late-born (38,9%) athletes were represented respectively (χ2 = 131,1, p < 0,001, ϖ = 0,24). The RAE is not significant in successful senior track and field athletes. Athletes born in the first half of the year are only 0.4% more prevalent than athletes born in the second half of the year (50,2% and 49,8%, respectively). Olympic Games and World Championship medalists are more often late-born athletes (44,1% and 55,9%, respectively) (p = 0,014, χ2 = 6,1, ϖ = 0,20). Conclusion: The RAE is only prevalent in successful young track and field athletes. The RAE was not observed in successful senior track and field athletes, regardless of gender, in any of the analyzed discipline groups. The RAE reverse was observed in successful senior track and field athletes.

Keywords: relative age effect, track, and field, talent identification, underdog effect

Procedia PDF Downloads 154

Authors: Eduard Marusic-Paloka

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The Darcy-Weisbach formula is used to compute the pressure drop of the fluid in the pipe, due to the friction against the wall. Because of its simplicity, the Darcy-Weisbach formula became widely accepted by engineers and is used for laminar as well as the turbulent flows through pipes, once the method to compute the mysterious friction coefficient was derived. Particularly in the second half of the 20th century. Formula is empiric, and our goal is to derive it from the basic conservation law, via rigorous asymptotic analysis. We consider the case of the laminar flow but with significant Reynolds number. In case of the perfectly smooth pipe, the situation is trivial, as the Navier-Stokes system can be solved explicitly via the Poiseuille formula leading to the friction coefficient in the form 64/Re. For the rough pipe, the situation is more complicated and some effects of the roughness appear in the friction coefficient. We start from the Navier-Stokes system in the pipe with periodically corrugated wall and derive an asymptotic expansion for the pressure and for the velocity. We use the homogenization techniques and the boundary layer analysis. The approximation derived by formal analysis is then justified by rigorous error estimate in the norm of the appropriate Sobolev space, using the energy formulation and classical a priori estimates for the Navier-Stokes system. Our method leads to the formula for the friction coefficient. The formula involves resolution of the appropriate boundary layer problems, namely the boundary value problems for the Stokes system in an infinite band, that needs to be done numerically. However, theoretical analysis characterising their nature can be done without solving them.

Keywords: Darcy-Weisbach law, pipe flow, rough boundary, Navier law

Procedia PDF Downloads 353

Authors: László Forró

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The organolead halide perovskite CH3NH3PbI3 and its derivatives are known to be very efficient light harvesters revolutionizing the field of solid-state solar cells. The major research area in this field is photovoltaic device engineering although other applications are being explored, as well. Recently, we have shown that nanowires of this photovoltaic perovskite can be synthesized which in association with carbon nanostructures (carbon nanotubes and graphene) make outstanding composites with rapid and strong photo-response. They can serve as conducting electrodes, or as central components of detectors. The performance of several miniature devices based on these composite structures will be demonstrated. Our latest findings on the guided growth of perovskite nanowires by solvatomorph graphoepitaxy will be presented. This method turned out to be a fairly simple approach to overcome the spatially random surface nucleation. The process allows the synthesis of extremely long (centimeters) and thin (a few nanometers) nanowires with a morphology defined by the shape of nanostructured open fluidic channels. This low-temperature solution-growth method could open up an entirely new spectrum of architectural designs of organometallic-halide-perovskite-based heterojunctions and tandem solar cells, LEDs and other optoelectronic devices. Acknowledgment: This work is done in collaboration with Endre Horvath, Massimo Spina, Alla Arakcheeva, Balint Nafradi, Eric Bonvin1, Andrzej Sienkievicz, Zsolt Szekrenyes, Hajnalka Tohati, Katalin Kamaras, Eduard Tutis, Laszlo Mihaly and Karoly Holczer The research is supported by the ERC Advanced Grant (PICOPROP670918).

Keywords: photovoltaics, perovskite, nanowire, photodetector

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Authors: Cristina Damian, Eduard Malcek, Ana Leahu, Sorina Ropciuc, Andrei Lobiuc

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Given the high rivalry nowadays, the food sector must offer the markets an attractive product, which at the same time has good quality and is safe from health aspects for the consumers. Known for their high content of antioxidant compounds, especially anthocyanins, which proven human health benefits, berries from the Rosaceae family plants have a significantly high level of phytochemicals: phenolic flavonoids, such as anthocyanins, ellagic acid (tannin), quercetin, gallic acid, cyanidin, pelargonidine, catechins, kaempferol and salicylic acid. Colour and bioactive compounds, such as vitamin C and anthocyanins, are important for the attractiveness of berries and their preserved products. The levels of bioactive compounds and sensory properties of the product as it reaches the consumer are dependent on raw material, i.e., berries used, processing, and storage conditions. In this study, four varieties of raspberry jam were analyzed, 3 of them purchased commercially; they were purchased at reasonable prices, precisely to include as large a sample of the consumer population as possible. The fourth assortment was made at home according to the traditional recipe without the addition of sweeteners or preservatives. As for the homemade red raspberry jam, it had a sugar concentration of 64.9%, being the most appreciated of all assortments. The homemade raspberry jam was most appreciated due to the taste and aroma of the product. The SCHWARTAU assortment was chosen in second place by the participants in the study (sensory analysis). The quality/price ratio is also valid this time, finding that a high-quality product will have a higher purchase price. Thus, the study had the role of presenting the preferences of the sample participating in the study by age categories.

Keywords: red raspberry, jam, antioxidant, colour, sensory analysis

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Authors: Tinatin Sabauri, Eduard Gelagutashvili, Salome Pataridze

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The COVID-19 pandemic presents a serious challenge to all sectors of the country. Particularly difficult and important was the rapid mobilization of educational institutions to ensure the continuous flow of the educational process and effective fulfillment of the transaction. Developed countries managed to overcome this challenge quickly because, before the pandemic, part of universities had implemented blended learning (a mixture of online and face-to-face learning). The article aims to evaluate the use of electronic platforms by non-Georgian-speaking students and their involvement in the e-learning process at Ilia State University. Based on the phenomenological research design, a comparative analysis has been conducted - what was the use of electronic systems by non-Georgian-speaking students before 2019, and what was it like during the COVID-19 pandemic? Concretely, the phenomenological design was used in the research to evaluate the efficiency of distance learning with non-Georgian speaking students at Ilia State University. Focus groups were created within the phenomenological design. In the focus groups, students answered a pre-designed semi-structured questionnaire. Based on the analysis of the questionnaires, it was revealed that online learning and access to electronic portals were not a particular difficulty for ethnic minorities. The following positive and negative aspects of e-learning were identified in the research. Students named as positive aspects: Enables joining online classes directly from home before the start of the lecture, It saves time and money on travel and accommodation (for some students). It was named as negative aspects: Learning a language online is more difficult than in face-to-face classrooms, lack of teamwork activity, lack of strong and stable internet connections, and audio problems. Based on the results of the research, it was shown that in the post-pandemic period, the involvement of non-Georgian speaking students has significantly increased; therefore, the use of electronic systems by non-Georgian speaking students.

Keywords: electronic system, distance learning, COVID-19, students

Procedia PDF Downloads 82

Authors: Maria Vargas Hernandez, Jose Rojas Suarez, Carmelo Dueñas Castell, Sandra Contreras, Camilo Bello, Diana Borre, Walter Anichiarico, Harold Vasquez, Eduard Perez, Jose Santacruz

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In the last two decades, there have been outbreaks of emerging infectious diseases, with an impact on both the general population and the obstetric population. These infections, which affect the general population, pose a high risk for adverse maternal and perinatal outcomes, taking into account that physiological and immunological changes that occur during pregnancy can increase their risk or severity. Among these, the pandemics of viral infections, Influenza A/H1N1 and SARS-CoV-2/COVID-19, stand out. In 2009, Influenza A/H1N1 infection (H1N1 2009pdm) affected approximately 3,110 obstetric patients, with data reported from 29 countries, including 1,625 (52.3%) cases that were hospitalized, 378 (23.3%) admissions to ICU and 130 (8%) deaths; and since the end of 2019, the Severe Acute Respiratory Syndrome - 2 (SARS-CoV-2) has been identified, causing the COVID-19 pandemic, with global mortality that is around 2-4% for the general population, and higher mortality in patients requiring admission to the intensive care unit. Its impact on the obstetric population is still unknown. Objectives: To evaluate the impact on maternal and perinatal outcomes of COVID-19 infection in reference to influenza A/H1N1 infection in the obstetric population. Methodology: Systematic review of the literature and meta-analysis. Results: Mortality from maternal infection with influenza A/H1N1 appears to be higher (8%) than mortality due to maternal infection with COVID-19 (3%). The rates of ICU admission, hospitalization, the requirement for invasive mechanical ventilation, and fetal death also appear to be higher in the maternal population with A/H1N1 infection, in reference to the maternal population with COVID-19 infection. Within perinatal outcomes, the admission to the neonatal ICU appears to be higher in the infants born to mothers with COVID-19 infection (28% vs. 15% for COVID-19 and A/H1N1, respectively). Conclusion: A/H1N1 infection in the obstetric population seems to be associated with a higher proportion of adverse outcomes in relation to COVID-19 infection. The actual impact of maternal influenza A/H1N1 infection on perinatal outcomes is unknown. More COVID-19 studies are needed to understand the impact of maternal infection on perinatal outcomes in this population.

Keywords: A/H1N1, COVID-19, maternal outcomes, perinatal outcomes

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Authors: Natalia Buderatskaya, Igor Ilyin, Julia Gontar, Sergey Lavrynenko, Olga Parnitskaya, Ekaterina Ilyina, Eduard Kapustin, Yana Lakhno

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Introduction: It is known that cryopreservation of oocytes has peculiar features due to the complex structure of the oocyte. One of the most important features is that mature oocytes contain meiotic division spindle which is very sensitive even to the slightest variation in temperature. Thus, the main objective of this study is to analyse the resulting euploid embryos obtained from thawed oocytes in comparison with the data of preimplantation genetic screening (PGS) in fresh embryo cycles. Material and Methods: The study was conducted at 'Medical Centre IGR' from January to July 2016. Data were analysed for 908 donor oocytes obtained in 67 cycles of assisted reproductive technologies (ART), of which 693 oocytes were used in the 51 'fresh' cycles (group A), and 215 oocytes - 16 ART programs with vitrification female gametes (group B). The average age of donors in the groups match 27.3±2.9 and 27.8±6.6 years. Stimulation of superovulation was conducted the standard way. Vitrification was performed in 1-2 hours after transvaginal puncture and thawing of oocytes were carried out in accordance with the standard protocol of Cryotech (Japan). Manipulation ICSI was performed 4-5 hours after transvaginal follicle puncture for fresh oocytes, or after defrosting - for vitrified female gametes. For the PGS, an embryonic biopsy was done on the third or on the fifth day after fertilization. Diagnostic procedures were performed using fluorescence in situ hybridization with the study of such chromosomes as 13, 16, 18, 21, 22, X, Y. Only morphologically quality blastocysts were used for the transfer, the estimation of which corresponded to the Gardner criteria. The statistical hypotheses were done using the criteria t, x^2 at a significance levels p<0.05, p<0.01, p<0.001. Results: The mean number of mature oocytes per cycle in group A was 13.58±6.65 and in group B - 13.44±6.68 oocytes for patient. The survival of oocytes after thawing totaled 95.3% (n=205), which indicates a highly effective quality of performed vitrification. The proportion of zygotes in the group A corresponded to 91.1%(n=631), in the group B – 80.5%(n=165), which shows statistically significant difference between the groups (p<0.001) and explained by non-viable oocytes elimination after vitrification. This is confirmed by the fact that on the fifth day of embryos development a statistically significant difference in the number of blastocysts was absent (p>0.05), and constituted respectively 61.6%(n=389) and 63.0%(n=104) in the groups. For the PGS performing 250 embryos analyzed in the group A and 72 embryos - in the group B. The results showed that euploidy in the studied chromosomes were 40.0%(n=100) embryos in the group A and 41.7% (n=30) - in the group B, which shows no statistical significant difference (p>0.05). The indicators of clinical pregnancies in the groups amounted to 64.7% (22 pregnancies per 34 embryo transfers) and 61.5% (8 pregnancies per 13 embryo transfers) respectively, and also had no significant difference between the groups (p>0.05). Conclusions: The results showed that the vitrification does not affect the resulting euploid embryos in assisted reproductive technologies and are not reflected in their morphological characteristics in ART programs.

Keywords: euploid embryos, preimplantation genetic screening, thawing oocytes, vitrification

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Authors: Julia Gontar, Natalia Buderatskaya, Igor Ilyin, Olga Parnitskaya, Sergey Lavrynenko, Eduard Kapustin, Ekaterina Ilyina, Yana Lakhno

Abstract:

Introduction: It is well known that oocytes obtained from older reproductive women have accumulated mitochondrial DNA mutations, which negatively affects the morphology of a developing embryo and may lead to the birth of a child with mitochondrial disease. Special techniques have been developed to allow a donor oocyte cytoplasmic transfer with the parents’ biological nuclear DNA retention. At the same time, it is important to understand whether the procedure affects the future embryonic chromosome sets as the nuclear DNA is the transfer subject in this new complex procedure. Material and Methods: From July 2015 to July 2016, the investigation was carried out in the Medical Centre IGR. 34 donor oocytes (group A) were used for the manipulation with the aim of donating cytoplasm: 21 oocytes were used for zygotes pronuclear transfer and oocytes 13 – for the spindle transfer. The mean age of the oocyte donors was 28.4±2.9 years. The procedure was performed using Nikon Ti Eclipse inverted microscope equipped with the micromanipulators Narishige system (Japan), Saturn 3 laser console (UK), Oosight imaging systems (USA). For the preimplantation genetic screening (PGS) blastocyst biopsy was performed, trophectoderm samples were diagnosed using fluorescent in situ hybridization on chromosomes 9, 13, 15, 16, 17, 18, 21, 22, X, Y. For comparison of morphological characteristics and euploidy, was chosen a group of embryos (group B) with the amount of 121 blastocysts obtained from 213 oocytes, which were gotten from the donor programs of assisted reproductive technologies (ART). Group B was not subjected to donor oocyte cytoplasmic transfer procedure and studied on the above mentioned chromosomes. Statistical analysis was carried out using the criteria t, x^2 at a significance levels p<0.05, p<0.01, p<0.001. Results: After the donor cytoplasm transfer process the amount of the third day developing embryos was 27 (79.4%). In this stage, the group B consisted of 189 (88.7%) developing embryos, and there was no statistically significant difference (SSD) between the two groups (p>0.05). After a comparative analysis of the morphological characteristics of the embryos on the fifth day, we also found no SSD among the studied groups (p>0.05): from 34 oocytes exposed to manipulation, 14 (41.2%) blastocysts was obtained, while the group B blastocyst yield was 56.8% (n=121) from 213 oocytes. The following results were obtained after PGS performing: in group A euploidy in studied chromosomes were 28.6%(n=4) blastocysts, whereas in group B this rate was 40.5%(n=49), 28.6%(n=4) and 21.5%(n=26) of mosaic embryos and 42.8%(n=6) and 38.0%(n=46) aneuploid blastocysts respectively were identified. None of these specified parameters had an SSD (p>0.05). But attention was drawn by the blastocysts in group A with identified mosaicism, which was chaotic without any cell having euploid chromosomal set, in contrast to the mosaic embryos in group B where identified chaotic mosaicism was only 2.5%(n=3). Conclusions: According to the obtained results, there is no direct procedural effect on the chromosome in embryos obtained following donor oocyte cytoplasmic transfer. Thus, the technology introduction will enhance the infertility treating effectiveness as well as avoiding having a child with mitochondrial disease.

Keywords: donor oocyte cytoplasmic transfer, embryos’ chromosome set, oocyte spindle transfer, pronuclear transfer

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Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

Abstract:

Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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