The Role of Vascular Endothelial Growth Factor and Antithrombin III In The Pathogenesis of the Ovarian Hyperstimulation Syndrome

Ovarian hyperstimulation syndrome (OHSS) is an iatrogenic complication of controlled ovarian hyperstimulation (COH). Its pathogenesis is not clarified yet. In the recent years a number of studies focused on the vascular endothelial growth factor (VEGF) and antihtrombin III (AT III) indicators. VEGF is homodimeric, heparin-binding glycoprotein, stimulating vascular permeability. Antithrombin III is protease inhibitor of activated clotting factors. This study aimed at examining the VEGF-A165 and AT III indicators with two OHSS patients. Two methods were used for the determination of the indicators of VEGF-A165 and AT III: ELISA for VEGF and chromogenic assay for ATT III. Kits of R/D Systems and American Diagnostica Inc. were used to estimate VEGF and AT III indicators in serum and plasma. There were higher indicators of VEGF-A165 (180pg/ml) and reduction of AT III indicators (48%) in the patient with a severe form of OHSS than in the control group while these indicators were normal in the patient with a moderate form of OHSS. Our results confirmed some published data concerning the importance of VEGF and AT III in the genesis of OHSS. This study should include a larger group of patients in order tofollow-up statistically and authentically the variations of the indicators of both factors and their importance for OHSS

Легочные осложнения у детей и подростков после трансплантации гемопоэтических стволовых клеток

Relevance. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) makes it possible to treat severe malignant and non-malignant hematopoietic disorders system. Pulmonary complications (PC) occur in 40–60 % of patients after allo-HSCT. However to date, the effect of HSCT on functional and morphological pulmonary changes in recipients remains insufficiently studied.The objective of current study was to evaluate risk factors affecting long-term survival in children and adolescents after allo-HSCT.Methods and materials. The current study was both retrospective and prospective. The analysis included 362 patients with acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML), aged 5 months to 18 years, who received allo-HSCT at Raisa Gorbacheva Memorial Research Institute for Pediatric Oncology, Hematology and Transplantation in 2000–2018. All the patients underwent chest computed tomography (CT). When detecting CT changes, we performed fibrobronchoscopy (FBS) with microbiological examination of bronchoalveolar lavage (BAL).Results. PC were diagnosed in 124 patients (64 %) who received allo-HSCT in 2014–2018. Decrease of overall survival (OS) is associated with PC development during the first year after allo-HSCT(р<0,001).The development of early PC in remission of the underlying disease significantly affected OS (p=0.001).The probability of PC development is 2.26 times higher in patients older than 9 years (p=0.006). When comparing the intensity of conditioning regimens (MACvsRIC) in remission of the underlying disease, we did not get significant differences in the incidence of PC (p>0.05). Graft source, donor type, HLA-compatibility, recipient gender did not affect the incidence of PC (p>0.05). When using graft-versus-host disease (GVHD) prophylaxis (ptCYvsATG), the 5-year OS in patients without PC was 78.8 % and 62.8 %respectively. The 5-year OS in patients with PC was 51.8 % and 42.4 % respectively (р=0.007). Decrease of OS in patients with PC is associated with chGVHD(58.3 %,) (р=0.03).Conclusion. Pulmonary complications (infectious and non-infectious) in allo-HSCT recipients are more likely to occur in the first year after transplantation. Among bacterial pathogens, the predominance of Gr(-) flora remains. The incidence of pulmonary complications was significantly lower when using ptCY as a prevention of GVHD. Введение. Аллогенная трансплантация гемопоэтических стволовых клеток (алло-ТГСК) позволяет излечить тяжелые злокачественные и незлокачественные заболевания системы крови. Легочные осложнения (ЛО) после алло-ТГСК встречаются у 40–60% пациентов. Однако до настоящего времени остается недостаточно изученным влияние ТГСК на функциональные и морфологические изменения в легких у реципиентов.Цель – изучить факторы риска, влияющие на долгосрочную выживаемость у детей и подростков после алло-ТГСК.Методы и материалы. Настоящее исследование было как ретроспективным, так и проспективным. В анализ включены 362 пациента с острым лимфобластным лейкозом (ОЛЛ) и острым миелоидным лейкозом (ОМЛ), в возрасте от 5 месяцев до 18 лет, получивших алло-ТГСК в НИИ ДОГиТ им. Р. М. Горбачёвой в период с 2000 по 2018 г. Всем пациентам проводилась компьютерная томография (КТ) грудной клетки. При обнаружении изменений на КТ мы выполняли диагностическую фибробронхоскопию (ФБС) с микробиологическим исследованием бронхоальвеолярного лаважа (БАЛ). Результаты. ЛО диагностированы у 124 (64 %) пациентов из 193 пациентов, получавших алло-ТГСК с 2014 по 2018 г. Снижение общей выживаемости (ОВ) ассоциировано с развитием ЛО в течение первого года после алло-ТГСК (р<0,001). Развитие ранних легочных осложнений в ремиссии основного заболевания значимо влияло на ОВ (р=0,001). Шанс развития ЛО в 2,26 раза выше у пациентов старше 9 лет (р=0,006). При сравнении интенсивности режимов кондиционирования (миелоаблативные (МАК) режимы и режимы со сниженной интенсивностью доз (РИК)) в ремиссии основного заболевания нами не было получено значимых различий в частоте возникновения легочных осложнений (р>0,05). Источник трансплантата, тип донора, совместимость по генам HLA-системы, пол реципиента не оказывали влияния на частоту развития легочных осложнений (р>0,05). При использовании режима профилактики реакции «трансплантат против хозяина» (РТПХ) (птЦФ против АТГ) 5-летняя ОВ у пациентов без ЛО составила 78,8 и 62,8 % соответственно. У пациентов с ЛО 5-летняя ОВ – 51,8 и 42,4 % соответственно (р=0,007). Ухудшение общей выживаемости у пациентов с ЛО ассоциировано с хРТПХ (58,3 %) (р=0,03).Заключение. Легочные осложнения (инфекционные и неинфекционные) у реципиентов алло-ТГСК чаще возникают в первый год после выполнения трансплантации. Среди бактериальных возбудителей сохраняется преобладание Гр(-)-флоры. Частота развития легочных осложнений была значительно ниже при использовании птЦФ в качестве профилактики РТПХ.

CRYPTOCURRENCIES LEGAL REGULATION

This article evaluates the legal framework of cryptocurrency in various countries. The new currency instrument is abstract currencies. They are currencies in the sense that they can be exchanged peer-to-peer. They are representations of numbers, i.e. abstract objects. An abstract currency system is a self-enforcing system of property rights over an abstract instrument which gives its owners the freedom to use and the right to exclude others from using the instrument. Cryptocurrency or virtual currency is a cryptographically protected, decentralized digital currency used as a means of exchange. Due to the development of new technologies and innovations, the rate of use of virtual currency is rapidly increasing throughout the globe, replacing not only cash payments and payments by bank transfer, but also electronic cash payments. Among the best-known representatives of cryptocurrencies are Bitcoin, Litecoin and Ethereum. Legal scholars have not yet reached a consensus regarding the nature and legal status of virtual currency. Virtual currency possesses the nature of obligations righ ts as well as property rights, since it may be both a means of payment and a commodity. Depending on the country, the approach to cryptocurrencies may be different. Today there is already an international cryptocurrency community that does not have a single coordinating center. Only progressive jurisdiction and state regulation of cryptocurrency activity will allow the creation of the conditions that will ensure the implementation of legitimate and safe cryptocurrency relations

Effect of cryoprotectants and male on motility parameters and fertilization rate in paddlefish (Polyodon spathula) frozen-thawed spermatozoa

A sperm cryopreservation method using different cryoprotectants and sperm from different males was developed. Different percentages of pure cryoprotectants (methanol and DMSO) were added to extender 1 or 2 (20 mM tris pH 8 + 30 mM sucrose + 0.5 mM KCl or 20 mM tris pH 8 + 50 mM sucrose + 0.5 mM KCl, dilution 1:1) or non-extended sperm and every 1 ml of mixture was transferred to a 2-ml cryotube. The cryotubes were directly transferred to a pre-programmed PLANER Kryo 10 series III freezer at 0 oC and cooled from 0 Celsius to –5 Celsius at a rate of 3 Celsius.min-1, from –5 Celsius to – 15 Celsius at a rate of 5 Celsius.min-1, from –15 Celsius to –25 Celsius at a rate of 10 Celsius.min(-1), from –25 Celsius to –80 Celsius at a rate of 20 Celsius.min(-1). Thereafter the samples were held for 5 min at -80 Ceslius and finally transferred into LN(2) until next morning. The sperm was then thawed in a water bath at 40 Celsius for 105 s. Fertilization rate of control sperm was 81.5% (kept unfrozen; samples tested after 24-h storage at 3 Celsius), indicating that the gametes were of good quality. Percentage and the velocity of motile sperm from were evaluated in fresh and post-thawed sperm using video frames and subsequent image analysis. The results on hatching rates were significantly correlated with post-thawed sperm motility (r=0.49, P=0.035) and velocity (r=0.55, P=0.014) and not correlated with velocity of post-thawed spermatozoa (r=0.32, P=0.177). The best fertilization rates were obtained with 64-75 % in post-thawed sperm (3.6.105 spermatozoa per egg) when sperm were treated either without any extender or with both extenders with methanol concentrations of 8 or 10 %. These results were not significantly different compared with those obtained using fresh sperm control samples. Hatching rate was very low, only 8-15 %, when sperm was frozen with 8 or 10 % DMSO. ANOVA showed a significant effect of males on sperm motility, velocity and fertilization rate in post-thawed sperm

DETERMINATION OF LEVEL EXPRESSION OF mRNA SPLICING VARIANTS FOR DR3 IN BLOOD CELLS IN INFECTIOUS MONONUCLEOSIS

The DR3 «death receptor» plays an important role in the initiation of apoptosis, proliferation, or inflammation. This receptor is shown to be involved in various diseases, including infectious conditions. Different variants of mRNA DR3 are formed as a result of alternative splicing. These variant transcripts encode membrane and soluble forms of the receptor which have different functions. Features of their expression and contribution of individual DR3 variants to the immune pathogenesis of infectious mononucleosis (IM) are poorely understood.The purpose of this work was to develop, validate and test the techniques of DR3 gene expression assays, as well as to evaluate the DR3 mRNA splice variants by means of real-time RT-PCR and RT-PCR in the IM patients.The original version of real-time RT-PCR allowed to determine relative amounts of DR3 mRNA, DR3 membrane variants (LARD1a + LARD8), and ratios of mRNAs encoding membrane and soluble forms of the receptor. The technique proved to be specific and sensitive (a semi-quantitative detection limit = 34-35 cycles) when tested in healthy volunteers and patients with acute infectious mononucleosis (AIM). Lower expression levels were shown for two alternative membrane variants of DR3 mRNA (LARD1b and DR3beta) thus regarding these isoforms as minor fractions. The relative levels of total DR3 mRNA expression were decreased in patients with AIM, as compared to healthy volunteers, whereas mRNA expression of membrane receptor variants did not differ between IM and controls.To determine a qualitative contribution of either LARD1a and LARD8 variants into the expression of membrane forms of DR3, a two-step «nested» version of RT-PCR has been developed. It was shown that, in majority of control and IM samples, both main LARD1a, and alternative LARD8 membrane forms are contributing to mRNA expression of membrane DR3 variants.The presented methods for evaluation of expression and occurrence of DR3 mRNA variants allow to characterize changes in the expression and function of this receptor associated with infectious mononucleosis. Further investigations are required to test the technique in larger number of samples