Lipidomic profile of seminal plasma in non-obstructive azoospermia with sperm maturation arrest

Introduction. The difference between obstructive and non-obstructive azoospermia with sperm maturation arrest is important for the choice of treatment tactics and adequate counseling of a married couple.Purpose of the study. The study aimed to assess the semen lipid profile in patients with sperm maturation arrest. Materials and methods. Samples of seminal plasma for lipid composition of 24 men with normozoospermia and 64 men with azoospermia were studied. Patients with azoospermia underwent microdissection testicular biopsy followed by the detection of testicular tissue pathology. Lipid extracts were analyzed by liquid chromatography with mass spectrometry. Lipid data were compared with the results of pathomorphological studies.Results. Comparison of two groups revealed a statistically significant concentration differences for 22 lipids detected in positive-ion mode and 11 lipids detected in negative-ion mode. Those lipids mainly belong to the classes hexosylceramides, sphingomyelins and phosphatidylcholines — simple ethers and oxidized lipids. In multivariate analysis, the following lipids were found to be statistically significant predictors of sperm maturation arrest: PC 16: 0_22: 6 lipid (β-coefficient: -0.73; 95% confidence interval (95% CI): -1.42 to -0.27; odds ratio (OR): 0.48; OR CI: 0.24 to 0.76; Wald's test: -2.58; p = 0.01), SM d20: 1/22:2 lipid (β-coefficient 4.96; 95% CI 2.29 to 9.13; OR: 142.31; OR CI: 9.90 to 9.22^103; Wald's test: 2.93; p = 0.003); PG 20:3_22: 6 lipid (β-coefficient 2.52; 95% CI 1.13 to 4.49; OR: 12.37; OR CI: 3.10 to 89.27; Wald's test: 3.02; p = 0.002); PC O- 16: 1/16:0 lipid (β-coefficient 1.96; 95% CI -4.12 to 0.27; OR: 0.14; OR CI: 0.02 to 0.76; Wald's test: -2.05; p = 0.04). The prediction model characteristics of sperm maturation arrest, obtained during cross-validation in the positiveion mode composed: sensitivity 91%, specificity 85%; in negative-ion mode: sensitivity 75%; specificity 81%.Conclusions. Even though early stages of spermatogenesis are equally preserved in both fertile men and men with homogeneous sperm maturation arrest, the semen in the studied group of patients differed in its lipid profile. Patients with non-obstructive azoospermia, associated with meiosis arrest, may have unique lipidomic characteristics of seminal plasma, which in the future will make it possible to differentiate various variants of severe male infertility using non-invasive methods

The introduction of modern educational technologies of discipline «Life safety, emergency medicine»

One of the most acute problems of the Russian higher education system is the real transition from the «knowledge» to the practice-oriented approach. The solution to this problem affects not only the content, but also the technological aspects of the educational process. Among the latter, the technologies of formation of students‘ competences are especially important. For realization of cognitive and educational activity of students in educational process the modern educational technologies giving the chance to improve quality of education are used, more effectively to use classroom time.Одной из острейших проблем системы российского высшего образования является переход парадигмы от «знаниевого» к практико-ориентированному. Решение этой проблемы затрагивает не только содержательные, но и технологические аспекты образовательного процесса. Среди последних особенно важны технологии формирования компетенций обучающихся. Для реализации познавательной и учебной активности студентов используются современные образовательные технологии, позволяющие повышать качество образования, более эффективно использовать аудиторное время

The role of educational films creating in the learning experiences

The students’ initiative in the educational films creation improves the assimilation of material on the "Disaster Medicine" stuff. Questioning of students after the creation of educational films has generated interest in scientific research in general and writing articles in particular.Творческая инициатива студентов при создании учебных фильмов способствует улучшению усвоения материала по предмету «Медицина катастроф». Анкетирование студентов после создания учебных фильмов породило интерес к научным исследованиям вообще и написанию статей в частности

Synthesis of γ-, δ-, and ε-Lactams by Asymmetric Transfer Hydrogenation of N-(tert-Butylsulfinyl)iminoesters

Highly enantiomerically enriched γ- and δ-lactams have been prepared by a simple and very efficient procedure that involves the asymmetric transfer hydrogenation of N-(tert-butylsulfinyl)iminoesters followed by desulfinylation of the nitrogen atom and spontaneous cyclization to the desired lactams during the basic workup procedure. Five- and six-membered ring lactams bearing aromatic, heteroaromatic, and aliphatic substituents have been obtained in very high yields and ee’s up to >99%. A slight modification of the procedure also allowed the preparation of ε-lactams in good yields and very high enantioselectivities. Both enantiomers of the final lactams could be prepared with equal efficiency by changing the absolute configuration of the sulfinyl chiral auxiliary

СРАВНИТЕЛЬНЫЙ ПРОТЕОМНЫЙ АНАЛИЗ КОНДЕНСАТА ВЫДЫХАЕМОГО ВОЗДУХА У ПАЦИЕНТОВ С РАКОМ ЛЕГКОГО МЕТОДОМ МАСС-СПЕКТРОМЕТРИИ ВЫСОКОГО РАЗРЕШЕНИЯ

Analysis of exhaled breath condensate (EBC) is a promising non-invasive method to diagnose respiratory diseases. Most researchers emphasize the importance of proteomic analysis of EBC for early diagnosis of certain respiratory diseases including lung cancer. This study was aimed at identification of potential biomarkers of neoplastic disorders in EBC of patients with lung cancer using high-performance liquid chromatography and high resolution mass-spectrometry. The study involved 26 patients with lung carcinoma (21 males, 5 females, mean age 57 ± 12 years) and 23 healthy non-smokers (19 males, 4 females, mean age 30 ± 7 years). EBC samples were collected using a disposable portable condenser R-Tube. The most of proteins identified (65 %) belonged to keratin family including type 1 (1; 2; 5 and 6А) and type 2 (9; 10; 14; 16 and 17) cytoskeletal keratins and transport proteins (albumin, lipocalin-1). Keratin family proteins (5, 6 and 14) prevailed in lung cancer patients compared to controls (p < 0.05). Other 6 proteins were also detected predominantly in lung cancer patients including b-subunit and a-subunit of haemoglobin, nuclear ubiquitous casein (NUCKS), high-mobility group proteins (HMG-I/HMG-Y), and lactoferrin. Most of these proteins could be used as a diagnostic panel to detect lung cancer. Further investigations are needed to estimate diagnostic values of these biomarkers and their role in pathogenesis of lung cancer.Анализ конденсата выдыхаемого воздуха (КВВ) является перспективным неинвазивным методом оценки состояния дыхательной системы. Многие исследователи указывают на важность анализа протеома КВВ для раннего выявления заболеваний респираторного тракта, в т. ч. диагностики рака легкого (РЛ). В исследовании, в которое были включены 2 группы доноров: основная – больные РЛ (n = 26; 21 мужчина, 5 женщин; средний возраст – 56,5 ± 11,5 года) и контрольная (n = 23; 19 мужчин, 4 женщины; средний возраст – 30,0 ± 7,0 года) – здоровые некурящие добровольцы, у больных РЛ методом высокоэффективной жидкостной хроматографии и тандемной масс-спектрометрии была проведена идентификация потенциальных белков-онкомаркеров в КВВ. Образцы КВВ были собраны с помощью одноразового портативного конденсора R-Tube. Основную часть (65 %)  идентифицированных белков составили белки кератиновой группы, в т. ч. кератины цитоскелетные 1-го (1, 2, 5 и 6А) и 2-го (9, 10, 14, 16 и 17) типов, а также группа транспортных белков (альбумин, липокалин-1). Было показано, что группа кератинов (5, 6 и 14) более значительно выражена (р < 0,05) у онкологических больных по сравнению со здоровым контролем. Также 6 белков были преимущественно определены в КВВ доноров основной группы, в т. ч. b- и a-субъединицы гемоглобина, ядерный убиквитиновый казеин (NUCKS), белки группы высокой мобильности (HMG-I/HMG-Y), лактоферрин. Большинство выявленных белков может быть предложено в качестве панели для диагностики РЛ. Однако необходимы дальнейшие исследования для определения диагностической значимости предложенных биомаркеров и их роли в патогенезе РЛ

Glycosylation of Erythrocyte Spectrin and Its Modification in Visceral Leishmaniasis

Using a lectin, Achatinin-H, having preferential specificity for glycoproteins with terminal 9-O-acetyl sialic acid derivatives linked in α2-6 linkages to subterminal N-acetylgalactosamine, eight distinct disease-associated 9-O-acetylated sialoglycoproteins was purified from erythrocytes of visceral leishmaniaisis (VL) patients (RBCVL). Analyses of tryptic fragments by mass spectrometry led to the identification of two high-molecular weight 9-O-acetylated sialoglycoproteins as human erythrocytic α- and β-spectrin. Total spectrin purified from erythrocytes of VL patients (spectrinVL) was reactive with Achatinin-H. Interestingly, along with two high molecular weight bands corresponding to α- and β-spectrin another low molecular weight 60 kDa band was observed. Total spectrin was also purified from normal human erythrocytes (spectrinN) and insignificant binding with Achatinin-H was demonstrated. Additionally, this 60 kDa fragment was totally absent in spectrinN. Although the presence of both N- and O-glycosylations was found both in spectrinN and spectrinVL, enhanced sialylation was predominantly induced in spectrinVL. Sialic acids accounted for approximately 1.25 kDa mass of the 60 kDa polypeptide. The demonstration of a few identified sialylated tryptic fragments of α- and β-spectrinVL confirmed the presence of terminal sialic acids. Molecular modelling studies of spectrin suggest that a sugar moiety can fit into the potential glycosylation sites. Interestingly, highly sialylated spectrinVL showed decreased binding with spectrin-depleted inside-out membrane vesicles of normal erythrocytes compared to spectrinN suggesting functional abnormality. Taken together this is the first report of glycosylated eythrocytic spectrin in normal erythrocytes and its enhanced sialylation in RBCVL. The enhanced sialylation of this cytoskeleton protein is possibly related to the fragmentation of spectrinVL as evidenced by the presence of an additional 60 kDa fragment, absent in spectrinN which possibly affects the biology of RBCVL linked to both severe distortion of erythrocyte development and impairment of erythrocyte membrane integrity and may provide an explanation for their sensitivity to hemolysis and anemia in VL patients

МЕДИКАМЕНТОЗНОЕ ЛЕЧЕНИЕ МУКОВИСЦИДОЗА В РОССИИ: АНАЛИЗ ДАННЫХ НАЦИОНАЛЬНОГО РЕГИСТРА (2014)

The aim of this study was to analyze drug therapy of patients with cystic fibrosis (CF) in Russia. Methods. Findings of the Russian National Register of CF patients, 2014, were used in this study. The 2014 Register included data for 2,131 patients (2,092 alive and 39 died) from 74 regions of Russia. The median age was 10.2 (15.2) years, 29.2% of patients were aged 18 years and older. We analyzed administration of bronchodilators, inhaled corticosteroids (ICS), systemic steroids, dornase alfa, hypertonic saline solution, macrolides, systemic and inhaled antibiotics, pancreatic enzymes, ursodeoxycholic acid, and fat-soluble vitamins. Results. Drug administration rates were as follows: bronchodilators, 65.9%; ICS, 21.7%; systemic steroids, 5.5%; macrolides, 32.4%; dornase-alfa, 92.8%; hypertonic saline solution, 45.9%; intravenous antibiotics, 62.3%; oral antibiotics, 73.3%; inhaled antibiotics, 41.3%; pancreatic enzymes, 93.3%; ursodeoxycholic acid, 91.4%; and fat-soluble vitamins, 88.3%. Conclusion. Compared to treatment of CF patients in the Western Europe, there is higher rate of administration of pancreatic enzymes, ursodeoxycholic acid, dornase alfa, hypertonic saline solution and intravenous antibiotics in Russia. Compared to treatment strategy in USA, there is higher rate of administration of pancreatic enzymes and lower rate of administration of bronchodilators, hypertonic saline solution and inhaled antibiotics in Russia. Муковисцидоз (МВ) – тяжелое мультисистемное заболевание, требующее комплексного медикаментозного и немедикаментозного лечения. Особенностям лекарственной терапии разнородных по числу и возрасту групп больных МВ, проживающих в разных регионах России, посвящено ограниченное число работ, однако общий анализ данных большой группы пациентов в рамках единого регистра ранее не проводился. Цель. Выявление особенностей медикаментозной терапии больных МВ в России по данным национального Регистра (2014). Материалы и методы. Использованы сведения о пациентах (n = 2 131; 2 092 живых и 39 умерших) из 74 регионов России, состоящих в национальном Регистре больных муковисцидозом (2014). Медиана возраста пациентов составила 10,2 (15,2) года, доля взрослых (не моложе 18 лет) – 29,2 %. Проанализирована частота назначения лекарственной терапии бронходилататорами, ингаляционными (иГКС) и системными (сГКС) глюкокортикостероидами (ГКС), дорназой альфа, гипертоническим раствором натрия хлорида, макролидами, системными и ингаляционными антибактериальными препаратами (АБП), панкреатическими ферментами, урсодезоксихолевой кислотой (УДХК), жирорастворимыми витаминами. Результаты. Частота применения медикаментозной терапии по России распределена следующим образом: бронходилататоры – 65,9 %; иГКС – 21,7 %; сГКС – 5,5 %; макролиды – 32,4 %; дорназа альфа – 92,8 %; гипертонический раствор натрия хлорида – 45,9 %; внутривенные АБП – 62,3 %; пероральные АБП – 73,3 %; ингаляционные АБП – 41,3 %; панкреатические ферменты – 93,3 %; УДХК – 91,4 %; жирорастворимые витамины – 88,3 %. Отмечено, что взрослым больным наиболее часто назначаются бронхолитические препараты, АБП и ГКС; реже применяются панкреатические ферменты и УДХК. Заключение. Среди особенностей терапии российских больных по сравнению со странами Западной Европы можно выделить высокую долю назначения панкреатических ферментов, УДХК, дорназы альфа, гипертонического раствора и внутривенных АБП. По сравнению с США чаще назначаются панкреатические ферменты, реже – бронходилататоры, гипертонический раствор натрия хлорида и ингаляционные АБП

Optimization of cultivation conditions in vitro and ex vitro of juvenile material of pedunculate oak

The application of biotechnological approaches for producingnumerous tree species, including pedunculate oak Quercus robur L., may become the only way to preserve and reproduce unique breeding objects, which is of great importance for practical forestry. It is now possible to clone individual trees with valuable properties and scientifically advantageous genotypes. In experiments on clonal micro propagation of oak, juvenile plant material (seedlings) was grown in the laboratory. The use of additional stage of rearing and micropropagation of explants of one-month seedlings on media with different volume of 6-benzylaminopurine (6-BAP) showed the expediency of this procedure. When applying ex vitro media, many cultures continued to develop axillary or adventive shoots, the height of which was 11–15 mm (depending on the concentration of 6-BAP). The results of this study will help to cope with the difficulties associated with the induction and preservation of the morphogenic potential of explants in the process of repeated subculturing. The highest rhizogenic response (73 %) was achieved for the micro buds of juvenile material on BTM medium with full composition of macronutrients and indolylbutyric acid (IBA) at a concentration of 0.3 mg/l. It was also found that the combination of IBA and 1-naphthalenacetic acid (NAA) in concentrations of 0.2 and 0.1 mg/l, respectively, completely suppresses the development of roots in micro shoots of pedunculate oak. Of the 4 tested substrates, the highest increase in oak seedlings was observed in the variant with forest oak soil and sand (5.0 cm). The use of two-stage adaptation of regenerative plants ensured their 100 % survival in the laboratory and greenhouse. The use of soil as a substrate, taken from natural oak plantations, will significantly reduce the cost of the process of growing planting material

Risk factors for recurrence of external genital endometriosis after surgical treatment: prospective cohortant study

Objectives of the study: improve early non-invasive diagnosis of external genital endometriosis (EGE) using the mass spectrometry method, and determine the risk factors for recurrence of EGE after surgical treatment. Material and methods: a prospective cohort study included 100 patients with EGE who underwent surgical treatment during the 2014 to 2016 period. The EGE diagnosis was made on the basis of diagnostic and therapeutic laparoscopic findings and finally confirmed at histological exam. All patients underwent blood sampling before and after surgical treatment to detect recurrence of endometriosis using direct mass spectrometry method. Results: recurrences of EGE were diagnosed in 22% of patients at 18–36 months postoperatively. High incidence of recurrent EGE was reported in the group of patients who did not receive post-surgical hormonal therapy. The hormone therapy with dienogest 2 mg for 6-9 months, as well as with goserelin 3.6 mg for 3-6 months showed the greatest efficacy for the prevention of recurrent EGE. Significant predictive factors of recurrence after surgical treatment for EGE include intensive acyclic pelvic pain; presence of surgical interventions for the history of EGE; infertility; stimulation of ovulation with hormonal drugs as part of the Assisted Reproductive Technologies Program; degree 3-4 disease spread; pronounced adhesions of the small pelvis; infiltrative forms of endometriosis; endometrioid cysts of large size; bilateral ovarian lesions. The plasma lipid profile including determination of phosphatidylcholines, phosphoethanolamines, diglyceride and sphingomyelin in the patients with endometriosis provides an approach for noninvasive early diagnosis of recurrence of this disease by mass spectrometry (sensitivity 93%, specificity 95%). Conclusion: This study developed an individual comprehensive approach to the prevention of recurrent EGE, including surgical treatment, hormonal therapy and diagnosis of the disease at an early stage of development. Timely administration of a-GnRH or dienogest 2 mg at the post-surgery stage is associated with a minimal or total absence of recurrent EGE. The administration of COCs showed no efficacy in preventing the recurrence of EGE, and therefore COCs cannot be recommended as a prophylactic drug, but exclusively for contraceptive use