Evaluation of strawberry cultivars as sources of high winter hardiness and productivity

Background. The realization of high productivity potential in strawberry plants significantly depends on their adaptability to soil and climate conditions of their cultivation. Winter hardiness is the most important trait in the central part of Russia. With this in view, the purpose of this study was to identify sources of high winter hardiness combined with high productivity and yield potential among the studied cultivars.Materials and methods. The research was carried out at the primary variety testing site of the All-Russian Research Institute of Fruit Crop Breeding (VNIISPK), Orel Province, in accordance with the published program and methodology of fruit, berry and nut variety studies. The testing covered 23 domestic and foreign strawberry cultivars. An Espec PSL-2KPH climate chamber (Japan) and a Polair refrigerator cabinet (Russia) were used for artificial freezing.Results. Winter hardiness of strawberry cultivars was assessed under artificial freezing conditions. Cvs. ‘Korona’, ‘Tsaritsa’, ‘Solovushka’ and ‘Sara’ were the most winter-hardy cultivars. The study of the cultivars in the field made it possible to identify the most productive and large-fruited ones: ‘Alba’, ‘Azia’, ‘Bereginya’ and ‘Tsaritsa’. Cv. ‘Tsaritsa’ combines high winter hardiness and high yield productivity and yield potential, being a valuable source of these traits in breeding practice. According to a set of indicators ‘Tsaritsa’ is promising for large-scale cultivation in the Central Black Earth Region of Russia

APPROACHES TO THE FUTURE ENGINEERS FOREIGN COMMUNICATIVE CULTURE FORMATION

Purpose: The main aim of the article is to define the approaches to the formation of future engineers’ communicative culture. The main research method used while working on the article is analysis of the domestic and foreign publication space for critical consideration of different ideas on the pedagogical problem of the future engineers’ foreign language communicative culture formation process. Methodology: In this study Content abstraction, generalization and the comparative method was applied. Result: The approaches (cultural, connectivism, technological, axiological, communicative, environmental approach) will help to the formation of the communicative culture of future engineers in the process of foreign language training. Applications: This research can be used for engineers and companies. Novelty/Originality: In this research, the model of approaches to future engineers' foreign communicative culture formation is presented in a comprehensive and complete manner

Lack of GAGA protein in Trl mutants causes massive cell death in Drosophila spermatogenesis and oogenesis

Drosophila protein GAGA (GAF) is a factor of epigenetic transcription regulation of a large group of genes with a wide variety of cellular functions. GAF is encoded by the Trithorax-like (Trl) gene, which is important for the formation of various organs and tissues at all stages of ontogenesis. In our previous works, we showed that this protein is necessary for the development of the reproductive system, both in males and females of Drosophila. Decreased expression of the Trl gene led to multiple disorders of spermatogenesis and oogenesis. One of the significant disorders was associated with massive degradation and loss of cells in the germline. In this work, we carried out a more detailed cytological study to determine what type of germ cell death is characteristic of Trl mutants, and whether there are disturbances or changes in this process compared to the norm. The results obtained showed that the lack of GAF protein causes massive germ cell death in both females and males of Drosophila, but this death manifests itself in different ways, depending on the sex. In Trl females, this process does not differ phenotypically from the norm. In the dying egg chambers, signs of apoptosis and autophagy were revealed, as well as morphological features that are characteristic of the wild type. In males, Trl mutations induce mass germ cell death through autophagy, which is not typical of Drosophila spermatogenesis, and has not been previously described, neither in the norm nor in other genes’ mutations. Thus, GAF lack in Trl mutants leads to increased germ cell death through apoptosis and autophagy. Ectopic cell death and germ line atrophy are probably associated with impaired expression of the GAGA factor target genes, among which there are genes that regulate both apoptosis and autophagy

Особенности прохождения фенологических фаз интродуцированных сортов земляники садовой в условиях Орловской области

Relevance. The study of the timing and duration of individual phases of the development of strawberry cultivars of different ecological and geographical origin in these climatic conditions is of great practical importance. The purpose of the study was the determination of the timing of the main phenophases of introduced strawberry cultivars in the Orel region, the compliance of the phenorhythms of the studied cultivars with climatic conditions, as well as the ranking of the cultivars by the terms of flowering and maturation.Methods and materials. The article presents the results of phenological observations for the period 2016-2019. 34 strawberry cultivars of domestic and foreign selection were studied. The research was carried out at the VNIISPK site of primary variety study.Results. Based on long - term observations, on the time of entry into the flowering and fruiting phase, the cultivars were ranked into early, medium and late-maturing. The influence of effective temperatures on the onset of phenophases was analyzed. The sum of effective temperatures necessary for the beginning of flowering of strawberry cultivars of different maturation periods in the conditions of the Orel region was determined. For the onset of strawberry flowering, it takes from126.68 to 260.37°effective temperatures. Early cultivars need the sum of effective temperatures of 126.68-197.55°, cultivars of middle maturation need 146.31-225.44°, late cultivars - 159.93-260.37 depending on the year of study. Fruiting occurs at the sum of effective temperatures in early cultivars from 351.73°C to 465.43°C, in middle-maturing cultivars from 390.96 to 535.44°C, in late-maturing cultivars from 450°C to 649°C.Актуальность. Изучение сроков наступления и продолжительности отдельных фаз развития сортов земляники садовой разного эколого-географического происхождения в данных климатических условиях имеет большое практическое значение.Целью исследования являлось определение сроков прохождения основных фенофаз интродуцированных сортов земляники в условиях Орловской области, соответствие феноритмов изучаемых сортов климатическим условиям, а также ранжирование сортов по срокам цветения и созревания.Материал и методы. В статье представлены результаты фенологических наблюдений за период 2016-2019 годов. Изучено 34 сорта земляники садовой отечественной и зарубежной селекции. Исследования выполнены на участке первичного сортоизучения ФГБНУ ВНИИСПК.Результаты. На основании многолетних наблюдений по времени вступления в фазу цветения и плодоношения сорта ранжированы на ранние, средне- и позднеспелые. Проанализировано влияние эффективных температур на сроки наступления фенофаз. Определена сумма эффективных температур, необходимая для начала цветения сортов земляники разных сроков созревания в условиях Орловской области. Для наступления цветения земляники требуется от 126,68 до 260,37° эффективных температур: ранним сортам – 126,68-197,55°, сренеспелым –146,31-225,44°, поздним – 159,93-260,37 в зависимости от года исследования. Плодоношение наступает при сумме эффективных температур у ранних сортов – от 351,73°С до 465,43°С, у среднеспелых – от 390,96 до 535,44°С, у позднеспелых – от 450°С, до 649°С

Формирование устойчивости к гипотермии сортов Fragaria x ananassa Duch. разного эколого-географического происхождения в осенний период

Relevance. One of the most important factors which can influence on the growth and development of strawberries is the negative temperature in winter and especially in its snowless period. We were interested to study special aspects of the oxidative stress, accumulation of protector’s conjunctions, antioxidant and water status in autumn period of varieties strawberry garden different ecologicalgeographical origin in the conditions of the Orel region.Methods. The objects of the research were the leaves of Fragaria ananassa of different ecological and geographical origin in autumn period: Tsaritsa, Alba, Sara, Korona, Kokinskaya Rannya, Solovushka, Urozhainaya TsGL.Results. As the result was shown that in autumn period, when plants began preprocessing for winter, strawberry varieties Solovushka, Tsaritsa, Sara and Korona have been marked by increased antioxidant activity (for example, the enzymes SOD and catalase), little formation of reactive oxygen species and, in the end, low structural and functional damage to cell membranes. Also it was shown an increase in bound and a decrease in free water in the leaves of plants when the temperature decreased. At the same time, in the end of autumn, on the background of high content of free proline (19.38-29.73 mg/kg) and sucrose (2.65-6.46 mg/g), the varieties Solovushka, Tsaritsa, Sara and Korona had the highest ratio of bound/free water (4.10-8.10) in comparison with the with Urozhainaya TsGL and Alba. Thus, the varieties Solovushka, Tsaritsa, Sara and Korona are characterized by high adaptability to the winter conditions in the Orel region. Актуальность. Одним из значительных факторов, влияющих на рост и развитие земляники садовой, является отрицательная температура зимой и, особенно в ее бесснежный период. Представляло интерес изучить особенности окислительного стресса, накопления протекторных соединений, антиоксидантного и водного статуса в осенний период у сортов земляники садовой разного эколого-географического происхождения в условиях Орловской области.Материал и методы. Объектами исследований в осенний период служили листья сортов Fragaria ananassa различного эколого-географического происхождения: Царица, Alba, Sara, Korona, Кокинская ранняя, Соловушка, Урожайная ЦГЛ.Результаты. В осенний период при подготовке к зиме у сортов земляники Соловушка, Царица, Sara и Korona отмечали повышенную антиоксидантная активность (на примере ферментов СОД и каталазы), незначительное образование активных форм кислорода и, как следствие, низкое структурно-функциональное повреждение клеточных мембран. Также показано, что по мере снижения температуры происходило увеличение связанной и снижение свободной воды в листьях растений. При этом к концу осени на фоне высокого содержания свободного пролина (19,38-29,73 мг/кг) и сахаров (2,65-6,46 мг/г) у сортов Соловушка, Царица, Sara и Korona по сравнению с Урожайной ЦГЛ и Alba наблюдался наибольший показатель отношения связанная/свободная вода (4,10-8,10). Таким образом, сорта Соловушка, Царица, Sara и Korona характеризуются высокой адаптационной способностью к зимним условиям Орловской обрасти.

Mutations in transcription factor as rare causes of diabetes in pregnancy

MODY1 and MODY3 represent rare causes of diabetes in pregnancy. Establishing a molecular diagnosis of MODY1 or MODY3 during pregnancy may be important for minimizing risk of perinatal complications and for improving glycemic control after pregnancy. The objective of the study was to evaluate the contribution of mutations in HNF4A and HNF1A genes in development of diabetes in pregnancy and to describe clinical characteristics of diabetes in pregnancy associated with these mutations. 230 pregnant women (20-43 years) with different type of glucose intolerance complicated during their current pregnancy were included in the study. A custom NGS panel targeting 28 diabetes causative genes was used for sequencing. Heterozygous mutations in HNF4A and HNF1A genes were detected in 3% of cases. Mutations p.I271T in HNF4A gene and p.L148F, p.Y265C, p.G288W in HNF1A gene were novel. This study includes a description of patients with pregnancy diabetes due to mutations in hepatocyte nuclear factors

Clinical, hormonal and molecular-genetic characteristics of monogenic diabetes mellitus associated with the mutations in the <i>INS</i> gene

Background: Currently more than 50 mutations of the INS gene are known to affect the various stages of insulin biosynthesis in the beta cells of the pancreas. However only individual cases of diabetes mellitus (DM) associated with heterozygous mutations in the coding region of the INS gene were reported in Russian Federation. We report a group of patients with a clinical manifestation of DM caused by mutations in both coding and non-coding regions of the INS gene. The patients with a mutation in the intron of the INS gene are reported for the first time in Russian FederationMaterials and methods: 60 patients with an isolated course of neonatal DM (NDM), 52 patients with a manifestation of DM at the age of 7–12 months and the absence of the main autoimmune markers of type 1 DM, 650 patients with the MODY phenotype were included in the study. NGS technology was used for molecular genetic research. Author’s panel of primers (Custom DNA Panel) was used for multiplex PCR and sequencing using Ion Ampliseq™ technology. The author’s panel “­Diabetes Mellitus” included 28 genes (13 candidate genes of MODY and other genes associated with DM).Results: 13 heterozygous mutations were identified in 16 probands and 9 relatives. The majority of mutations were detected in patients with PNDM (18.75%) and in patients with an onset of DM at the age of 7–12 months (9.6%). Mutations in the INS gene were detected in 2 patients (0.3%) in the group with the MODY phenotype. Mutations in the INS gene were not detected in patients with transient NDM (TNDM). Analysis of clinical data in patients with PND and onset of diabetes at the age of 7–12 months did not show significant differences in the course of the disease. The clinical characteristics of the cases of MODY10 and diabetes caused by a mutation in the intron of the INS gene are reported in details.Conclusion: The role of INS gene mutations in NDM, MODY, and DM with an onset at the age of 7–12 months was analyzed in a large group of patients. The clinical characteristics of DM due to a mutation in the intron of the INS gene are reported for the first time in the Russian Federation

Разработка оптимальных условий нанофильтрации в технологии производства иммуноглобулина G человека нормального для внутривенного введения

Scientific relevance. Medicinal products based on immunoglobulin class G (IgG) from human plasma are widely used in clinical practice to treat bacterial and viral infections, primary and secondary immunodeficiencies, and autoimmune diseases. Nanofiltration is a way to mitigate the risk of in-process contamination of raw materials with various pathogens, including viruses. Therefore, it is relevant to investigate the development and implementation of additional viral inactivation and/or elimination steps.Aim. This study aimed to develop and validate optimum nanofiltration conditions and to scale up the nanofiltration step for the manufacturing of human IgG for intravenous administration.Materials and methods. The study used a solution of  IgG  from  plasma  fractions  II  and  III. The authors paired nanofilters manufactured by Planova 20N and BioEx (Asahi Kasei, Japan), Viresolve Pro (Merck Millipore, USA), Virosart HC and HF (Sartorius, Germany), and Pegasus  SV4 and Prime (Pall, USA) with Sartopore polyethersulphone prefilters by Sartorius (Germany), Virosart MAX polyamide prefilters by Sartorius (Germany), and EKX-P regenerated cellulose prefilters by Pall (Germany). Virus reduction validation studies were performed with model viruses (human immunodeficiency virus type 1, porcine transmissible gastroenteritis virus, porcine parvovirus, murine encephalomyocarditis virus, and bovine viral diarrhoea virus) in the laboratories of the N.F. Gamaleya centre. The sample data analysis involved calculating mean values with 95% confidence intervals.Results. For all the selected combinations of prefilters and filters, the maximum nanofiltration throughput depended on the IgG concentration in the test solution. With the combination of an EKX-P filter with a Pegasus SV4 nanofilter, the maximum throughput and the IgG yield reached 6300 g/m2 and 95%, respectively. When combined with a Planova 20N nanofilter, EKX-P and Sartopore (polyethersulphone) filters provided a maximum throughput of up to 2980 g/m2 and an IgG yield of almost 100%, provided that the test solution had an IgG concentration of 10 g/L. With different filter combinations, virus reduction levels ranged from 4.00±0.05 to 4.75±0.04 log10  for human immunodeficiency virus type 1, from 4.30±0.04 to 4.55±0.06 log10 for porcine transmissible gastroenteritis virus, from 5.38±0.08 log10  to 5.57±0.04 log10  for murine encephalomyocarditis virus, 5.12±0.10 log10 to 5.25±0.08 log10 for porcine parvovirus, and exceeded 5.00 log10 for bovine viral diarrhoea virus. The virus reduction levels achieved were not statistically associated with prefilter brands.Conclusions. The study demonstrated that nanofiltration was effective at removing  viruses with various virion sizes and physicochemical characteristics, including viruses as small as parvovirus B19. The levels of virus reduction exceeded 4 log10 and met the acceptance criteria.The laboratory-scale nanofiltration parameters and the corresponding filtration times, as well as IgG yields, did not change when the process was scaled up. Therefore, nanofiltration is an effective and productive technique that helps eliminate various types of viruses and considerably improve viral safety without affecting the quality of biological medicinal products.Актуальность. Лекарственные препараты на основе иммуноглобулина G (IgG) из плазмы крови человека широко применяются в терапии бактериальных и вирусных инфекций, первичных и вторичных иммунодефицитов, аутоиммунных заболеваний. Снизить риск производственной контаминации сырья различными патогенами, в том числе вирусами, позволяет процесс нанофильтрации. Для повышения вирусной безопасности необходимы исследования по разработке и внедрению дополнительных этапов инактивации и (или) элиминации вирусов.Цель. Разработка оптимальных условий процесса нанофильтрации, валидация и масштабирование данной стадии для  производства  лекарственного  препарата  иммуноглобулина G человека для внутривенного введения.Материалы и методы. Раствор IgG из фракции II+III плазмы крови, нанофильтры Planova — 20N и BioEx (Asahi Kasei, Япония), Viresolve Pro (Merck Millipore, США), Virosart — HC и HF (Sartorius, Германия), Pegasus — SV4 и Prime (Pall, США), предфильтры Sartopore из полиэфирсульфона, Virosart MAX (Sartorius,  Германия)  из  полиамида,  EKX-P  (Pall,  Германия) из регенерированной целлюлозы. Лабораторные исследования по валидации вирусной редукции выполняли с модельными тест-вирусами (ВИЧ-1, трансмиссивного гастроэнтерита (коронавируса) свиней, парвовирус свиней,  вирус  энцефаломиокардита  мышей,  вирус бычьей диареи) на базе ФГБУ «НИЦЭМ им. Н.Ф. Гамалеи» Минздрава России. Анализ данных по выборке проводили с помощью среднего значения при 95% доверительном интервале.Результаты. Установлено, что производительность нанофильтрации для всех выбранных комбинаций «предфильтр — нанофильтр» зависит от концентрации IgG в испытуемом растворе. Максимальная пропускная способность и выход продукта составили:  предфильтр (фильтр) ЕКХ-P с нанофильтром Pegasus SV4 —  6300  г/м2  (выход  IgG  более  95%); EKX-P или Sartopore (полиэфирсульфон) с Planova 20N — до 2980 г/м2 (выход IgG — практически 100% при условии проведения  процесса  только  при  концентрации  IgG  10  г/л).  Для разных комбинаций фильтров уровень редукции соответствовал критериям приемлемости: ВИЧ-1 —  от  4,00±0,05  до  4,75±0,04  log10;  коронавирус  свиней  —  от  4,30±0,04 до 4,55±0,06 log10; вирус энцефаломиокардита мышей — от 5,38±0,08 до 5,57±0,04 log10; вирус бычьей диареи — более 5,00 log10; парвовирус свиней — от 5,12±0,10 до 5,25±0,08 log10. Статистически достоверного различия в зависимости уровня редукции от марки предфильтров не выявили.Выводы. Подтверждена эффективность противовирусной нанофильтрации для вирусов  с различными размерами вириона и физико-химическими характеристиками, включая мелкий парвовирус В19: уровень редукции вирусов для всех комбинаций составил более 4 log10, что соответствует критериям приемлемости. Разработанные в лабораторных условиях параметры и соответствующая им длительность нанофильтрации, а также выход целевого продукта для всех комбинаций исследованных фильтров не изменились при масштабировании. Нанофильтрация может служить эффективным и высокопродуктивным инструментом удаления различных типов вирусов, который не влияет на качество продукта и значительно повышает вирусную безопасность биологических препаратов

Spermatozoal sensitive biomarkers to defective protaminosis and fragmented DNA

Human sperm DNA damage may have adverse effects on reproductive outcome. Infertile men possess substantially more spermatozoa with damaged DNA compared to fertile donors. Although the extent of this abnormality is closely related to sperm function, the underlying etiology of ensuing male infertility is still largely controversial. Both intra-testicular and post-testicular events have been postulated and different mechanisms have been proposed to explain the presence of damaged DNA in human spermatozoa. Three among them, i.e. abnormal chromatin packaging, oxidative stress and apoptosis, are the most studied and discussed in the present review. Furthermore, results from numerous investigations are presented, including our own findings on these pathological conditions, as well as the techniques applied for their evaluation. The crucial points of each methodology on the successful detection of DNA damage and their validity on the appraisal of infertile patients are also discussed. Along with the conventional parameters examined in the standard semen analysis, evaluation of damaged sperm DNA seems to complement the investigation of factors affecting male fertility and may prove an efficient diagnostic tool in the prediction of pregnancy outcome