MOLECULAR AND GENETIC CHARACTERISTICS OF SURFACE AND NONSTRUCTURE PROTEINS OF PANDEMIC INFLUENZA VIRUSES A(H1N1)PDM09 IN 2015-2016 EPIDEMIC SEASON

The aim of the present study was identifying of molecular and genetic changes in hemaglutinin (HA), neuraminidase (NA) and non-structure protein (NS1) genes of pandemic influenza A(H1N1)pdm09 strains, that circulated in Ukraine during 2015-2016 epidemic season. Samples (nasopharyngeal swabs from patients) were analyzed using real-time polymerase chain reaction (RTPCR). Phylogenetic trees were constructed using MEGA 7 software. 3D structures were constructed in Chimera 1.11.2rc software.Viruses were collected in 2015-2016 season fell into genetic group 6B and in two emerging subgroups, 6B.1 and 6B.2 by gene of HA and NA. Subgroups 6B.1 and 6B.2 are defined by the following amino acid substitutions. In the NS1 protein were identified new amino acid substitutions D2E, N48S, and E125D in 2015-2016 epidemic season. Specific changes were observed in HA protein antigenic sites, but viruses saved similarity to vaccine strain. NS1 protein acquired substitution associated with increased virulence of the influenza virus

Resistance of old winter bread wheat landraces to tan spot

Background. The most effective and environmentally safe way to combat wheat diseases is to produce cultivars resistant to their pathogens. For this purpose, old landraces are often used as genetically diverse sources of traits important for breeding. In the process of wheat breeding for resistance to tan spot caused by the fungus Pyrenophora tritici-repentis (Died.) Drechs. (abbr. Ptr), selection is carried out against the dominant allele of Tsn1, the gene of sensitivity to the toxin Ptr ToxA, which induces necrosis and represents the main pathogenicity factor of Ptr controlled by the ToxA gene. The aim of the study was to characterize a set of bread wheat (Triticum aestivum L.) accessions from the VIR collection for resistance to various Ptr populations, genotype these accessions using Xfcp623 – a DNA marker of the Tsn1 gene, and identify sources of tan spot resistance.Materials and methods. Sixty-seven accessions of winter bread wheat landraces were studied. Seedling resistance to two Ptr populations was assessed using a 5-point scale adopted at VIZR. The allelic state of Tsn1 was identified by PCR.Results. Dominant alleles of Tsn1 were found for 55% of the studied accessions. Seventeen accessions were resistant or moderately resistant to two Ptr populations and an isolate from Krasnodar Territory previously used for their characterization. Nine of them had the tsn1tsn1 genotype, and 8 had Tsn1Tsn1. The accessions mainly belonged to three agroecological groups proposed by N. I. Vavilov: “steppe winter bread wheat (Banatka wheats)”, “North European forest awnless bread wheats (Sandomirka wheats)”, and “Caucasian mountain winter bread wheat”.Conclusion. The identified 17 accessions resistant to Ptr are potential breeding sources of resistance. In the studied set of accessions, no significant relationship was found between the allelic state of the Tsn1 gene in the accession and its response to the infection with pathogen populations, including isolates with the ToxA gene

Bioethics In The Context Of Socio-Economic, Socio-Political And Cultural Development Of Society

Біоетика – не лише новий напрямок в науці, але й інша сфера філософського пізнання життя та його збереження на Землі. В роботі розглянуто проблеми етико-правового регулювання сучасних біомедичних досліджень, де одним із методів вивчення та опису об’єктів навколишнього середовища є експеримент. Він є ключовим інструментом практики, без якої неможлива теорія. Враховуючи темпи розвитку сучасної науки та загалом світу, актуальною є тема вивчення принципів регулювання біомедичних досліджень та їх удосконалення. У статті також висвітлено основні принципи використання людей та тварин у біомедичних дослідженнях. Пояснено чому так важливо посилити гуманізацію цього процесу. Висновки: дуже важливо запобігати жорстокому поводженню з будь-якими біологічними об’єктами в контексті вирішення біоетичних та правових проблем, пов’язаних із реалізацією принципу гуманізму та захистом прав живих організмів. Сучасна наука та практика доклінічних та клінічних випробувань повинні бути залучені до цієї проблеми з метою вдосконалення та універсалізації процесу медико-біологічних досліджень in vitro. Мета статті – вивчити основні міжнародно-правові принципи використання організмів у біомедичних експериментах, а також визначити основні проблеми з цього питання та окреслити шляхи їх вирішення. Оглянути сучасний стан етико-правового регулювання, принципів та правил проведення експериментальних медико-біологічних досліджень. На сьогоднішній день міжнародне співтовариство прийняло достатньо правових актів, що регулюють належне проведення біомедичних доклічнічних та клінічних досліджень з біологічними об’єктами, в тому числі спеціально вирощених для таких досліджень організмів (в лабораторії). Останнім часом як у національному законодавстві європейських країн, так і в міжнародному законодавстві спостерігаються тенденції посилення їх захисту у галузі біомедичних досліджень. В статті приведено приклади знущання, і як наслідок прямого порушення основних принципів біоетики – поваги до автономності та гідності людини, Доведено, що основна задача біоетики – збереження життя та здоров’я кожного організму. Обов’язковим є як на міжнародному, так і на національному рівнях прийняття програм розвитку та підтримки наукового вивчення впровадження біосистем in vitro у практику досліджень. На національному рівні необхідно криміналізувати порушення міжнародних стандартів проведення біомедичних досліджень з використанням організмів, посилити інші види відповідальності та створити спеціальну систему державних органів, відповідальних за політику їх використання у біомедичних цілях та здійснення контролю в цій області.The paper considers the problems of ethical and legal regulation of modern biomedical research, where one of the methods of studying and describing environmental objects is an experiment. It is a key tool of practice, without which theory is impossible. Given the pace of development of modern science and the world as a whole, the topic of studying the principles of regulation of biomedical research and their improvement is relevant. The article highlights the basic principles of the use of humans and animals in biomedical research. It was explained why it is so important to strengthen the humanization of this process. The conclusions are the following: it is very important to prevent cruelty of any biological objects in the context of resolving bioethical and legal problems associated with the implementation of the principle of humanism and the protection of the rights of living organisms. Modern science and the practice of preclinical and clinical tests should be involved into serious challenge in order to improve and universalize the process of medical and biological research technology in vitro. The purpose of the article is to study the main international legal principles of using living organisms in biomedical experiments, as well as to determine the main problems on this issue and to outline the ways to resolve them. To examine the current state of ethical and legal regulation, principles and rules for conducting experimental biomedical research. Nowadays the international community has adopted enough legal acts regulating the proper behavior with biological objects, including those specially cultivated for such studies (laboratory) when conducting biomedical experiments. Recently, both, in the national legislation of European countries and international legislation, there have been trends to strengthen their protection in the field of biomedical research. The article provides examples of bullying, and as a result of a direct violation of the basic principles of bioethics – respect for the autonomy and dignity of a person. It is proved that the main task of bioethics is to preserve the life and health of every organism. It is imperative, both at the international and national levels, to adopt programs for development and support of scientific study of the in vitro biosystems introduction into research practice. At the national level, it is necessary to criminalize the violation of international standards for conducting biomedical research using organisms. Both, international and national legislation should impose a direct ban on the use of in vivo technologies in case of the possibility of using in vitro technologies.Биоэтика – не только новое направление в науке, но и другая сфера философского познания жизни и его сохранения на Земле. В работе рассмотрены проблемы этико-правового регулирования современных биомедицинских исследований, где одним из методов изучения и описания объектов окружающей среды является эксперимент. Он является ключевым инструментом практики, без которой невозможна теория. Учитывая темпы развития современной науки и в целом мира, актуальной является тема изучения принципов регулирования биомедицинских исследований и их усовершенствованию. В статье также отражены основные принципы использования людей и животных в биомедицинских исследованиях. Объяснено почему так важно усилить гуманизацию этого процесса. Выводы: очень важно предотвращать жестокого обращения с любыми биологическими объектами в контексте решения биоэтических и правовых проблем, связанных с реализацией принципа гуманизма и защитой прав живых организмов. Современная наука и практика доклинических и клинических испытаний должны быть привлечены к этой проблеме с целью совершенствования и универсализации процесса медико-биологических исследований in vitro. Цель статьи – изучить основные международно-правовые принципы использования организмов в биомедицинских экспериментах, а также определить основные проблемы по этому вопросу и наметить пути их решения. Осмотреть современное состояние этико-правового регулирования, принципов и правил проведения экспериментальных медико-биологических исследований. На сегодняшний день международное сообщество приняло достаточно правовых актов, регулирующих надлежащее проведение биомедицинских доклинических и клинических исследований с биологическими объектами, в том числе специально выращенных для таких исследований организмов (в лаборатории). В последнее время как в национальном законодательстве европейских стран, так и в международном законодательстве наблюдаются тенденции усиления их защиты в области биомедицинских исследований. В статье приведены примеры издевательства, и как следствие прямого нарушения основных принципов биоэтики – уважения к автономности и достоинства человека, Доказано, что основная задача биоэтики – сохранение жизни и здоровья каждого организма. Обязательным является как на международном, так и на национальном уровнях принятия программ развития и поддержки научного изучения внедрения биосистем in vitro в практику исследований. На национальном уровне необходимо криминализировать нарушение международных стандартов проведения биомедицинских исследований с использованием организмов, усилить другие виды ответственности и создать специальную систему государственных органов, ответственных за политику их использования в биомедицинских целях и осуществления контроля в этой области

Influence of allelic polymorphism in the 3’ untranslated region of the <i>StTCP23</i> gene on the tolerance of potato cultivars to the potato spindle tuber viroid

Background. It is known that the pathological phenotype of potato plants can be mediated by complementary interactions between the genomic RNA of PSTVd and mRNA of some regulatory genes, which consequently lead to RNA interference, the synthesis of small interfering RNAs (vd-sRNA PSTVd), and impaired morphogenesis. At the same time, symptoms caused by the viroid may vary in different potato cultivars. Here we predict the interactions between the 3’ UTRs of various alleles of the StTCP23 transcription factor gene and the complementary regions in PSTVd genomic RNA.Materials and methods. We selected eight commercial potato cultivars with different symptoms of viroid infection and disease. For each cultivar, six clones of each cDNA amplicon of StTCP23 with a 3’ UTR were identified, and the allelic compositions of the target regions within their 3’ UTRs were characterized.Results. In total, 11 types of alleles of the 3’ UTR StTCP23 segment complementary to the vd-sRNA PSTVd were identified. Cultivars with the A allele (‘Gala’, ‘Colomba’, ‘Favorit’, and ‘Fioletovy’) identical to the reference genome or a high dose of the C allele with a deletion of four nucleotides (cv. ‘Impala’) were characterized by high susceptibility already at the primary (firstyear) infection with the PSTVd. Cvs. ‘Krepysh’, ‘Labadia’ and ‘Riviera’, classified as tolerant during primary inoculation, on the contrary, were characterized by the absence of the A allele and the presence of cultivar-specific mutant alleles.Conclusion. A high degree of polymorphism in the target site (3’ UTR region) of StTCP23 indicates a possible selection pressure on this locus. It can be assumed that cultivars with shorter alleles, which have fewer bases complementary to vd-sRNA in hypothetical duplexes and therefore less likely to induce target gene silencing, are more tolerant to the PSTVd upon primary viroid infection

The selection pressure on the neuraminidase gene of influenza viruses isolated in Ukraine from 2009 to 2015

A broad range of naturally occurring antigenic variants of the influenza virus is caused by its rapid evolutionary variability. The survival of viable influenza virus variants occurs through natural selection. The treatment of influenza infection with modern antiviral drugs – neuraminidase (NA) inhibitors – leads to the occurrence of mutations in the NA gene, which thereby result in the emergence of virus resistance to these drugs. The goal of this study was to determine the selection pressure on the NA protein of influenza viruses isolated in Ukraine from 2009 to 2015. The main method for assessing the selection pressure on proteins is to quantify the ratio of substitution rates at nonsynonymous (dN) and synonymous (dS) sites. With the help of this method, we showed that only a few codons in the NA gene were under positive selection resulting in mutations at the following sites: for influenza A viruses of the A(H1N1)pdm09 subtype – site 40, for viruses of the A(H3N2) subtype – sites 93 and 402, for Influenza B viruses of the B/Yamagata lineage – sites 74, 99, and 268, and for the viruses of the B/Victoria lineage – sites 358, 288, and 455. These sites are not associated with the NA active site, transmembrane domain, or the antigenic sites of this protein. We concluded that NA inhibitors are not a significant factor in the process of selection of the influenza viruses in Ukraine because the sites associated with the resistance of influenza viruses to NA inhibitors were not affected by positive selection. This finding could be explained by the limited use of NA inhibitors for the treatment of influenza infections in Ukraine. A broad range of naturally occurring antigenic variants of the influenza virus is caused by its rapid evolutionary variability. The survival of viable influenza virus variants occurs through natural selection. The treatment of influenza infection with modern antiviral drugs – neuraminidase (NA) inhibitors – leads to the occurrence of mutations in the NA gene, which thereby result in the emergence of virus resistance to these drugs. The goal of this study was to determine the selection pressure on the NA protein of influenza viruses isolated in Ukraine from 2009 to 2015. The main method for assessing the selection pressure on proteins is to quantify the ratio of substitution rates at nonsynonymous (dN) and synonymous (dS) sites. With the help of this method, we showed that only a few codons in the NA gene were under positive selection resulting in mutations at the following sites: for influenza A viruses of the A(H1N1)pdm09 subtype – site 40, for viruses of the A(H3N2) subtype – sites 93 and 402, for Influenza B viruses of the B/Yamagata lineage – sites 74, 99, and 268, and for the viruses of the B/Victoria lineage – sites 358, 288, and 455. These sites are not associated with the NA active site, transmembrane domain, or the antigenic sites of this protein. We concluded that NA inhibitors are not a significant factor in the process of selection of the influenza viruses in Ukraine because the sites associated with the resistance of influenza viruses to NA inhibitors were not affected by positive selection. This finding could be explained by the limited use of NA inhibitors for the treatment of influenza infections in Ukraine.

Potato resistance to quarantine diseases

The casual agent of potato wart Synchytrium endobioticum (Schilb.) Perc. and potato golden nematode (PGN) Globodera rostochiensis (Wollenweber) Behrens are the quarantine species causing the most widespread and destructive diseases of potato in the Russian Federation and other countries of the world. The potato pale nematode Globodera pallida (Stone) Behrens is not found in Russia, although in the European Union it is found everywhere. The review provides information on the harmfulness of S. endobioticum and PGN. To date, 43 pathotypes of S. endobioticum and 5 pathotypes of PGN have been revealed in the world. In the Russian Federation, only the first (D1) pathotype of potato wart and pathotype Ro1 of PGN have been found. Modern sets of differentials for S. endobioticum and PGN and methods of pathotype composition determination, including efforts to develop molecular markers (SSR) to determine the race of S. endobioticum, are presented. Data on the resistance of commercial potato cultivars to these quarantine diseases and methods for resistance determination are reviewed. Modern data on the genetics of potato resistance to S. endobioticum, G. rostochiensis and G. pallida, including mapping and cloning of R-genes, are presented. Available literature data on molecular markers of R-genes for marker assisted selection and the evaluation of their effectiveness are presented. The use of multiplex systems allows the presence of several genes for resistance to one or more pathogens to be analyzed at once. Mechanisms of potato quantitative resistance to S. endobioticum and PGN and adaptation processes in pathogens populations resulting in overcoming resistance of host are discussed. Cultivation of cultivars poorly susceptible to PGN can stimulate the adaptive variability of the pathogen and induce virulent pathotypes for 2–3 pathogen generations

Transmission of potato spindle tuber viroid between <i>Phytophthora infestans</i> and host plants

Potato spindle tuber viroid (PSTVd) is a naked, circular, single-stranded RNA (356–363 nucleotides in length) which lacks any protein-coding sequences. It is an economically important pathogen and is classified as a high-risk plant quarantine disease. Moreover, it is known that PSTVd is mechanically transmitted by vegetative plant propagation through infected pollen, and by aphids. The aim of this study is to determine the possibility of viroid transmission by potato pathogen Phytophthora infestans (Mont.) de Bary. PSTVd-infected (strain VP87) potato cultivars Gala, Colomba, and Riviera were inoculated with P. infestans isolate PiVZR18, and in 7 days, after the appearance of symptoms, re-isolation of P. infestans on rye agar was conducted. RT-PCR diagnostics of PSTVd in a mixture of mycelia and sporangia were positive after 14 days of cultivation on rye agar. The PSTVd-infected P. infestans isolate PiVZR18v+ was used to inoculate the healthy, viroid-free plants of potato cv. Gala and tomato cv. Zagadka. After 60 days, an amplification fragment of PSTVd was detected in the tissues of one plant of tomato cv. Zagadka by RT-PCR with the primer set P3/P4, indicating successful transmission of PSTVd by P. infestans isolate PiVZR18v+. This result was confirmed by sequencing of the RT-PCR amplicon with primers P3/P4. The partial sequence of this amplicon was identical (99.5 %) to PSTVd strain VP87. RT-PCR showed the possibility of viroid stability in a pure culture of P. infestans isolate PiVZR18v+ after three consecutive passages on rye agar. PSTVd was not detected after the eighth passage on rye agar in P. infestans subculture. These results are initial evidence of potato viroid PSTVd being bidirectionally transferred between P. infestans and host plants

Quarantine nematode species and pathotypes potentially dangerous for domestic potato production: populations diversity and the genetics of potato resistance

The review considers quarantine species and nematode pathotypes potentially dangerous for domestic potato production. Potatoes are affected by more than 30 types of parasitic nematodes, but the review focuses on the most harmful representatives of genera that cause great damage to potato production: Globodera, Ditylenchus, Nacob bus and Meloidogyne. Phytopathological and molecular methods of identification of species and pathotypes and the main achievements in studying the population variability of parasitic potato nematodes were analyzed. It was shown that due to the peculiarities of the life cycle of nematodes and lability of their genomes, the genetic variability of these organisms is very high, which creates a threat of forming new pathogenic genotypes of the parasites. The information about the intra- and interpopulation variability of nematodes is important for studying the ways of introduction and distribution of separate species, as well as for searching for the correlations of molecular markers with the pathotype. Phylogenetic studies based on modern data on genetic variability of populations have allowed to reveal species complexes in Globodera pallida (Stone) Behrens and Nacobbus aberrans (Thorne) Thorne &amp; Allen (sensu lato), including cryptic species. The main components of successful protection preventing a wide distribution of parasitic nematodes are quarantine measures, agricultural techniques, biological methods of protection and cultivation of resistant cultivars. Special attention in the review is paid to the breeding of potato cultivars with durable resistance to various nematode pathotypes, because the cultivation of such varieties is the most ecologically safe and economically advantageous way to prevent epiphytoties. Currently, significant progress has been made in the genetic protection of potato cultivars, especially against cyst-forming nematodes. The review provides data on sources of potato resistance to parasitic nematodes identified in collections of wild and cultivated species. Data on identified R-gens and QTL of resistance that have been introduced into breeding varieties using different methods and approaches are analyzed. The literature data on the study of structural and functional organization of genes for resistance to potato cyst nematodes are given. The results of molecular research on revealing the polymorphisms of loci involved in the control of resistance to cyst and gall nematodes, the development of molecular markers of certain genes and their use in marker-assisted selection for developing of new resistant cultivars, including those with group resistance, are considered