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JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Scientific Tracks & Abstracts Day 1
Euro Virology 2018 & Influenza 2018
Page 47 Session-1 Day 1 July 02, 2018 Human Virology and Hepatitis|HIV, AIDS and other Emerging Viruses |Molecular Virology Medical Virology |Plant Virology | Current Focus in Virology Research | Viral Oncology Current and Novel Approach for Influenza vaccines | Influenza Vaccines and Vaccination Pathogenesis of Influenza Virus | Influenza Viruses -Advance in Detection & Differentiation Session Chair Session Co-Chair Joseph C. Glorioso Alfredo Berzal-Herranz University of Pittsburgh, USA Institute of Parasitology and Biomedicine, Spain Session Introduction Title: Role of Herpesviruses dUTPases in the immune dysregulation associated with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome and Gulf war illness Maria Eugenia Ariza, The Ohio State University, USA Title: Generation of monoclonal antibodies against foot-and-mouth disease virus SAT 2 and development of lateral flow strip test for virus detection Ming Yang, Canadian Food Inspection Agency, Canada Title: Proteins candidates for complementary diagnosis of goats naturally infected by Lentivirus Angela Maria Xavier Eloy, Brazilian Agricultural Research Corporation, Brazil Title: Protein binders mimicking surface glycoprotein epitopes recognized by broadly neutralizing antibodies as a new platform for identification of peptide-prints as tools for development of more protective vaccines Petr Maly, Czech Academy of Sciences, Czech Republic Title: Long-read sequencing uncovers a hidden complexity of the herpesvirus transcriptome Zsolt Boldogkoi, University of Szeged, Hungary Title: AIDS process from the perspective of evolution Zajac Vladimir, Cancer Research Institute BMC SAS, Slovakia Title: A.I.R vaccines – A synthetic self-amplifying RNA-based vaccine against Influenza Annette B Vogel, Biopharmaceutical New Technologies (BioNTech) Corporation, Germany Title: Velvet bean severe mosaic begomovirus DNA-A encoded RNA silencing suppressor proteins Narayan Rishi, Amity University, India Title: Pathogenesis of the adapted a(h1n1)pdm09 influenza viruses in different organs of infected mice Prokopyeva Elena, Novosibirsk State University, Russia Title: Comprehensive analysis of Vaccinia virus transcriptome by third-generation sequencing Dora Tombacz, University of Szeged, Hungary Title: The protective efficacy of zika polyclonal antibodies against lethal zika virus infection in ifnar1-/- mouse model Shantha Kodihalli, Emergent BioSolutions Canada, Canada Title: Multiplex detection of Aspergillus fumigatus mycoviruses Selin Ozkan-Kotiloglu, Ahi Evran University, Turkey Title: Metagenomic Analysis of Viral Diversity in Respiratory samples from patients with Respiratory Tract Infections in Kuwait Nada Madi, Kuwait University, Kuwait Title: Persistent TGEV infection enhances ETEC K88 adhesion by promoting epithelialmesenchymal transition in intestinal epithelial cells Qian Yang, Nanjing Agricultural University, China Title: A novel pathway of virus dissemination within the host: spread of porcine YuchenEuro Li, Nanjing Virology Agricultural University, 2018 China & Influenza 2018
Page 48 Maria Eugenia Ariza et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Role of Herpesviruses dUTPases in the immune dysregulation associated with myalgic encephalomyelitis/chronic fatigue syndrome and Gulf war illness Maria Eugenia Ariza1,2, Marshall V Williams1,2, Brandon Cox2 and Conner Nealer2 1Wexner Medical Center - The Ohio State University, USA 2Institute for Behavioral Medicine Research, USA
yalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Gulf war illness (GWI) are debilitating diseases Mpresenting with complex immune, endocrine and neurological symptoms. Annual health care costs are estimated at $24 billion. Diagnosis is based on exclusion and there are currently no validated tests for definitive diagnosis of either syndrome. While there is accumulating evidence supporting the premise that some herpesviruses may act as possible triggers in ME/CFS, the mechanism by which they contribute to the pathogenesis of ME/CFS remains unclear. Our studies are the first to demonstrate that the deoxyuridine triphosphate nucleotidohydrolase (dUTPase) encoded by the human herpesviruses represents a new class of pathogen-associated molecular pattern (PAMP) proteins, which alter immune and neurocognitive functions. In this study, we demonstrate that ME/CFS and GWI patients’ sera exhibit reactivation of multiple herpesviruses, differential antibody expression patterns to the herpesviruses-encoded dUTPase early protein as well as increased autoantibodies to the human nuclear dUTPase. A significant increase in IL-21 levels was also observed in a cohort of ME/CFS patients. Interestingly, IL-21 is produced at high levels by CD4+ follicular helper T cell (TFH) and regulates germinal center (GC) B cell survival and plasma- cell differentiation. Further in vitro studies in primary human cells demonstrated that the EBV-dUTPase induced activin A secretion by dendritic cells, which lead to the increased formation of CD4+ follicular helper T cells (TFH) and subsequent production of IL-21 and CXCL13 by TFH. Our data suggest a role for the herpesviruses dUTPase proteins in the immune dysregulation and pathophysiology observed in these patients possibly by altering the GC reaction and antibody responses as well as inducing the production of pleiotropic cytokines. Thus, screening for the presence of anti-herpesvirus dUTPase antibodies in these patients may serve as useful diagnostic biomarkers for the selection of appropriate treatments. Recent Publications 1. Saito T, Miyagawa K, Chen S Y, Tamosiuniene R, Wang L, Sharpe O, Samayoa E, Harada D, Grow E, Moonen J R, Cao A, Chen P I, Hennigs J K, Gu M, Li C G, Leib R D, Li D, Adams C M, del Rosario, P A, Bill M, Haddad F, Montoya J G, Robinson W H, Fantl W J, Nolan G, Zamanian R T, Nicolls M R, Chiu C Y, Ariza M E and Rabinovitch M (2017) Upregulation of HERV-K is linked to immunity and inflammation in pulmonary arterial hypertension. Circulation 136(20):1920-1935. 2. Halpin P, Williams M, Klimas N G, Fletcher M A, Barnes Z and Ariza M E (2017) Differential antibody patterns to the herpesviruses-encoded dUTPases in myalgic encephalomyelitis/chronic fatigue syndrome and Gulf war illness. Journal of Medical Virology 89:1636–1645. 3. Williams M, Cox B and Ariza M E (2017) Herpesviruses dUTPases: A new family of pathogen-associated molecular pattern (PAMP) proteins with implications for human disease. Pathogens DOI: 10.3390/pathogens6010002. 4. Young N A, Williams M V, Jarjour W N, Bruss M S, Bolon B, Parikh S, Satoskar A and Ariza M E (2016) Epstein-Barr virus (EBV) encoded dUTPase exacerbates the immune pathology of lupus nephritis in vivo. International Journal of Immunology and Immunotherapy 3(2):023. 5. Aubrecht T G, Weil Z M, Salloum B A, Ariza M E, Williams M, Reader B, Glaser R, Sheridan J and Nelson R J (2015) Chronic physical stress does not interact with Epstein-Barr virus (EBV) encoded dUTPase to alter the sickness response. J Behavioral Brain Science 5:513-523.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 49 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Biography Maria Eugenia Ariza received her PhD degree in Medical Microbiology and Immunology and is currently a Research Assistant Professor in the Department of Cancer Biology and Genetics, College of Medicine, at The Ohio State University. She has a long-standing passion for understanding how viruses contribute to the pathophysiology of human diseases, specifically the role that the human herpesviruses and human endogenous retroviruses (HERVs) have in the development of autoimmune diseases and cancer. Her studies were the first to demonstrate that the dUTPases from the human herpesviruses and HERV-K represent a new class of pathogen-associated molecular pattern (PAMP) proteins, which contribute to the immune pathology associated with several immune-mediated diseases, including chronic fatigue syndrome (CFS), SLE, psoriasis and pulmonary arterial hypertension. Her studies have identified this family of viral dUTPases as potential disease biomarkers and provide novel molecular targets for the development of alternative therapeutic agents/interventions for CFS, SLE and psoriasis.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 50 Ming Yang et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Generation of monoclonal antibodies against foot-and-mouth disease virus SAT 2 and development of lateral flow strip test for virus detection Ming Yang1, Kaye Quizon1, Boitumelo Mudabuka2 and Charles Nfon1 1National Centre for Foreign Animal Disease, Canada 2Botswana Institute for Technology Research and Innovation, Botswana
oot-and-mouth disease (FMD) remains one of the world’s most widespread epizootic and highly contagious animal Fdiseases affecting a wide host range species. More than 100 countries worldwide are not yet accepted as FMD free by the World Organisation for Animal Health. FMD virus (FMDV) is recognized as seven serotypes: O, A, C, Asia 1, SAT 1, SAT 2 and SAT 3. Several FMD outbreaks due to SAT 2 had been reported from 1990 to 2012. The development of a rapid and easily performed test for FMD detection is critical for controlling FMD outbreaks and containing its spread. The aim of the project was to generate FMDV/SAT2 specific monoclonal antibodies (mAbs) and develop a lateral flow immuno chromatographic (LFI) strip test for the rapid detection of FMDV/SAT 2. A total of eight mAbs were generated and examined for their reactivity and specificity using ELISAs. The mAb #10 was selected as the capture mAb because it reacted with all tested SAT 2 isolates. The LFI strip test was developed using two mAbs. The LFI strip test was able to identify SAT 2 isolates (n=23) in culture supernatants. The calculated diagnostic specificities were 100% and 98% for the strip test and ELISA, respectively. Thirty four of 50 FMDV/SAT 2 PCR-positive tissue suspensions from experimental inoculated animals without application were identified as positive by the LFI strip test. While, sixteen samples were positive using an ELISA. Diagnostic sensitivity for LFI strip test and ELISA were 67% and 33%, respectively calculated based on the fifty samples. In conclusion, a lateral flow strip test for detection of FMDV/SAT 2 was developed. The performance of the strip test in terms diagnostic specificity and sensitivity was higher than the ELISA. The ability of strip tests to generate rapid results would be useful for the early diagnosis on-site during FMD outbreaks.
Figure 1: Diagram of the LFI strip test. A biotin-binding protein was sprayed on the test line that captures biotinylated capture mAb. An antibody that binds the mouse antibody was sprayed on the control line. Samples and antibodies were mixed in the running buffer. gRAD strips were dipped into the tube. After 30 minutes, results were determined by visualization. A positive result is demonstrated by reddish-purple bands at both lines (test and control). A negative result is demonstrated by single visible band at the control line only.
Recent Publications 1. Ferris N P, Nordengrahn A, Hutchings G H, Reid S M, King D P, Ebert K, Paton D J, Kristersson T, Brocchi E, Grazioli S and Merza M (2009) Development and laboratory validation of a lateral flow device for the detection of foot-and-mouth disease virus in clinical samples. Journal of Virological Methods 155(1):10-17. 2. Yang M, Goolia M, Xu W, Bittner H and Clavijo A (2013) Development of a quick and simple detection methodology for foot−and−mouth disease virus serotypes O, A and Asia 1 using a generic rapid assay device. Virology Journal 10:125. 3. Yang M, Nigel Caterer, Wanhong Xu and Melissa Goolia (2015) Development of a multiplex lateral flow strip test for foot- and-mouth disease virus detection using monoclonal antibodies. Journal of Virological Methods 221:119−26.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 51 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Biography Ming Yang is an Immunologist at the National Center for Foreign Animal Diseases/Canadian Food Inspection Agency, where she produces and characterizes monoclonal antibodies against vesicular disease viruses such as foot-and-mouth disease (FMD) virus, vesicular stomatitis (VS) virus and swine vesicular disease (SVD) virus. She developed several immune assays for the diagnoses of vesicular diseases, such as ELISA and lateral flow strip tests. She has published close to 40 manuscripts in the peer-reviewed journals. She graduated from University of Manitoba with MSc degree in Immunology and PhD in Microbiology. [email protected]
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 52 Angela Maria Xavier Eloy, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Proteins candidates for complementary diagnosis of goats naturally infected by lentivirus Angela Maria Xavier Eloy Brazilian Agricultural Research Corporation, Brazil
he caprine arthritis encephalitis (CAE) is a disease caused by Lentivirus genus, Orthoretrovirinae subfamily, and Retroviridae Tfamily belonging to the same human immunodeficiency virus (HIV) family. This viral disease is widespread in the world, affecting especially goat dairy, being easily transmitted by contact among animals secretion, milk ingestion and also through contaminated ejaculate. Has no cure and is difficult to control, since the tests in use are not accurate due to virus latency, providing false negative results. The present study aimed to identify the major seminal plasma protein profile of goats chronically infected by CAE. Two groups containing five males each, aging 4 to 5 years were used. The first group was composed by naturally and chronically CAE-infected animals and the control by seronegative, both confirmed by two blood tests of Western blotting (WB) and by polymerase chain reaction (PCR). The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. The proteins of high expression identified only in seropositive animals play an important role in the viral infection, such as the protease arylsulfatase A, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. These results show that seminal plasma proteins are involved on reproductive process protection in chronically infected goats by CAE. As the arylsulfatase A enzyme participates in the physiological events of fertilization in bulls and sheep, and it is absent in seronegative goat to CAEV, probably the main function of this enzyme in goats can be related to metabolism control of sulfatides, involved to virus control.
Recent Publications 1. Bezerra Junior R G, Eloy A M X, Furtado J R, Pinheiro R R, Andrioli A P, Moreno F B M, Lobo M D P, Monteiro-Moreira A C O Moreira, R A, Pinto T M F and Teixeira M F S (2017) A panel of protein candidates for comprehensive study of caprine arthritis encephalitis (CAE) infection. Tropical Animal Health and Production 50(1):43-48. 2. Bezerra Junior R, Eloy A M X, Pereira E P, Furtado J R, Souza k C, Lima A R, Pinheiro R R, Andrioli A and Teixeira M F S (2015) Avaliação das metaloproteinases de matriz no sangue de reprodutores caprinos naturalmente infectados com Artrite Encefalite Caprina na Região Semiárida do Brasil. Acta Scientiae Veterinariae 43:1-7. 3. Eloy A M X, Galiza Y S, Pinheiro R R and Andrioli A P (2017) Activation of MMPs during experimental infection of goats by CAEV. In: 5th International Congress on Analytical Proteomics, Caparica. ICAP 2017 Proceedings Book. Lisboa: Proteomass, 1:43-317.
Biography Angela Maria Xavier Eloy, Graduate in Veterinary Medicine, PhD from the University of Leeds, England. She is one of the first researchers to study the relationship between caprine arthritis encephalitis (CAE), a viral disease caused by lentivirus, and proteomics, involving the identification of proteins and the behavior of metalloproteinases (MMPs). She is in search of markers aiming to complement the diagnosis of CAE, since there is still no safe test to control this virus disease.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 53 Petr Maly et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Protein binders mimicking surface glycoprotein epitopes recognized by broadly neutralizing antibodies as a new platform for identification of peptide-prints as tools for development of more protective vaccines Petr Maly1, Milan Kuchar1, Michal Maly1, Petr Kosztyu2, Lucia Barkocziova2, Jaroslav Turanek3 and Milan Raska2 1Institute of Biotechnology, Czech Academy of Sciences, v.v.i., BIOCEV Research Center, Vestec, Czech Republic 2Institute of Immunology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Czech Republic 3Veterinary Research Institute, Czech Republic
arbohydrates-based immunogens are generally less effective in generation of long-lasting antibody responses and Cneutralizing epitopes of surface glycoproteins are poorly immunogenic. Therefore, proteins mimicking glycan epitopes represent a promising alternative for development of more protective vaccines. Highly complex combinatorial libraries derived from scaffolds of small and robust protein domains represent an excellent tool for the identification of protein binders mimicking surface glycopeptide epitopes of viruses or bacteria that are recognized by broadly neutralizing antibodies. We use our established concept of a highly complex combinatorial library derived from scaffold of 46 amino acid albumin- binding domain (ABD) and, in combination with ribosome display, we target broadly neutralizing(bn) IgG to identify unique binding candidates recognizing antigen-binding-domain of the tested bn-IgG. In our proof-of-concept study we target glycan epitopes carried by gp120/gp41 protein complex of the HIV-1 Env.ABD variants as potential (glyco)peptide mimetics are currently being characterized for the stimulation of HIV-1 gp120-specific neutralizing antibody response. Thus, ABD-derived recombinant mimotopes could serve as a useful molecular clue for generation of more efficient HIV-1 vaccine and provide a platform for development of other viral or bacterial disease-preventing vaccines. The project was supported by Czech Ministry of Health grant AZV MZ 15-32198A and Czech Ministry of Education, Youth, and Sport grant CEREBIT CZ.02.1.01/0.0/0.0/ 16_025/0007397. Recent Publications 1. Lucie Křížová, Milan Kuchař, Hana Petroková, Radim Osička, Marie Hlavničková, Ondřej Pelák, Jiří Černý, Tomáš Kalina, Petr Malý (2017). p19-targeted ABD-derived protein variants inhibit IL-23 binding and exert suppressive control over IL- 23-stimulated expansion of primary human IL-17+ T-cells. Autoimmunity 2017 Mar 19;50(2):102-113. 2. Jan Maly, Ondrej Stanek, Jan Frolik, Marek Maly, Franka Ennen, Dietmar Appelhans, Alena Semeradtova, Dominika Wrobel, Marcel Stofik, Tereza Knapova, Milan Kuchar, Lucie Cervenkova Stastna, Jan Cermak, Peter Sebo, Petr Maly (2016). Biocompatible Size-Defined Dendrimer-Albumin Binding Protein Hybrid Materials as a Versatile Platform for Biomedical Applications. Macromol Biosci 2016 Apr 8;16(4):553-66 3. Lucie Marečková, Hana Petroková, Radim Osička, Milan Kuchař, Petr Malý (2015) Novel binders derived from an albumin- binding domain scaffold targeting human prostate secretory protein 94 (PSP94). Protein Cell 2015 Oct; 6(10):774-9. 4. Milan Kuchař, Lucie Vaňková, Hana Petroková, Jiří Cerný, Radim Osička, Ondřej Pelák, Hana Sípová, Bohdan Schneider, Jiří Homola, Peter Sebo, Tomáš Kalina, Petr Malý (2014). Human interleukin-23 receptor antagonists derived from an albumin-binding domain scaffold inhibit IL-23-dependent ex vivo expansion of IL-17-producing T-cells. Proteins 2014 Jun 23;82(6):975-89.
Biography Petr Maly is head of Laboratory of Ligand Engineering at the Institute of Biotechnology, Czech Academy of Sciences in Vestec, Czech Republic. He studied at Department of Biochemistry, Faculty of Science, Charles University in Prague, Czech Republic(1980-1985) and completed doctorate at the Institute of Molecular Genetics ASCR (IMG) in Prague. He spent postdoctoral fellowship (1992-1995) at Department of Pathology and Howard Hughes Medical Institute, The University of Michigan Medical School, Ann Arbor, USA, in the laboratory of Prof. John B. Lowe where he published several substantial papers related to in vivo role of mammalian glycosyltransferases. Since 1998 to 2005 he was a research group leader at the IMG in Prague. As a visiting scientist he also worked at Department of Biochemistry and Molecular Biology, College of Medicine, University of Oklahoma, USA. He also was participating investigator of Consortium for Functional Glycomics (USA, 2001–2008) and Member of Editorial Board (2001-2005) and Editor (since 2003) of the Czech journal “Biologicke listy” (Biological Letters). Since 2008, he has been working on the development of combinatorial protein libraries derived from small protein scaffolds andconstruction of novel high-affinity protein binders with therapeutic and diagnostic potential.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 54 Zsolt Boldogkoi, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Long-read sequencing uncovers a hidden complexity of the herpesvirus transcriptome Zsolt Boldogkoi University of Szeged, Hungary
erpesviruses are large, enveloped, double-stranded DNA viruses. Although, the members of the three subfamilies (α-, β-, Hand γ-herpesviruses) differ in tissue tropisms and in many aspects of molecular pathogenesis, the basic mechanisms of their DNA replication and transcription are largely conserved. The herpesvirus transcriptome have already been investigated by various techniques, including Northern-blot and microarray analyses, as well as short-read sequencing supplemented with other techniques such as primer extension, or S1 nuclease analyses. However, these techiques are inefficient for the detection of embedded RNAs, transcript isoforms, polycistronic RNAs, and transcritional overlaps. Long-read sequencing can circumvent these problems. We used two sequencing platforms, the RS II and Sequel methods from Pacific Biosciences and the cDNA and direct RNA sequencig methods from Oxford Nanopore Technology for studying the transcriptome of various herpesviruses including herpes simplex virus type 1, varicella-zoster virus, pseudorabies virus, and human cytomegalovirus. Our investigations revealed a much more complex transcritional landscape than it has been known earlier. We identified novel protein-coding genes, which are embededd into longer host genes. Our analysis detected a number of novel non-coding RNAs, polycistronic transcripts and various transcript isoforms including splice variants as well as transcript start sites and transcript end sites variants. Additionally, our studies uncovered a genome-wide meshwork of transcritional overlaps. This latter phenomenon suggests the existence of a transcriptional interference network controlling the global gene expression through the interactions between the transcritional machineries at the ovelapping regions.
Biography Zsolt Boldogkoi has completed his PhD from Szent Itvan University and Postdoctoral studies from University of Bonn, School of Medicine. He is the Director of the Department of Medical Biology, University of Szeged. He has published close to 100 papers in reputed journals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 55 Zajac Vladimir, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
AIDS process from the perspective of evolution Zajac Vladimir Cancer Research Institute BMC- SAS, Slovakia
espite great success in the diagnostics and therapy of AIDS, there are many unanswered questions. Without giving the Danswers to these questions more successful treatment of patients can not be expected. The strong argument for this prediction is a fact that it is not possible to stop the worldwide spread of AIDS, especially in Africa. The data leading to the conclusion that HIV alone is the etiologic agent responsible for AIDS is generally accepted. According to this claim, virus was transferred to humans from monkeys in Africa through random contacts 35-50 and according to recent reports even 100 years ago. This claim, which turned into dogma, however, has not been sufficiently confirmed and is unacceptable from epidemiological, statistical point of view and also by common sense. If we want to move forward the analysis of AIDS process we need to go deeper into the history of mankind and try to identify how was evolve the state of health of humankind. From the evolutionary point of view, the biggest changes in the human health condition occurred during a series of epidemics, particularly in Europe and the adjacent parts of Asia and Africa. The largest epidemic of plague - the Black Death - was caused by the bacterium Yersinia pestis. Started in 1346 in Italy from where it spread throughout Europe and then to Asia. Population after epidemic decreased by 30 to 50%. In 2001, epidemiologists S. Scott, Ch. Duncan and S. Kohn proposed a theory according to which the „Black Death“ may have been caused by hemorrhagic viruses. This version corresponds to the means of human-to-human transmission, speed and intensity of the epidemic. Based on our results, we assume that HIV is very likely an inseparable part of man since the beginning of our existence. These results and the subsequent analysis have led us to propose a theory that the "Black Dead" epidemic in the 14th century was attended, in addition to Yersenia pestis and other factors, and therefore, in our opinion, it could be HIV. This epidemic took place in Europe, parts of Asia and North Africa, but not in America and sub-Saharan Africa. The victims of the Black Death epidemic were individuals with a damaged immune system due to violation of symbiosis between the prokaryotic and eukaryotic kingdom in their body. The epidemic was so devastating, because resulted also in the elimination of HIV carriers. Those who survived had delta 32 mutation in the CCR5 co-receptor, which is predominantly expressed in T cells, macrophages, dendritic cells, and eosinophils. Mutation CCR5-Δ32 protect participants from Yersenia pestis infection, but the smallpox virus and HIV infection, as well. The “Black Dead” epidemic results in an increase in the number of CCR5 delta 32 mutations in the Caucasus population to 10%, in some areas to 15-20%. Statistically, it is confirmed that locations with a higher frequency of CCR5-Δ32 allele are much lower “Black Death” mortality. This epidemic on the other side as "sanitary process" led to the restoration of balance between the two kingdoms in the human body and to the recovery of most of the human population. In Sub-Saharan Africa, this epidemic and subsequent "sanitation process" has not taken place, that’s why HIV-related genetic information has not been eliminated in the population. Therefore, there is no CCR5Δ32 mutation in this population and the level of HIV genetic information is much higher than in other parts of the world. Options to remedy this situation in Sub-Saharan Africa are under discussion. Confirmation of the presented hypothesis can bring new insight into AIDS, especially in Africa, and open up new possibilities in diagnostics and therapy of this syndrome. Recent Publications 1. Duncan S R, Scott S and Duncan C J (2005) Reappraisal of the historical selective pressures for the CCR5-D32 mutation. Journal of Medical Genetics 42:205–208. 2. Parker Z F, Iyer S S, Wilen C B, Parrish N F, Chikere K C, Lee F H et al. (2013) Transmitted/founder and chronic HIV-1 envelope proteins are distinguished by differential utilization of CCR5. Journal of Virology 87(5):2401-11. 3. Plantier J C, Leoz M, Dickerson J E, De Oliveira F, Cordonnier F, Lemée V et al. (2009) A new human immunodeficiency virus derived from gorillas. Nature Medicine 15(8):871-72. 4. Zhen A and Kitchen S (2013) Stem-cell-based gene therapy for HIV infection. Viruses 6(1):1-12. 5. Kay M A and Walker B D (2014) Engineering cellular resistance to HIV. The New England Journal of Medicine 370(10):968-9.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 56 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Biography Vladimir Zajac has completed his PhD in 1982 at the Cancer Research Institute of Slovak Academy of Sciences in Bratislava (Slovakia), where he worked as the Head of Department of Cancer Genetics from 1996 to 2010. He joined the Medical Faculty of the Comenius University as Associate Professor of Genetics in 2007. He has published 72 papers mostly in reputed journals and he was editor of the book Bacteria, viruses and parasites in AIDS process (InTech, 2011).
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 57 Annette B. Vogel et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
A.I.R vaccines – A synthetic self-amplifying RNA-based vaccine against Influenza Annette B. Vogel1, Stephanie Erbar1, Tim Beissert2, Heinrich Haas1, Kerstin Walzer1, Lena Wicke1, John S. Tregoning3, Sebastian Kreiter1,2, U. Sahin1,2 1Biopharmaceutical New Technologies (BioNTech) Corporation, Germany 2TRON-Translational Oncology at the University Medical Center of the Johannes Gutenberg University, Germany 3Imperial College, UK
accines are the most effective method of controlling infectious diseases. However, todays vaccine production is facing Vdifficult challenges, as the concepts are often not fast and flexible enough to allow quick responses for the efficient control and prevention of global outbreaks of newly emerging and re-emerging viruses and the adaption to new antigenic drifts. To address this challenge, BioNTech RNA Pharmaceuticals GmbH is developing an innovative synthetic Amplified Immune Response (A.I.R) vaccine platform against Infectious Diseases that is characterized by short manufacturing times, high flexibility and the feasible production of at least 100,000 RNA-based human vaccination doses per week based on the low concentration needed. The administration of in vitro transcribed self-amplifying RNA (saRNA) results in higher antigen expression than delivery of comparable amounts of mRNA, correlating rather to the final subgenomic transcript copy number than to initially transferred RNA amounts. However, antigen expression is still transient as innate immunity effectively prevents persistent replication. Against influenza virus, both B and T cell-responses are induced. We were able to show high antibody titres in mice for over a one year period and protection against live viral challenge after prime-boost as well as after single vaccination by using submicrogram quantities of saRNA. In summary, A.I.R vaccines give equivalent protection against Influenza to mRNA vaccines but at much lower doses. Recent Publications 1. Vogel, Annette & Lambert, Laura & Kinnear, Ekaterina & Busse, David & Erbar, Stephanie & C. Reuter, Kerstin & Wicke, Lena & Perkovic, Mario & Beissert, Tim & Haas, Heinrich & T. Reece, Stephen & Sahin, Ugur & S. Tregoning, John. (2017). Self-Amplifying RNA Vaccines Give Equivalent Protection against Influenza to mRNA Vaccines but at Much Lower Doses. Molecular Therapy. 26. 10.1016/j.ymthe.2017.11.017. 2. Annette B. Vogel. (2016). An mRNA Vaccine Encoding Rabies Virus Glycoprotein Induces Protection against Lethal Infection in Mice and Correlates of Protection in Adult and Newborn Pigs. https://doi.org/10.1371/journal.pntd.0004746. 3. Kägebein D1, Gutjahr M, Große C, Vogel AB, Rödel J, Knittler MR. (2014). Chlamydia trachomatis-infected epithelial cells and fibroblasts retain the ability to express surface-presented major histocompatibility complex class I molecules. Infect Immun. 2014 Mar;82(3):993-1006. doi: 10.1128/IAI.01473-13. 4. Khoufache, K., Berri, F., Nacken, W., Vogel, A. B., Delenne, M., Camerer, E., Riteau, B. (2013). PAR1 contributes to influenza A virus pathogenicity in mice. The Journal of Clinical Investigation, 123(1), 206–214. 5. Benjamin Petsch, Margit Schnee, Annette B. Vogel, Elke Lange, Bernd Hoffmann, Daniel Voss, Thomas Schlake, Andreas Thess, Karl-Josef Kallen, Lothar Stitz, et al. Nat Biotechnol. 2012 Dec; 30(12): 1210–1216. Published online 2012 Nov 25. doi: 10.1038/nbt.2436.
Biography Annette B. Vogel is presently the Head of Infectious Disease Vaccines, BioNTech AG, Germany. She has been associated with Friedrich-Loeffler-Institut, Federal Reserch Institute for Animal Health as scientist. She has done her education from Georg-August-University Goettingen and completed her DSc from Eberhard-Karls-University Tubingen. She has many publications to her name like “PAR1 contributes to influenza A viruspathogenicity in mice”, “Influenza virus infection aggravates stroke outcome” to name a few. Her Research interest spans around cell culture, animal models, infectious disease, flow cytometry, virology, vaccination, serology and genomics.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 58 Narayan Rishi et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Velvet bean severe mosaic begomovirus DNA-A encoded RNA silencing suppressor proteins Narayan Rishi and Aruna Jangid, Vanita Chandel Amity University Noida, India
eminiviruses (family Geminiviridae) are classified into nine genera and unassigned viruses on the basis of host range, Ginsect vector and genome organization. These genera are: Becurtovirus, Begomovirus, Capulavirus, Curtovirus, Eragrovirus, Grablovirus, Mastrevirus, Topocuvirus and Turncurtovirus. Begomoviruses are divided into two categories i.e. bipartite or monopartite (old world) bipartite (new world). Viruses have evolved to encode unique proteins to counter RNA silencing known as RNA silencing suppressors (RSS). Several begomovirus and their associated betasatellite-encoded viral proteins have been identified as RSSs. A large number of structural such as coat protein (CP) and non-structural viral proteins possessing suppressor activity have been involved in crucial viral functions, including viral movement, viral replication etc. Mucuna pruriens (L) DC (Velvet bean/“magic bean”) of family Fabaceae is used in Ayurvedic medicines for Parkinson’s disease, liver dysfunction, blood-related diseases, snake bite, endocrine and male reproductive disorders. Velvet bean severe mosaic virus (VbSMV) is a bipartite DNA virus infecting Mucuna pruriens (Velvet bean) belongs to the genus Begomovirus, family Geminiviridae. In this study VbSMV was identified from velvet bean. For the identification of suppressor proteins, primers were designed for all genes of VbSMV by adding appropriate site for restriction enzymes for inframe cloning in the vector. Out of the two assays reported for the identification of suppressor genes we used Agrobacterium co-infiltration assay. It was delineated that proteins encoded by VbSMV viz. AV2 (pre-coat protein), AC2 (TrAP), AV1 (coat protein) are suppressors of RNA silencing as identified through Agrobacterium co-infiltration assay using Nicotiana benthamiana as a host plant. AV2 showed strong suppressor activity whereas AC2 and AV1 were found to be weak suppressors. This is the first report on identification of suppressor of RNA silencing encoded by VbSMV infecting a medicinal plant. Identified suppressor proteins are being used to develop virus resistant transgenic plants and understanding RNA silencing pathway.
Biography Research interests of Narayan Rishi are: (i) Developed DAC&DAS-ELISA based detection of citrus yellow mosaic Badnavirus using expressed recombinant VAP; (ii) Identified strains of PVS&PVY inducing partial resistance to potato late & early blight; (iii) Genetic variability in cotton leaf curl begomovirus (CLCuV),cloning & sequencing of coat protein & movement genes of CLCuV;(iv) Developed CLCuV resistant GM cotton the first in world;(v) Sequencing and diversity in DNA β associated with monopartite begomoviruses; (vi)First report of association of Mycoplasma (Phytoplasma) with grassy shoot disease of sugarcane; (vii)First report of identification of strains A&F of sugar cane mosaic virus (SCMV) in India; (viii)Purification of SCMV & raising high titer antisera that was used in ELISA and other serological tests in India. President, Indian Virological Society; Member ICTV and IUMS; Member Quinquennial Review Team, Indian Council of Agricultural Research; Awarded Prof SN Das Gupta- and Prof MV Naydu- Memorial Lectures and Prof Kameshwar Sahai Bhargava Oration Awards.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 59 Prokopyeva Elena, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Pathogenesis of the Adapted А(H1N1)pdm09 Influenza Viruses in different organs of infected Mice Prokopyeva Elena Novosibirsk State University, Russia
andemic A (H1N1) pdm09 virus has caused substantial morbidity and mortality globally and continues to circulate, which Pmay lead to an increase the pathogenic features of viruses by adaptation to the human. To address this problem, we studied changes of biological properties of the pandemic viruses during adaptation to experimental mammals and analyzed cellular localization of positive-stranded A(H1N1)pdm09 RNA in the inner organs of infected mice. To increase the virulence of pandemic H1N1 isolates in mice, we produced mouse-adapted variants of A(H1N1)pdm09 strain by serial lung-to- lung passages in BALB/c mice. After total of 7 passages we got the lethal strains to BALB/c mice (BALB/c-MA) with meaning of 50% lethal dose 1,2 lgTCID50/ml. Hematoxylin-eosin staining, immunohistochemistry for type A influenza nucleoprotein antigen, and real-time reverse transcription-PCR assay for viral RNA were performed. Complete genome sequences of the wild-type and mouse-adapted A (H1N1)pdm09 influenza viruses revealed 19 amino acid substitutions in different viral proteins (HA, NA, NS2, NS1, PB2, PB1, NP). In lung tissue under the influence of not adapted and mouse-adapted variants of A(H1N1)pdm09 influenza viruses developed interstitial pneumonitis, but it is noted the greatest degree of inflammation in case of infection of the strain BALB/c-MA. Comparative analysis revealed accumulation of viral titers in the brain (3,75±0,22 lgTCID50/ml), liver (2,5±0,5 lgTCID50/ml) and the kidney (0,74±0,48 lgTCID50/ml) in case of infection only of the strain BALB/c-MA. Immunohistochemistry staining of viral antigens was demonstrated in the lung pneumocytes and mucous glands under influence of both wild-type and mouse-adapted viruses. But only in case of infection with BALB/c-MA immunostaining was detected also in the brain, liver, kidney and in the intestine. This study demonstrates cellular localization of positive-stranded A(H1N1)pdm09 RNA in the lungs, brain, liver, kidney and in the intestine that suggests viral replication of the mouse adapted variants of A(H1N1)pdm09 influenza virus in these tissues. The study was supported by a grant from the Russian Scientific Foundation (project No.17-44-07001). Recent Publications 1. Prokopyeva EA, Sobolev IA, Prokopyev MV, Shestopalov AM (2016). Adaptation of influenza A(H1N1)pdm09 virus in experimental mouse models. Infect Genet Evol. 2016 Apr;39:265-271. doi: 10.1016/j.meegid.2016.01.022. 2. Prokopyeva EA, Romanovskaya AA, Sharshov KA, Zaykovskaya AV, Alekseev AY, Shestopalov AM (2017) Pathogenicity assessment of wild-type and mouse-adapted influenza A(H1N1) pdm09 viruses in comparison with highly pathogenic influenza A(H5N1) virus. Histol Histopathol. 2017 Oct;32(10):1057-1063. doi: 10.14670/HH-11-866. 3. Yarushkin AA, Kazantseva YA, Prokopyeva EA, Markova DN, Pustylnyak YA, Pustylnyak VO. (2016) Constitutive androstane receptor activation evokes the expression of glycolytic genes. Biochem Biophys Res Commun. 2016 Sep 23;478(3):1099-105. doi: 10.1016/j.bbrc.2016.08.075.
Biography Prokopyeva Elena has completed her PhD at the age of 29 years from FBRI State Research Center of Virology and Biotechnology ‘Vector’(Koltsovo, Russia). Dissertation title: «Phenotypic and genotypic properties of pandemic influenza A(H1N1)pdm09 virus during adaptation to mice of different genotypes». She conduct postdoctoral studies from Novosibirsk State University and Research Center of Experimental and Clinical Medicine (Novosibirsk, Russia). She is the researcher of the Laboratory experimental simulation and pathogenesis of infectious diseases, and the teacher of the course “Embryology” and “ General Histology” at Novosibirsk State University (NSU). She has published more than 25 papers in reputed journals. She has been conducting supervision of undergraduates at NSU since 2016. Also Prokopyeva Elena has obtained different honors and awards in the field of Virology, Pathology and Medical Microbiology.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 60 Dora Tombacz et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Comprehensive analysis of vaccinia virus transcriptome by third-generation sequencing Dora Tombacz1, Istvan Prazsak1, Gabor Torma1, Attila Szucs1, Michael Snyder2 and Zsolt Boldogkoi1 1University of Szeged, Hungary 2Stanford University, USA
he Vaccinia virus (VACV) is a prototype member of the Poxviridae family, it has a relatively large double-stranded DNA Tgenome containing about 220 protein coding genes. The VACV encodes DNA and RNA polymerases, transcription factors, enzymes for capping and polyadenylation, which allows VACV to replicate in the cytoplasm, rather than in the nucleus of the infected cell. The VACV is a historically important virus: it has been successfully applied as a vaccine for immunization against human smallpox, which had been eradicated in 1980 as a result of global vaccination. The most virulent strain of VACV, the Western Reserve (WR) was used in this study. For the thorough analysis of VACV transcriptome, we utilized the currently available third-generation sequencers (TGS) such as the Pacific Biosciences (PacBio) RSII and the Sequel, as well as the Oxford Nanopore Technologies (ONT) MinION, which in contrast to the next-generation sequencing approaches enables to identify full-length RNA isoforms, and distinguis between overlapping RNAs. We have characterized the static VACV transcriptome by using the RSII and MinION, while the Sequel and Minion was applied for the detailed analysis of the transcriptional dynamics of the viral RNAs. Our detailed study redefined the VACV transcriptome. Our data showed that the VACV transcriptome profile is much more complex than it was previously known. We have identified hundreds of novel transcripts and isoforms. We have described hundreds of bi-, polycistronic, and complex transcripts. These forms were not known before our study. These RNA molecules together represent very long transcriptional overlaps.
Biography Dora Tombacz has completed her MSc in Biology (2006) and PhD in Medical Sciences (2010) from the University of Szeged, Hungary. She is working in the Department of Medical Biology as an Assistant Professor at the same university in the Genomics & Gene Technology group. She has published more than 30 papers in reputed journals. Her primary field of interest is transcriptomics, the analysis of different organisms at the RNA level. She is currently working with next- and 3rd generation sequencing techniques at the University of Szeged, and as a Visiting Assistant Professor at the Stanford University, USA.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 61 Shantha Kodihalli et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
The protective efficacy of Zika polyclonal antibodies against lethal Zika virus infection in Ifnar1-/- mouse model Shantha Kodihalli1, Ayo Yila Simon1, Emilie Branch2, Hongyu Qiu1 and Sujan Shresta2 1Emergent BioSolutions, Canada 2La Jolla Institute for Allergy & Immunology, USA
ika virus (ZIKV) infection during pregnancy has become a global public health concern due to its ability to cause severe Zcongenital infections. There are no licensed vaccines or therapeutics available for ZIKV infections. To address this need, a human polyclonal Zika Immune Globulin product (ZIKV-IG) is being developed for prophylaxis of ZIKV infection in at-risk populations, including women of childbearing potential and pregnant women. To evaluate the efficacy, groups of Ifnar1-/- mice were infected with lethal doses ZIKV (FSS13025 strain) and treated with various doses of ZIKV-IG. Mice were monitored daily for body weights, clinical signs, and mortality for 21 days. In another separate study, the dose-ranging effect of ZIKV-IG on the viral load in target tissues was analyzed on days 3 and 7 using qRT-PCR and focus forming assay. ZIKV-IG administered after lethal infection provided a significant survival benefit in a dose-dependent manner. Mice treated with higher doses of ZIKV-IG (10, 50 mg/kg) provided a statistically significant survival of 87.5 to 100% in comparison to 0% in PBS controls. A similar response was observed in the viral load analyses with the highest dose providing significant reductions in target tissues including the brain, kidney, liver, sciatic nerve, serum and spleen. The 2.0 mg/kg and 0.5 mg/kg (lower dose levels) did not confer any reasonable protection in terms of survival or reduction in viral load. The efficacy of ZIKV-IG has been successfully demonstrated in a well-characterized model of Zika disease. Results of these studies demonstrate that ZIKV-IG given at 50 mg/kg in a post-exposure setting significantly enhanced survival over control. Additionally, the treatment prevented virus dissemination into target tissues. These results clearly demonstrate the potential of ZIKV-IG for post-exposure prophylaxis of human ZIKV infections.
Biography Shantha Kodihalli has completed her PhD at University of Minnesota and Postdoctoral studies at St. Jude Children’s Research Hospital, Memphis, TN USA. She is the Director of Preclinical Research at Emergent Bio Solutions. She has over 19 years of experience in the development of therapeutics for infectious diseases. She has been at Emergent for 15 years and has extensive experience in developing, managing, and executing non-clinical studies involving select agents for product development under the “Animal Rule”. She has published more than 20 papers in peer-reviewed journals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 62 Selin Ozkan Kotiloglu et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Multiplex detection of Aspergillus fumigatus mycoviruses Selin Ozkan Kotiloglu1, 2 and Robert H A Coutts1, 3 1Imperial College London, UK 2Ahi Evran University, Turkey 3University of Hertfordshire, UK
ycoviruses are viruses that naturally infect and replicate in fungi. They are widespread in all major fungal groups Mincluding plant and animal pathogenic fungi. Different dsRNA mycoviruses have been reported in Aspergillus fumigatus. Multiplex PCR amplification is a version of PCR, which enables amplification of different targets simultaneously. This technique has been widely used for detection and differentiation of viruses especially plant viruses such as those which infect tobacco, potato and garlic. For rapid detection, multiplex RT-PCR was developed to screen new isolates in terms of presence of A. fumigatus mycoviruses. AfuCV, AfuPV-1 and AfuTmV-1 dsRNAs were amplified in separate reactions using a mixture of multiplex primer pairs. It was demonstrated that in the presence of a single infection, primer pair mixtures only amplify the corresponding single virus infection. Mixed infections using dual or triple combinations of dsRNA viruses were also amplified simultaneously using multiplex RT-PCR. Up until now methods for the rapid detection, Aspergillus mycoviruses have been restricted to small scale dsRNA extraction approaches which are laborious and for large numbers of samples not as sensitive as RT-PCR. The multiplex RT-PCR assay developed here will be useful for the studies on determining the incidence of A. fumigatus mycoviruses. Moreover, it could be useful to detect mycovirus infection rapidly for further studies on the impact of mycoviruses on fungal pathogenicity. This is the first report on multiplex detection of A. fumigatus mycoviruses.
Biography Selin Ozkan Kotiloglu was awarded a Turkish Government Scholarship and earned her PhD in Molecular Biology/Molecular Mycology from Imperial College London. She is currently working as an Assistant Professor at Ahi Evran University, Department of Molecular Biology and Genetics. Her main research areas are gene expressions and polymorphism in various diseases caused both by biological and chemical agents; RNA silencing; profiling of small RNAs and microRNAs using bioinformatics; silencing pathways in various organisms.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 63 Nada Madi, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Metagenomic analysis of viral diversity in respiratory samples from patients with respiratory tract infections in Kuwait Nada Madi Kuwait University, Kuwait
metagenomic approach based on target independent next-generation sequencing has become a known method for the Adetection of both known and novel viruses in clinical samples. This study aimed to use the metagenomic sequencing approach to characterize the viral diversity in respiratory samples from patients with respiratory tract infections. We have investigated 86 respiratory samples received from various hospitals in Kuwait between 2015 and 2016 for the diagnosis of respiratory tract infections. A metagenomic approach using the next-generation sequencer to characterize viruses was used. According to the metagenomic analysis, an average of 145, 019 reads were identified, and 2% of these reads were of viral origin. Also, metagenomic analysis of the viral sequences revealed many known respiratory viruses, which were detected in 30.2% of the clinical samples. Also, sequences of non-respiratory viruses were detected in 14% of the clinical samples, while sequences of non-human viruses were detected in 55.8% of the clinical samples. The average genome coverage of the viruses was 12% with the highest genome coverage of 99.2% for respiratory syncytial virus, and the lowest was 1% for Torque teno midi virus 2. Our results showed 47.7% agreement between multiplex real-time PCR and metagenomics sequencing in the detection of respiratory viruses in the clinical samples. Though there are some difficulties in using this method to clinical samples such as specimen quality, these observations are indicative of the promising utility of the metagenomic sequencing approach for the identification of respiratory viruses in patients with respiratory tract infections. Recent Publications 1. Madi N, Al Nakib W, Mustafa A S, Khan M and Habibi N (2018) Metagenomic analysis of viral diversity in respiratory samples from patients with respiratory tract infections in Kuwait. Journal of Medical Virology 90(3):412-420. 2. Chehadeh W, Al Qaseer M, Albaksami O, Altawalah H, Suhail Ahmad, Madi N, John SE and Al Nakib W (2015) Phylogenetic analysis of HIV-1 subtypes and drug resistance profile in treatment-naïve people from Kuwait. Journal of Medical Virology 87(9):1521-6. 3. Madi N, Al Qasser M, Abdul Khalik D, Edan R and Al Nakib W (2015) Clinical utility of viral load management of CMV infection in SOT patients in Kuwait. Transplantation Proceedings 47(6):1802-7. 4. Al Tawalah H, Madi N and Al Qaseer M (2014) Prevalence of blood borne viruses in the dialysis unit, Mubarak Al-Kabeer Hospital. Kuwait Medical Journal 47(1):30-32. 5. Madi N, Al Tawalah H and Al Nakib W (2014) Clinical presentations of HHV-6 infection in infants and children in Kuwait: A retrospective study. Advances in Microbiology 4(15):1088-11094.
Biography Nada Madi is an Assistant Professor in the Department of Microbiology, Faculty of Medicine at Kuwait University where she has been a Faculty Member since January 2015. She completed her PhD and MSc at Faculty of Medicine, Kuwait University and her Undergraduate study at Faculty of Science, Kuwait University. Her research interest lies in the area of developing advanced techniques in viral diagnostics such as metagenomics approach for the detection of viruses causing different diseases such as respiratory tract infections and gastroenteritis.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 64 Yuchen Li et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
A novel pathway of virus dissemination within the host: Spread of porcine epidemic diarrhea virus (PEDV) from the nasal cavity to the intestinal mucosa in swine Yuchen Li and Weigang Nanjing Agricultural University, China
orcine epidemic diarrhea (PED) has made catastrophic impacts on the global pig industry since 2011. The causative Pagent, porcine epidemic diarrhea virus (PEDV), was a typical intestinal coronavirus and transmitted by the generally acknowledged fecal-oral route. However, high infectivity of airborne PEDV and quickly spread between pig farm (even over the far distance) indicated that airborne transmission may make contributions to the rapid spread of PEDV. This study demonstrated that PEDV could cause typical diarrhea in piglets through nasal spraying and the exact mechanisms involved has been well studied in vitro and in piglets. At first, PEDV was detected by immunohistochemistry test in nasal epithelium at early stages of the infection. Then, the results were further verified by establishing air liquid interface culture of pig’s nasal epithelial cells (NECs) in vitro. Moreover, PEDV captured by dendritic cells (DCs) in nasal passage were observed in nasal passage and DC/NECs co-culture system, demonstrating that PEDV could recruit DCs to the nasal epithelial cells (ECs) and form transepithelial dendrites (TEDs) to capture luminal viruses. Subsequently, PEDV carried DCs could form firm adhesion with T cells and transmit the virus to CD3+ T cells via virological synapse. Additionally, the virus loaded CD3+ T lymphocyte could enter the blood circulation through the lymphocyte recirculation and reach the intestinal mucosa. Finally, virus caused infection in intestinal epithelium (Vero cells, susceptible cells for PEDV) by CD3+ T cells medicated transfer infection. Our finding is the first to demonstrate a novel pathway of PEDV dissemination within host and illustrated the mechanism of it transport from entry site to pathogenic site, which sheds light on prevention measures and pathogenic mechanism for viruses with the same characteristics.
Recent Publications 1. Li Y, Wang G, Wang J, Man K and Yang Q (2017) Cell attenuated porcine epidemic diarrhea virus strain Zhejiang08 provides effective immune protection attributed to dendritic cell stimulation. Vaccine 35(50):7033-41. 2. Li Y, Cai Y, Tao J, Kang X, Jiao Y, Guo R, Wang G, Pan Z and Jiao X (2016) Salmonella isolated from the slaughterhouses and correlation with pork contamination in free market. Food Control 59:591-600. 3. Li Y C, Pan Z M, Kang X L, Geng S Z, Liu Z Y, Cai Y Q and Jiao X A (2014) Prevalence, characteristics, and antimicrobial resistance patterns of Salmonella in retail pork in Jiangsu province, eastern China. Journal of Food Protection 77(2):236-45
Biography Yuchen Li has been studying in Nanjing Agricultural University as a PhD candidate since 2015. Currently, he is studying in the major of preventive veterinary medicine under supervision of Professor Qian Yang. His research focuses on the pathogenic mechanism of PEDV. He has published three research articles and interested in the research process.
[email protected] Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 65 Qian Yang et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Persistent TGEV infection enhances ETEC K88 adhesion by promoting epithelial-mesenchymal transition in intestinal epithelial cells Qian Yang and Weigang Nanjing Agricultural University, China
ransmissible gastroenteritis virus (TGEV) is a coronavirus, characterized by diarrhea, high morbidity, and the mortality is T100% in piglets less than 2 weeks old. Pigs infected with TGEV often suffer from secondary infection with other pathogens, which aggravates the severity of diarrhea, but the mechanisms remain unknown. Here, we hypothesized that persistent TGEV infection stimulates the epithelial–mesenchymal transition (EMT), thereby generating cells that more easily adhere to enterotoxigenic Escherichia coli (ETEC). Intestinal epithelial cells are the primary targets of TGEV and ETEC infection. We found that TGEV can persistently infect porcine intestinal columnar epithelial cells (IPEC-J2), and cause EMT, consistent with multiple changes in key cell characteristics. Infected cells display fibroblast-like shapes, exhibit increases in mesenchymal markers with a corresponding loss of epithelial markers, have enhanced expression of IL-1β, IL-6, IL-8, TGF-β, and TNF-α mRNAs, and demonstrate increases in migratory and invasive behaviors. Additional experiments showed that activation of the PI3K/Akt and ERK signaling pathways via TGF-β are critical for the TGEV-mediated EMT process. Cellular uptake is also modified in cells that have undergone EMT. TGEV-infected cells have higher levels of integrin α5 and fibronectin and exhibit enhanced adhesion of ETEC K88. Reversal of EMT reduces ETEC K88 adhesion and inhibits the expression of integrin α5 and fibronectin. Overall, these results suggest that TGEV infection induces EMT in IPEC-J2 cells, increasing the adhesion of ETEC K88 in the intestine and facilitating dual infection.
Figure 1: Schematic diagram showing a persistent TGEV infection induces EMT and promotes bacterial adhesion. TGEV infection induces the production of TGF-β, activates the P13K/Akt and ERK signaling pathways and induces the EMT markers. During TGEV- induced EMT, the expression of integrin α5 and fibronectin is enhanced, leading to increased bacterial adherence.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 66 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Recent Publications 1. Yin Yinyan, Qin Tao, Wang Xiaoqing, Lin Jian, Yu Qinghua and Yang Qian (2015) CpG DNA assists the whole inactivated H9N2 influenza virus in crossing the intestinal epithelial barriers via transepithelial uptake of dendritic cell. Mucosal Immunology 8(4):799-814.
2. Liu H, Xu W, Yu Q, Yang Qian et al. (2017) 4,4′-Diaponeurosporene-producing Bacillus subtilis increased mouse resistance against Salmonella typhimurium infection in a CD36-dependent manner. Frontiers in Immunology 8:483.
3. Xia L, Dai L, Yu Q, et al. (2017) Persistent TGEV infection enhances ETEC K88 adhesion by promoting epithelial- mesenchymal transition in intestinal epithelial cells. Journal of Virology DOI:10.1128/JVI.01256.
4. Qin Tao, Yin Yinyan, Yu Qinghua, Huang Lulu, Wang Xiaoqing, Lin Jian and Yang Qian (2015) CpG oligodeoxynucleotides facilitate delivery of whole inactivated H9N2 influenza virus via transepithelial dendrites of dendritic cells in nasal mucosa. Journal of Virology 89(11):5904-5918.
Biography Qian Yang worked as a Professor in College of Veterinary Medicine. Her research work focuses on the mucosal immunity in domestic animals. At first, several mucosal adjuvants were studied to increase the efficiency of inactivated viruses (influenza). Secondly, some delivery vehicles (Lactobacillus and Bacillus subtilis) for antigens for oral immunization have been studied because of degradation of the antigen by gastric acid and proteases present in the gastrointestinal tract. The other fields includes: the interaction between epithelium and pathogens and the relevant pathogenic mechanism. She has published more than 80 original and review articles in scientific journals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 67 conferenceseries.com
JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Scientific Tracks & Abstracts Day 2
Euro Virology 2018 & Influenza 2018
Page 72 Sessions: Day 2 July 03, 2018 Bacterial Virology |Plant and Agricultural Virology |HIV, AIDS and other Emerging Viruses Veterinary Virology |Current Focus in Virology Research | Viral Vaccines | Zoonotic Diseases: Global Infectious Disease Burden | Evolution and Epidemiological Aspects of Influenza and Zoonotic Diseases Antiviral treatment for flu and public health policy Session Chair Session Co-Chair Bryan R. Cullen John A Walsh Duke University Medical Center, USA University of Warwick, UK
Session Introduction Title: Isolation, characterization and application for phage biocontrol of bacteriophages infecting Acidovorax citrulli, the causal agent of bacterial fruit blotch Tae-Jin Choi, Pukyong National University, South Korea Title: Differentiation of Subtypes for Influenza Surveillance using a Peptide-Based Detection Platform (FluType) Henry Memczak, Stanford University School of Medicine, USA Title: The various roles of CsRDR1 family genes in cucumber defense against different viruses Diana Leibman, The Volcani Center, Israel Title: Novel platform (wEB) to study flu virus evolution and predict vaccine efficacy Slobodan Paessler, University of Texas Medical Branch, USA Title: Nonsynonymous mutation in integrase catalytic core domain affects feline foamy viral DNA integration Cha-Gyun Shin, Chung-Ang University, Republic of Korea Title: Characterization of a Rsv3 Gene that confers Strain-Specific Resistance to Soybean Mosaic Virus Kook-Hyung Kim, Seoul National University, Republic of Korea Title: Pathogenesis of infectious pulmonary bronchiolitis associated with flu related viral respiratory illness and the drastic impact on global resources S.E. Morgan, University of Chicago Medicine, USA Title: Global pandemic influenza vaccine preparedness: Progress under the Global Action Plan for Influenza Vaccines and next steps Christopher Chadwick, World Health Organization, Switzerland Title: ORF1a of RVRp22, a ribavirin-resistant attenuated phenotype of PRRSV, plays an important role on viral virulence and genetic stability assessed in pigs Amina Khatun, Chonbuk National University (CBNU), Republic of Korea Title: Characterization of a granulovirus from the fall webworm, hyphantria cunea Remziye Nalcacioglu, Karadeniz Technical University, Turkey Title: Mimivirus on a bad hair day Yael Fridmann Sirkis, Weizmann Institute of Science, Israel Title: Agaricus brasiliensis sulfated polysaccharide prevents DENV-2 NS1 induced endothelial barrier dysfunction in vitro Francielle Tramontini Gomes de Sousa, University of California –Berkeley, USA Title: Chinese Surveillance Results: Was the Influenza Activity Level of 2017-2018 Influenza Season Close to the 2009 Influenza Pandemic? Chen Tao, Chinese National Influenza Center, China Euro Virology 2018 & Influenza 2018
Page 73 Tae Jin Choi et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Isolation, characterization and application for phage bio control of bacteriophages infecting Acidovorax citrulli, the causal agent of bacterial fruit blotch Tae Jin Choi, Aryan Rahimi Midani, Yelin Kim and Min Jeong Kim Pukyong National University, South Korea
Statement of Problems: Bacterial fruit blotch (BFB) is an economically important bacterial disease that has caused huge economic losses in melon and watermelon crops around the world. There is no commercially available cultivars resistance to this disease caused by bacteria Acidovorax citrulli. Mainly, two genotypes (genotype I and II) are reported in A. citrulli, in which genotype II is the main causal agent of BFB in water melon that is major problem in Korea. Methodology & Theoretical Orientation: We have isolated more than 50 bacteriophages infecting A. citrulli, from watermelon leaf samples which were collected from different parts of Korea. Two of the isolated phages with large plaque size named as ACP17 and ACPHW were further characterized and used for phage biocontrol. Findings: Based on electron microscope observations, ACP17 belongs to Myoviridae family with head diameter 100±5 nm and a tail length of 150±5 nm while ACPHW has a head size of 60±5 nm and tail size 180±5 nm which belongs to Siphoviridae family. Among forty A. citrulli strains, ACP17 can lyse 27 strains of which most belongs to genotype I, and ACPHW can lyse 39 strains containing group I and II. In planta assay showed that the germination rate of watermelon seeds coated with the bacteriophages was up to 80% in the presence of A. citrulli contrast to untreated seed showing no germination. Also, these germinated plants showed 100% survival in A. citrulli treated soil. Conclusion & Significance: These results suggest the possible use of these phages as an effective bio control agent for BFB.
Recent Publications 1. Rahimi Midani A, Lee S, Kang S W, Kim M K and Choi T J (2018) First isolation and molecular characterization of bacteriophages infecting Acidovorax citrulli, the causal agent of bacterial fruit blotch. The Plant Pathology Journal 34(1): 59–64. 2. Frampton R A, Pitman A R and Finerann P C (2012) Advances in bacteriophage-mediated control of plant pathogens. International Journal of Food Microbiology Article ID 326452:1-11. 3. Zivanovic M and Walcott R R (2016) Further characterization of genetically distinct groups of Acidovorax citrulli strains. Phytopathology 107(1):29-35. 4. Borah P K, Jindal J K and Verma J P (2000) Integrated management of bacterial leaf spot of mungbean with bacteriophages of Xav and chemicals. J. Mycol. Plant Pathol 30:19-21.
Biography Tae Jin Choi has completed his PhD from University of California, Berkeley in 1993 and 2 years Postdoctoral studies from University of Wisconsin, Madison. Since 1995 he is working as a Professor in Pukyong National University in Busan Korea. He had published over 70 papers on the viruses of plant, fish, shrimp and vaccines for aquatic viruses. Recently he is working on the development of microalgae as bioreactor for recombinant protein production and bacteriophage biocontrol of water melon.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 74 Henry Memczak et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Differentiation of subtypes for influenza surveillance using a peptide-based detection platform (FluType) Henry Memczak, Marc Hovestadt, Bernhard Ay, Sandra Sanger, Jan Grzegorzewski, Matthias Konig, Thorsten Wolff and Frank F Bier Stanford University, USA
he only cost-effective protection against influenza is vaccination. Due to rapid mutation continuously new subtypes appear, Twhat requires annual reimmunization. For a correct vaccination recommendation, the circulating influenza strains have to be detected promptly and exactly and characterized regarding their antigenic properties. Due to recurring incidents of vaccine mismatches, there is a great need to speed up the process chain from identifying the right vaccine strains to their administration. The monitoring of subtypes as part of this process chain is carried out by national reference laboratories within the WHO Global Influenza Surveillance and Response System (GISRS). To this end, thousands of viruses from patient samples (e.g. throat smears) are isolated and analyzed each year. Currently this analysis involves complex and time-intensive (several weeks) animal experiments to produce specific hyperimmune sera in ferrets, which are necessary for the determination of the antigen profiles of circulating virus strains. These tests also bear difficulties in standardization and reproducibility, which restricts the significance of the results. To replace this test a peptide-based assay for influenza virus subtyping is developed. The differentiation of the viruses takes place by a set of specifically designed peptidic recognition molecules which interact differently with the different influenza virus subtypes. The differentiation of influenza subtypes is performed by pattern recognition guided by machine learning algorithms, without any animal experiments. Recent Publications 1. Memczak, Henry & Lauster, Daniel & Kar, Parimal & Di Lella, Santiago & Volkmer, Rudolf & Knecht, Volker & Herrmann, Andreas & Ehrentreich-Förster, Eva & Bier, Frank & F. M. Stöcklein, Walter. (2016). Anti-Hemagglutinin Antibody Derived Lead Peptides for Inhibitors of Influenza Virus Binding. PLoS ONE. 11. 10.1371/journal.pone.0159074. 2. Memczak, Henry & Stöcklein, W & Ehrentreich-Förster, E & Lauster, Daniel & Bier, Frank & Herrmann, Andreas. (2014). Peptides for binding of influenza viruses and inhibition of influenza infections. 3. Hoppe, Sebastian & Memczak, Henry. (2015). Biomarkers: From Discovery to Commercialization. Biomarker Validation: Technological, Clinical and Commercial Aspects. 183-205. 10.1002/9783527680658.ch10. 4. Siemeling, Ulrich & Memczak, Henry & Bruhn, Clemens & Vogel, Florian & Träger, Frank & E Baio, Joe & Weidner, Tobias. (2012). Zwitterionic dithiocarboxylates derived from N-heterocyclic carbenes: Coordination to gold surfaces. Dalton transactions (Cambridge, England : 2003). 41. 2986-94. 10.1039/c2dt11976e. 5. Hovestädt, Marc & Memczak, Henry & Pleiner, Dennis & Zhang, Xin & Rappich, Joerg & Bier, Frank & F. M. Stöcklein, Walter. (2014). Characterization of a new maleimido functionalization of gold for surface plasmon resonance spectroscopy. Journal of Molecular Recognition. 27. 10.1002/jmr.2396.
Biography Henry Memczak studied nanotechnology at the University of Kassel, Germany and completed his PhD in biochemistry in 2014 at the University of Potsdam, Germany and the Fraunhofer Institute for Cell Therapy and Immunology, Germany. He has worked on the development of analytical biosensors for influenza detection and methods for peptide-based biointeraction analysis for several years, published several papers, holds two patents and co-founded the company qpa bioanalytics GmbH for the commercialization of novel peptide biochips. For his dedicated translational research he received several awards and scholarships.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 75 Diana Leibman et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
The various roles of CsRDR1 family genes in cucumber defense against different viruses Diana Leibman, Dalia Woolf and Amit Gal-On Agricultural Research Organization, Israel
NA-dependent RNA polymerases (RDR) play an important role in virus protection and plant gene control. Their activity Ris based on the synthesis of double stranded RNA that is a template for the gene silencing system. Four RDR1 (RDR1a, b, c1, c2) genes were identified in cucumber having different expression levels before and after infection by various viruses. The CsRDR1a, b, c genes have a homology of 60-58% at the amino acid level, while CsRDR1c1 and c2 are almost identical (97% homology), but have different promoter sequences. A high expression level of CsRDR1b was characterized in cucumber strains with partial resistance to a number of viruses, while CsRDR1c1 and c2 not expressed in healthy plants. Cucumber plants infected by several different viruses showed a moderate expression level ofCsRDR1b and a dramatic level of CsRDR1c1, c2. The expression level of CsRDR1a, CsRDR2 and CsRDR6 did not increase following virus infection. The differential expression of CsRDR1b and CsRDR1c1, c2 indicates dissimilar control of these genes. The relationship between the high levels of expression of CsRDR1b in resistant cucumber cultivars reinforces the assumption that this gene is unique for virus resistance. Using the CRISPR/Cas9 system, we created a variety of mutants in the rdr1b and rdr1c1 and c2 genes. Knockdown of these genes increased virus accumulation. Plants with a homozygous mutation in the rdr1b genes (deletion of 34 nucleotides) and rdr1c (2 and 1 nucleotides in the rdr1c1 and rdr1c2 genes respectively) showed increased susceptibility to Cucumber mosaic virus 4 dpi, especially rdr1c mutants plants which collapse 6 days after infection. The viral symptoms increased also after Cucurbit vein yellowing virus infection, whereas the susceptibility to Zucchini yellow mosaic virus infection was less. Notably, CsRDR1b expression in healthy rdr1b mutants was less than in non-mutant plants. The control of CsRDR1b and CsRDR1c1, c2 is under study in our laboratory to help understand the silencing mechanism of plants defense. Recent Publications 1. Leibman D, Kravchik M, Wolf D, Haviv S, Weissberg M, Ophir R, Paris H S, Palukaitis P, Ding S W, Gaba V and Gal- On A (2017) Differential expression of cucumber RNA-dependent RNA polymerase 1 genes during antiviral defense and resistance. Molecular Plant Pathology 19(2):300-312. 2. Chandrasekaran J, Brumin M, Wolf D, Leibman D, Klap C, Pearlsman M, Sherman A, Arazi T and Gal-On A (2016) Development of broad virus resistance in non-transgenic cucumber using CRISPR/Cas9 technology. Molecular Plant Pathology 17(7):1140-53. 3. Leibman D, Wolf D, Saharan V, Zelcer A, Arazi T, Shiboleth M Y, Gaba V, and Gal-On A (2011) A high level of transgenic viral small RNA is associated with broad potyvirus resistance in cucurbits. Molecular Plant-Microbe Interactions 24(10):1220– 1238.
Biography Diana Leibman completed her PhD in 2012 from the Hebrew University in Jerusalem. She works as a Research Engineer in the Agricultural Research Organization, The Volcani Center. The research topics in which she is involved are: The role of RNA-dependent RNA polymerase 1 in plant defense against viruses; CRISPR/Cas9 genome editing technology for crop improvement; development of resistance to RNA and DNA viruses in cucurbits and tomato by transgenic approaches; identification of plant genes associated with disease symptom development to plant virus infection and; genetic engineering of attenuated ZYMV-AG as a plant virus vector, for gene expression and epitope presentation.
[email protected] Notes:
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 76 Slobodan Paessler et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Novel platform (wEB) to study flu virus evolution and predict vaccine efficacy Slobodan Paessler1 and Veljko Veljkovic2 1Galveston National Laboratory, USA 2Biomed Protection, USA
lu epidemics and potential pandemics pose great challenges to public health institutions, scientists and vaccine producers. FCreating right vaccine composition for different parts of the world is not trivial and has been historically very problematic. This often resulted in decrease in vaccinations and reduced trust in public health officials. To improve future protection of population against flu we urgently need new methods for vaccine efficacy prediction and vaccine virus selection. Recently, novel bioinformatics platform based on electronic biology was successfully utilized for real-time monitoring of influenza A viruses as well as for prediction of vaccine efficacy in Australia and USA in 2017 and 2018. Here we present wEB platform and its usage in identifying functional biological changes in haemagglutinin protein of influenza A viruses and how this knowledge can be applied for vaccine design, prediction of future vaccine efficacy and real-time virus evolution monitoring.
Biography Slobodan Paessler, is a Professor in the Department of Pathology and Director of Galveston National Laboratory Preclinical Studies Core. Dr. Paessler is a co-principle investigator on the Universal Influenza Vaccine project funded by an NIAID grant at Etubics Corporation. He serves as the Director of Animal Biosafety Level 3 for the Institute of Human Infections and Immunity. He has been Member of Scientific Advisory Board at Etubics Corporation since July 2015. He serves as a Member of the Center for Biodefense & Emerging Infectious Diseases. He received a Dr. Med Vet (D.V.M) at Ludwig-Maximilian University and a Ph.D in Experimental Pathology from UTMB.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 77 Cha Gyun Shin et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Nonsynonymous mutation in integrase catalytic core domain affects feline foamy viral DNA integration Cha Gyun Shin, Ga Eun Lee and Jinsun Kim Chung-Ang University, South Korea
ntegrase is the retroviral protein responsible for incorporating a double-stranded viral DNA copy into the host chromosome. ISingle amino acid substitution in integrase (IN) active site is replication-defective, and reduces viral infectivity. This study is to investigate whether mutation in DD(35)E motif and residues near the active site of catalytic core domain is critical for feline foamy viral integration. In vitro enzymatic activities of IN mutant proteins were analyzed by using p32-radiolabeled oligonucleotide substrates and 15% polyacrylamide gels. DDE mutation almost lost their IN activities. But Q165A, Y191A, and S195A showed reduced effects. Although DDE mutants produced replication-defective virions by one cycle transfection, Q165A, Y191A, and S19A mutants had infectivity on their natural host cells. Known as the immature virions containing mutated IN executed integration aberrantly and had trouble in producing viral DNAs for new virion particles, in the case of DDE mutants no integrated viral DNAs were detected at 24 h and 48 h post infected host chromosomal DNAs. However, integration of viral DNA whose virions have IN mutants in the amino acid residues present near the active site such as Q165A and Y191A were detected, and then quantitated by competitive PCR. Nonsynonymous substitutions in highly conserved region of feline foamy viral IN resulted in viruses with replication-defective. Defects in viral DNA synthesis, viral production, and integration processing were observed for all of the replication-defective mutants. Especially, feline foamy viral natural host cells are not infected with replication-defective DDE mutant viruses.
Biography Cha Gyun Shin has completed his PhD from The Ohio State University and Postdoctoral studies from Dana Faber Cancer Institute. He is a Professor at Department of Systems Biotechnology, Chung-Ang University, Republic of Korea.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 78 Kook Hyung Kim et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Characterization of a RSV3 gene that confers strain-specific resistance to soybean mosaic virus Kook Hyung Kim1, Phu Tri Tran1, Kristin Widyasari1 and Jang Kyun Seo2 1College of Agriculture and Life Sciences - Seoul National University, South Korea 2Institutes of Green Bio Science and Technology - Seoul National University, South Korea
oybean mosaic virus, a member of the genus Potyvirus, significantly reduces soybean production worldwide. Rsv3, which Sconfers strain-specific resistance to SMV, was previously mapped between the markers A519F/R and M3Satt in chromosome 14 of the soybean [Glycine max (L.) Merr.] genotype L29. Analysis of the soybean genome database revealed that five different NBS-LRR sequences exist between the flanking markers. Among these candidate Rsv3 genes, the full-length cDNA of the Glyma.14g204700 was successfully cloned from L29. Over-expression of Glyma.14g204700 in leaves inoculated with SMV inhibited viral infection in a soybean genotype lacking Rsv3. In addition, the transient silencing of the candidate gene caused a high accumulation of a virulent strain in L29 carrying Rsv3. Our results therefore provide additional line of evidence to support that Glyma.14g204700 is likely Rsv3 gene that confers strain-specific resistance to SMV.
Recent Publications 1. Seo JK, Kang SH, Seo BY, Jung JK, Kim KH (2010) Mutational analysis of interaction between coat protein and helper component-proteinase of soybean mosaic virus involved in aphid transmission. Molecular Plant Pathology 11: 265-276. 2. Seo JK, Kwon SJ, Cho WK, Choi HS, Kim KH (2014) Type 2C protein phosphatase is a key regulator of antiviral extreme resistance limiting virus spread. Scientific Reports 4: 5905. 3. Seo JK, Lee SH, Kim KH (2009) Strain-specific cylindrical inclusion protein of soybean mosaic virus elicits extreme resistance and a lethal systemic hypersensitive response in two resistant soybean cultivars. Molecular Plant-Microbe Interaction 22: 1151-1159. 4. Seo JK, Vo Phan MS, Kang SH, Choi HS, Kim KH (2013) The charged residues in the surface-exposed C-terminus of the soybean mosaic virus coat protein are critical for cell-to-cell movement and virion assembly. Virology 446: 95-101. 5. Tran PT, Widyasari K, Kim KH (2018) Isolation and validation of a candidate Rsv3 gene from a soybean genotype that confers strain-specific resistance to soybean mosaic virus. Virology 513: 153-159. 6. Tran PT, Widyasari K, Park JY, Kim KH (2017) Engineering an auto-activated R protein that is in vivo activated by a viral protease. Virology 510: 242-247.
Biography Dr. Kim has completed his PhD from the Department of Plant Pathology, North Carolina State University (NCSU) and postdoctoral studies from NCSU Department of Biochemistry. He is the director of Plant Clinic, Seoul National University. He has published more than 100 papers in reputed journals and has been serving as an editorial board member of Virology, Virus Research, and Scientific Reports.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 79 S E Morgan et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Pathogenesis of infectious pulmonary bronchiolitis associated with flu related viral respiratory illness and the drastic impact on global resources S E Morgan, S Mosakowski, G V Mathai, J R Lyerly, LT Grays-Walton, U Dyamenahalli and J P Kress University of Chicago Medicine, USA
he mechanics of flu related respiratory illness is not completely implicit as it includes; influenza, zoonotic and non- Tinfluenza pathogens. Precise diagnosis is difficult as it often mimics asthma out of control which has perplexed researchers for decades. This has led to treatment confusion and an underestimation that the primary cause of breathing difficulties is related to bronchiolitis-bronchiectasis. A microbiology respiratory viral panel (RVP) test via polymerase chain reaction (PCR) can identify whether there is a co-existing viral lung infection that may worsen the lung function. Viral flu-related respiratory infections are highly transmittable and may increase morbidity and mortality in patients with premorbid pulmonary disease and weakened immune systems. The symptoms of flu include dyspnea and coughing; after usual treatment with steroids and asthma medications, continue to have worsening symptoms causing re-hospitalization. Chest radiography for patients with respiratory distress due to flu are notable for; bronchial wall thickening, bronchiectasis and sub-segmental atelectasis, related air-flow obstruction. Rhinoviruses (RV) – enterovirus (EV) for example is under recognized as the leading cause of hospitalization for viral outbreaks. Respiratory Enterovirus is responsible for 10 to 15 million hospitalizations annually. Enterovirus (D-68) was attributed to 14 deaths in 2014 in the United States (USA) and 70 deaths in the 2011 Philippines D68 outbreak. Ever since the 2014 D68 outbreak, there has been a drastic increase in the number of patients hospitalized and re- hospitalized for flu symptoms associated with severe acute respiratory distress on the pediatric and oncology wards. Zoonotic agents such as coronavirus (HCoV) are passed bi-directionally between animals and humans and capable of joining with other viral agents. All this has created undefined burden on global clinical resources. More research is needed to understand the pathogenesis of viral bronchiolitis and bronchiectasis related respiratory illness to assist clinicians with recognition and treatment of this highly morbid disease. Recent Publications 1. Morgan, Sherwin & Mosakowski, Steve & Ostrawski, Stephanie & Logan, Richard & Garrity, Edward. (2017). Use of High Flow Nasal Cannula and Aerosolized Epoprostenol as a Bridge to Lung Transplantation. 2. Morgan, Sherwin. (2016). Forty Percent Heliox as an Adjunctive Therapy to Mechanically Ventilate a Child with Rhinovirus/ Enterovirus Related Respiratory Failure. 10.13140/RG.2.2.21314.38086. 3. Morgan, Sherwin & Mosakowski, Steve & Solano, Patti & Hall, Jesse & Tung, Avery. (2015). High-Flow Nasal Cannula and Aerosolized Agonists for Rescue Therapy in Children With Bronchiolitis: A Case Series. Respiratory care. 60. 10.4187/ respcare.03996. 4. Morgan, Sherwin & Vukin, Kirissa & Mosakowski, Steve & Solano, Patti & Stanton, Lolita & Lester, Lucille & Lavani, Romeen & Hall, Jesse & Tung, Avery. (2014). Use of Heliox Delivered via High-Flow Nasal Cannula to Treat an Infant With Coronavirus-Related Respiratory Infection and Severe Acute Air-Flow Obstruction. Respiratory care. 59. 10.4187/ respcare.02728. 5. Tung, Avery & L Drum, Melinda & Morgan, Sherwin. (2005). Effect of inspiratory time on tidal volume delivery in anesthesia and intensive care unit ventilators operating in pressure control mode. Journal of clinical anesthesia. 17. 8-15. 10.1016/j.jclinane.2004.02.005.
Biography S E Morgan completed his respiratory care training from Malcolm X College of Respiratory Care in Chicago, IL. He is an advanced respiratory care practitioner with the National Board for Respiratory Care in the United States. He is Clinical Practice and Development /Educator/Research Coordinator for the Department of Respiratory Care Services, Section of Pulmonary and Critical Care Medicine at the University of Chicago Medicine. He has published more than 25 peer review papers in multiple medical journals. He has designed, engineered, and collaborated with a number of research studies with the pulmonary medicine department.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 80 Christopher Chadwick, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Global pandemic influenza vaccine preparedness: Progress under the global action plan for influenza vaccines and next steps Christopher Chadwick World Health Organization, Switzerland
Purpose: The World Health Organization’s Global Action Plan for Influenza Vaccine (GAP) was a 10-year initiative dedicated to reducing the global shortage and inequitable access to influenza vaccines in the event of an influenza pandemic. The overarching goal of the GAP was to develop the capacity to produce enough vaccines to immunize 70% of the global population with two doses of vaccines. The GAP aimed to achieve this goal by increasing evidence based seasonal influenza vaccine use; developing influenza vaccine production and regulatory capacity in 14 low and middle income countries (LMICs) and; encouraging the development of improved influenza vaccines. Methods: Between 2006 and 2016, the WHO collaborated with member states and key stakeholders to address the global shortage of and increase equitable access to pandemic influenza vaccines in the event of an outbreak. Results: The outcomes of the GAP include: A dramatic increase in countries with a seasonal influenza policy in place (115 member states by 2014 from a baseline of 74 in 2006); the development of 8 licensed pandemic influenza vaccines and 3 licensed seasonal influenza vaccines in 6 LMICs and; A global expansion of pandemic vaccine production capacity, especially in LMICs (potential global capacity of 6.4 billion doses estimated in 2015). Discussion: Following the conclusion of the GAP in 2016, priorities for influenza vaccine preparedness moving forward are to sustain the production capacity of influenza manufacturers in LMICs, promote and stimulate innovative influenza vaccine research and development, identify root causes of influenza vaccine hesitancy, generate more evidence on vaccine effectiveness in specific risk groups, and identify innovative ways of addressing global pandemic influenza preparedness. Recent Publications 1. Nannei C, Chadwick C, Fatima H, Goldin S, Grubo M, Ganim A. (2016) Considerations for sustainable influenza vaccine production in developing countries. Vaccine. 2016 Oct 26;34(45):5425-5429. doi: 10.1016/j.vaccine.2016.08.056 2. Petrie MJ, Blakey CM, Chadwick C, Davies HG, Blundell CM, Davies MB (2018) A new and reliable classification system for fractures of the navicular and associated injuries to the midfoot. Bone Joint J. 2018 Feb;100-B(2):176-182. doi: 10.1302/0301-620X.100B2.BJJ-2017-0879.R1. 3. Loewenstern J, Hernandez CM, Chadwick C, Doshi A, Banik R, Sarkiss CA, Bederson J, Shrivastava RK. (2018) Optical Coherence Tomography in the Management of Skull Base Fibrous Dysplasia with Optic Nerve Involvement. World Neurosurg. 2018 Jan;109:e546-e553. doi: 10.1016/j.wneu.2017.10.018.
Biography Christopher Chadwick is a Technical Officer in the Global Action Plan for Influenza Vaccines Secretariat at World Health Organization. Previously, he was a Global Health Officer in the office of Global Affairs at the US Department of Health and Human Services. He received a Master of Science degree in Public Health with a concentration in microbiology and emerging infectious diseases from Milken Institute School of Public Health at George Washington University and a Bachelor of Science in Microbiology from Louisiana State University.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 81 Amina Khatun et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
ORF1a of RVRp22, a ribavirin-resistant attenuated phenotype of PRRSV, plays an important role on viral virulence and genetic stability assessed in pigs Amina Khatun1, Nadeem Shabir1, 2, Salik Nazki1, Chang Gi Jeong1 and Won Il Kim1 1Chonbuk National University, Specialized Campus, South Korea 2Sher-e-Kashmir University of Agricultural Sciences and Technology, India
ow genetic fidelity and limited cross-protection are two major hurdles faced in porcine reproductive and respiratory Lsyndrome virus (PRRSV) vaccinology, which is the most challenging threat to the swine industries worldwide. MLV vaccines are commonly used for homologous protection though there have been safety concerns as vaccine viruses reverted to virulence reported in fields. RVRp22, a ribavirin-resistant attenuated phenotype of PRRSV had high genetic stability during sequential replication in pigs reported in previous study. Wherein, ORF1a of RVRp22 genome was assumed to be involved in enhanced genetic stability and viral virulence. Therefore, the present study constructed four chimeric viruses based on ORF1a, 1b, 1ab and nsp2 regions of RVRp22 genome named as RVRp22-1a, 1b, 1ab and nsp2, respectively into VR2332 backbone to see the potential role of ORF1a in viral virulence and genome stability during serial passages in pigs following in vitro evaluation in MARC-145 cells and PAMs. In results, the replication of RVRp22-1a, 1ab and nsp2 was significantly suppressed in PAMs but replicated efficiently in MARC-145 like RVRp22 (attenuated strain). RVRp22-1b replicated in both of PAMs and MARC-145 cells with the similar trend as found in VR2332 (prototype strain). Consistently, RVRp22-1a, 1ab and nsp2 challenged pigs had lower viral loads in sera and lungs than RVRp22-1b, which was maintained even after third passage. Furthermore, RVRp22-1a, 1ab and nsp2 showed significantly a lower mutation frequency in nsp2 (most variable region in PRRSV genome) compared to RVRp22-1b. In conclusion, the present study indicated that ORF1a of RVRp22 genome might be critically involved in PRRSV virulence and the attenuated replication was maintained during serial passages in pigs which were enhanced with high genetic stability.
Biography Amina Khatun has completed her PhD in Veterinary Medicine (Microbiology) from Chonbuk National University (CBNU), South Korea. Currently, she is working as a Postdoctoral Researcher in the Department of Veterinary Immunology, College of Veterinary Medicine, CBNU. Her major research mostly focuses on functional genomics of PRRSV largely prospecting of viral pathogenesis to address two major hurdles in PRRSV vaccinology i.e. low genetic fidelity and limited cross-protection. Previously, she has completed her DVM and MS (Veterinary Pathology) from SAU, Sylhet, Bangladesh. She has published more than 10 papers in reputed journals. She has also been serving as a Reviewer of Ecology and Evolution Journal.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 82 Remziye Nalcacioglu et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Characterization of a granulovirus from the fall webworm, Hyphantria cunea (Drury) (Lepidoptera: Arctiidae) Remziye Nalcacioglu, Zeynep Bayramoglu, Hacer Muratoglu, Zihni Demirbag and Ismail Demir Karadeniz Technical University, Turkey
broad survey of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) populations in agricultural and forested Aareas in the central black sea region of Turkey led to the detection of granuloviruses (GVs). Thirty insect cadavers were collected from different locations, 8 of which contained GVs. All of them were determined to be Hyphantria cunea granulovirus (HycuGV) by tissue PCR and sequence analysis. A selected isolate (Hc1) was characterized and tested against third instar larvae of H. cunea. Electron microscopy confirmed typical GV morphology with ovoid granules of approximately 318-546 nm × 174-240 nm. Each granule contained a single rod-shaped virion with a mean size of 35-51 nm × 202-341 nm. The genome was estimated to be 123 kb by restriction endonuclease analysis. Partial sequencing of the granulin ,late expression factor-8 (lef-8) and late expression factor-9 (lef-9) genes also confirmed the identity of the virus as HycuGV. A phylogenetic analysis based on these conserved genes, HycuGV-Hc1 grouped together with the previous HycuGV isolate (A5-1) and Estigmene acrea granulovirus (EsacGV) isolate from the same family. The LC50 of Hc1 isolate was 2.6×104 occlusion bodies (OBs/ml) against third instar H. cunea larvae. HycuGV-Hc1 caused 80.17% mortality with 109 OBs/ml in pot experiments performed in a greenhouse. This is the first study to describe a novel Turkish HycuGV-Hc1 isolate and preliminary data suggest the virus have a significant potential as an effective biopesticide for H. cunea control.
Biography Remziye Nalcacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses, iridoviruses.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 83 Yael Fridmann Sirkis, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Mimivirus on a bad hair day Yael Fridmann Sirkis Weizmann Institute of Science, Israel
canthamoeba polyphaga mimivirus (APMV) is the first giant virus discovered almost 15 years ago. It has uncommon Acharacteristics such as a stargate shape in one of its vertices through which its dsDNA is released into the cytoplasm. It also has an external thick fibril layer that was shown to be important for adhesion. This finding was based on mimivirus strain that suffered a drastic reduction in the number of its fibrils after a third of its genes lost activity during continuous passaging in germ-free amoebae (PMID:21646533). Here, we subcultured mimiviruses under normal conditions and continually passed them through 0.45 μm filter. Thus, a population of hair deficient (HD) viruses was enriched and particles were cloned and imaged. Genomic analysis of the filtered viruses revealed three mutations that affected only three genes. One of the mutations showed an in-frame deletion in L71 gene, a collagen-like protein that eliminated almost all of its collagen motif sequences. The resulting HD viruses revealed a significant reduction in their infection titer as well as substantially reduced virus yield. HD- infected amoebae also burst less readily.
Figure 1: Transmission electron microscopy sectioning of mimiviruses. Left: Hair-deficient mimivirus. Right: Wild-type.
Recent Publications 1. Fridmann Sirkis Y, et al. (2016) Efficiency in complexity: composition and dynamic nature of mimivirus replication factories. Journal of Virology 90:10039-10047.
Biography Yael Fridmann Sirkis has her expertise in Structural Biology as well as characterizing protein-protein interactions. She is currently working in the protein analysis unit at the Weizmann Institute of Science in Israel.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 84 Francielle Tramontini Gomes de Sousa et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Agaricus brasiliensis sulfated polysaccharide prevents DENV-2 NS1 induced endothelial barrier dysfunction in vitro Francielle Tramontini Gomes de Sousa1, 2, Henry Puerta Guardo2, Camila Malta Romano1, Eva Harris2 and Ester Cerdeira Sabino1 1University of Sao Paulo, Brazil 2University of California, Berkeley, USA
Introduction: Vascular leakage is an adverse outcome in response to dengue virus (DENV) infection, resulting in depleted intravascular volume and hypotension, which may evolve into hypovolemic shock. Dengue non-structural protein 1 (DENV NS1) has been shown to contribute to pathogenesis by directly triggering endothelial glycocalyx layer (EGL) degeneration, in a cytokine independent pathway, as well as by inducing the release of vasoactive cytokines from PBMCs, both leading to plasma leakage. Agaricus brasiliensis is a basidiomycete fungus native to Brazil widely consumed and studied due its therapeutic properties, most of them related to its polysaccharidic content. A (1→6)-(1→3)-beta-D-glucan isolated from A. brasiliensis fruiting bodies (FR) was chemically modified to produce its corresponding sulfated derivative (FR-S). Since there is no specific treatment for DENV and the current available Sanofi dengue vaccine does not confer full protection to the disease, reversing or preventing EGL degeneration has therapeutic potential on severe dengue. Methodology & Theoretical Orientation:FR-S was tested against DENV NS-1 induced endothelial barrier disruption by measuring trans-endothelial electrical resistance (TEER). The effect of FR-S on NS-1 binding to endothelial cells was evaluated by immunohistochemistry. Findings: Figure 1 show that FR-S completely inhibited TEER reduction induced by DENV-2 NS1 treatment on HPMECs at the two higher concentrations tested (0.25 and 0.12 μg/mL). At the two lower tested concentrations (0.06 and 0.03 μg/mL), no protection against NS1-induced TEER reduction was observed. Results in figure 2 show that the mechanism by which FR-S prevents endothelial barrier dysfunction includes inhibition of NS1 binding to HPMECs at a concentration dependent manner. Conclusion & Significance: The findings indicate that FR-S inhibits NS1 binding to endothelial cells and prevents NS1 induced endothelial dysfunction. FR-S may have an anti-vascular leak effect since NS1 is in part responsible for the plasma leakage occurring in patients with severe dengue.
Figure 1: Effect of FR-S on endothelial dysfunction induced by DENV- Figure 2: Effect of FR-S on DENV-2 NS1 binding to HPMECs. Human 2 NS1. Human pulmonar microvascular endothelial cells (HPMECs) pulmonar microvascular endothelial cells (HPMECs) monolayers monolayers in contact or not with 10 µg/mL of DENV-2 NS1 were grown in gelatin coated coverslips were treated or not with two different treated with FR-S at four different concentrations. Trans-Endothelial concentrations ofFR-S (0.25 and 0.025 ug/mL) for 1 h. Then 10 ug/mL Electrical Resistance (TEER) was measured at sequential 2-hour time of DENV-2 NS1 histidine tag was added to pre-chilled cells. After 1 h of points following the treatments. The groups were compared through contact with NS1, monolayers were fixed and stained with Hoechst (blue) two-way ANOVA + Dunnett’s test. *Groups NS1+media, NS1+FR-S and anti-His Tag Alexa 547 (red). Fluorescence intensity, measured by 0.06 µg/mL, NS1+FR-S 0.03 µg/mL displayed statistically significant ImageJ software, indicate that FR-S 0.25 and 0.025 ug/mL treatments lower TEER values when compared to the other groups (p<0.05) with respectively reduced 93.05 % and 81.97 % of NS1 binding to HPMECs no differences among them. when compared to untreated controls (No FR-S).
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 85 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Recent Publications 1. Martina B E E, Koraka P and Osterhaus A D M E (2009) Dengue virus pathogenesis: An integrated view. Clinical Microbiology Reviews 22(4):564-81. 2. Avirutnan P, Punyadee N, Noisakran S, Komoltri C, Thiemmeca S, Auethavornanan K, et al. (2006) Vascular leakage in severe dengue virus infections: a potential role for the nonstructural viral protein NS1 and complement. The Journal of infectious diseases 193(8):1078-88. 3. Cardozo F T G S, Camelini C M, Cordeiro M N S, Mascarello A, Malagoli B G, Larsen I et al. (2013) Characterization and cytotoxic activity of sulfated derivatives of polysaccharides from Agaricus brasiliensis. International Journal of Biological Macromolecules 57:265-72.
Biography Francielle Tramontini Gomes de Sousa has her expertise on dengue pathogenesis and evaluation of antiviral activity and mechanism of action of natural and semi-synthetic compounds. She has developed an in vitro co-culture model of endothelial cells and monocytes to screen compounds with therapeutic potential for management of endothelial extravasation in severe dengue. In a previous work, she had shown that serum from acute severe dengue patients differentially affects endothelial cells barrier function in vitro and found some correlations between immunomediators and vascular leakage. Her current interests include studying the pathogenic mechanism of dengue vascular leak as well as searching for therapeutic candidates against dengue virus by using in vitro (endothelial cells) and in vivo (murine) models.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 86 conferenceseries.com
JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Scientific Tracks & Abstracts Day 3
Euro Virology 2018 & Influenza 2018
Page 92 Sessions: Day 3 July 04, 2018 Current Focus in Virology Research | HIV, AIDS and other Emerging Viruses | Molecular Virology | Veterinary Virology | General Virology Session Chair Session Co-Chair Vsevolod A. Zinserling Zsolt Boldogkoi Saint-Petersburg University, Russia University of Szeged, Hungary
Session Introduction Title: Developing a bioinformatic package for fast identification of viral transcripts using long-read sequencing methods Attila Szucs, University of Szeged, Hungary Title: Virology in the biomedical field: Biotechnological tool for the treatment of cystic fibrosis Antonio Alberto Rodríguez Sousa, Complutense University of Madrid, Spain Title: Why Cytomegalovirus(CMV) infection is still important in patients with HIV-infection and CD4-counts < 100 x 106/mL Sven Grützmeier, Karolinska Institutet, Sweden Title: Spectrum of opportunistic infections and predictors of hospitalized hiv-infected patients in sichuan, China Wenwen Pang, Sichuan University, China Title: Full-genome sequence analysis of an uncommon norovirus from South Korea Jae Woong Lee, The Catholic University of Korea, Republic of Korea Title: Chilo iridescent virus (CIV) encodes two functional metalloproteases Remziye Nalcacioglu, Karadeniz Technical University, Turkey Title: Pure red cell Aplasia secondary to parvovirus B19 infection HIV positive patients, clinical epidemiological description and observation in a third level hospital in Colombia Larry Luber Martinez R, La Maria Hospital, Colombia Title: Evaluation of pathogenicity and immunity of Type 2 Porcine Reproductive and Respiratory Syndrome virus in pigs Salik Nazki, Chonbuk National University (CBNU), Republic of Korea Title: Comparative pathogenicity of Type 1 and Type 2 Porcine Reproductive and Respiratory Syndrome virus in pregnant sows Chang-Gi Jeong, Chonbuk National University (CBNU), Republic of Korea Title: Characterization of the AcMNPV Transcriptome Using Long-read Real-time Sequencing Norbert Moldovan, University of Szeged, Hungary Title: Analysis of the vip3 genes in local Bacillus thuringiensis kurstaki strains and their insecticidal activity Remziye Nalcacioglu, Karadeniz Technical University, Turkey Title: Serum TNF alpha as a biomarker of Temporal Mesial Epilepsy associated with HHV6/HHV7 neuroinfections in humans Dmytro Maltsev, Bogomolets National Medical University, Ukraine Title: Replication of EHV-1 after modulation of autophagy process – in vitro research Izabela Serafinska,Warsaw University of Life Sciences, Poland
Euro Virology 2018 & Influenza 2018
Page 93 Attila Szucs et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Developing a bio informatic package for fast identification of viral transcripts using long-read sequencing methods Attila Szucs, Zsolt Balazs, Dora Tombacz and Zsolt Boldogkoi Department of Medical Biology, Faculty of Medicine, Szeged, Hungary
The Oxford Nanopore Technologies MinION and the Pacific Biosciences Sequel, RSII long-read sequencing platforms are capable to sequence full length transcripts, although several limitations are still occurred. The sequencing depths are much lower compared to short-read sequencing methods. One of the main problem is to distinguish between the degraded mRNAs molecules and full-length transcripts. This problem can be relatively easily solved for the abundant genes, but it is difficult for low abundant ones. Furthermore, not a trivial task to distinguish between the sequences which conatin false and true intronic sites. The false introns are generated by the strand switching effect of the reverse transcriptase. False prime sites in the genome are another source of the sequencing error. The currently available mapping softwares not always provide the most significant matches. To circumvent the above mentioned problems, we have written several routines. These routine scripts are able to correct the alignments at the end of mapped reads and remove extra false exons. Our scripts add extra parameters to the BAM files, which contain information about the positions and similarity of adaptor sequence. These routins use statistic methods to distinguish between the real and the false transcription start and end sites (TSS and TES). The program removes the potential TSS and TES sites derived from false priming. Moreover, the reads which contain false introns are also removed by our routines. Finally, our programs are able to collect the “real” transcripts and their abundance.
Biography Attila Szucs graduated in Biology and defended his PhD thesis at the University of Szeged, Faculty of Science. His area of research is in the analysis of high-throughput DNA/RNA data. His focus is on characterization of mRNAs using both quantitative and qualitative methods and combining data from various source into databases. He has published 36 papers on these subjects in reputed journals up to now.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 94 Antonio Alberto Rodriguez Sousa, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Virology in the biomedical field: Biotechnological tool for the treatment of cystic fibrosis Antonio Alberto Rodriguez Sousa Complutense University of Madrid, Spain
Statement of the Problem: Cystic fibrosis is the autosomal recessive genetic disorder with the highest incidence in the Caucasian population, being caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Although the current treatments focus on increasing the quality of life of the patient, a gene therapy of viral origin that could involve the regeneration of damaged alveolar parenchyma is located in the experimental phase. The purpose of the present work is to establish a theoretical framework of the disease from which clinical work can be carried out aimed at the development of new therapeutic techniques. Methodology & Theoretical Orientation: A dynamic simulation model was developed on patients' hope and quality of life, taking the number of pulmonary alveoli as an estimator of respiratory function. The model was calibrated in face of different clinical situations: healthy individual; untreated sick individual; sick individual with conventional treatments; and sick individual treated with gene therapy. Findings: The study showed that the life expectancy of sick individuals was significantly reduced when compared with healthy individuals. On the other hand, while the application of conventional treatments reflected an improvement in the quality of life of the simulated patients, the administration of a viral-type gene therapy was consolidated as an option in which the treated individual’s hope of life was not affected due to the disease. Conclusion & Significance: By using viruses as a biotechnological tool, being transmission vectors of functional copies of the CFTR gene that would be inserted into the host's DNA, it would be possible to correctly resume the cellular processes altered by the disease. Therefore, the research in virology and its application in gene therapies are essential to develop curative treatments that suppose a new clinical horizon.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 95 conferenceseries.com JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Recent Publications 1. Davies J C, Geddes D M and Alton E W (2001) Gene therapy for cystic fibrosis. The Journal of Gene Medicine 3(5):409-417. 2. Ferrari S, Geddes D M and Alton E W (2002) Barriers to and new approaches for gene therapy and gene delivery in cystic fibrosis. Advanced Drug Delivery Reviews 54(11):1373-1393. 3. Lane S, Rippon H J and Bishop A E (2007) Stem cells in lung repair and regeneration. Regenerative Medicine 2(4):407-415. 4. Martinez Garcia M A, Soler Cataluna J J, Perpiná Tordera M, Román Sánchez P and Soriano J (2007) Factors associated with lung function decline in adult patients with stable non-cystic fibrosis bronchiectasis. Chest Journal 132(5):1565-1572. 5. Zabner J, Couture L A, Gregory R J, Graham S M, Smith A E and Welsh M J (1993) Adenovirus-mediated gene transfer transiently corrects the chloride transport defect in nasal epithelia of patients with cystic fibrosis. Cell 75(2):207-216.
Biography Antonio Alberto Rodríguez Sousa has an extensive experience in systems dynamics and modeling. With biological training, he develops part of his personal work at Complutense University of Madrid on the simulation of the effects presented by different treatments on alveolar development in people with cystic fibrosis, a disease in which he has a high degree of specialization on the lung involvement in adults. In this sense, he has evaluated how the application of different drugs contributes to improve the hope and quality of life of patients. Specifically, and under the appropriate assumptions, it tries to make the scientific and medical sector aware that the application of a gene therapy using a virus as a transmission vector would contribute to a reversion of cystic fibrosis when the affected cellular processes are resumed. Finally, research in this area is essential to be able to propose curative treatments for diseases of genetic transmission.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 96 Sven Grutzmeier, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Why cytomegalovirus infection is still important in patients with HIV- infection and CD4-counts < 100x106/mL Sven Grutzmeier Karolinska Institute, Sweden
Background: Before the era of combination therapy (c-ART) more than 90% of the patients with HIV-infection died of one or more opportunistic infections (OI). We and others noted early on that cytomegalovirus (CMV) was an important pathogen in these patients. We investigated: all OIs and opportunistic cancers (OCs) in patients who died with CD4+ counts below 100x106/mL; the prevalence of CMV encephalitis (CMV-E) and Korsakoff syndrome; CMV retinitis (CMV-R) in relation to CMV-E; CMV adrenalitis (CMV-A) and its relation to CMV-R and CMV-E; the correlation between CMV disease and other OIs and OC and; the interaction between CMV, Epstein Barr virus (EBV) and other human herpes viruses in a case of anaplastic large cell lymphoma (ALCL). Material & Methods: We followed all patients died at Venhälsan from 1989-1996, with intensive blood testing, X-rays, CT scans, Synacthen tests, neurological examinations, and ophthalmologic examinations to. Results: Of all 219 patients died with CD4+ < 100x106/mL, 87% showed signs of reactivated CMV-infection. CMV-R was found in 84, CMV-E in 65, CMV-A in 41 and CMV in the gastrointestinal tract in 21. Mycobacterial infection was found in 87 and toxoplasmosis in 29. Kaposi’s sarcoma was the most common tumor (68 cases) followed by 22 patients with malignant lymphoma and 20 with CNS-lymphoma. CMV-reactivation was seen in most. A case of primary CMV-infection leading to a malignant lymphoma by interaction with two other herpes viruses (EBV and HHV-8) was also seen. Conclusion: CMV-infection was the main OI in AIDS-patients during the pre-c-ART era and the main cause of death by itself or together with other OIs. Reactivation of CMV was found in 87%. The most important CMV manifestations were CMV-R, CMV-E and CMV-A that seemed to occur at the same time. This is still today important in patients with CD4+ <100x106/mL without access to modern HIV-treatment. These findings reveal the intimate interaction between HIV and CMV which should be considered in all co-infected patients also today. Recent Publications 1. Grützmeier S, Porwit A, Schmitt C, Sandström E, Åkerlund B and Ernberg I (2016) Fulminant anaplastic large cell lymphoma (ALCL) concomitant with primary cytomegalovirus (CMV) infection, and human herpes virus 8 (HHV-8) infection together with Epstein-Barr-virus (EBV) reactivation in a patient with asymptomatic HIV-infection. Infectious Agent Cancer DOI: 10.1186/s13027-016-0094-5. 2. Pirskanen Matell R, Grützmeier S, Nennesmo I, Sandström E and Ehrnst A (2009) Impairment of short-term memory and Korsakoff syndrome are common in AIDS patients with cytomegalovirus encephalitis. European Journal of Neurology 16(1):48-53.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 97 Wenwen Pang, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Spectrum of opportunistic infections and predictors of hospitalized HIV-infected patients in sichuan, China Wenwen Pang Sichuan University, China
IDS is an ever-growing public health concern in China. Some HIV-infected patients get admitted because of severe Aopportunistic infections (OIs) which are the significant complication of HIV infection. By the end of 2014, there were 501,000 reported cases of people living with HIV/AIDS across China, with 66,035 people in Sichuan province alone. However, the prevalence and spectrum of OIs among Chinese HIV-infected patients are poorly understood. In the present study, 2298 cases of HIV infection in Sichuan were retrospectively investigated at the Public Health Clinical Center of Chengdu. We found that bacterial pneumonia (25.8%) was the most common OIs, followed by candida (18.3%), PCP (11.9%), tuberculosis (11.5%), infectious diarrhoea (9.3%), cryptococcus (7.3%), CMV (4.9%), toxoplasmosis (4.6%), HCV (4.0%), NTM (2.2%), and PM (0.3%). A noteworthy observation in this study is that CD4+ T cell count was found to be a predictor of some OIs. The specific pathogens causing bacterial pneumonia and/or candida infections and an effect of TB on CD4+ T cell count were also analysed between HIV-infected and non-HIV-infected patients. Understanding the spectrum of OIs in Sichuan could help us develop successful and efficient public health strategies. Such information could also help clinicians diagnose and initiate proper treatment more rapidly in hard-hit areas with limited resources.
Biography Wenwen Pang is studying for a PhD in West China School of Public Health, Sichuan University. She has published 3 papers in journals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 98 Jae Woong Lee et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Full-genome sequence analysis of an uncommon norovirus from South Korea Jae Woong Lee, Lae Hyung Kang, Yu Jung Won, Hyun Kim and Soon Young Paik The Catholic University of Korea, South Korea
oroviruses (NoVs) are major causal agents of acute gastroenteritis in humans. NoV GII.4 is the predominant genotype Nglobally. However, uncommon and minor types of NoVs are consistently detected and some have been shown to dominate over GII.4. Therefore, the prevalence of dominant and uncommon NoVs makes the identification of these viruses important for the prediction and prevention of pandemics. In this study, the full-genome sequence of a NoV (strain JW) detected in Korea was extensively characterized. The full-length genome was 7510 nucleotides long, and phylogenetic analysis based on the whole-genome sequences, including open reading frame (ORF)1, ORF2, and ORF3, indicated that it belonged to the GII.21 genotype. Strain JW showed maximum identity with strain YO284; however, comparison of the amino acid sequence of ORF2, which functions as an antigen, showed substitutions in several amino acids. GII.21 is not a prevalent epidemiological agent of acute gastroenteritis in humans, but it is consistently found in gastroenteritis patients from several countries. The present study provides the first full-genome sequence analysis of NoV GII.21 isolated from a patient in Korea. Our findings provide not only valuable genome information but also data for epidemiology studies, epidemic prevention, and vaccine development strategies.
Biography Jae Woong Lee has completed his Master’s from Jun-Nam University, South Korea. He is pursuing his PhD from Catholic-University, South Korea. His major is Molecular- biology and Virology.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 99 Remziye Nalcacioglu et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Chilo iridescent virus (CIV) encodes two functional metalloproteases Remziye Nalcacioglu, Aydin Yesilyurt, Hacer Muratoglu and Zihni Demirbag Karadeniz Technical University, Turkey
he genome of Chilo iridescent virus (CIV) has two open reading frames (ORFs) with matrix metalloproteinase (MMP) Tdomains. The proteins encoded by 136R and 165R ORFs contain 178 amino acids with over 40% amino acid sequence identity to hypothetical metallopeptidases of other viruses and 264 amino acids with over 40% amino acid sequence identity to metallopeptidases of a large group of organisms including primarily variety of Drosophila species, respectively. These proteins possess conserved zinc-binding motifs in their catalytic domains. In this study, we focused on the functional analysis of these ORFs. These ORFs were cloned into the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) Bac- to-Bac baculovirus expression-vector system, expressed in insect Sf9 cells with an N-terminal His tag and purified after 96 hours post infection to homogeneity by using Ni-NTA affinity chromatography. Western blot analyses of purified 136R and 165R proteins with histidine tags resulted in 24 and 34 kDa protein bands, respectively. Biochemical assays with the purified proteins, performed using dye-impregnated collagen (Azocoll) and Azo-casein as substrates, showed that both proteins have protease activity. The enzymatic activities were inhibited by metalloproteinase inhibitor EDTA. Effects of these proteins were also investigated on Galleria mellonella larvae. Insecticidal activities were carried out by injecting the larvae with the AcMNPV carrying 136R and 165R ORFs. Results showed that the baculoviruses harboring the iridoviral metallopeptidases caused early death of the larvae compared to control group which was performed with only wild type AcMNPV. All these data suggest that the CIV 136R and 165R ORFs encode functional metalloproteinases which can be utilized in biological control of lepidopteran pests.
Biography Remziye Nalcacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses and iridoviruses.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 100 Larry Luber Martinez Rosado Md, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Pure red cell aplasia secondary to parvovirus b19 infection in hiv positive patients. Clinical epidemiological description and observations in a third level hospital in colombia. Larry Luber Martinez Rosado Md La María Hospital, Medellín – Colombia Argentine Catholic University Santa Maria de los Buenos Aires
arvovirus B19 is a global infection that can cause serious and life threatening disorders in susceptible patient groups. [1] PViruses of the family Parvoviridae, are among the smallest viruses described. The first pathogenic human parvovirus was discovered and named B19 from the coding of a serum sample, number 19 in panel B, that gave anomalous results during testing for hepatitis B. [2]. Parvovirus B19 genotype 1, has a worldwide distribution. Genotypes 2 and 3 tend to be found in Europe and Africa.[3]. We present the clinical and epidemiological description of cases of aplasia of the red series without affecting hematimetric indices in HIV positive patients with positive serology to Parvovirus B19 admitted to hospitalization between April 2016 to April 2017. Sixteen cases were documented, 11 men (69%) and 5 women (31%) with an average age of 40.7 and 44.4 years respectively, 5 patients (31.25%) had positive IgM levels without IgG activity documented in the same sample and 4 of these 5 patients had abandoned treatment for their HIV (80%), the average in grams per deciliter of hemoglobin and hematocrit at the time of sampling was 8.92 g / dl and 28.6 g / dl respectively. All the patients included had IgG titles but only 6 had positive titles with a positive reference value> 11 (37.5%) and an average CD4 / ul cells count of 115 for men and 187.2 for women. Draws attention the most prevalent opportunist in the sampling is mycobacterium tuberculosis. High viremia could represent a great risk in plasma derivatives, [4-6] and all our patients required transfusions of red blood cells units (1.4 and 4.7 times, for women and men respectively). In patients with a clearly disturbed immunity, its relevant a deep molecular investigation to define real implications, epidemiology and distribution of this agent in our country.
Recent Publications 1. Bernstein, D.I., et al., Safety and immunogenicity of a candidate parvovirus B19 vaccine. Vaccine, 2011. 29(43): p. 7357-63. 2. Ururahy, E, K., etimologia Parvovirus B19, Emerg Infect Dis, 2018. Vol. 24, No. 2. 3. NAIDES, S.J., Parvovirus B19. En Manual of Molecular and Clinical Laboratory Immunology, in En Manual of Molecular and Clinical Laboratory Immunology, . 2016, 2016: American Society of Microbiology. p. 591-597 4. Jia, J., et al., Prevalence of human parvovirus B19 in Chinese plasma pools for manufacturing plasma derivatives. Virol J, 2015. 12: p. 162. 5. He, M., et al., Human immunodeficiency virus/human parvovirus B19 co-infection in blood donors and AIDS patients in Sichuan, China. Blood Transfus, 2012. 10(4): p. 502-14. 6. Hourfar, M.K., et al., Recipients potentially infected with parvovirus B19 by red blood cell products. Transfusion, 2011. 51(1): p. 129-36.
Biography Larry Martinez is a doctor specialized in internal medicine and infectious diseases, with experience in the management of vulnerable population since he was a medical student as a volunteer of the Colombian Red Cross. He is currently number member of de Infectology Colombian Asociation, working in third level hospitals ans clinics in the city of Medellín of Colombia through the HIV / AIDS and Tuberculosis program. Focused on advancing studies and observations regarding the incidence of viral agents and other groups of opportunists in HIV / AIDS patients. It aims to demonstrate the real incidence and prevalence of some germs and their role as coinfectants in patients with HIV / AIDS.
Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 101 Salik Nazki et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Evaluation of pathogenicity and immunity of type 2 porcine reproductive and respiratory syndrome virus in pigs Salik Nazki, Amina Khatun, Chang Gi Jeong, Sameerul Salam, Suna Gu, Sang Myeong Lee and Won Il Kim Chonbuk National University Specialized Campus, South Korea
orcine reproductive and respiratory syndrome (PRRS) virus causes significant economic losses to swine industries. To Pdevelop an effective vaccine against PRRSV is a major challenge due to huge genetic variation in PRRSV isolates. However, modified live virus (MLV) vaccines are widely used for homologous protection though there have been safety issues as vaccine viruses reverted to virulent. Besides, the existence of both genotypes together in field facilitated to emerge new strains. Therefore, regular characterization of prevalent isolates is essential to enhance protective-efficacy against PRRSV isolates. In the present study, seven type 2 PRRSV strains (NA4, NA8, NA10, NA31, NA45, NA73 and 10D415) prevalent in Korea were characterized for their immunomodulatory effects and pathogenicity in pigs. Eight piglets were infected with each strain including VR2332 (prototype of type 2 PRRSV) and kept upto 28 dpi. Serum viremia and body weight were measured weekly. Pigs from each group were euthanized at 14 and 28 dpi. Pathological evaluation was conducted and various samples were collected. Various T cell responses were analyzed in peripheral blood mononuclear cells (PBMCs) and tissues collected from the pigs. In results, NA10 and 10D415 showed highest levels of virulence as they induced high body temperature, reduced weight gain and showed high mortality in challenged pigs though 10D415 induced lower viral loads in serum and nasal fluids. On the other hand, NA8 which shares highest sequence homology with VR2332 showed lowest virulence. Similarly, the viruses evaluated in the current study showed various levels of γδ-T, Th1, Th17 and CTL responses in PBMCs or tissues. In conclusion, the present study suggested that type 2 PRRSVs in Korea showed a wide range of pathogenicity and immune responses and the information might be helpful in designing efficient vaccine platform against PRRSV infection.
Biography Salik Nazki is currently pursuing PhD from Chonbuk National University, South Korea. He has completed DVM and Master’s in Veterinary Microbiology and Immunology from SKUAST-Kashmir, India. The major focus of his research is on PRRSV and is well versed with the virological and immunological techniques. Previously, during his Master’s degree, his research work was based on anaerobic bacteria and also has experience on other viruses. He has recently published some research articles in reputed journals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 102 Chang Gi Jeong et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Comparative pathogenicity of type 1 and type 2 porcine reproductive and respiratory syndrome virus in pregnant sows Chang Gi Jeong, Amina Khatun , Salik Nazki and Won Il Kim Chonbuk National University Specialized Campus, South Korea
orcine reproductive and respiratory syndrome (PRRS) is a challenging threat to the swine industries caused by PRRS virus Pand characterized by reproductive failures in pregnant and respiratory distress in piglets. As an RNA virus, PRRSV mutated quickly and evolved continuously, which caused huge genetic and antigenic variation within genotypes or even in the same viruses. Furthermore, the biological properties of PRRSV responsible for viral pathogenesis and host-immune responses have not been characterized clearly. Many previous studies have been conducted by using a respiratory disease model in weaned piglets for convenience but the reproductive disease caused by PRRSV is still poorly understood. Therefore, the present study was aimed to demonstrate the pathogenicity and immune response following infections with both type 1 (D40 and CBNU0495) and type 2 (K07-2273 and K08-1054) PRRSV strains in pregnant sows. Two pregnant sows were infected with each virus at 93 days of gestation. At 21 days after infection, all sows and their fetuses were euthanized for pathological evaluation. In addition, viral loads were measured in serum samples from the sows and various tissues collected from the sows and their fetuses and fetal weights were recorded. In results, CBNU-0495 and K08-1054 showed higher virulence as compared with other viruses as they exhibited higher levels of viral loads in sera and tissues of sows and fetuses and also caused higher levels of weight losses in fetuses.
Biography Chang Gi Jeong has completed his MS in Veterinary Medicine (Microbiology) from Chonbuk National University (CBNU), South Korea. During his Master’s course, he worked on the Clostridium novyi and studied the genome characteristics of isolates using Next Generation Sequencing (NGS). Currently, he is pursuing PhD course at the same university. His major research focuses on PRRSV and Japanese encephalitis virus. He is well-adapted in animal studies mostly dealing with farm animals.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 103 Norbert Moldovan et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Characterization of the ACMNPV transcriptome using long-read real-time sequencing Norbert Moldovan1, Zsolt Csabai1, Attila Szucs1, Michael Snyder2, Dora Tombacz1, 2 and Zsolt Boldogkoi1 1University of Szeged, Hungary 2Stanford University, USA
he Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is an insect virus belonging to the Baculoviridae Tfamily. The 134 kbp long double-stranded circular DNA of the virus encompasses 150 tightly packed open reading frames. Genes of the AcMNPV are expressed in three phases: early, late and very late. Promoters of early genes are recognized by the host RNA polymerase, and usually consist of a canonical TATA motif located upstream of the transcriptional start site. At the same time some early transcripts start from the arthropod initiator element (CAGT). Late and very late genes are transcribed by the viral RNA polymerase, which recognizes a late initiator sequence (TAAG). A short-read sequencing technique was used previously to elucidate the structure of the AcMNPV transcripts; however short-read sequencing cannot tackle the highly complex, overlapping nature of the viral transcriptome. In this study we found and annotated four novel putative protein coding, four non-coding transcripts and forty-seven novel length isoforms of previously annotated transcripts using the Oxford Nanopore Technologies’ MinION and the Pacific Biosciences Sequel platforms. We demonstrated that the canonical promoters and initiators of a transcript are in concordance with the motifs found upstream of its isoforms. We also discovered the extensive overlapping nature of the viral transcriptome. Additionally we determined the expression characteristics of novel transcript isoforms and revised it for the already annotated transcripts using long-read real-time sequencing of the AcMNPV transcriptome.
Biography Norbert Moldovan graduated in Biology from Babes-Bolyai University and now is a PhD student at the University of Szeged, Faculty of General Medicine. His area of research is in molecular genetics of human and non-human pathogenic viruses. His main focus is in structural and expressional characterization of viral mRNAs using third-generation sequencing. He has published seven papers on these subjects in reputed journals during his two years of doctoral studies.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 104 Remziye Nalcacioglu et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Analysis of the vip3 genes in local Bacillus thuringiensis kurstaki strains and their insecticidal activity Remziye Nalcacioglu, Aysun Adiguzel, Ebru Guney, Zihni Demirbag and Kazim Sezen Karadeniz Technical University, Turkey
novel group of insecticidal proteins named vegetative insecticidal proteins (Vıp) are produced by Bacillus thuringiensis A(Bt) bacteria during its vegetative stage. In this study we characterised the vip3 genes of two local Bt isolates (BnBt, MnD). Firstly, partial-purified Vip3 proteins of some local Bt isolates were tested against the Spodoptera littoralis larvae. After obtaining good insecticidal activity with BnBt and MnD isolates, Vip3 proteins of these bacteria were purified from supernatants of bacterial cultures by ion exchange chromatography. Purified proteins were subjected to SDS-PAGE analysis and 90 kDa band of proteins were determined. These purified proteins were tested against S. littoralis larvae. Results showed that Vip3 proteins of BnBt and MnD produced 86.66% and 83.33% insecticidal activity on S. littoralis larvae, respectively. The lethal concentrations
(LC50) of BnBt and MnD were determined as 41.860 ng and 55.154 ng, respectively. These results suggest that vip3 genes of our local isolates may be alternatives for preventing resistance in various insect–pest species. Also these proteins may be used at developing bio-pesticides.
Biography Remziye Nalcacioglu has completed her PhD in 2003 from Karadeniz Technical University, in Turkey. She has been working in the same universirty as an academician. She is working on insect viruses including baculoviruses, iridoviruses and Bacillus thuringiensis bacteria.
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Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018
Page 105 Maltsev, Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria
Serum TNF alpha as a biomarker of Temporal Mesial Epilepsy associated with HHV6/HHV7 neuroinfections in humans Maltsev Immunology and Molecular Biology Laboratory of Research Institute of Experimental and Clinical Medicine, Ukraine
ccording to results another researches in this study mesial temporal lobe epilepsy (MTLE) is shown to be associated with Ahuman herpes virus 6 and 7 types (HHV6/HHV7) neuroinfections. We also demonstrate that the epileptic process is associated with an systemic inflammatory reaction, and that the proinflammatory cytokine, the tumor necrosis factor-alpha (TNF-α), is able to potentiate the reproduction of the herpes viruses. The study group (SG) included 43 patients between 16 and 60 years with MTLE and HHV neuroinfections, diagnosed according to the PCR of the cerebrospinal fluid (CSF), serum or abnormal serum/CSF IgG ratio. The control group (CG) included 30 patients of similar age with MTLE, but without the HHV neuroinfections. The concentration of TNF-α in the serum was determined by enzyme-linked immunosorbent assay ("VektorBEST" RF; N=0-50 pg/ml). Patients of the SG had high concentrations of TNF-α in serum (288±44.7 pg/ml), that were significantly higher than in the CG (p<0.05; Z Notes: Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018 Page 106 Izabela Serafinska et al., Virol Mycol 2018, Volume 7 conferenceseries.com DOI: 10.4172/2161-0517-C2-026 JOINT EVENT 10th International Virology Summit & 4th International Conference on Influenza & Zoonotic Diseases July 02-04, 2018 | Vienna, Austria Replication of EHV-1 after modulation of autophagy process – in vitro research Izabela Serafinska, Joanna Cymerys, Anna Golke, Marcin Chodkowski, Joanna Brzezicka, Anna Słońskaand Marcin W Bańbura Warsaw University of Life Sciences, Poland utophagy is an evolutionary conservative, intracellular process. It plays an important role in maintaining homeostasis Ain cell through degradation of their own components such as proteins or organelles via lysosomes. Viral infection can modulate this process. During infection autophagy may be stimulated or inhibited, because virions can be degraded in autophagolysosomes. Some viruses can utilize this process for replication. To investigate the influence of this process on the level of viral replication, stimulators or inhibitors of autophagy can be applied. Primary cultures of murine neurons were treated with autophagy inductors (rapamycin and temozolomide) and inhibitors (wortmannin and chloroquine) for 24 h. After incubation neurons were infected with equine herpesvirus type 1 (EHV-1) for 2, 24 and 48 h. In experiments non- neuropathogenic Jan-E EHV-1 strain was used. Level of viral replication was analyzed using real-time PCR. Presence and localization of viral antigens and LC3B protein (autophagy marker) were detected using confocal microscopy. Results obtained from real-time PCR showed an increase of level of viral DNA in control. At 2 h p.i., after chemical treatment, increase in EHV- 1 DNA amount was observed. At 24 h p.i. this level was decreased and then at 48 h p.i. increased again, except sample treated with rapamycin. In this cells level of viral DNA also decreased at 48 h p.i. Immunofluorescence staining showed that LC3B protein was presented inside neurons during EHV-1 infections. In neurons at 2 and 48 h p.i. higher number of viral particles was observed in comparison to 24 h p.i. Results showed changes in replication kinetics during EHV-1 infection after using autophagy modulators which can be useful in antiviral therapy. Biography Izabela Serafinska is a PhD candidate in the Division of the Veterinary Medicine in Warsaw University of Life Sciences, Poland. Her main research topic is autophagy in neurons during EHV-1 infection. Her scientific interests include neuroinfections and microscopy. This work was supported by grant from the KNOW (Leading National Research Centre) Scientific Consortium „Healthy Animal – Safe Food", decision of Ministry of Science and Higher Education No. 05-1/KNOW2/2015 Notes: Virology & Mycology Volume 7 ISSN: 2161-0517 Euro Virology 2018 & Influenza 2018 July 02-04, 2018 Page 107