10^th International Virology Summit 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.

Abstract:

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) and Gulf war illness (GWI) are debilitating diseases presenting 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 (T_FH) 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 (T_FH) and subsequent production of IL-21 and CXCL13 by T_FH. 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.

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.

Abstract:

Foot-and-mouth disease (FMD) remains one of the world’s most widespread epizootic and highly contagious animal diseases 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.

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.

Abstract:

The caprine arthritis encephalitis (CAE) is a disease caused by Lentivirus genus, Orthoretrovirinae subfamily, and Retroviridae family 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.

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 vivorole 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.

Abstract:

Carbohydrates-based immunogens are generally less effective in generation of long-lasting antibody responses and neutralizing 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.

Biography:

Zsolt Boldogkoi has completed his PhD from Szent Itván 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.
 

Abstract:

Herpesviruses are large, enveloped, double-stranded DNA viruses. Although, the members of the three subfamilies (α-, β-, and 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:

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 

Abstract:

Despite great success in the diagnostics and therapy of AIDS, there are many unanswered questions. Without giving the answers 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.

Biography:

Dr Annete 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 virus pathogenicity 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.

Abstract:

Vaccines are the most effective method of controlling infectious diseases. However, todays vaccine production is facing difficult 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.

Biography:

Research interests of :(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.

Abstract:

Geminiviruses (family Geminiviridae) are classified into nine genera and unassigned viruses on the basis of host range, insect 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:

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.

Abstract:

Pandemic A (H1N1) pdm09 virus has caused substantial morbidity and mortality globally and continues to circulate, which may 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

Biography:

Abstract:

The Vaccinia virus (VACV) is a prototype member of the Poxviridae family, it has a relatively large double-stranded DNA genome 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:

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.

Abstract:

Zika virus (ZIKV) infection during pregnancy has become a global public health concern due to its ability to cause severe congenital 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:

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.

Abstract:

Mycoviruses are viruses that naturally infect and replicate in fungi. They are widespread in all major fungal groups including 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:

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.

Abstract:

A metagenomic approach based on target independent next-generation sequencing has become a known method for the detection 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.

Biography:

Dr. Qian Yang worked as a professor in College of Veterinary Medicine.The focus of her research work is on the mucosal immunity in domestic animal. Firstly 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 had 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 relvant pathogenic mechanism. She has published more than 80 original and review articles in scientific journals.

Abstract:

Transmissible gastroenteritis virus (TGEV) is a coronavirus, characterized by diarrhea, high morbidity, and the mortality is 100% in piglets less than 2 weeks old. Pigs infected with TGEV are often suffer 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.

Biography:

 Yuchen Li has been studied in Nanjing Agricultural University as a PhD candidate since 2015. Now I am studying in the major of preventive veterinary medicine under supervisor of professor Qian Yang. My research focuses on the pathogenic mechanism of PEDV. I am working hard and have published three research article. I am really interested in the research process, which is amazing and attractive. I hope could get the chance to show my research subject in the conference.

Abstract:

Porcine epidemic diarrhea (PED) has made catastrophic impacts on the global pig industry since 2011. The causative agent, 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.

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.

Abstract:

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.

Biography:

Abstract:

The only cost-effective protection against influenza is vaccination. Due to rapid mutation continuously new subtypes appear, what 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.

Biography:

Abstract:

RNA-dependent RNA polymerases (RDR) play an important role in virus protection and plant gene control. Their activity is 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 of CsRDR1b 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.

Biography:

Dr. 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.

Abstract:

Flu epidemics and potential pandemics pose great challenges to public health institutions, scientists and vaccine producers. Creating 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:

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.

Abstract:

Integrase is the retroviral protein responsible for incorporating a double-stranded viral DNA copy into the host chromosome. Single 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:

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.

Abstract:

Soybean mosaic virus, a member of the genus Potyvirus, significantly reduces soybean production worldwide. Rsv3, which confers 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.

Biography:

Sherwin 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.

Abstract:

The mechanics of flu related respiratory illness is not completely implicit as it includes; influenza, zoonotic and non-influenza 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.

Biography:

Abstract:

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.

Biography:

Abstract:

Low genetic fidelity and limited cross-protection are two major hurdles faced in porcine reproductive and respiratory syndrome 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:

Abstract:

A broad survey of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) populations in agricultural and forested areas 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:

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.

Abstract:

Acanthamoeba polyphaga mimivirus (APMV) is the first giant virus discovered almost 15 years ago. It has uncommon characteristics 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 mm 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.

Biography:

Abstract:

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.

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.

Abstract:

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:

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.

Abstract:

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.

Biography:

Abstract:

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.

Biography:

Abstract:

AIDS is an ever-growing public health concern in China. Some HIV-infected patients get admitted because of severe opportunistic 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:

Abstract:

Noroviruses (NoVs) are major causal agents of acute gastroenteritis in humans. NoV GII.4 is the predominant genotype globally. 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:

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.

Abstract:

The genome of Chilo iridescent virus (CIV) has two open reading frames (ORFs) with matrix metalloproteinase (MMP) domains. 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:

Larry Martínez 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.

Abstract:

Parvovirus B19 is a global infection that can cause serious and life threatening disorders in susceptible patient groups. [1]  Viruses 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

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

Abstract:

Porcine reproductive and respiratory syndrome (PRRS) virus causes significant economic losses to swine industries. To develop 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:

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.

Abstract:

Porcine reproductive and respiratory syndrome (PRRS) is a challenging threat to the swine industries caused by PRRS virus and 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:

Abstract:

The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is an insect virus belonging to the Baculoviridae family. 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:

Remziye Nalçacioglu 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.

Abstract:

A novel group of insecticidal proteins named vegetative insecticidal proteins (Vıp) are produced by Bacillus thuringiensis (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:

Abstract:

According to results another researches in this study mesial temporal lobe epilepsy (MTLE) is shown to be associated with human 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<Z_0.05). Serum concentrations of TNF-α greater than 100 pg/ml were associated with the severe general condition of the patients, more severe epileptic syndrome, a long history of illness, deep organic brain damage, low sensitivity to anticonvulsant drugs, overall with a poor prognosis. In patients with MTLE and HHV6/HHV7 neuroinfections marked systemic inflammatory response syndrome was noted, which affected the severity of the symptoms in the patient. TNF-α, therefore, can be used as a biomarker for an objective assessment of the severity and prognosis of the disease in patients with MTLE induced by HHV6/HHV7.

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.

Abstract:

Autophagy is an evolutionary conservative, intracellular process. It plays an important role in maintaining homeostasis in 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:

In Hyuk Baek studied Biotechnology in Saarland University in Saarbrücken, Germany.  Currently, he is doing his PhD course under supervision of Prof. Dr. Helms and working with Dr. Youngjun Kim in the environmental safety group in the Kist-Europe (Korea Institute of Science and Technology Europe branch). His main topic of his Master’s thesis is metagenomic analysis of the viral communities in human feces: molecular approaches to discovery and characterization of novel viruses. He has also participated in several topics followings with 3 SCI (+1 submitted), 3 SCIE papers, 2 presentations and 4 registered patents.

Abstract:

Background: Combined 3D cell culture in vitro assay with microenvironments mimicking systems are effective for cell based drug and chemicals toxicity screening test to close tissue mimicking micro-environmental systems that was contributed in single or multiple compounds to check cell migration, angiogenesis, metastasis and morphogenesis. The angiogenesis structure of endothelial cells in the ECM was confirmed as 3D and regulated by surface treatment of the microfluidic channels. Filamentous bacteriophages are a member of the family Inoviridae. ï»¿These are used for material science, drug delivery, tissue engineering, energy and biosensor. Genetically modified bacteriophages could deliver therapeutic molecules or genes to specific cancer tissues or organs.

Objectives: In this research, multi-functionalized biocompatible bacteriophage and ECM were applied for mimicking angiogenic micro-environmental systems.

Materials & Methods: E. coli MC1061 and K91BK were used for plasmid and phage amplification respectively. Phage preparation was performed by PEG/NaCl precipitation. The function of bacteriophages was tested by RT-qPCR, ELISA assay. The angiogenesis factors (e.g. CD31 and VE-cadherin) were confirmed by using confocal microscopy on commercialized lab-on-a-chip.

Conclusion: In this study, we demonstrated that endothelial cells contacted biocompatible bacteriophages which are found to migrate and spout into the ECM at the tube like structures. The function of angiogenesis was confirmed by angiogenesis factor of CD31 and VE-cadherin staining images. Our results suggested that biocompatible bacteriophage and ECM might continuously contribute to stimulate microenvironment for in vitro angiogenesis models. Also, we described that the functionalized bacteriophages can be used for feasible biomaterials on biomedical engineering fields. In the future, these studies are potentially applied for angiogenic matrix at the tissue engineering in vitro assay.

Biography:

Lamyaa Al-Dalawi has completed her Graduation in College of Veterinary Medicine, at University of Baghdad, Iraq. She worked as a Veterinarian in the Surgery Department of the Veterinary Medicine, University of Baghdad, Iraq. She has completed her Master degree in Internal and Preventive Medicine in the Veterinary College at the University of Baghdad. She later moved to the Medical Institute as a Lecturer in University of Kirkuk, Iraq. She obtained a scholarship to study PhD at Nottingham University. In October 2014, she started her PhD in the Department of Infection and Immunity/School of Veterinary Medicine and Science at the University of Nottingham. 

Abstract:

The influenza virus infection is influenced by a number of host cell factors, including host cell lipids. These lipids make up the bilayer membranes for both virus particles and host cells. The objective of this study is to determine the biophysical importance of lipids in terms of infectivity by pre-treating Influenza A viruses; avian influenza H2N3 virus, equine Influenza H3N8 virus and pandemic influenza H1N1 virus with various types of phospholipids. 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) had no significant impact on virus infectivity. However, 1, 2-dipalmitoyl-Sn-glycero-3-phospho-(1'-rac-glycerol) (DPPG) had a significant impact on H2N3, H3N8 and H1N1 infectivity. Treating the influenza viruses with lyso-analogues: 1-palmitoyl-2-hydroxy-sn- glycero-3-phospho-(1'-rac-glycerol) (LPG) and 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC) produced significant inhibition of influenza virus infection using MDCK cells and A549, that was dose-dependent. TEM images showed H2N3 and H3N8 without lipid pre-treatment are mostly spherical or filamentous, respectively. Incubating these viruses with lipids impact their morphology. Investigations of avian influenza H2N3 binding assay by flow cytometry demonstrates a high impact of the negatively charged phospholipids; i.e. either DPPG or LPG, blocking virus binding to cells significantly. Moreover, incubating influenza viruses with negatively charged phospholipids reduce cytokines expression especially IL-8. Overall, pre-incubating the virus with phospholipids seems to have an impact on the ability of the virus to bind to cells. So, specific lipids can be considered as a potential new inhibiting factor for influenza.

Biography:

Yu-Ju Lin is the Section Chief of the Division of Preparedness and Emerging Infectious Diseases, Centers for Disease Control in Taiwan (TCDC). She currently leads TCDC’s influenza pandemic preparedness and response activities, and her office is responsible for national and local readiness, the regulation of emergency operations, and managing the nation’s stockpile of related emergency countermeasures. She has more than 15 years of continuous service as a TCDC Officer; during her long career, she has taken part in a wide range of infectious disease outbreak investigations and responses, including SARS outbreak of 2003 in Taiwan, strategic development for the control of seasonal influenza/novel influenza A virus infections.

Abstract:

Background: The threat of a human influenza pandemic has prompted, thus, urgent development of national preparedness plans in Taiwan since 2003. We reviewed these plans to assess Taiwan’s preparedness for pandemic influenza.

Methods: The ‘National Influenza Pandemic Preparedness Plan’ and the ‘Strategic Plan’ of Taiwan, published in 2015 and 2011 respectively, were evaluated by using a checklist containing five criteria of preparation, surveillance, prevention and containment, case investigation and treatment, and risk communication (there are 68 indicators in total). This checklist was developed using the latest WHO guidelines, and in consultation with influenza experts of the WHO Eastern Mediterranean Office.

Results: The average score for aggregate completeness is 72.5%. For the five included criteria, the on average scores are 83.3% for preparation, 83.3% for surveillance, 57.4% prevention and containment, 71.4% case investigation and treatment, and 58.3% risk communication. Among the individual indicators, 31 (45.6%) indicators scored 3, 20 (29.4%) indicators scored 2, 15 (22.1%) indicators scored 1, and 2 (2.9%) indicators scored 0. Both of the 2 indicators scored 0 due to the lack of mention triage system and strategy for storage and disposal of corpses.

Conclusion: Taiwan’s preparedness plans are satisfactory, the plan scores particularly well on surveillance system and mobilization of resources such as health care facilities, personal protective equipment, antivirals, and vaccines. Moreover, gaps in preparedness planning remain; especially those operational guidelines for the implementation of related prevention and containment measures should be detailed or addressed in the operational documents.

Biography:

Abstract:

Equine herpesvirus type 1 (EHV-1) is a main cause of respiratory disease, abortion and myeloencephalopathy in horses. Similarly to other alphaherpesviruses EHV-1 is neurotropic and causes latent infection in the neurons of natural host. Despite the fact that many studies have been devoted to the pathogenesis of various clinical forms of EHV-1 infection, mechanisms of the neuronal damage are not fully understood. In the present work, we examined one of the aspects of these disorders- tau-mediated neurodegeneration as a consequence of its hyperphosphorylation and sequestration in aggregates in infected primary murine neurons. Previous reports mainly focus on those HHV-1 infections which proved that tau protein (a microtubule-associated protein) in important in the neurodegenerative process. It is expressed in central nervous system neurons, where it plays a powerful role in regulating the dynamics of microtubule polymerization. Furthermore, it takes part in regulating axonal diameter, in axonal transport and in neurogenesis. When infected with HHV-1, tau protein may undergo modification, mainly via phosphorylation. As a result, phosphorylated tau protein isoforms aggregate and form neurofibrillary tangles, typical of neurodegenerative diseases called tauopathies. We describe, for the first time, that EHV-1 is capable of intensely modifying in the phosphorylation state of tau and that infection with EHV-1 leads to the accumulation of phosphorylated tau in the cytoplasm of neurons, especially during late hours after infection. This accumulation is dependent on the type of tau phosphorylation and on the time of infection.

Biography:

J Brzezicka is a PhD Student in the Department of Preclinical Science, Faculty of Veterinary Medicine, Warsaw University of Life Sciences. Her scientific interests include the impact of viral infection on the cytoskeleton, molecular virology and neuroinfections.

Abstract:

Adenoviruses are nonenveloped, double-stranded DNA viruses. Human adenoviruses (HAdV) are ubiquitous in populations worldwide. HAdV are classified into seven species (A to G). Due to the different tissue tropism, adenoviruses can be an etiological factor of infections of upper respiratory tract, digestive tract, urinary tract, eyes and the central nervous system. Children and adults with impaired immunity are particularly susceptible for infection. The aim of this study was to assess the changes in the actin cytoskeleton in A549 cells (adenocarcinomic human alveolar basal epithelial cells) after infection with different types of HAdVs. In the current study, three types of HAdVs were used: HAdV4, HAdV5 and HAdV7. Filament structures of actin were visualized using TRITC-phalloidin conjugate. Polyclonal antiserum ADENO MAB conjugated to FITC was used to detect viral antigens. Cell nuclei were stained with Hoechst 33258. Infected cells exhibited morphological changes, followed by cell lysis at the final step of infection. In A549 cells infected with HAdV4, 5 and 7 (at 12, 24 and 48 h p.i.), CPE consisted of disintegration and degradation of a nucleus, changes in a cell shape and rearrangements of the actin filaments. Furthermore, HAdVs used in this study caused actin accumulation in the nuclei of infected cells. Cells which did not undergo lysis showed high amounts of viral antigens in the cytoplasm. In the present study, we demonstrate that all used types of HAdVs are able to infect A549 cells, without the need for initial adaptation. The infection causes changes in cell morphology and cytoskeleton rearrangements. That may indicate that actin cytoskeleton is crucial for penetration into the cells, viral transport and transcription of viral genome.

Biography:

Abstract:

The body's ability to fight viral infection depends on the effective activation of the innate immune response. Activation of specific pattern recognition receptors (PRRs), including toll-like receptors (TLRs) triggers the production of pro-inflammatory cytokines, including interferons, which should lead to inhibition of viral replication. Because viruses have developed a number of mechanisms to interfere with TLR signaling, synthetic TLR activators can be considered as potential antiviral drugs. In the present study we have been investigating the effect of TLR synthetic ligands on the replication of equine herpesvirus type 1 (EHV-1) in ED cell line (equine dermal). Two different TLR ligands were used: TLR2/TLR6 ligand - Pam2CSK4 (0.1-100 μg/ml) and TLR9 ligand - ODN D-SL03 (1-10 μM/ml). The effect of TLR ligands on EHV-1 replication was assessed basing on the cytopathic effect and the quantitative evaluation of the number of viral DNA copies in the cells and the culture medium (real-time PCR). Both, on the basis of the cytopathic effect evaluation and the number of viral DNA copies quantification the antiviral activity of Pam2CSK4 at concentrations 10 μg/ml and 100 μg/ml was observed. No significant effect of ODN D-SL03 on EHV-1 replication was observed. However, preliminary observations based on the cytopathic effect evaluation showed that Pam2CSK4 at a concentration of 100 μg/ml in combination with ODN D-SL03 at a concentration of 10 μM/ml inhibited EHV-1 replication more efficiently than each of them individually. This stays with agreement with other studies, which show that it is necessary to stimulate both, TLR2 and TLR9 to trigger an effective immune response against herpes viral infections.

Biography:

M Chodkowski is a PhD candidate in the Division of the Veterinary Medicine in Warsaw University of Life Sciences. His main research topic is: Changes in the mitochondrial network in primary murine neurons infected with EHV-1. He is interested in molecular virology, neurovirology and novel treatment approaches using viruses e.g. oncolytic viruses.

Abstract:

Mitochondria have emerged as one of the key organelles in the maintenance of cellular homeostasis, innate immunity metabolism, aging, innate immunity, metabolism, apoptosis and other signaling pathway. In the last decade, work on the mitochondria has expanded our knowledge of its roles in cellular homeostasis in many parallel ways. There are not many report about interaction mitochondria – virus. Although all mitochondria have the same architecture, they vary greatly in shape and size. The mitochondria are composed of outer mitochondrial membrane, inner mitochondrial membrane, inter membrane space (space between outer and inner membrane), and matrix (space within inner mitochondrial membrane). The outer membrane is a smooth phospholipid bilayer, with different types of proteins imbedded in it. Mitochondria in eukaryotic cell formed network and they are distributed throughout the cells. In this study we examined changes of the mitochondrial network in HaCat cells during herpesvirus infection. For immunofluorescent staining, cells were plated onto laminin-coated coverslips. After 24, 48 h.p.i., infected cultures were fixed in 3.7% paraformaldehyde/PBS (Sigma Chemicals) for 30 min at room temperature. The cells were permeabilized in 0.5% Tween/PBS for 5 min, washed in PBS and blocked with PBS containing 1% bovine serum albumin (BSA) (Sigma Chemicals). Mitochondria were visualized using MitoRed (300 nM; Sigma Chemicals) and the cell nuclei were stained with Bisbenzimide /Hoechst 33258 according to the manufacturers recommendations. Results were evaluated under confocal microscope (Fluoview FV10i, Olympus). Due to infection with HHV-1 and HHV-2, the mitochondrial network is reorganized, both in the early and late stages of infection. Changes are manifested mainly through fragmentations of the mitochondrial network.

Biography:

Abstract:

Hepatitis C virus (HCV) infection is a major public health problem with an estimated 3-4 million people infected each year worldwide. 20–30% of individuals acutely infected with HCV will spontaneously clear the virus, with the remaining 70–80% developing persistent HCV infection. The interplay between the virus and host innate and adaptive immune responses determine the outcome of HCV infection (Rauch et al., 2009). The present study aims to determine the level of cellular immune subsets in responders and non-responders HCV-infected patients as a result of the standard treatment (PEG-IFN and ribavirin) and to correlate the results with the major HCV genotypes in Kuwait. Data of the immunophenotyping for cellular subsets include 30 healthy controls and

genotype-4 HCV-infected patients (39 responders vs. 21 non-responders) at baseline and after treatment. The immunophenotyping was evaluated by flow cytometry using antibodies specific to mature T cells, T cytotoxic cells, regulatory T cells, T helper cells, activated T cells, natural killer cells, NKT cells and pan B cells. There were significant differences in the mean values of percentages for T helper cells, T cytotoxic cells, B cells, NK cells, NKT cells and activated T cells between HCV-responder vs. HCV-non-responder patients. Also, significant differences were noticed in the mean values of the absolute counts for T helper cells, B cells, NK cells, and T cells. Cellular subsets of the immune system play an important role in the pathogenesis, progression, and clearance of HCV. The screening for multiple cellular markers in the present study showed significant variations in the absolute counts and percentages of essential immune cellular subsets. These findings could lead to new possibilities for immune-based interventions and/or vaccine development with the aim of restoring functional antiviral T cell responses combined with improved viral clearance

Biography:

Abstract:

Equine herpesvirus type 1 (EHV-1) is a member of the subfamily Alphaherpesvirinae of the family Herpesviridae. In its target host, it induces mild respiratory diseases, abortion, neonatal foal death, and neuropathogenic disorders. EHV-1 has a broad host range in vitro, allowing for study of the mechanisms of productive viral infection, including endocytic pathways or intracellular transport in various cell cultures. The productive infection of astrocytes has been described for HHV-1, HHV-5 and HHV-6; however, there were no data about the ability of EHV-1 to infect the astrocytes. Recently, we reported for the first time that primary murine astrocytes were permissive to EHV-1 infection. Similarly to HHV-1, EHV-1 productively infected astrocytes and displayed cytopathic effect that resulted in the death of a portion of cell population. In the current study we investigated the mechanisms by which EHV-1 enters primary murine astrocytes. Using drugs that inhibit clathrin-dependent (chlorpromazine) or caveola-dependent endocytosis (nystatin), we showed that EHV-1 entry into murine astrocytes require clathrin, but not caveolae. The treatment of the cells with nystatin did not affect the replication of EHV-1. However, the use of chlorpromazine caused a significant decrease in the level of replication of EHV-1 detected by real-time PCR. In conclusion, EHV-1 efficiently entered and replicated in primary murine astrocytes. According to our results, we can assume that the principal pathway of EHV-1 entry into astrocytes appears to be caveolin-dependent and clathrin-independent.

Biography:

Jing An graduated from Chinese Medical University in 1982 in Shenyang of China. She got her Master’s degree and PhD in Clinical Medicine in 1989 at the Third Military Medical University in Chongqing (China), followed by a Postdoctoral position and Research Fellow position at the Department of Microbiology and Immunology in Tokyo (Japan) at the Metropolitan Institute for Neuroscience. In autumn 2000, she returned to China and occupies since the position of a Principal Investigator and Head of the Department of Microbiology at the Capital Medical University of Beijing (China). Her research focuses on the interaction between mosquito-borne flaviviruses and host as well as prevention of dengue virus infection.

Abstract:

Recently, Zika virus (ZIKV) causes millions of infections which emerge as a new dangerous member of the genus of Flavivirus. Unlike other well-known flaviviruses, ZIKV can be transmitted sexually and infect testes in murine models. However, the exact susceptible cells are not entirely clear. To investigate these issues, we infected interferon α/β and γ receptors deficient AG6 mice with ZIKV and examined the outcomes of infection. Infected mice displayed signs of reproductive system disorder, altered androgen levels in serum, and high viral load in semen and testes. Seminiferous tubules showed atrophy, accompanied by positive staining of ZIKV antigens on Sertoli cells. Viral particles and vacuole changes were observed within Sertoli cells, whose susceptibility to ZIKV was further validated in vitro study using cell lines. The disruption of tight junctions within testis and altered sperm morphology were also observed in ZIKV infected mice. Our results therefore demonstrated that Sertoli cells are susceptible to ZIKV infection, which results in the disruption of tight junctions in testis and causes abnormal spermatogenesis in mice. These results also imply that long-term impact of ZIKV infection on human male reproductive system requires close monitoring.

Biography:

Abstract:

In Asia, Chinese cabbage (Brassica rapa) is mainly grown and utilized in various ways as a healthful food source. However, increasing virus damage resulting from changes in trade of agriculture products including seedlings and seeds, as well as climate change and repeated cultivation, has reduced Chinese cabbage production. According to recent research, three plant viruses - Turnip mosaic virus (TuMV), Cucumber mosaic virus (CMV), and Youcai mosaic virus (YoMV) - are reported to affect Chinese cabbage yields. Recently we detected new isolates of YoMV in Korean radish fields, and full-length infectious clones of two isolates were generated in the dual 35S /T7 promoter driven pJY binary vector. Four amino acid differences (V383I, M492I in 125kDa, T1245M in 182kDa and M17T in CP) between two isolates resulted in either severe or mild symptom development in Nicotiana benthamiana. In order to reveal the amino acids related to severe pathogenesis, four hybrid constructs were generated through by gene exchanges between the isolates. Hybrid constructs maintaining CP residue 17 as threonine in the MP/CP overlap region developed severe symptoms. Further analysis expressing CP_M(17)T from a Potato virus X vector produced differential symptoms in N. tabacum cv. Xanthi, inducing HR (T17) and mild symptoms (M17) respectively.

Biography:

Hyun Kim has a Master's course from Catholic University School of Medicine. Mainly her study is waterborne viruses.

Abstract:

Severe fever with thrombocytopenia syndrome (SFTS) is a newly emerging infectious disease and caused by SFTS virus (SFTSV), a tick-borne phlebovirus in family Bunyaviridae. SFTSV was reported in China in 2011, more recently in Japan and South Korea. In this study, samples were collected from patients with SFTS in Jeju, South Korea in 2017, and its identity was confirmed as SFTSV by RT-PCR, and nucleotide sequencing and alignment analysis. The whole-genome of the SFTSV strains sequence analysis revealed three segments comprising the whole genome: L segment (6,368 bp), M segment (3,378 bp), and S segment (1,744 bp). The SFTS strain showed amino acid identities with Korea reference strains of 98.30-99.83%, 98.51-99.44%, and 99.38-99.57% in the S segment, M segment, and L segment, respectively. Additional, it showed 98.94-99.49%, 98.42-98.70%, and 99.09-99.42% similarity with Japan reference strains and similarities with reference strains in China were 98.39-99.28%, 97.49-98.23%, and 99.14-99.47%. Of the total amino acids, 28 mutations were identified, among 11 of which was special substitution mutations found only in our strains. This research will be used as a full-length SFTSV sequence standard for future comparison studies. Also, it may prove useful to the field of public health by facilitating the diagnosis and the prediction of new emerging variants.

Biography:

Abstract:

High-throughput sequencing methods have revolutionized genomics, including RNA-sequencing; however the widespread short read-sequencing techniques cannot distinguish between the different transcript isoforms and overlaps. The third-generation, long-read sequencers (TGS) are enabled to identify full-length isoforms. For the deep analysis herpesvirus transcriptomes, we utilized the Pacific Biosciences (PacBio) RSII and the Sequel TGS machines, as well as the Oxford Nanopore Technologies (ONT) MinION device. In this project, we characterized the RNA profiles of the following viruses: human herpesvirus-1 (HSV-1), pseudorabies (PRV), varicella zoster virus (VZV), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV). For the analysis of the static transcriptome of HSV, PRV and HCMV the PacBio IsoSeq protocol was used. For the analysis of the dynamic properties of PRV transcript isoforms we applied the PacBio non-amplified method. To obtain a more comprehensive picture about the RNA variants, the ONT cDNA and direct RNA sequencing protocols, as well as a CAP-selection method combined with ONT cDNA sequencing were also utilized. These ONT approaches were applied for the analysis of EBV and VZV transcriptomes. Our surveys redefined the herpesvirus transcriptomes profiles. We discovered that the herpesvirus genomes are much more complex and the transcriptional-read-throughs between the genes are more common than it was known. Our data revealed that these read-throughs can generate very long RNAs including some containing oppositely oriented ORFs, which have been classified as complex transcripts. The applied TGS techniques have been shown that the extensive RNA variations are common among herpesviruses

Biography:

Hyoun Sub Lim was trained for his PhD in University of Illinois at Urbana-Champaign and he continued Postdoctoral studies in University of California at Berkeley. His researches have mainly focused on plant viral movement in plant cell for more than 20 years and more than fifty published papers proved his field in Plant Virology. Currently he is Professor in Chungnam National University, Korea and has worked for an Editorial Board Member of Plant Pathology journal.

Abstract:

As trading of agricultural products with neighboring countries has increased new plant diseases have been reported in Korea. Specifically, virus-contaminated imported seeds have damaged vegetable and fruit production. Little seed is now produced in Korea, and it is therefore very important to ensure that imported seed is not a source of new viruses. In order to investigate seed transmitted viruses we surveyed pepper fields nationwide and detected Pepper mild mottle virus (PMMoV) in the main pepper production regions of Sangcheng and Jeongson. We have generated full length clones of two isolates, named Sangcheong 47 (S-47, KX399390) and Jeongsong 76 (J-76, KX399389) respectively, in a T7 promoter-driven vector; sequencing revealed that these isolates shared ~99% nucleotide and amino acid identity, and are closely related to Japanese and Chinese isolates at the nucleotide level. Amino acid sequence comparisons revealed 99.73, 99.81, 98.44, and 100% identity in ORF1, ORF2, MP, and CP, respectively, between S-47 and J-76. Both isolates induced severe symptoms in Nicotiana benthamiana, but mild symptoms in Capsicum annuum. Each ORF was expressed as a GFP fusion from a binary vector, and no differences in subcellular localization were detected except for the 126 kDa proteins; the J-76 126 kDa clearly formed intracellular aggregates not observed with S-47 126 kDa protein. In addition, seed transmission rates from J-76 and S-47 infected Capsicum annuum showed 66% and 34% germinated plants from 500 harvested seeds respectively. Despite indistinguishable symptoms, J-76 showed a two times higher seed transmission rate that may result from some of the substitutions (S-47>J-76) of R(142)K, D(583)N, and V(931)I in 126 kDa and K(134)R, V(192)A, N(226)D, L(250)S in MP function with respect to S-47 and J-76.

Biography:

Abstract:

Apple is an economically important fruit crop, covering an area of 23,355 ha with an annual production of about 545,000 tonnes in South Korea. Apple trees are affected by at least six viruses and viroid diseases, which cause economic losses depend on the plant species and virus strains. Among these viruses, Apple chlorotic leafspot virus (ACLSV), Apple stem grooving virus (ASGV) and Apple stem pitting virus (APSV) commonly occur in Korean commercial apple orchards. In 2017, we observed several apple trees (Malus domestica Borkh.)  showing mosaic patterns on leaves cv. Shinano sweet or dappling and crinkling on fruits cv. Gamhong. To apply next generation sequencing (NGS) for apple virome of symptomatic apple trees, total RNA was extracted and used for constructing a cDNA library followed by high throughput sequencing (HTS). From the HTS data, ACLSV, ASGV, ASPV and Apple scar skin viroid (ASSVd) were identified, all of which have been previously reported in South Korea. In addition, Apple necrotic mosaic virus (ApNMV) recently described a novel ilarvirus was identified. To confirm the presence of ApNMV, RT-PCR was performed using specific primer pairs ApNMV-F and ApNMV-R. The sequence showed 94% identity with the CP region of the previously published ApNMV (LC108995) from the samples showing leaf mosaic symptoms. The ApNMV positive apple trees were also co-infected with ACLSV, ASGV and ASPV. The symptomatic fruit samples showing dappling and crinkling were infected with ACLSV, ASGV, ASPV and ASSVd. ASSVd is recognized as the causal agent of the dapple apple disease. ASSVd showed 99% nt. sequence identity with apple reference variants. NGS was a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees.

Biography:

Shu Hui Lin has her expertise in gene mutation test, cancer diagnosis and cancer research. She is a Leader of Molecular Pathology Department of Changhua Christian Hospital in Taiwan. She has PhD of Institute of Medicine. She is committed to cytological diagnosis and establishment technical of gene mutations test. Her research field focuses on oral squamous cell carcinoma and she hopes that she can find novel therapeutic targets in oral cancer.

Abstract:

Background: Oral cancer is the fourth highest incidence of malignancy in males and the seventh highest in the general population of Taiwan. Oral squamous cell carcinoma (OSCC) is the main subtype of oral cancer, which accounts for >95% of all cases of oral cancer. Discoidin domain receptors (DDRs), a collagen receptor tyrosine kinase, play a major role in cancer progression. DDR2 has been suggested as a prognostic marker in several cancer types; however, the correlation between DDR2 expression and clinical outcome of oral cancer patients in Taiwan population has not been investigated.

Material & Methods: In the present study we sought to determine the clinical significance of discoidin domain receptor tyrosine kinase 2 (DDR2) expression in OSCC patients. We examined DDR2 expression in OSCC specimens by immunohistochemistry and then we analyzed the association of DDR2 expression with clinicopathological factors in OSCC.

Result: We divided 254 OSCC cases into two groups based on DDR2 expression levels and compared with several clinicopathological factors and their overall survival. The group with high DDR2 expression had significantly higher frequencies of lymph node metastasis (P=0.0094) and AJCC stage (P=0.0058) compared to the group with low DDR2 expression. Furthermore, the lymph node metastasis oral cancer patients with high DDR2 expression had low survival rate than low DDR2 group (P=0.0458).

Conclusions: Our data indicate that DDR2 is a potent biomarker that can be used as an effective therapeutic target for treating OSCC patients with lymph node metastasis.

Biography:

Abstract:

Introduction: Hepatitis C virus (HCV) replication is closely linked to lipid metabolism. Therefore, lipidomic analysis of HCV infected hepatic cells can offer insights into the pathogenesis of HCV infection and identify molecular targets that can serve as potential targets for new treatments.

Aim: The aim of this project was to study the effects of HCV infection on intracellular lipid homeostasis and turnover in hepatic cells. We have previously reported HCV-induced changes in neutral lipids; so now present data on phospholipids.

Methods: Huh7 cells were infected with HCV (JFH1 strain) and cultured until they were 90% infected. Cellular lipids were separated by high performance thin layer chromatography (HP-TLC) to measure changes in the major phospholipid species. The rate limiting enzymes of phosphatidylcholine metabolism were knocked down and the effects on HCV replication were measured. 

Results: HP-TLC showed increased amounts of phosphatidylcholine in lipid extracts from whole cells and from ER fractions of JFH1 infected Huh-7 cells. PC was the only phospholipid species detected in purified lipid droplets, and was significantly increased in infected cells. PYTC1 (CTP: phosphocholine cytidylyltransferase) and PEMT (phosphatidylethanolamine N-methyltransferase) are the rate limiting enzymes of PC biosynthesis in hepatocytes. Silencing PYTC1 had no effect on HCV replication or infectivity. However, when PEMT was silenced, both viral replication and infectivity were decreased by more than 50%, and less lipids accumulation was observed.

Conclusion: Our previous data reveal global changes in lipid abundance, particularly in the ER, which are predicted to impact the HCV life cycle and pathogenesis. We now report increased PC content in the ER and in lipid droplets. Our data suggest that in HCV infected cells the minor PC synthesis pathway is most important, as inhibiting PEMT inhibits replication and production of infectious virus.

Biography:

Boyoun Moon completed her phD thesis in college of veterinary medicine of Seoul National Univeirsity. She is charge of the diagnosis and prevention of companion animal viral disesases in animal disease diagnostic division(ADDD), Animal and plant qurantine agency (APQA) in South Korea.

Abstract:

Canine parvovirus2 (CPV2) is an etiological agent that causes acute hemorrhagic enteritis and fatal myocarditis in carnivore species. CPV2 evolved from feline parvoviurs by host transmission and has contineously been replaced by the genetic variants of capsid protein (VP2). In this study, it was investigated that the genetic diversity and phylogenetic relatedness of the VP2 gene encoding capsid protein in CPV2 obtained from symptomatic domestic dogs obtained ranging from 2011 to 2017 in Korea. Positively selected sites on VP2 of CPV2 variants during a decade was identified and the multiple proteins were evaulated with the theoretical homology modeling using SWISS-MODEL and PyMOL. To investigate the subtype of CPV circulating in Korea, a total of 28 partial VP2 sequences were analyzed. Both CPV2a (15/28, 53.6%), and  CPV2b (12/28, 42.9%) were dominant subtypes and Korea CPV2c in 2017 was first detected in this study. Of them, eighteen complete VP2 nucleotide sequences (1755bp) and the amino acid were constructed with phylogenetic tree. A clustering of our variants was closely related to China and Vietnam strains, but in different groups from US, Eruope and vaccine strains.Non-synonymous mutations, Phe267Ala, Tyr324Ile and Thr440Ala have been increased during last decade and minor 10 mutations have been also observed. A codon-based maximum likelihood analysis of dN/dS (ω) revealed that specific amino acid 324 (ω=3.518), 426 (ω=3.517) and 440 (ω=3.459) in VP2 gene was under strong positive selection of Korea CPV2. These results support that dynamic process of CPV2 in Korea results in the locally adaptation.

Biography:

Mu Kuan Chen is currently the Superintendent of Changhua Christian Hospital. He is a prolific author in Head and Neck Medicine. He has served on numerous Editorial Boards of several international journals, been invited to be the book chapter writer. His main interests are focused on development of new techniques of minimally invasive surgery in otorhinolaryngology, head and surgery including endoscopic skull base surgery, endoscopic nasopharyngectomy, endoscope assisted parotidectomy and ablation of submandibular gland, and endoscopic sinonasal tumor excision. Besides, he has published many articles which focused on basic research of oral cancer and nasopharyngeal cancer

Abstract:

Oral squamous cell carcinoma (OSCC) is the most common primary malignancy occurring in the head and neck. It is the fourth most common male cancer and the fourth leading cause of male cancer death in Taiwan. Metastatic tumors are extremely common in the late stages of cancer. Treatment for metastatic cancer aims to decelerate the growth or spread of cancer. The extracellular matrix degradation involves various proteases such as matrix metalloproteinase (MMPs), are calcium-dependent zinc-containing endopeptidases. MMP-2 is type-IV collagenases of MMPs, and correlated with lymph node metastasis and advanced tumor stage groups. Pinosylvin, a preinfectious stilbenoid toxin, is used to study its properties as a fungi toxin and therapeutic agent. Pinosylvin is used as a representative stilbene to study its biological actions and therapeutic value in processes such as cell survival, apoptosis and cell mobility. However, the pharmacological activities of pinosylvin in anti-metastasis remain unclear. In this study, we used wound closure assay and transwell assay to determine the effects of pinosylvin on oral cancer cell migration and invasion. Pinosylvin treatment significantly inhibited the migration and invasion abilities of oral cancer cells in vitro. Gelatin zymography results revealed that pinosylvin inhibited MMP-2 activity. In addition, pinosylvin suppressed carcinoma-associated epithelial–mesenchymal transition in oral cancer cells. Pinosylvin inhibits the invasion of human oral cancer cells and is a potential chemopreventive agent against oral cancer metastasis.