Antiviral Drug Discovery and Development
Biography: Ljudmila Stojanovich received her Ph.D. in Medicine, with the thesis “Neuropsychiatric manifestations in patients with Systemic Lupus Erythematosus” in 1999. She is the scientific director in the Bezhanijska Kosa, University Medical Center of Belgrade University, where she is currently a Research Professor. Dr. Stojanovich’s research focuses on Systemic Lupus Erythematosus, Antiphospholipid Syndrome, and Vaccination in patients with Autoimmune Rheumatic diseases. She is an author of three monographs and of about 250 articles on various aspects of Autoimmune Rheumatic disorders, published in international and domestic journals and in conference proceedings. She is in Editorial Boards (Editorial Boards LUPUS (LONDON). /Reviewer in the “CURRENT CONTENSTS” or “Science citation index”, like LUPUS REWIEWER DATABAS, Cellular and Molecular Neurobiology, The Journal of Vaccine
Abstract: Objectives: Compared to the healthy population, patients suffering from autoimmune rheumatic diseases have a significantly increased risk of various infections. The issue of vaccinating against the seasonal flu in these patients is still surrounded by numerous dilemmas about its efficiency and the possible harmful effects of exacerbation of the underlying disease. We aim to assess the protective role of the influenza vaccine by analyzing the association between respiratory infections occurrence and humoral response to influenza A (H1N1) infection in patients with autoimmune rheumatic diseases.
Methods: Our study includes three groups of patients (99 in total) with stable underlying diseases status, suffering from: 30 patients with systemic lupus erythematosus (SLE), 37 with rheumatoid arthritis (RA) and 32 with Sjögren's Syndrome (SjS). In November 2011. 46 patients were immunized with an inactivated trivalent split vaccine (15 µg HA A/California/7/2009 (H1N1), 15 µg HA A/Pert/16/2009 (H3N2) and 15 µg / HA B Brisbane / 60/2008) whereas 52 patients did not accept the proposed vaccination.These three groups of patients were divided into two subgroups depending on vaccination: vaccinated - SLE1 (19), RA1 (15) and SjS1 (14), and unvaccinated - SLE2 (11), RA2 (22), SjS2 (18). During the following year disease activity parameters (SLEDAI for SLE), presence of viral and bacterial infections and concentration of A H1N1 antibodies were monitored in vaccinated and unvaccinated patients. Previous respiratory infections from 2006-2011 were regarded as a potentially significant predictor of a more frequent future onset of influenza and secondary bacterial complications.
In the following six months parameters of disease activity (from the date of vaccination until April 2013) and the titer of antibodies against influenza A H1N1 were monitored. We used hemagglutination inhibition test (according to the method of the Center for Disease Control and Prevention (CDC) with antigen A/California/7/2009 influenza virus (H1N1), and turkey erythrocytes for the detection of antibodies against the A H1N1. Value of seroprotective titer (ST) was defined as ≥ 32. Importance of predisposing factors for influenza occurrence (i.e. previous respiratory infections and vaccinations in last five years) was also analyzed.
Results: The incidence of viral and bacterial infections among vaccinated patients (primarily influenza) was significantly lower, compared to the non-vaccinated group. Influenza occurrence was significantly associated with previous respiratory infections (p=0.001). The mean titer of antibodies was highest in SLE patients and significantly higher in all vaccinated patients (p=0.018). Mid-level antibody titer was significantly related to last vaccination in all patients (p= 0.001) and has not been proven after removal of last vaccination effects (p=0.227). Similar results were obtained for patients with SLE, while in RA and SjS these correlations were not significant. ST levels for all vaccinated patients (84.17) were significantly higher than in non-vaccinated patients (8.80) (p=0.008) and were associated with last vaccination in all patients and in SLE group (p=0.012, p=0.039 respectively). Highest levels were observed in vaccinated SLE patients (141.05) and were significantly higher than in non-vaccinated patients (p=0.002).Seroprotective rate for all vaccinated patients was 48% compared to 15% in unvaccinated (p=0.014) and it was highest among SLE patients (53%) (p= 0.049). In RA and SjS groups, these differences were not statistically significant. There was no significant difference between three diseases regarding the mean ranks of antibody titer (p=0.418). Previous bronchitis and pneumonia increased influenza risk significantly.
Based on results to date, it is our opinion that overall, influenza vaccination for patients suffering from SLE, RA and Sjögren’s Syndrome is safe, efficient and sufficiently immunogenic. Based on several years of monitoring respiratory infections in our patients, it is clearly visible that a high risk for exacerbation of the underlying disease was linked to viral or bacterial infection, and practically never to the vaccination itself
Biography: Divocha Valentinain 1967 shegraduated from I. I. Mechnikov OdessaState University, Faculty of Biology (Department of Virology). In 1973 continued herpostgraduate study ate OdessaInstitute of Virologyand Epidemiology (specialty virology).In 1974 she was awarded her candidate degree with the thesis"Interaction ofCoxsackie Bviruseswith sensitivecell culturesand theirantigenicrelationships." In 2009 she was awarded her doctoral degree with the thesis entitled"Biological basis antiproteinasetherapyof influenza". Under her leadershipperformed a doctoraland twomaster's theses. Scientificexperienceis 35 years.I have more than190 scientific publications,3monographs, textbook "Virology" (2012), 10 patents, 3 innovations.
I am currently workingas the head ofthe Laboratory of ExperimentalandClinical Pathology for UkrainianResearch Instituteof Transport Medicine, is the supervisor ofthe nineresearch programsin virologyand biochemistry.
Abstract: Now preventive maintenance of a flu by means of vaccination is conventional and is supported by experts of all world. vaccination is recommended by WHO as a unique and obligatory measure of the prevention of a flu and its consequences for persons from groups of the raised risk on development of complications and death as a result of disease caused by viruses.
Keywords: trypsin-like proteinase, an inhibitor, vaccines, immunobiological blood preparations.
Objective – to check presence of trypsin-like proteinase and its inhibitor in antiflu and other vaccines and in immunobiological blood preparations of domestic and foreign manufacture.
Material and methods. In work following commercial preparations have been used:" Interferon leukocytic human"," the Immunoglobulin of human placental, donor 10 % "(Biofarma, Kiev), a gonococcal vaccine (Biolek, Kharkov) a herpetic vaccine (the Odessa factory of bacterial preparations, Odessa), vaccines for preventive maintenance of a flu, a season 2002/2003 -"Influvac" which consists of hemagglutinins and a neuraminidase of a virus of a flu, strains: А/Moscow/10/99 (H3N2), А/New Caledonia/20/99 (H/N), B/Hong Kong/330/2001 (Solvay Pharmaceuticals B.V. The Netherlands), "Fluarix" which consists of hemagglutinins of strains (H1N1) A/New Caledonia (H3N2), А/Panama and В/Shandont 17/97 (Smith Klein Bichem biologicals, Belgium) and "Vaxigrip" which consists of three strains of a flu virus (Pasteur Mere Konnot, France), a vaccine for preventive maintenance of a hepatitis A - "Avaxim" (Pasteur Mere Konnot, France), a blood preparation received from a heparin (the antifactor of Ha) - "Fraxiparine" (Sanofi-Сhinoin, France), a preparation from a blood of calfs for a hemodialysis - "Solcoseryl" (Solco, Switzerland). Preparations were investigated before the termination of a period of validity. All preparations have been bought in drugstores of Odessa.
Activity of tripsyn-like proteinase and its inhibitor defined by the method of A. P.Levitsky in our updating , the general protein - O.Lowry's method.
Resuits.Work is devoted to study presence of components of a cell-owner and its inhibitor in vaccines and blood preparations and to define presence trypsin-like proteinase and its inhibitor in vaccines and blood preparations. It is revealed that anti influenza vaccines (influvac, vaxigrip, fluarix), herpetic and tularemic vaccines contained an inhibitor of trypsin-like proteinase in considerable quantity. Commercial preparations from a human donor blood (an immunoglobulin, interferon, fraxiparine and solcoseryl) contained as trypsin-like proteinase, and its inhibitor. The immunoglobulin contained in 4,0 times more inhibitor, than interferon.
Conclsions. Hence, the modern vaccines applied to prophilaxis and treatment, are insufficiently cleared. Presence of cellular components (enzymes and inhibitors) could lead to allergization and follow complication which is not very known.
Therapeutic Approaches and Targets for Viral Infections
Biography: Dr. Dimitrov graduated and completed his PhD at the University of Sofia, Sofia, Bulgaria, and thereafter he worked in the Bulgarian Academy of Sciences where he defended his ScD and was Professor of Biophysics until he joined the National Cancer Institute (NCI) of the National Institutes of Health (NIH), USA, in 1990. There he was tenured as Senior Investigator and appointed at the Senior Biomedical Research Service. His research group includes molecular biologists who are experts in display/screening/libraries methodologies, antibody engineering, and protein biochemistry, and a structural biologist. His major long-term goal is the development of clinically useful therapeutics and vaccines based on human monoclonal antibodies including engineered antibody domains. He has authored or coauthored more than 370 articles cited more than 20,000 times, and is inventor or coinventor of more than 60 inventions, patent applications or patents. See also his web site https://ccr.cancer.gov/Cancer-and-Inflammation-Program/dimiter-s-dimitrov
Abstract: Our group work on the identification, characterization and engineering of human monoclonal antibodies (mAbs) in IgG1, Fab, scFv, VH and CH2 formats, and as antibody drug conjugates, chimeric antigen receptors (CARs) and bispecific antibodies. Our most important engineered antibody and CD4 domains are those against HIV-1 with the goal of HIV-1 eradication – will present recent unpublished data from animal studies demonstrating the exceptional potency of our bispecific multivalent fusion protein based on one-domain soluble CD4 and the antibody domain m36.4 targeting coreceptor binding site. I will also discuss our full-size mAbs against emerging and biodefense-related viruses with an emphasis on henipaviruses and coronaviruses mostly MERS-CoV especially for prophylaxis and therapy of humans.
Biography: Elham O Mahgoub was graduated her B.Sc in Animal Sciences at Alneelain University, Sudan in 1999 work as teaching assistant at Alneelain University.In 2005 she had her M.Sc. in Virology from University Putra Malaysia, Malaysia. In 2006 to 2010 she was a fellowship student at University Putra Malaysia, faculty of biotechnology. She graduated her PhD in Molecular immunologyin 2013 from Alneelain University. She spent four years as research fellow in Supreme Council of health of Qatar. In 2015 she works as a research fellow at Weill Cornell College in Qatar. In 2016 she works as research fellow at Qatar university until present day.
Abstract: The development and evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of serum antibody to chicken anaemia virus (CAV) are described.This test depends on the availability of CAV polyclonal antibodies present in convalescent chicken serum to react with the VP1 antigen and adsorbed to the ELISA plate. In the methods,VP1 gene is translated to be thecapsid protein that hold most the receptor responsible of diagnosis of chicken anaemia virus (CAV). VP1 antigen was manufactured to use as coating protein on absorbent face of indirect ELISA.As start, the VP1 gene was inserted intopRSET–B plasmid. The pRSET–B plasmid together with inserted VP1 was transformed into E. coli top10 competent cells. The VP1 protein expression was then detected when anti-VP1 monoclonal antibodywas used Batch fermentation was used to scale up production of the protein in E. colhost. The recombinant VP1 protein was successfully expressed during high cell density culture. The use ofTangential Flow Filtration (TFF)filtration step for dialysis and desalting increasedboth the specific activity and the final yield of the purifiedfraction. The protein expressed has been tested as an antigen for detection of antibody to CAV in infected chicken. An enzyme-linked immunosorbent assay (ELISA) of the chicken anaemia virus was prepared for the detection of serum antibody to CAV.sera samples from 60 (positive CAV chicken sera) were tested. Statistical analysis methods were applied to measure ELISA specificity, sensitivity,p-value andt-value. In the results and discussions: a band of 50 kDa was showed in western blot test as a proof of the VP1 protein expression.Theindirect ELISA specificity was 93.3% and sensitivity was 100%. A t test produced a t-value of 15.805 for the indirect ELISA and revealed a significant differencebetween CAV-positive serum and CAV-negative serum (p-value of 0.001). For the second variable. the t-test yielded a t-value of 5.063, which revealed a significant differencebetween CAV-positive serum and CAV-negative serum (p-value of 0.015). In conclusions: This indicate that the indirect ELISA approach using VP1 fusion protein has many advantages compared to the commercial indirect ELISA. However, the present study can be a base for the development of ELISA assay for CAV that could reduce the test cost of indirect ELISA method for diagnosis and increase its reliability.
Biography: Segundo Mesa Castillo. As Specialist in Neurology, he worked for 10 years in the Institute of Neurology of Havana, Cuba. He has worked in Electron Microscopic Studies on Schizophrenia for 32 years. He was awarded with the International Price of the Stanley Foundation Award Program and for the Professional Committee to work as a fellowship position in the Laboratory of the Central Nervous System Studies, National Institute of Neurological Diseases and Stroke under Dr. Joseph Gibbs for a period of 6 months, National Institute of Health, Bethesda, Maryland, Washington D.C. USA, June 5, 1990.
Abstract: There is increasing evidences that favor the prenatal beginning of schizophrenia. These evidences point toward intra-uterine environmental factors that act specifically during the second pregnancy trimester producing a direct damage of the brain of the fetus. The current available technology doesn't allow observing what is happening at cellular level since the human brain is not exposed to a direct analysis in that stage of the life in subjects at high risk of developing schizophrenia. Methods. In 1977 we began a direct electron microscopic research of the brain of fetuses at high risk from schizophrenic mothers in order to finding differences at cellular level in relation to controls. Results. In these studies we have observed within the nuclei of neurons the presence of complete and incomplete viral particles that reacted in positive form with antibodies to herpes simplex hominis type I [HSV1] virus, and mitochondria alterations. Conclusion. The importance of these findings can have practical applications in the prevention of the illness keeping in mind its direct relation to the aetiology and physiopathology of schizophrenia. A study of amniotic fluid cells in women at risk of having a schizophrenic offspring is considered. Of being observed the same alterations that those observed previously in the cells of the brain of the studied foetuses, it would intend to these women in risk of having a schizophrenia descendant, previous information of the results, the voluntary medical interruption of the pregnancy or an early anti HSV1 viral treatment as preventive measure of the later development of the illness.
Biography: Hua Zhu is from Department of Microbiology, Biochemistry and Molecular Genetics.
Abstract: Reactivation of human cytomegalovirus (HCMV) in transplant recipients can cause lifethreatening
disease. Consequently, for transplant recipients, killing latently infected cells
could have far-reaching clinical benefits. In vivo, myeloid cells and their progenitors are an
important site of HCMV latency, and one viral gene expressed by latently infected myeloid
cells is US28. This viral gene encodes a cell surface G protein-coupled receptor (GPCR) that
binds chemokines, triggering its endocytosis. We show that the expression of US28 on the
surface of latently infected cells allows monocytes and their progenitor CD34þ cells to be
targeted and killed by F49A-FTP, a highly specific fusion toxin protein that binds this viral
GPCR. As expected, this specific targeting of latently infected cells by F49A-FTP also robustly
reduces virus reactivation in vitro. Consequently, such specific fusion toxin proteins could
form the basis of a therapeutic strategy for eliminating latently infected cells before
haematopoietic stem cell transplantation.
Biography: TourajA.Farzani has been earned his DVM from Urmia veterinary medicine school of Iran. After graduation and 2 years stint in the Navy as a veterinary surgeon, he started his virology PhD in the Virology Department of Veterinary Faculty of Ankara University, Turkey. During his doctorate studentship there, he has been involved in Prof. Dr. AykutÖzkul's project to develop a new strategies including DNA and viral vector (adenoviral vectors) against CCHFV. His doctorate thesis is to create BHV-4-BAC system to use in future vaccination and gene therapy under the mentorship of Prof. Dr. Seval Bilge Dağalp. During these activities, he learned numerous laboratory techniques including recombinant DNA cloning, cell culture and homologous recombination in eukaryotic and prokaryotic cells. His main focus is on the viral vector systems in vaccination and gene therapy.
Abstract: BoHV-4 is a member of the family of Herpesviridae, subfamily Gammaherpesvirinae in the Rhadinovirus genus, and it is found worldwide among cattle populations (1, 2).
According to the previous researches, Bovine herpesvirus type 4(BoHV-4) is sparkling member of the attractive new emerging viral vectors in the vaccination and gene therapy fields. Several biological characteristics of bovine herpesvirus 4 (BoHV-4) make it a good candidate as a gene delivery vector for vaccination purposes. These characteristics include little or no pathogenicity, unlikely oncogenicity, the capability to accommodate large amounts of foreign genetic material, the ability to infect several cell types coming from different animal species, and the ability to maintain transgene expression in both undifferentiated and differentiated cells (3).
In addition, compared to other herpesviruses BoHV-4 has less complex genome with some determined gene areas to insert the desired gene. Being episomal in the target cells including macrophages,B and T cells makes this virus as a new candidate in the field of viral vectors (4).
To develop a BoHV-4-BAC vector in this research, we have selected the gene area among gp3 and gp4 which has been previously shown to besuitable for introduce of foreign DNA and after insertion of the BAC cassette the resulting virus is stable and able to replicate in vitro and in vivo(5).
For this, BAC cassette from pBeloBAC11vector that has been ligated to CMV-EGFP-Neo cassette(from pEGFP-C1) flanked by loxp has been inserted between gp3 (Bo2) and gp4 (Bo3) of Movar33/63 strain of BoHV-4. The extrachromosomal homologous recombination has been performed in the MDBK cell line after electroporation of circular plasmid containing homologous arms and BAC cassette after inoculation of the virus. After three rounds of G418 selection in BEK cells (700µg/ml) and one plaque purification, the recombinant virus DNA has been extracted by Hirt methodand used for transformationof DH10B cells by electroporation(6). After 20 serial passages in the bacteria, the stability of BoHV-4-BAC has been proved.
This BoHV-4-BAC vector system will be utilized in the future researches including viral vaccination and gene therapy.
Retrovirus-HIV/AIDS-Basic Sciences and Implications
Biography: Dr. De received Bachelor of Science degree with honors in chemistry and Master of Science in biochemistry from Calcutta University, India. He worked for few years at a National Cancer Institute in India before coming to US for post- graduate education in 1975. After completion of Ph.D in biochemistry from University of Maryland in 1980, he underwent two postdoctoral trainings in basic cancer research under the supervisions of Dr. Leon Heppel, National Academy of Sciences, at Cornell University, Ithaca, NY and Dr. Stanley Cohen, Nobel laureate, Medicine 1986, at Vanderbilt University, Nashville, TN. Gradually, he became highly interested in Clinical Chemistry. In 1993, he took a fellowship in clinical chemistry and toxicology under the supervisions of both Dr. Amadeo Pesce and Dr. Bradley Copeland at University of Cincinnati Medical Center, Cincinnati, OH. Following completionhe joined University of Mississippi Medical Center in 1995 as the Associate Director of clinical chemistry and toxicology. In 2005, Dr. De was appointed the Director of clinical chemistry and toxicology in the department of Pathology, University of Arizona Health Sciences Center. He is a board certified clinical chemist from the American Board of Clinical Chemistry, a board certified toxicologist from the National Registry of Clinical Chemistry and a fellow of the National Academy of Clinical Biochemistry. Since 2010, Dr. De has been serving as a member of the board of directors in the American Board of Clinical Chemistry, Toxicology and Molecular Biology. Last year he is elected to be the chair of the nomination committee, American Board of Clinical Chemistry, Toxicology and Molecular Pathology. He has published more than 68 peer reviewed articles and meeting abstracts in basic and clinical sciences. He is a reviewer of both national and international journals and received recognitions as a Clinical Chemist from the American Association of Clinical Chemistry. In 2013 he was honored for the “Paul Finley Chemist Award” from University of Arizona Health Sciences Center. He is the co-principal investigator of a Center of Disease Control (CDC) sponsored project on HIV screening and diagnosis.
Abstract: Acquired immunodeficiency syndrome (AIDS) is caused by two types of human immunodeficiency viruses HIV-1 and HIV-2, collectively knownas HIV. HIV infection is spreading globally, particularly in developing countries. HIV-1 is the prevalent infection in US and Western European countries. HIV-1 infection goes through several stages during which an individual can be diagnosed with HIV. Following initial viral transmission, the first stage is known as an “acute early phase”where the immune system has no specific antibody defenseand the viral load is high,making a person highly infectious. Once the immune system begins producing antibodies, during the later acute phasefollowingseroconvertion,the HIV entersa chronic phase, known as established infection. Subsequently, there is a latency period, during which the immune system is slowly degraded and after which the individual becomes immunodeficient. At this stage CD4 cell count is below 200 per cubic mm regardless of the presence or absence of symptoms, and is designated as having AIDS. Presently, the majority of HIV diagnosis involves use of testing that involves anti-body only detection, and do not detect the acute phase infection during which both the viral RNA and p24 antigen are expressed. These less sensitive tests (both immunoassays and western blots) detect antibodies to viral antigens which are typically seroconverted about three weeks later in the disease process following acute infection. These antibodies are detected in both asymptomatic HIV-infected individuals as well as AIDS patients. Studies indicate that early diagnosis and treatment of HIV infection canimprove survival, reduce medical costs, and reduce transmission of HIV to new uninfected partners. Newer 4th generation combination antigen/antibody tests are highly sensitive and specific for detection of acute and established HIV infection (HIV1 and HIV2) enabling immediate linkage to care. The CDC (Center of Disease Control, USA) recently recommended an algorithm involving three different tests to screen and diagnose acute and established infections of HIV-1 and HIV-2 in a general population. Initially a 4th generation combo test detects a viral antigen p24 and specific antibodies against HIV -1 and HIV-2 envelope proteins. If the test becomes positive it is followed by a second test, known as a differentiation assay, which detects antibodies against specific HIV-1 and HIV-2 envelope proteins confirming established infection of either HIV-1 or HIV-2. If this differentiation assay is negative,then another test is performed that measures viral load confirming an acute HIV-1 infection.The turnaround time of this new 4th gen. test algorithm is significantly faster than 3rd generation test algorithm tests practiced in many healthcare centers. Studies in the U.S. indicate that this new algorithm of tests are rapid enough to be employed in the Emergency Department setting for HIV screening and linkage to ongoing care. This approachshould effectively reduce HIV transmissionthrough immediate treatment and patient education, following early diagnosis by screening.
Biography: Rongtuan Lin is associate professor from Lady Davis Institute-Jewish General Hospital, Department of Medicine, McGill University, Montréal, Canada.
Abstract: Oncolytic viruses (OVs) are novel anticancer agents that infect and effectively kill cancer cells but not normal cells. Although tumor growth is delayed or eliminated in numerous animal models following treatment with OVs, several cancer models remain partially or completely resistant to viral oncolysis. To overcome this resistance, experimental strategies are now combining OVs with different cytotoxic compounds to improve OV efficacy. Our laboratory has previously demonstrated that OV replication can be bolstered by co-administration of other chemical agents such as Triptolide, a natural molecule derived from the medicinal herb.In the current study, we investigated the capacity of sulforaphane (SFN) -an anti-cancer compound naturally occurred in cruciferous vegetables, with demonstrated potent antioxidant and possible anti-inflammatory actions- to enhance vesicular stomatitis virus (VSV) oncolysis in OV-resistant cancer cells. We show that the manipulation of the anti-oxidant network via transcription factor Nrf2 augments vesicular stomatitis virus Δ51 (VSVΔ51) replication and sensitizes cancer cells to viral oncolysis. Activation of Nrf2 signaling by sulforaphane (SFN) leads to enhanced VSVΔ51 spread in OV-resistant cancer cells and improves the therapeutic outcome in different murine syngeneic and xenograft tumor models. Furthermore, chemoresistant lung cancer cells displaying constitutive dominant hyperactivation of Nrf2 signaling are the most susceptible to VSVΔ51 oncolysis. Mechanistically, enhanced Nrf2 signaling stimulates viral replication in cancer cells and disrupts the type I IFN response via increased autophagy. This study reveals a previously unappreciated role for Nrf2 in the regulation of autophagy and innate cellular antiviral response that complements the therapeutic potential of VSV-directed oncolysis against multiple types of OV-resistant or chemoresistant cancer.
Biography: Zhanqiu Yang has completed his MD at the age of 35 years from Wuhan University School of Medicine. He is the director of Institute of Medical Virology, Wuhan University. He has published more than 150 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract: Arbidol Hydrochloride (ARB) is an indole-derivative molecule with broad-spectrum antiviral activity. Herpes simplex virus type 1(HSV-1) causes significant human diseases from skin lesions to encephalitis, especially in neonates and immunocompromised hosts. The discovery of novel anti-HSV-1 drugs with low toxicity is in demand for public health In this study, we evaluated the antiviral effects of ARB against HSV-1 infection using in vitro and in vivo models. The results showed that ARB presents potent inhibitory effect on HSV-1 plaque formation and generation of progeny virus with EC50 value of 5.39 μg/ml (10.49μM) and 2.26 μg/ml (4.40μM), respectively. Moreover, time-of-addition and -removal assays further suggested that ARB showed viral inhibitory effects if added up to 12 h p.i., which could be further corroborated by determining the expression of viral immediate early (ICP4, ICP22, ICP27), early (ICP8, UL42) and late genes (gB, gD, gH, VP1/2, VP16) by qRT-PCR and the expression of viral protein ICP4 and ICP8 at 6 and 12 h p.i . The results of in vivo study showed that ARB could reduce the guinea pigs skin lesion caused by HSV-1 infection. Conclusively, this report offers new perspectives in the search for therapeutic measures in the treatment of HSV-1 infection.
Biography: Mulatu Biru is from Department of Health Sciences, Faculty of Medicine, Lund University, Sweden.
Abstract: Introduction: Attrition from antiretroviral therapy (ART) programs is a critical challenge for children living with HIV. However, only few studies examined rates and predictors of attrition through a prospective cohort design. Our aim was therefore to determine the rates and predictors of attrition from treatment follow up among children on ART in selected health facilities in Ethiopia.
Materials & Method: We conducted prospective cohort study among children aged between
3 months and 14 years on ART from December 2014 to September 2016 in eight health facilities in Ethiopia. Study outcome was attrition due to death and/or loss to follow-up. We used Cox proportional hazards models to examine the association between predictors and times to attrition.
Result: Out of the 309 children approached, 304 eligible children were included in the study of whom 52% were male and their median age was 9 years (IQR 6-12). Their baseline median CD4 was 362 cells/mm3 and 74.3 % had WHO stage 1 or 2 disease. During 287.7 person-years of observation (PYO), 24 attritions were recorded yielding an attrition rate of 8.3 per 100 PYO. Of these, six attritions were due to death with five of the deaths occurring during the first eight months of follow-up. Child age below three years [aHR]= 5.14 (95% CI: 2.07-12.96) and baseline Hgb < 10 g/dl [aHR]= 5.68 (95% CI: 2.03-6.23) independently predicted attrition. On the other hand, baseline Hgb < 10 g/dl (aHR= 16.63, 95% CI: 1.64, 168.4) and WHO stage III or IV (aHR= 12.25, 95% CI: 1.26, 119.05) were found to predict the death of a child. Surprisingly, five out of six deaths occurred among parents coming from urban areas and most of the attrition (71%) occurred among children whose biological parents were alive.
Conclusion: Younger children and those with anaemia were at increased risk of attrition, with most of the deaths occurring during the early months of treatment. More attention should be given to at-risk children during the early months of treatment. Further studies are needed to better understand the underlying reasons for the higher attrition.
Key words: Children, HIV, attrition, antiretroviral, treatment, outcomes, Ethiopia
Biography: Zaki Monawar Eisa is from head of Virology and Molecular Diagnosis
King Fahd Hospital, Jazan, KSA.
Abstract: Hepatitis C Virus (HCV) is a serious problem of chronic liver disease. It may lead to development of liver cirrhosis and hepatocellular carcinoma. Therapy used to treat patients with chronic HCV infection includes treatment with interferon either on its own or in combination with ribavirin. Some infection cases do not respond to treatment and an alternative treatment is mandatory.
RNA interference (RNAi) is an antiviral mechanism which is present in
plants that induces double-stranded RNA degradation. Oligonucleotides corresponding to different parts of the HCV NS3 coding region were ligated into pSUPER, and expressed as small interfering RNAs (siRNAs) after transfection. Fluorescent Activated Cell Staining (FACS), northern blotting, Western blotting, and Real Time-PCR demonstrated that one of these siRNAs was able to ablate HCV RNA by RNAi.
RNAi may represent a new approach for the treatment of HCV infection.
Biography: Decheng Yang, PhD, Professor is in Department of Pathology and Laboratory Medicine, University of British Columbia. He is Principal Investigator, The Center for Heart and Lung Innovation, St. Paul’s Hospital, Vancouver, British Columbia, Canada. Research Interest: picornaviral pathogenesis, microRNA, signal transduction and antiviral drug development.
Abstract: Coxsackievirus 3 (CVB3) is a primary causal agent of viral myocarditis. Emodin is a natural compound isolated from certain plant roots. In the present study, we found that emodin inhibited CVB3 replication in vitro and in mice, and now we report an unrecognized mechanism by which emodin inhibits CVB3 replication through suppression of viral protein translation via multiple pathways. On one hand, emodin treatment inhibited Akt/mTOR (mammalian target of rapamycin) signalling and activated 4EBP1 (eukaryotic initiation factor 4E-binding protein 1), leading to suppression of translation initiation of ribosomal protein L32. On the other hand, emodin treatment differentially regulated multiple signal cascades, including Akt/mTORC1/p70S6K (p70 S6 kinase), ERK1/2 (extracellularsignal-regulated kinase 1/2)/p90RSK (p90 ribosomal S6 kinase) and Ca2 + /calmodulin, leading to activation of eEF2K (eukaryotic elongation factor 2 kinase) and subsequent inactivation of eEF2 (eukaryotic elongation factor 2), resulting in inhibition of CVB3 VP1 (viral protein 1) synthesis. These data imply that eEF2K is a major factor mediating cross-talk of different arms of signalling cascades in this signal network. This notion was verified by either overexpressing eEF2K or treating the cells with siRNAs or eEF2K inhibitor A484954. We showed further that the emodin-induced decrease in p70S6K phosphorylation plays a dominant positive role in activation of eEF2K and in turn in conferring the antiviral effect of emodin. This finding was further solidified by expressing constitutively active and dominant-negative Akt. Collectively, our data reveal that emodin inhibits viral replication through impairing translational machinery and suppression of viral translation elongation.