Search results for: human immunodeficiency virus
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 8465

Search results for: human immunodeficiency virus

8345 Efficacy of Pooled Sera in Comparison with Commercially Acquired Quality Control Sample for Internal Quality Control at the Nkwen District Hospital Laboratory

Authors: Diom Loreen Ndum, Omarine Njimanted

Abstract:

With increasing automation in clinical laboratories, the requirements for quality control materials have greatly increased in order to monitor daily performance. The constant use of commercial control material is not economically feasible for many developing countries because of non-availability or the high-cost of the materials. Therefore, preparation and use of in-house quality control serum will be a very cost-effective measure with respect to laboratory needs.The objective of this study was to determine the efficacy of in-house prepared pooled sera with respect to commercially acquired control sample for routine internal quality control at the Nkwen District Hospital Laboratory. This was an analytical study, serum was taken from leftover serum samples of 5 healthy adult blood donors at the blood bank of Nkwen District Hospital, which had been screened negative for human immunodeficiency virus (HIV), hepatitis C virus (HCV) and Hepatitis B antigens (HBsAg), and were pooled together in a sterile container. From the pooled sera, sixty aliquots of 150µL each were prepared. Forty aliquots of 150µL each of commercially acquired samples were prepared after reconstitution and stored in a deep freezer at − 20°C until it was required for analysis. This study started from the 9th June to 12th August 2022. Every day, alongside with commercial control sample, one aliquot of pooled sera was removed from the deep freezer and allowed to thaw before analyzed for the following parameters: blood urea, serum creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), potassium and sodium. After getting the first 20 values for each parameter of pooled sera, the mean, standard deviation and coefficient of variation were calculated, and a Levey-Jennings (L-J) chart established. The mean and standard deviation for commercially acquired control sample was provided by the manufacturer. The following results were observed; pooled sera had lesser standard deviation for creatinine, urea and AST than commercially acquired control samples. There was statistically significant difference (p<0.05) between the mean values of creatinine, urea and AST for in-house quality control when compared with commercial control. The coefficient of variation for the parameters for both commercial control and in-house control samples were less than 30%, which is an acceptable difference. The L-J charts revealed shifts and trends (warning signs), so troubleshooting and corrective measures were taken. In conclusion, in-house quality control sample prepared from pooled serum can be a good control sample for routine internal quality control.

Keywords: internal quality control, levey-jennings chart, pooled sera, shifts, trends, westgard rules

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8344 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells

Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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8343 John Cunningham Virus Interaction with Multiple Sclerosis Disease Progression

Authors: Sina Mahdavi

Abstract:

Background and Objective: Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the central nervous system (CNS) that affects the myelination process in the CNS. Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially the John Cunningham virus (JCV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on JCV infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis", " John Cunningham virus ", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2019 and 2022 were searched, and 12 articles were chosen, studied, and analyzed. Results: MS patients are candidates for natalizumab therapy, which inhibits lymphocyte migration and increases the risk of progressive multifocal leukoencephalopathy (PML), a rare lytic infection of glial cells caused by JCV. Oligodendrocytes may be the target of JCV infection in the central nervous system (CNS). Conclusion: There is a high expression of JCV during the natalizumab treatment period for MS patients, suggesting that the virus may play a role in the development of MS by inducing an inflammatory state. Therefore, it is necessary to evaluate anti-JCV antibody serum as an important risk factor for the development of PML before deciding on the treatment course for these patients.

Keywords: multiple sclerosis, John Cunningham virus, central nervous system, autoimmunity

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8342 A Description Analysis of Mortality Rate of Human Infection with Avian Influenza A(H7N9) Virus in China

Authors: Lei Zhou, Chao Li, Ruiqi Ren, Dan Li, Yali Wang, Daxin Ni, Zijian Feng, Qun Li

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Background: Since the first human infection with avian influenza A(H7N9) case was reported in China on 31 March 2013, five epidemics have been observed in China through February 2013 and September 2017. Though the overall mortality rate of H7N9 has remained as high as around 40% throughout the five epidemics, the specific mortality rate in Mainland China varied by provinces. We conducted a descriptive analysis of mortality rates of H7N9 cases to explore the various severity features of the disease and then to provide clues of further analyses of potential factors associated with the severity of the disease. Methods: The data for analysis originated from the National Notifiable Infectious Disease Report and Surveillance System (NNIDRSS). The surveillance system and identification procedure for H7N9 infection have not changed in China since 2013. The definition of a confirmed H7N9 case is as same as previous reports. Mortality rates of H7N9 cases are described and compared by time and location of reporting, age and sex, and genetic features of H7N9 virus strains. Results: The overall mortality rate, the male and female specific overall rates of H7N9 is 39.6% (608/1533), 40.3% (432/1072) and 38.2% (176/461), respectively. There was no significant difference between the mortality rates of male and female. The age-specific mortality rates are significantly varied by age groups (χ²=38.16, p < 0.001). The mortality of H7N9 cases in the age group between 20 and 60 (33.17%) and age group of over 60 (51.16%) is much higher than that in the age group of under 20 (5.00%). Considering the time of reporting, the mortality rates of cases which were reported in the first (40.57%) and fourth (42.51%) quarters of each year are significantly higher than the mortality of cases which were reported in the second (36.02%) and third (27.27%) quarters (χ²=75.18, p < 0.001). The geographic specific mortality rates vary too. The mortality rates of H7N9 cases reported from the Northeast China (66.67%) and Westeast China (56.52%) are significantly higher than that of H7N9 cases reported from the remained area of mainland China. The mortality rate of H7N9 cases reported from the Central China is the lowest (34.38%). The mortality rates of H7N9 cases reported from rural (37.76%) and urban (38.96%) areas are similar. The mortality rate of H7N9 cases infected with the highly pathogenic avian influenza A(H7N9) virus (48.15%) is higher than the rate of H7N9 cases infected with the low pathogenic avian influenza A(H7N9) virus (37.57%), but the difference is not statistically significant. Preliminary analyses showed that age and some clinical complications such as respiratory failure, heart failure, and septic shock could be potential risk factors associated with the death of H7N9 cases. Conclusions: The mortality rates of H7N9 cases varied by age, sex, time of reporting and geographical location in mainland China. Further in-depth analyses and field investigations of the factors associated with the severity of H7N9 cases need to be considered.

Keywords: H7N9 virus, Avian Influenza, mortality, China

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8341 Influenza Virus Circulation among the Population of Kazakhstan in 2012-2014

Authors: N. G. Klivleyeva, T. I. Glebova, G. V. Lukmanova, S. B. Bayseit, S. Z. Taubaeva, M. K. Kalkozhaeva

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The role of viral diseases in the general infectious disease incidence increases every year and requires special attention to the problem of interpreting the etiology of infectious agents. Influenza and acute respiratory viral infections are one of the most pressing public health issues. In the period 2012-2014, collection of 419 nasal swabs and 150 blood sera has been carried out in the patient care institutions of the various Kazakhstan regions from patients with symptoms of ARVI and pneumonia. Primary identification of biosamples for the presence of influenza viral antigens in enzyme immunoassay on nitrocellulose membrane gave positive results in 125 swabs (29.8%). Biosample screening in immunofluorescence test revealed the presence of influenza viral antigens against A/H1 in 63 samples (15.0%), A/H3 – in 70 samples (16.7%) and type B – in 9 samples (2.1%). As a result of primary infection, and successive passages in chick embryos and MDCK cell cultures, 38 HAAg were isolated from 419 samples with a clear cytopathic effect and hemagglutination titre in MDCK cell culture within 1:2-1:4, in CE - 1:8-1:256. The infectivity of isolates in chicken embryos were 3.5-6.5 lg EID50/0.2, in MDCK cell culture – 2.5-6.5 lg PFU/ml. Identification of 28 isolates was carried out in inhibition reactions of hemagglutinating activity and neuraminidase activity, showed their belonging to the influenza virus: 26 strains to A/H1N1, one - to A/H3N2, and one - to type B. Serological examination of blood sera for the presence of specific antibodies being an indirect evidence of the performed isolation and contributing to the timely interpretation of the disease etiology in the epidemics takes an important place in the comprehensive study of influenza viruses circulating among people. Serological analyzes were carried out in HAI assay using a kit consisting of 12 reference strains obtained from the WHO centre for reference and research on Influenza (CDC, Atlanta, USA) and three Kazakhstan (A/Almaty/347/09 (H1N1v), A/Almaty/462/11 (H3N2) and B/Almaty/414/10) human influenza viruses that are stored in the laboratory collection. The results of serological analysis of 150 blood sera showed that antihaemagglutinins against the A/H3N2 virus serosubtype were found in 46 samples (49.4%) out of 93 sera collected in 2012-2013. The antibody titres were within 1:160-1:320. 19 sera (20.4%) were seropositive against influenza A/H1N1 virus, the antibodies were observed in titres of 1:20-1:40. Six sera (6.4%) were positive against the influenza A/H1N1+A/H3N2 virus (mixed infection); the antibodies were recorded in titres of 1:20-1:40. Antihaemagglutinins against influenza type B virus were detected only in five sera (5.4%). The results of analysis of 57 sera collected in 2014 showed that antihaemagglutinins against A/H3N2 virus subtype were detected in 32 blood sera (56.1%) in titres of 1:160-1:640. Ten sera (17.5%) were seropositive against A/H1N1 virus; antihaemagglutinins against influenza type B virus were not detected. Therefore, virological and serological studies have shown that in Kazakhstan, as well as in the world, the influenza viruses A/H1N1, A/H3N2 and influenza B viruses were actively circulating during the epidemic seasons in 2012-2014.

Keywords: influenza, MDCK cell, serological analysis, virus

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8340 Representative Concentration Pathways Approach on Wolbachia Controlling Dengue Virus in Aedes aegypti

Authors: Ida Bagus Mandhara Brasika, I Dewa Gde Sathya Deva

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Wolbachia is recently developed as the natural enemy of Dengue virus (DENV). It inhibits the replication of DENV in Aedes aegypti. Both DENV and its vector, Aedes aegypty, are sensitive to climate factor especially temperature. The changing of climate has a direct impact on temperature which means changing the vector transmission. Temperature has been known to effect Wolbachia density as it has an ideal temperature to grow. Some scenarios, which are known as Representative Concentration Pathways (RCPs), have been developed by Intergovernmental Panel on Climate Change (IPCC) to predict the future climate based on greenhouse gases concentration. These scenarios are applied to mitigate the future change of Aedes aegypti migration and how Wolbachia could control the virus. The prediction will determine the schemes to release Wolbachia-injected Aedes aegypti to reduce DENV transmission.

Keywords: Aedes aegypti, climate change, dengue virus, Intergovernmental Panel on Climate Change, representative concentration pathways, Wolbachia

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8339 Serological and Molecular Detection of Alfalfa Mosaic Virus in the Major Potato Growing Areas of Saudi Arabia

Authors: Khalid Alhudaib

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Potato is considered as one of the most important and potential vegetable crops in Saudi Arabia. Alfalfa mosaic virus (AMV), genus Alfamovirus, family Bromoviridae is among the broad spread of viruses in potato. During spring and fall growing seasons of potato in 2015 and 2016, several field visits were conducted in the four major growing areas of potato cultivation (Riyadh-Qaseem-Hail-Hard). The presence of AMV was detected in samples using ELISA, dot blot hybridization and/or RT-PCR. The highest occurrence of AMV was observed as 18.6% in Qaseem followed by Riyadh with 15.2% while; the lowest infection rates were recorded in Hard and Hail, 8.3 and 10.4%, respectively. The sequences of seven isolates of AMV obtained in this study were determined and the sequences were aligned with the other sequences available in the GenBank database. Analyses confirmed the low variability among AMV isolated in this study, which means that all AMV isolates may originate from the same source. Due to high incidence of AMV, other economic susceptible crops may become affected by high incidence of this virus in potato crops. This requires accurate examination of potato seed tubers to prevent the spread of the virus in Saudi Arabia. The obtained results indicated that the hybridization and ELISA are suitable techniques in the routine detection of AMV in a large number of samples while RT-PCR is more sensitive and essential for molecular characterization of AMV.

Keywords: Alfamovirus, AMV, Alfalfa mosaic virus, PCR, potato

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8338 Critical Role of Lipid Rafts in Influenza a Virus Binding to Host Cell

Authors: Dileep Kumar Verma, Sunil Kumar Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Influenza A Virus (IAV) surface protein hemagglutinin is known to play an important role in viral attachment to the host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Selective nature of Influenza A virus to utilize rafts micro-domain for efficient virus assembly and budding has been explored in depth. However, the detailed mechanism of IAV binding to host cell membrane and entry into the host remains elusive. In the present study we investigated the role of lipid rafts in early life cycle events of IAV. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol by Methyl-β-Cyclodextrin. Using GM1, a well-known lipid raft marker, we were able to observe co-localization of IAV on lipid rafts on the host cell membrane. This experiment suggests a direct involvement of lipid rafts in the initiation of the IAV life cycle. Upon disruption of lipid rafts by Methyl-b-cyclodextrin, we observed a significant reduction in IAV binding on the host cell surface indicating a significant decrease in virus attachment to coherent membrane rafts. Our results provide proof that host lipid rafts and their constituents play an important role in the adsorption of IAV. This study opens a new avenues in IAV virus-host interactions to combat infection at a very early steps of the viral lifecycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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8337 Virus Diseases of Edible Seed Squash (Cucurbita pepo L.) in Aksaray Province

Authors: Serkan Yesil

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Cucurbits (the Cucurbitaceae family) include 119 genera and 825 species distributed primarily in tropical and subtropical regions of the world. The major cultivated cucurbit species such as melon (Cucumis melo L.), cucumber (Cucumis sativus L.), squash (Cucurbita pepo L.), and watermelon (Citrullus lanatus (Thunb) Matsum.&Nakai) are important vegetable crops worldwide. Squash is grown for fresh consuming, as well as its seeds are used as a snack in Turkey like some Mediterranean countries and Germany, Hungary, Austria and China. Virus diseases are one of the most destructive diseases on squash which is grown for seeds in Aksaray province. In this study, it was aimed to determine the virus infections in major squash growing areas in Aksaray province. Totally 153 plant samples with common virus symptoms like mosaic, curling, blistering, mottling, distortion, shoestring, stunting and vine decline were collected from squash plants during 2014. In this study, DAS-ELISA method is used for identifying the virus infections on the plant samples. According to the results of the DAS-ELISA 84.96 % of plant samples were infected with Zucchini yellow mosaic Potyvirus (ZYMV), Watermelon mosaic Potyvirus-2 (WMV-2), Cucumber mosaic Cucumovirus (CMV), Papaya ringspot Potyvirus-watermelon strain (PRSV-W) and Squash mosaic Comovirus (SqMV). ZYMV was predominant in the research area with the ratio of 66.01 %. WMV-2 was the second important virus disease in the survey area, it was detected on the samples at the ratio of 57.51 %. Also, mixed infections of those virus infections were detected commonly in squash. Especially, ZYMV+WMV-2 mixed infections were common. Cucumber green mottle mosaic Tobamovirus (CGMMV) was not present in the research area.

Keywords: Aksaray, DAS-ELISA, edible seed squash, WMV-2, ZYMV

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8336 Serological Screening of Barrier Maintained Rodent Colony

Authors: R. Posia, J. Mistry, K. Kamani

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The health screening of laboratory rodents is essential for ensuring animal health and the validity of biomedical research data. Routine health monitoring is necessary to verify the effectiveness of biosecurity and the specific pathogen free (SPF) status of the colony. The present screening was performed in barrier maintained rat (Rattus norvegicus) colony. Rats were maintained under a controlled environment and strict biosecurity in the facility. The screening was performed on quarterly bases from randomly selected animals from breeding and or maintenance colonies. Selected animals were subject to blood collection under isoflurane anaesthesia. Serum was separated from the collected blood and stored samples at -60 ± 10 °C until further use. A total of 88 samples were collected quarterly bases from animals in a year. In the serological test, enzyme-linked immunosorbent assay (ELISA) was used for screening of serum samples against sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV). ELISA kits were procured from XpressBio, USA. Test serum samples were run along with positive control, negative control serum in 96 well ELISA plates as per the procedure recommended by the vendor. Test ELISA plate reading was taken in the microplate reader. This screening observed that none of the samples was observed positive for the sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV), indicating that effectiveness of biosecurity practices followed in the rodent colony. The result of serological screening helps us to declare that our rodent colony is specifically pathogen free for these pathogens.

Keywords: biosecurity, ELISA, specific pathogen free, serological screening, serum

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8335 The Role of Public Representatives and Legislatures in Strengthening HIV and AIDS Prevention Strategies: The Case of South Africa

Authors: Moses Mncwabe

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Both Public Representatives and Legislatures have an imperative role towards strengthening interventions to reduce and cease Sexual Transmitted Infections (STIs) specifically the Human Immunodeficiency Virus (HIV). Scaling-up constituency work in support of interventions earmarked for mitigating the compromising socio-economic impacts of advanced HIV is extremely essential. Though the antiretroviral treatment (ART) has saved million lives that would have perished without it, the Joint United Nations Programme on HIV/AIDS (2012) states that more efforts should be redirected to prevention strategies to close the tap of new infections. It is against this backdrop that Legislatures as law making institutions have undisputed role to play in HIV alleviation because of the position they occupy in the society. Furthermore, Public Representatives are arguably idolised by young people for the role they play hence it is incumbent upon them to use their moral and political responsibility to aid the interventions for HIV prevention (Inter-Parliamentary Union, Joint United Nations Programme on HIV/AIDS & United Nations Development Programme, 2007). Moreover, the continuous HIV infection and its devastating effects specifically in Southern African region has brought closer the disease to public representatives and demanded calculated interventions warranting both public representatives and legislatures to be more visible in various ways such as taking HIV counselling and testing publicly, oversight, reducing stigma and discrimination, partnering with civil society organisations (CSOs) and facilitating debates on HIV across parliamentary and social platforms. The effects of advanced HIV yearn for public representatives to be seen, accessed, felt, engaged, partnered and lobbied for pro-human rights legislations and ideal oversight to coerce the executive to deliver on their core responsibilities like providing basic services to the electorates (AIDS Law Project (2003). The National Democratic Institute for International Affairs and the Southern African Development Community Parliamentary Forum (2004) assert that the omission of Public Representatives and Legislatures in the HIV prevention agenda is a serious deficiency in the fight against HIV and AIDS. In light of this, this paper argues the innovative and legislative ways in which both the Public Representative and the Legislatures should play in HIV prevention.

Keywords: legislature, public representative, oversight, HIV and AIDS, constituency, service delivery

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8334 A Novel Epitope Prediction for Vaccine Designing against Ebola Viral Envelope Proteins

Authors: Manju Kanu, Subrata Sinha, Surabhi Johari

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Viral proteins of Ebola viruses belong to one of the best studied viruses; however no effective prevention against EBOV has been developed. Epitope-based vaccines provide a new strategy for prophylactic and therapeutic application of pathogen-specific immunity. A critical requirement of this strategy is the identification and selection of T-cell epitopes that act as vaccine targets. This study describes current methodologies for the selection process, with Ebola virus as a model system. Hence great challenge in the field of ebola virus research is to design universal vaccine. A combination of publicly available bioinformatics algorithms and computational tools are used to screen and select antigen sequences as potential T-cell epitopes of supertypes Human Leukocyte Antigen (HLA) alleles. MUSCLE and MOTIF tools were used to find out most conserved peptide sequences of viral proteins. Immunoinformatics tools were used for prediction of immunogenic peptides of viral proteins in zaire strains of Ebola virus. Putative epitopes for viral proteins (VP) were predicted from conserved peptide sequences of VP. Three tools NetCTL 1.2, BIMAS and Syfpeithi were used to predict the Class I putative epitopes while three tools, ProPred, IEDB-SMM-align and NetMHCII 2.2 were used to predict the Class II putative epitopes. B cell epitopes were predicted by BCPREDS 1.0. Immunogenic peptides were identified and selected manually by putative epitopes predicted from online tools individually for both MHC classes. Finally sequences of predicted peptides for both MHC classes were looked for common region which was selected as common immunogenic peptide. The immunogenic peptides were found for viral proteins of Ebola virus: epitopes FLESGAVKY, SSLAKHGEY. These predicted peptides could be promising candidates to be used as target for vaccine design.

Keywords: epitope, b cell, immunogenicity, ebola

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8333 Urban Development Criteria with a Focus on Resilience to Pandemics: A Case Study of Corona Virus (Covid-19)

Authors: Elham Zabetian Targhi, Niusha Fardnava, Saba Saghafi

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Urban resilience to Corona Virus has become a major concern for cities these days. Our country also has not been safe from the destructive effects of this virus in social, economic, physical, governance, and management dimensions; and according to official statistics, hundreds of thousands of people in Iran have been infected with this virus and tens of thousands have died so far. Therefore, to measure urban resilience to this pandemic, some criteria and sub-criteria were developed based on the authors’ documentary and field studies, and their significance or weights were determined using analytical-comparative research method using a questionnaire of paired or L-Saati comparisons from the viewpoint of experts in urban sciences and urban development using AHP hierarchical analysis in EXPERT CHOICE software. Then, designing a questionnaire with a five-point Likert scale, the satisfaction of Tehran residents with the extracted criteria and sub-criteria was measured and the correlation between the important criteria in each dimension was assessed using correlation tests in SPSS16 software. According to the obtained results of AHP analysis and the scores of each sub-criterion, the weight of all criteria was normal. In the next stage, according to the pairwise correlation tests between the important criteria in each dimension from the viewpoint of urban science experts and Tehran residents, it was concluded that the reliability of the correlation between the criteria is 99%. In all the cases, the P-value or the same significance level was less than 0.05, which indicated the significance of the pairwise relations between the variables.

Keywords: Urban Resilience, Pandemics, Corona Virus (Covid-19), Criteria.

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8332 Development of Nanoparticulate Based Chimeric Drug Delivery System Using Drug Bioconjugated Plant Virus Capsid on Biocompatible Nanoparticles

Authors: Indu Barwal, Shloka Thakur, Subhash C. Yadav

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The plant virus capsid protein based nanoparticles are extensively studied for their application in biomedical research for development of nanomedicines and drug delivery systems. We have developed a chimeric drug delivery system by controlled in vitro assembly of separately bioconjugated fluorescent dye (as reporting molecule), folic acid (as receptor binding biomolecule for targeted delivery) and doxorubicin (as anticancer drug) using modified EDC NHS chemistry on heterologously overexpressed (E. coli) capsid proteins of cowpea chlorotic mottle virus (CCMV). This chimeric vehicle was further encapsidated on gold nanoparticles (20nm) coated with 5≠ thiolated DNA probe to neutralize the positive charge of capsid proteins. This facilitates the in vitro assembly of modified capsid subunits on the gold nanoparticles to develop chimeric GNPs encapsidated targeted drug delivery system. The bioconjugation of functionalities, number of functionality on capsid subunits as well as virus like nanoparticles, structural stability and in vitro assembly were confirmed by SDS PAGE, relative absorbance, MALDI TOF, ESI-MS, Circular dichroism, intrinsic tryptophan fluorescence, zeta particle size analyzer and TEM imaging. This vehicle was stable at pH 4.0 to 8.0 suitable for many organelles targeting. This in vitro assembled chimeric plant virus like particles could be suitable for ideal drug delivery vehicles for subcutaneous cancer treatment and could be further modified for other type of cancer treatment by conjugating other functionalities (targeting, drug) on capsids.

Keywords: chimeric drug delivery vehicles, bioconjugated plant, virus, capsid

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8331 Ultrasensitive Hepatitis B Virus Detection in Blood Using Nano-Porous Silicon Oxide: Towards POC Diagnostics

Authors: N. Das, N. Samanta, L. Pandey, C. Roy Chaudhuri

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Early diagnosis of infection like Hep-B virus in blood is important for low cost medical treatment. For this purpose, it is desirable to develop a point of care device which should be able to detect trace quantities of the target molecule in blood. In this paper, we report a nanoporous silicon oxide sensor which is capable of detecting down to 1fM concentration of Hep-B surface antigen in blood without the requirement of any centrifuge or pre-concentration. This has been made possible by the presence of resonant peak in the sensitivity characteristics. This peak is observed to be dependent only on the concentration of the specific antigen and not on the interfering species in blood serum. The occurrence of opposite impedance change within the pores and at the bottom of the pore is responsible for this effect. An electronic interface has also been designed to provide a display of the virus concentration.

Keywords: impedance spectroscopy, ultrasensitive detection in blood, peak frequency, electronic interface

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8330 Human Security and Human Trafficking Related Corruption

Authors: Ekin D. Horzum

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The aim of the proposal is to examine the relationship between human trafficking related corruption and human security. The proposal suggests that the human trafficking related corruption is about willingness of the states to turn a blind eye to the human trafficking cases. Therefore, it is important to approach human trafficking related corruption in terms of human security and human rights violation to find an effective way to fight against human trafficking. In this context, the purpose of this proposal is to examine the human trafficking related corruption as a safe haven in which trafficking thrives for perpetrators.

Keywords: human trafficking, human security, human rights, corruption, organized crime

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8329 Isolation and Culture of Keratinocytes and Fibroblasts to Develop Artificial Skin Equivalent in Cats

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

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The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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8328 Stability Analysis of Endemic State of Modelling the Effect of Vaccination and Novel Quarantine-Adjusted Incidence on the Spread of Newcastle Disease Virus

Authors: Nurudeen Oluwasola Lasisi, Abdulkareem Afolabi Ibrahim

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Newcastle disease is an infection of domestic poultry and other bird species with virulent Newcastle disease virus (NDV). In this paper, we study the dynamics of modeling the Newcastle disease virus (NDV) using a novel quarantine-adjusted incidence. We do a comparison of Vaccination, linear incident rate, and novel quarantine adjusted incident rate in the models. The dynamics of the models yield disease free and endemic equilibrium states. The effective reproduction numbers of the models are computed in order to measure the relative impact for the individual bird or combined intervention for effective disease control. We showed the local and global stability of endemic equilibrium states of the models, and we found that stability of endemic equilibrium states of models are globally asymptotically stable if the effective reproduction numbers of the models equations are greater than a unit.

Keywords: effective reproduction number, endemic state, mathematical model, Newcastle disease virus, novel quarantine-adjusted incidence, stability analysis

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8327 Neuroblastoma in Children and the Potential Involvement of Viruses in Its Pathogenesis

Authors: Ugo Rovigatti

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Neuroblastoma (NBL) has epitomized for at least 40 years our understanding of cancer cellular and molecular biology and its potential applications to novel therapeutic strategies. This includes the discovery of the very first oncogene aberrations and tumorigenesis suppression by differentiation in the 80s; the potential role of suppressor genes in the 90s; the relevance of immunotherapy in the millennium first, and the discovery of additional mutations by NGS technology in the millennium second decade. Similar discoveries were achieved in the majority of human cancers, and similar therapeutic interventions were obtained subsequently to NBL discoveries. Unfortunately, targeted therapies suggested by specific mutations (such as MYCN amplification –MNA- present in ¼ or 1/5 of cases) have not elicited therapeutic successes in aggressive NBL, where the prognosis is still dismal. The reasons appear to be linked to Tumor Heterogeneity, which is particularly evident in NBL but also a clear hallmark of aggressive human cancers generally. The new avenue of cancer immunotherapy (CIT) provided new hopes for cancer patients, but we still ignore the cellular or molecular targets. CIT is emblematic of high-risk disease (HR-NBL) since the mentioned GD2 passive immunotherapy is still providing better survival. We recently critically reviewed and evaluated the literature depicting the genomic landscapes of HR-NBL, coming to the qualified conclusion that among hundreds of affected genes, potential targets, or chromosomal sites, none correlated with anti-GD2 sensitivity. A better explanation is provided by the Micro-Foci inducing Virus (MFV) model, which predicts that neuroblasts infection with the MFV, an RNA virus isolated from a cancer-cluster (space-time association) of HR-NBL cases, elicits the appearance of MNA and additional genomic aberrations with mechanisms resembling chromothripsis. Neuroblasts infected with low titers of MFV amplified MYCN up to 100 folds and became highly transformed and malignant, thus causing neuroblastoma in young rat pups of strains SD and Fisher-344 and larger tumor masses in nu/nu mice. An association was discovered with GD2 since this glycosphingolipid is also the receptor for the family of MFV virus (dsRNA viruses). It is concluded that a dsRNA virus, MFV, appears to provide better explicatory mechanisms for the genesis of i) specific genomic aberrations such as MNA; ii) extensive tumor heterogeneity and chromothripsis; iii) the effects of passive immunotherapy with anti-GD2 monoclonals and that this and similar models should be further investigated in both pediatric and adult cancers.

Keywords: neuroblastoma, MYCN, amplification, viruses, GD2

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8326 Metamorphic Computer Virus Classification Using Hidden Markov Model

Authors: Babak Bashari Rad

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A metamorphic computer virus uses different code transformation techniques to mutate its body in duplicated instances. Characteristics and function of new instances are mostly similar to their parents, but they cannot be easily detected by the majority of antivirus in market, as they depend on string signature-based detection techniques. The purpose of this research is to propose a Hidden Markov Model for classification of metamorphic viruses in executable files. In the proposed solution, portable executable files are inspected to extract the instructions opcodes needed for the examination of code. A Hidden Markov Model trained on portable executable files is employed to classify the metamorphic viruses of the same family. The proposed model is able to generate and recognize common statistical features of mutated code. The model has been evaluated by examining the model on a test data set. The performance of the model has been practically tested and evaluated based on False Positive Rate, Detection Rate and Overall Accuracy. The result showed an acceptable performance with high average of 99.7% Detection Rate.

Keywords: malware classification, computer virus classification, metamorphic virus, metamorphic malware, Hidden Markov Model

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8325 Quality of Life among Female Sex Workers of Selected Organization of Pokhara: A Methodological Triangulation

Authors: Sharmila Dahal Paudel

Abstract:

Background: There are around twenty-four thousand to twenty-eight thousand Female Sex Workers in Nepal. FSWs are the vulnerable groups for sexually transmitted infections (STIs) and human immunodeficiency virus (HIV) infections which directly and indirectly ease to reduce the quality of life of such groups. Due to their highly marginalized status, FSWs in Nepal have limited access to information about reproductive health and safe sex practices. The objectives of the study are to assess the quality of life of female sex workers and the factors affecting them. Materials and Methods: A descriptive cross-sectional study with methodological triangulation was conducted among 108 FSWs on the basis of service record of selected organization of Pokhara valley. The complete enumerative sampling was used to select FSWs. Structured interview schedule, WHOQOL-BREF and in-depth questionnaire were used to collect the data. The descriptive and inferential statistics were used to interpret the result. Results: The mean age of participants were 23.44 years and the mean quality of life score was 174.06 ranging from 56.54 to 370.78. Among the domain scores, the mean score is highest in social domain (55.89) followed by physical (45.42), psychological (39.27) and the environmental (34.23). Regarding the association of QOL with socio-demographic, occupation and health-related variables, the multi-linear regression suggests that the satisfaction with occupation was highly significant with the total QOL score (B=-50.50, SE=10.46; p= <0.001) and there is negative relation between QOL and feeling of exploitation and facing STI problems. This means those who feels exploited have significantly less QOL comparing with those who did not feel the same. In correlation analysis, all the domains are positively co-related with each domain which is found to be significant at 1% level of significance. Conclusion: The highest mean score was in social domain, and the lowest is in environmental domain which suggests that the items included in environmental domains could not be utilized or hindrance were there.

Keywords: FSWs, HIV, QOL, WHOQOL-BREF

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8324 Caregiver’s Perception Regarding Diagnosis Disclosure to Children Living with Human Immunodeficiency Virus in Resource-Limited Settings: Observational Study from India

Authors: Ramesh Chand Chauhan, Sanjay Kumar Rai, Shashi kant, Rakesh Lodha, Nand Kumar

Abstract:

Background: With a better understanding of HIV pathogenesis and availability of antiretroviral therapy more children are growing and entering in teenage group; informing children of their own HIV status has become an important aspect of long-term disease management. There is little evidence of how and when this type of disclosure takes place in a resource-limited setting. Methods: A cross-sectional study was conducted from June 2010 to May 2011 among a dyads of 156 HIV-infected children and their caregivers, those were visiting pediatric clinic at a tertiary care hospital in Delhi, India. The study protocol was approved by the Institute Ethics Committee. After taking written informed consent; pretested structured questionnaire was administered to caregivers during routine clinic visits. Information regarding socio-demographic characteristics, awareness of HIV infection status among children and their perception regarding disclosure was collected. Mean and frequencies were calculated and chi-square and logistic regression test were applied. Results: The mean age of children was 8.4 ±3.45 years. Among them 73.7% were male and 39.1% were orphans. Among 156 enrolled children, 74.4% (n=116) were of ≥ 6 years and were assessed for disclosure. Only 18.1% (n=21) children had been informed of their HIV status. Of those under 9 years, 6.4% knew their status, whereas 18.4% of 9-11 years and 35.5% of 12-14 years children knew they had HIV. Awareness among males (23.3%) was higher than females (3.3%). Both age and sex of child were significantly (p<0.01) associated with disclosure status. Other factors favoring disclosure were orphan-hood, non-perinatal mode of transmission (OR = 4.32; 95% CI 1.01-7.12), ART initiation (OR = 4.21; 95% CI 1.03-6.98), and caregiver educated beyond primary level (OR = 1.89; 95% CI 1.03-3.26). Repeated enquiry regarding the visit to clinic was the most common reason (66.6%) for disclosure. In 52.4% children disclosure was done with the involvement of other family members. 82.5% caregivers felt the age of > 10 years is appropriate for disclosing the HIV infection status to the child. Conclusion: Detailed guidelines on disclosure are required focusing on children of school-going age with perinatal infection who are not on ART and with caregivers of low educational status.

Keywords: HIV, children, India, disclosure

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8323 Preparedness and Control of Mosquito-Borne Diseases: Experiences from Northwestern Italy

Authors: Federica Verna, Alessandra Pautasso, Maria Caramelli, Cristiana Maurella, Walter Mignone, Cristina Casalone

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Mosquito-Borne Diseases (MBDs) are dangerously increasing in prevalence, geographical distribution and severity, representing an emerging threat for both humans and animals. Interaction between multiple disciplines is needed for an effective early warning, surveillance and control of MBDs, according to the One Health concept. This work reports the integrated surveillance system enforced by IZSPLV in Piedmont, Liguria and Valle d’Aosta regions (Northwestern Italy) in order to control MDBs spread. Veterinary services and local human health authority are involved in an information network, to connect the surveillance of human clinical cases with entomological surveillance and veterinary monitoring in order to implement control measures in case of outbreak. A systematic entomological surveillance is carried out during the vector season using mosquitoes traps located in sites selected according to risk factors. Collected mosquitoes are counted, identified to species level by morphological standard classification keys and pooled by collection site, date and species with a maximum of 100 individuals. Pools are analyzed, after RNA extraction, by Real Time RT-PCR distinctive for West Nile Virus (WNV) Lineage 1 and Lineage 2, Real Time RT-PCR USUTU virus (USUV) and a traditional flavivirus End-point RT-PCR. Positive pools are sequenced and the related sequences employed to perform a basic local alignment search tool (BLAST) in the GenBank library. Positive samples are sent to the National Reference Centre for Animal Exotic Diseases (CESME, Teramo) for confirmation. With particular reference to WNV, after the confirmation, as provided by national legislation, control measures involving both local veterinary and human health services are activated: equine sera are randomly sampled within a 4 km radius from the positive collection sites and tested with ELISA kit and WNV NAT screening of blood donors is introduced. This surveillance network allowed to detect since 2011 USUV circulation in this area of Italy. WNV was detected in Piedmont and Liguria for the first time in 2014 in mosquitoes. During the 2015 vector season, we observed the expansion of its activity in Piedmont. The virus was detected in almost all Provinces both in mosquitoes (6 pools) and animals (19 equine sera, 4 birds). No blood bag tested resulted infected. The first neuroinvasive human case occurred too. Competent authorities should be aware of a potentially increased risk of MBDs activity during the 2016 vector season. This work shows that this surveillance network allowed to early detect the presence of MBDs in humans and animals, and provided useful information to public authorities, in order to apply control measures. Finally, an additional value of our diagnostic protocol is the ability to detect all viruses belonging to the Flaviviridae family, considering the emergence caused by other Flaviviruses in humans such as the recent Zika virus infection in South America. Italy has climatic and environmental features conducive to Zika virus transmission, the competent vector and many travellers from Brazil reported every year.

Keywords: integrated surveillance, mosquito borne disease, West Nile virus, Zika virus

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8322 Detection and Distribution Pattern of Prevelant Genotypes of Hepatitis C in a Tertiary Care Hospital of Western India

Authors: Upasana Bhumbla

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Background: Hepatitis C virus is a major cause of chronic hepatitis, which can further lead to cirrhosis of the liver and hepatocellular carcinoma. Worldwide the burden of Hepatitis C infection has become a serious threat to the human race. Hepatitis C virus (HCV) has population-specific genotypes and provides valuable epidemiological and therapeutic information. Genotyping and assessment of viral load in HCV patients are important for planning the therapeutic strategies. The aim of the study is to study the changing trends of prevalence and genotypic distribution of hepatitis C virus in a tertiary care hospital in Western India. Methods: It is a retrospective study; blood samples were collected and tested for anti HCV antibodies by ELISA in Dept. of Microbiology. In seropositive Hepatitis C patients, quantification of HCV-RNA was done by real-time PCR and in HCV-RNA positive samples, genotyping was conducted. Results: A total of 114 patients who were seropositive for Anti HCV were recruited in the study, out of which 79 (69.29%) were HCV-RNA positive. Out of these positive samples, 54 were further subjected to genotype determination using real-time PCR. Genotype was not detected in 24 samples due to low viral load; 30 samples were positive for genotype. Conclusion: Knowledge of genotype is crucial for the management of HCV infection and prediction of prognosis. Patients infected with HCV genotype 1 and 4 will have to receive Interferon and Ribavirin for 48 weeks. Patients with these genotypes show a poor sustained viral response when tested 24 weeks after completion of therapy. On the contrary, patients infected with HCV genotype 2 and 3 are reported to have a better response to therapy.

Keywords: hepatocellular, genotype, ribavarin, seropositive

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8321 Isolation and Characterization White Spot Syndrome Protein Envelope Protein 19 from Black Tiger Shrimp (Penaeus monodon)

Authors: Andi Aliah Hidayani, Asmi Citra Malina A. R. Tassakka, Andi Parenrengi

Abstract:

Vanname Shrimp is one of the high yielding varieties that are more resistant to virus attacks. However, now this shrimp more death due to virus attack such as white spot disease caused by white spot syndrome virus (WSSV). Various efforts have done to prevent the disease, like immunostimulatory, probiotics, and vaccine. White spot syndrome virus (WSSV) envelope protein VP19 gene is important because of its involvement in the system infection of shrimp. This study aimed to isolate and characterize an envelope protein VP19 – encoding gene of WSSV using WSSV infected Vanname Shrimp sample from some areas in South Sulawesi (Pangkep, Barru and Pinrang). The genomic of DNA were isolated from shrimp muscle using DTAB-CTAB method. Isolation of gene encoding envelope protein VP19 WSSV ws successfully performed with the results of the length of DNA fragment was 387 bp. The results of homology analysis using BLASTn homology suggested that these isolates genes from Barru, Pangkep and Pinrang have closest relationship with isolates from Mexican.

Keywords: vanname, shrimp, WSSV, viral protein 19

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8320 Multidrug Therapies For HIV: Hybrid On-Off, Hysteresis On-Off Control and Simple STI

Authors: Magno Enrique Mendoza Meza

Abstract:

This paper deals with the comparison of three control techniques: the hysteresis on-off control (HyOOC), the hybrid on-off control (HOOC) and the simple Structured Treatment Interruptions (sSTI). These techniques are applied to the mathematical model developed by Kirschner and Webb. To compare these techniques we use a cost functional that minimize the wild-type virus population and the mutant virus population, but the main objective is to minimize the systemic cost of treatment and maximize levels of healthy CD4+ T cells. HyOOC, HOOC, and sSTI are applied to the drug therapies using a reverse transcriptase and protease inhibitors; simulations show that these controls maintain the uninfected cells in a small, bounded neighborhood of a pre-specified level. The controller HyOOC and HOOC are designed by appropriate choice of virtual equilibrium points.

Keywords: virus dynamics, on-off control, hysteresis, multi-drug therapies

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8319 Induction of Cellular and Humoral Immune Responses in BALB/c Mice Immunized With rB2L and rF1L Proteins of Orf Virus Adjuvanted With Alumina Nanoparticles

Authors: Alhaji Modu Bukar, Faez Firdaus Abdullah Jesse, Che Azurahanim Che Abdullah, Mustapha M. Noordin, Mohd-Lila Mohd Azmia

Abstract:

Orf virus (ORFV) is the causative agent of a proliferative skin lesion known as contagious ecthyma in sheep and goats. Currently used live attenuated vaccines against ORFV infection have been reported to cause severe outbreaks in vaccinated animals. In this study, we investigated the immunogenicity of the B2L and F1L proteins of the virus, which are thought to elicit a protective immune response The 6-week-old 50 female mice were divided into 8 groups: seven experimental groups and one control group. Each animal in the experimental group received an initial immunisation with the nanoparticles or proteins coated with the nanoparticles, followed by two booster immunizations with the same products 14 days apart. Ten days after the last booster inoculation, the mice were either humanely killed or lethally challenged with UPM /HSN-2-ORFV at a dose of 106 TCID50/mL in a volume of 50 μl. The spleen was examined for histopathological changes and quantification of T cells by flow cytometry. On the other hand, the degree of protection of mice from the lethal virus was evaluated by lesion size, weight loss, and histopathological examination of skin and liver. The results showed that mice immunised with rB2L alone, rB2L-Al₂O₃-NPs, rB2L/rF1L, and rB2L/rF1L-Al₂O₃-NPs elicited statistically higher levels of anti-rB2L and/or rF1L-specific IgA/IgG and CD4/CD8 cell immune responses than mice in the control groups (p < 0.01). The vaccine candidate did not exhibit severe skin damage after monitoring histopathology, morbidity, and mortality. Overall, the results suggest that recombinant rB2L and rF1L antigens may be useful universal vaccine candidates against ORFV infections.

Keywords: orf virus, antigen nanoparticles, virus, nanoparticles

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8318 Expression of Gro-El under Phloem-Specific Promoter Protects Transgenic Plants against Diverse Begomovirus-Beta Satellite Complex

Authors: Muhammad Yousaf Ali, Shahid Mansoor, Javeria Qazi, Imran Amin, Musarrat Shaheen

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Cotton leaf curl disease (CLCuD) is the major threat to the cotton crop and is transmitted by whitefly (Bemisia tabaci). Since multiple begomoviruses and associated satellites are involved in CLCuD, approaches based on the concept of broad-spectrum resistance are essential for effective disease control. Gro-El and G5 are two proteins from whitefly endosymbiont and M13 bacteriophage origin, respectively. Gro-El encapsulates the virus particle when it enters the whitefly and protects the virus from the immune system of the whitefly as well as prevents viral expression in it. This characteristic of Gro-El can be exploited to get resistance against viruses if expressed in plants. G5 is a single-stranded DNA binding protein, expression of which in transgenic plants will stop viral expression on its binding with ssDNA. The use of tissue-specific promoters is more efficient than constitutive promoters. Transgenics of Nicotiana benthamiana for Gro-El under constitutive promoter and Gro-El under phloem specific promoter were made. In comparison to non-transgenic plants, transgenic plants with Gro-El under NSP promoter showed promising results when challenged against cotton leaf curl Multan virus (CLCuMuV) along with cotton leaf curl Multan beta satellite (CLCuMB), cotton leaf curl Khokhran virus (CLCuKoV) along with cotton leaf curl Multan beta satellite (CLCuMB) and Pedilenthus leaf curl virus (PedLCV) along with Tobacco leaf curl beta satellite (TbLCB).

Keywords: cotton leaf curl disease, whitefly, endosymbionts, transgenic, resistance

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8317 Serological Assay and Genotyping of Hepatitis C Virus in Infected Patients in Zanjan Province

Authors: Abdolreza Esmaeilzadeh, Maryam Erfanmanesh, Sousan Ghasemi, Farzaneh Mohammadi

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Background: Hepatitis C Virus (HCV), a public health problem, is an enveloped, single-stranded RNA virus and a member of the Hepacivirus genus of the Flaviviridae family. Liver cancer, cirrhosis, and end-stage liver are the outcomes of chronic infection with HCV. HCV isolates show significant heterogeneity in genetics around the world. Therefore, determining HCV genotypes is a vital step in determining prognosis and planning therapeutic strategies. Materials and Methods: Serum samples of 136 patients were collected and analyzed for anti-HCV antibodies using ELISA (The enzyme-linked immunosorbent assay) method. Then, positive samples were exposed to RT-PCR, which was performed under standard condition. Afterwards, they investigated for genotyping using allele-specific PCR (AS-PCR), and HCV genotype 2.0 line probe assay (LiPA). Results: Samples indicated 216 bp bands on 2% agarose gel. Analyses of the results demonstrated that the most dominant subtype was 3a with frequency of 38.26% in Zanjan Province followed by subtypes of 1b, 1a, 2, and 4 with frequencies of 25.73%, 22.05%, 5.14%, and 4.41%, respectively. The frequency of unknown HCV genotypes was 4.41%. Conclusions: According to the results, it was found that HCV high prevalent genotype in Zanjan is subtype 3a. Analysis of the results provides identification of certain HCV genotypes, and these valuable findings could affect the type and duration of the treatment.

Keywords: anti-HCV antibody, Hepatitis C Virus (HCV), genotype, RT-PCR, AS-PCR

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8316 New Test Algorithm to Detect Acute and Chronic HIV Infection Using a 4th Generation Combo Test

Authors: Barun K. De

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Acquired immunodeficiency syndrome (AIDS) is caused by two types of human immunodeficiency viruses, collectively designated HIV. HIV infection is spreading globally particularly in developing countries. Before an individual is diagnosed with HIV, the disease goes through different phases. First there is an acute early phase that is followed by an established or chronic phase. Subsequently, there is a latency period after which the individual becomes immunodeficient. It is in the acute phase that an individual is highly infectious due to a high viral load. Presently, HIV diagnosis involves use of tests that do not detect the acute phase infection during which both the viral RNA and p24 antigen are expressed. Instead, these less sensitive tests detect antibodies to viral antigens which are typically sero-converted 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 can reduce medical costs, improve survival, and reduce spreading of infection 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 is 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 HIV-1 or HIV-2. However if it is negative then another test is performed that measures viral load confirming an acute HIV-1 infection. Screening results of a Phoenix area population detected 0.3% new HIV infections among which 32.4% were acute cases. Studies in the U.S. indicate that this algorithm effectively reduces HIV infection through immediate treatment and education following diagnosis.

Keywords: new algorithm, HIV, diagnosis, infection

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