The Scientific World Journal: Virology The latest articles from Hindawi Publishing Corporation © 2014 , Hindawi Publishing Corporation . All rights reserved. Voltage Dependent Anion Channel Is Redistributed during Japanese Encephalitis Virus Infection of Insect Cells Thu, 10 Jul 2014 07:59:54 +0000 Despite the availability of an effective vaccine, Japanese encephalitis remains a significant cause of morbidity and mortality in many parts of Asia. Japanese encephalitis is caused by the Japanese encephalitis virus (JEV), a mosquito transmitted flavivirus. Many of the details of the virus replication cycle in mosquito cells remain unknown. This study sought to determine whether GRP78, a well-characterized flavivirus E protein interacting protein, interacted with JEV E protein in insect cells, and whether this interaction was mediated at the cell surface. GRP78 was shown to interact with JEV E protein by coimmunoprecipitation, and was additionally shown to interact with voltage dependent anion protein (VDAC) through the same methodology. Antibody inhibition experiments showed that neither GRP78 nor VDAC played a role in JEV internalization to insect cells. Interestingly, VDAC was shown to be significantly relocalized in response to JEV infection, and significant levels of colocalization between VDAC and GRP78 and VDAC and ribosomal L28 protein were seen in JEV infected but not uninfected cells. This is the first report of relocalization of VDAC in response to JEV infection and suggests that this may be a part of the JEV replication strategy in insect cells. Chanida Fongsaran, Narumon Phaonakrop, Sittiruk Roytrakul, Chutima Thepparit, Atichat Kuadkitkan, and Duncan R. Smith Copyright © 2014 Chanida Fongsaran et al. All rights reserved. Evaluation and Prediction of the HIV-1 Central Polypurine Tract Influence on Foamy Viral Vectors to Transduce Dividing and Growth-Arrested Cells Mon, 09 Jun 2014 08:32:30 +0000 Retroviral vectors are potent tools for gene delivery and various biomedical applications. To accomplish a gene transfer task successfully, retroviral vectors must effectively transduce diverse cell cultures at different phases of a cell cycle. However, very promising retroviral vectors based on the foamy viral (FV) backbone lack the capacity to efficiently transduce quiescent cells. It is hypothesized that this phenomenon might be explained as the inability of foamy viruses to form a pre-integration complex (PIC) with nuclear import activity in growth-arrested cells, which is the characteristic for lentiviruses (HIV-1). In this process, the HIV-1 central polypurine tract (cPPT) serves as a primer for plus-strand synthesis to produce a “flap” element and is believed to be crucial for the subsequent double-stranded cDNA formation of all retroviral RNA genomes. In this study, the effects of the lentiviral cPPT element on the FV transduction potential in dividing and growth-arrested (G1/S phase) adenocarcinomic human alveolar basal epithelial (A549) cells are investigated by experimental and theoretical methods. The results indicated that the HIV-1 cPPT element in a foamy viral vector background will lead to a significant reduction of the FV transduction and viral titre in growth-arrested cells due to the absence of PICs with nuclear import activity. Sergey Shityakov, Carola Förster, Axel Rethwilm, and Thomas Dandekar Copyright © 2014 Sergey Shityakov et al. All rights reserved. Dengue Outbreak in a Hilly State of Arunachal Pradesh in Northeast India Tue, 21 Jan 2014 13:14:51 +0000 Dengue has been reported from plains as well as hilly regions of India including some parts of Northeast India. In July-August 2012, outbreak of fever with unknown origin (FUO) indicative of Dengue was reported in Pasighat, East Siang district of Arunachal Pradesh (AP) state. Serum samples (n = 164) collected from patients from Health Training and Research Centre General Hospital, Pasighat, were tested for NS1 antigen and IgM antibodies. NS1-positive samples were analyzed by RT-PCR assay and entomological surveys were carried out. The majority of suspected cases reported NS1 antigen positivity. Females and young adults were mostly affected. The majority of the amplified NS1-positive samples showed Dengue serotype 3 infection. Aedes (Stegomyia) albopictus, known as semiurban breeding mosquitoes, was the only potential vector species identified from the affected areas of Pasighat which single handedly contributed to the outbreak. Thus, the present work identifies Dengue as an emerging arboviral infection in hilly state of AP along with a looming risk of its spread to neighbouring areas. Siraj A. Khan, Prafulla Dutta, Rashmee Topno, Monika Soni, and Jagadish Mahanta Copyright © 2014 Siraj A. Khan et al. All rights reserved. Using Targeted Virotherapy to Treat a Resistant Ewing Sarcoma Model: From the Bedside to the Bench and Back Sun, 12 Jan 2014 13:05:38 +0000 Metastatic Ewing sarcoma (EWS) is often resistant to current multimodal chemotherapeutic regimens. Oncolytic virus therapy (OV) is a novel therapeutic platform whereby viruses can selectively infect as well as replicate in and kill tumor cells, while sparing normal tissues. The purpose of this study is to investigate the efficacy of the biotherapeutic oncolytic agent, vesicular stomatitis virus (VSVΔM51), to kill EWS cells that are resistant to conventional therapy. Our hypothesis is that systemic delivery of VSVΔM51 can demonstrate tumor-specific killing of resistant EWS cells, as well as a significant decrease of tumor burden in EWS bearing mice. Methods. A biopsy sample was obtained from a patient with metastatic EWS and was used to establish a novel EWS cell line. In vitro assays evaluated the oncolytic effect of vesicular stomatitis virus (VSVΔM51) on this cell line. EWS xenograft mice model bearing either lung or subcutaneous tumors was established to evaluate the antitumor specific oncolytic effect of VSVΔM51 after local and systemic delivery. Results. The established EWS cell line shared similar molecular and genetic traits to the patient’s original tumor specimen. VSVΔM51 effectively infected and killed EWS cells in vitro. In vivo, VSVΔM51 selectively infected and killed EWS and led to significant delay in tumor growth. Conclusion. This study has been designed to implement a translational link between the bedside and the bench, where a specific challenging clinical scenario guided this basic science research. This research demonstrated that a sarcoma, which is resistant to current conventional standard therapies, is still susceptible to an alternative therapeutic platform, such as OV. Adding OV to the armamentarium of sarcoma treatment can enhance the future therapeutic approach towards these cancer patients. Hesham Abdelbary, Christopher W. Brown, Joel Werier, and John Bell Copyright © 2014 Hesham Abdelbary et al. All rights reserved. Molecular and Virological Investigation of a Focal Chikungunya Outbreak in Northern India Tue, 24 Dec 2013 18:06:34 +0000 Chikungunya (CHIK) fever is one of the most important arboviral infections of medical significance. The objective of the present study is to identify and characterize the etiology of a focal febrile arthritis outbreak from Gwalior, northern India, during October-November 2010. A detailed virological (isolation) and molecular (end-point RT-PCR, quantitative RT-PCR, and nucleotide sequencing) investigation of this outbreak was carried out by collecting and studying 52 clinical samples and 15 mosquito pools from the affected region. The investigation revealed the presence of CHIK viral RNA in 29% of clinical samples and 13% mosquito pool by RT-PCR. The quantification of CHIK viral RNA in samples varied from 102.50 to 106.67 copies/mL, as demonstrated through quantitative RT-PCR. In addition, six CHIK viruses were isolated from RT-PCR positive samples. The nucleotide sequences of partial E1 gene of five representative CHIK viruses were deciphered, which revealed that all the viral strains from this outbreak belong to the recently emerging ECS African genotype. Identification of Chikungunya virus ECSA African genotype as the etiology of the present outbreak confirms the continued circulation of the novel genotype, since 2006, in India. The identification of CHIK virus in Aedes aegypti also confirmed it as the major vector in northern India. Manisha Soni, Anil Kumar Singh, Shashi Sharma, Ankita Agarwal, Natarajan Gopalan, P. V. Lakshmana Rao, Manmohan Parida, and Paban Kumar Dash Copyright © 2013 Manisha Soni et al. All rights reserved. Retracted: Recent Advances in DENV Receptors Mon, 18 Nov 2013 08:52:49 +0000 The Scientific World Journal Copyright © 2013 The Scientific World Journal. All rights reserved. Laboratory Diagnosis of Human Rabies: Recent Advances Thu, 14 Nov 2013 12:31:09 +0000 Rabies, an acute progressive, fatal encephalomyelitis, transmitted most commonly through the bite of a rabid animal, is responsible for an estimated 61,000 human deaths worldwide. The true disease burden and public health impact due to rabies remain underestimated due to lack of sensitive laboratory diagnostic methods. Rapid diagnosis of rabies can help initiate prompt infection control and public health measures, obviate the need for unnecessary treatment/medical tests, and assist in timely administration of pre- or postexposure prophylactic vaccination to family members and medical staff. Antemortem diagnosis of human rabies provides an impetus for clinicians to attempt experimental therapeutic approaches in some patients, especially after the reported survival of a few cases of human rabies. Traditional methods for antemortem and postmortem rabies diagnosis have several limitations. Recent advances in technology have led to the improvement or development of several diagnostic assays which include methods for rabies viral antigen and antibody detection and assays for viral nucleic acid detection and identification of specific biomarkers. These assays which complement traditional methods have the potential to revolutionize rabies diagnosis in future. Reeta Subramaniam Mani and Shampur Narayan Madhusudana Copyright © 2013 Reeta Subramaniam Mani and Shampur Narayan Madhusudana. All rights reserved. Dengue Virus Type 2: Protein Binding and Active Replication in Human Central Nervous System Cells Mon, 04 Nov 2013 14:10:24 +0000 An increased number of dengue cases with neurological complications have been reported in recent years. The lack of reliable animal models for dengue has hindered studies on dengue virus (DENV) pathogenesis and cellular tropism in vivo. We further investigate the tropism of DENV for the human central nervous system (CNS), characterizing DENV interactions with cell surface proteins in human CNS cells by virus overlay protein binding assays (VOPBA) and coimmunoprecipitations. In VOPBA, three membrane proteins (60, 70, and 130 kDa) from the gray matter bound the entire virus particle, whereas only a 70 kDa protein bound in white matter. The coimmunoprecipitation assays revealed three proteins from gray matter consistently binding virus particles, one clearly distinguishable protein (~32 kDa) and two less apparent proteins (100 and 130 kDa). Monoclonal anti-NS3 targeted the virus protein in primary cell cultures of human CNS treated with DENV-2, which also stained positive for NeuH, a neuron-specific marker. Thus, our results indicate (1) that DENV-2 exhibited a direct tropism for human neurons and (2) that human neurons sustain an active DENV replication as was demonstrated by the presence of the NS3 viral antigen in primary cultures of these cells treated with DENV-2. Ma Isabel Salazar, Marissa Pérez-García, Marisol Terreros-Tinoco, María Eugenia Castro-Mussot, Jaime Diegopérez-Ramírez, Alma Griselda Ramírez-Reyes, Penélope Aguilera, Leticia Cedillo-Barrón, and María Martha García-Flores Copyright © 2013 Ma Isabel Salazar et al. All rights reserved. Simultaneous Detection of Group A Rotavirus in Swine and Rat on a Pig Farm in Brazil Thu, 16 May 2013 14:45:32 +0000 This study investigated the occurrence of rotavirus in porcine and Rattus norvegicus, at the same time, on a pig farm in the city of Jaguariúna, São Paulo, Brazil. Swine () and rat () fecal samples were analyzed by nested RT-PCR assay. Rotavirus occurred in seven porcine and two rat samples. A total of three pig and one rat samples were further submitted to genetic sequencing. The partial NSP5 gene phylogeny showed that all strains were segregated in the genotype H1. These results point toward a cross-species transmission between rats and pigs on the surveyed farm and represent the first detection of rotavirus in Rattus norvegicus in Brazil. Paloma de Oliveira Tonietti, Aline Santana da Hora, Fernanda Dornellas F. Silva, Karen Linares Ferrari, Paulo Eduardo Brandão, Leonardo José Richtzenhain, and Fabio Gregori Copyright © 2013 Paloma de Oliveira Tonietti et al. All rights reserved. Recent Advances in DENV Receptors Tue, 23 Apr 2013 09:20:03 +0000 Dengue is an old disease caused by the mosquito-borne dengue viruses (DENVs), which have four antigenically distinct serotypes (DENV1–4). Infection by any of them can cause dengue fever (DF) and/or a more serious disease, that is, dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS). In recent decades, incidence of dengue disease has increased 30-fold, putting a third to half of the world’s population living in dengue-endemic areas at high infection risk. However, the pathogenesis of the disease is still poorly understood. The virus binding with its host cell is not only a first and critical step in their replication cycle but also a key factor for the pathogenicity. In recent years, there have been significant advances in understanding interactions of DENVs with their target cells such as dendritic cells (DC), macrophages, endothelial cells, and hepatocytes. Although DENVs reportedly attach to a variety of receptors on these cells, consensus DENV receptors have not been defined. In this review, we summarize receptors for DENVs on different cells identified in recent years. Shuyu Fang, Yanhua Wu, Na Wu, Jing Zhang, and Jing An Copyright © 2013 Shuyu Fang et al. All rights reserved. Hepatitis C Virus Adaptation to T-Cell Immune Pressure Mon, 11 Mar 2013 14:15:06 +0000 Replication of the hepatitis C virus (HCV) is an error-prone process. This high error rate results in the emergence of viral populations (quasispecies) within hosts and contributes to interhost variability. Numerous studies have demonstrated that both viral and host factors contribute to this viral diversity, which can ultimately affect disease outcome. As the host’s immune response is an important correlate of infection outcome for HCV, many of these viral variations are strongly influenced by T-cell immune pressure and accordingly constitute an efficient strategy to subvert such pressures (viral adaptations). This paper will review the data on viral diversity observed between and within hosts infected with HCV from the acute to the chronic stage of infection and will focus on viral adaptation to the host’s T-cell immune response. A. Plauzolles, M. Lucas, and S. Gaudieri Copyright © 2013 A. Plauzolles et al. All rights reserved. New Tools in HCV Diagnosis, in Light of the Enhanced Awareness and the New Drugs for Treatment: SMARTube and Stimmunology Thu, 14 Feb 2013 15:02:43 +0000 With improved HCV therapy, challenges regarding HCV diagnosis, such as seronegative window period, false positive readings, and differentiation between recent, chronic, and resolved infections, are of increasing importance. To address these challenges an innovative device—SMARTube HIV & HCV—was used. Blood samples were tested for anti-HCV antibodies before and after incubation in the SMARTube, which promotes the in vitro stimulation of in vivo HCV primed lymphocytes, thus enhancing levels of anti-HCV antibodies. Comparing antibody levels, in concordant samples before and after SMARTube, yielded the Stimulation Index (SI). Among 5888 fresh blood samples, from various populations and regions worldwide, 641 were seropositive using plasma, while SMARTube processing (yielding enriched plasma, termed SMARTplasma) enabled diagnosis of 10 additional carriers in high-risk cohorts, that is, earlier detection. Using SMARTplasma eliminated all false positive results, using the current assays. In addition we show that SI calculation may serve as an important tool for differentiating between those who recently seroconverted, carriers of long-term infection, and those who have cleared the virus. SMARTube and the SI could lead to better, more informative diagnosis of HCV infections and play an important role in changing the way we treat both the infected individuals and the epidemic as a whole. Svetlana Gorodin, Serhat Unal, Youchun Wang, Mikhail I. Mikhaylov, Ludmila Bigbulatova, and Tamar Jehuda-Cohen Copyright © 2013 Svetlana Gorodin et al. All rights reserved. Chloroquine Inhibits Dengue Virus Type 2 Replication in Vero Cells but Not in C6/36 Cells Thu, 31 Jan 2013 09:47:22 +0000 Dengue viruses are the most important arthropod-borne viruses in terms of morbidity and mortality in the world. Since there is no dengue vaccine available for human use, we have set out to investigate the use of chloroquine as an antiviral drug against dengue. Chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal pH, was used in the in vitro treatment of Vero and C6/36 cells infected with dengue virus type 2 (DENV-2). Real-time RT-PCR and plaque assays were used to quantify the DENV-2 load in infected Vero and C6/36 cells after chloroquine treatment. Our results showed that a dose of 50 μg/ml of chloroquine was not toxic to the cells and induced a statistically significant inhibition of virus production in infected Vero cells when compared to untreated cells. In C6/36 cells, chloroquine does not induce a statistically significant difference in viral replication when compared to untreated cells, showing that this virus uses an unlikely pathway of penetration in these cells, and results were also confirmed by the plaque assay (PFU). These data suggest that the inhibition of virus infection induced by chloroquine is due to interference with acidic vesicles in mammalian cells. Kleber Juvenal Silva Farias, Paula Renata Lima Machado, and Benedito Antônio Lopes da Fonseca Copyright © 2013 Kleber Juvenal Silva Farias et al. All rights reserved. Influence of Healthcare-Associated Factors on the Efficacy of Hepatitis C Therapy Thu, 27 Dec 2012 11:35:43 +0000 Hepatitis C infection is a complex entity associated with sizable morbidity and mortality, with great social and economic consequences that put a heavy potential burden on healthcare systems allover the world. Despite the great improvement of hepatitis C virus (HCV) therapy and its high clinical efficacy, major influencing factors are still hindering and diminishing the effectiveness of hepatitis C treatment. This minimizes the quality of life of the infected patients and reduces the outcome of such therapy, particularly in certain groups of patients such as intravenous drug users and patients coinfected with human immune deficiency virus (HIV). A variety of factors were evolved either at patient individual level, healthcare providers, community surrounding levels, or healthcare setting systems. Analyzing and understanding these factors could help to improve HCV interventions and, thus, reduce the burden of such infection. The objectives of this paper were to highlight such factors and outline the holistic approaches that could be used to overcome such factors. Mohamed A. Daw, Aghynya A. Dau, and Mohamed M. Agnan Copyright © 2012 Mohamed A. Daw et al. All rights reserved. HIV-1 Replication in HIV-Infected Individuals Is Significantly Reduced When Peripheral Blood Mononuclear Cells Are Superinfected with HSV-1 Sun, 02 Sep 2012 12:28:53 +0000 Herpes simplex virus (HSV) can cause generalized infection in human immunodeficiency virus- (HIV-) infected patients leading to death. This study investigated HSV-1 replication in PBMCs from 25 HIV-infected individuals and 15 healthy donors and the effects of HSV-1 superinfection on HIV-1 production. Herpes viral entry mediator (HVEM) receptor on T lymphocytes was also evaluated. Our results confirmed that the number of activated (CD3+ and CD38+) T lymphocytes in HIV-infected individuals (46.51±17.54%) was significantly higher than in healthy donors (27.54±14.12%, P value = 0.001) without any significant differences in HVEM expression. Even though the percentages of HSV-1 infected T lymphocytes between HIV-infected individuals (79.25±14.63%) and healthy donors (80.76±7.13%) were not different (P value = 0.922), yet HSV-1 production in HIV-infected individuals (47.34±11.14×103 PFU/ml) was significantly greater than that of healthy donors (34.17±8.48×103 PFU/ml, P value = 0.001). Moreover, HSV-1 virions were released extracellularly rather than being associated with the cells, and superinfection of HSV-1 at a multiplicity of infection (MOI) of 5 significantly decreased HIV production (P value < 0.001). Taneth Yamsuwan, Chintana Chirathaworn, Pokrath Hansasuta, and Parvapan Bhattarakosol Copyright © 2012 Taneth Yamsuwan et al. All rights reserved. Risk Factors for Infection with Different Hepatitis C Virus Genotypes in Southern Brazil Tue, 22 May 2012 15:58:15 +0000 Objectives. To investigate the proportion of different genotypes in countryside microregions in southern Brazil, and their association with risk factors. Methods. Cross-sectional study including a convenience sample of patients who tested positive for HCV-RNA and were referred to a regional health center for genotyping, from December 2003 to January 2008. Data were obtained through the National Disease Surveillance Data System, from laboratory registers and from patient charts. Identification of genotypes was carried out using the Restriction Fragment Length Polymorphism “in house” technique. Independent associations with genotypes were evaluated in multinomial logistic regression and prevalence rates of genotypes were estimated with modified Poisson regression. Results. The sample consisted of 441 individuals, 41.1±12.0 years old, 56.5% men. Genotype 1 was observed in 41.5% (95% CI 37.9–48.1) of patients, genotype 2 in 19.3% (95% CI 15.0–23.6), and genotype 3 in 39.2% (95% CI 35.6–43.0). HCV genotype was significantly associated with gender and age. Dental procedures were associated with higher proportion of genotype 2 independently of age, education, and patient treatment center. Conclusions. The hepatitis C virus genotype 1 was the most frequent. Genotype 2 was associated with female gender, age, and dental procedure exposition. Marisa Lúcia Romani Paraboni, Marina Dallagasperina Sbeghen, Fernando Herz Wolff, and Leila Beltrami Moreira Copyright © 2012 Marisa Lúcia Romani Paraboni et al. All rights reserved. Imunocompetent Mice Model for Dengue Virus Infection Tue, 15 May 2012 17:44:32 +0000 Dengue fever is a noncontagious infectious disease caused by dengue virus (DENV). DENV belongs to the family Flaviviridae, genus Flavivirus, and is classified into four antigenically distinct serotypes: DENV-1, DENV-2, DENV-3, and DENV-4. The number of nations and people affected has increased steadily and today is considered the most widely spread arbovirus (arthropod-borne viral disease) in the world. The absence of an appropriate animal model for studying the disease has hindered the understanding of dengue pathogenesis. In our study, we have found that immunocompetent C57BL/6 mice infected intraperitoneally with DENV-1 presented some signs of dengue disease such as thrombocytopenia, spleen hemorrhage, liver damage, and increase in production of IFNγ and TNFα cytokines. Moreover, the animals became viremic and the virus was detected in several organs by real-time RT-PCR. Thus, this animal model could be used to study mechanism of dengue virus infection, to test antiviral drugs, as well as to evaluate candidate vaccines. Denise Gonçalves, Rafael de Queiroz Prado, Eric Almeida Xavier, Natália Cristina de Oliveira, Paulo Marcos da Matta Guedes, João Santana da Silva, Luiz Tadeu Moraes Figueiredo, and Victor Hugo Aquino Copyright © 2012 Denise Gonçalves et al. All rights reserved. Evolutionary Pattern of Asian HIV-1 Subtype B from 1990 to 2007: In Silico Analysis Based on Envelop Protein Thu, 03 May 2012 08:37:51 +0000 HIV-1 envelop gene is a major target for vaccine development. Envelop protein and its V3 loop is shown to be important determinant of HIV-1 pathogenecity. Herein, the evolutionary pattern of most prevalent HIV-1 subtype B in Asia is determined by analyzing envelop protein and V3 domain based on the 40 randomly selected sequences of HIV-1 from database (Los Alamos), divided into four groups since 1990–2007. Construction of envelop protein phylogeny by using MEGA 5 exhibit the active mutation pattern, increase in potential N-glycosylation sites which were predicted by using online software SignalP-NN. An online available tool Drawgram was used for multiple sequence alignment (MSA) of HIV-1 subtype B envelop region and V3 loop while the alignment was rechecked by using CLUSTAL W and further was analyzed for GPGX motif and conserved region in V3 loop. Variation at fourth position of the GPGX motif and 60% conservation was found in V3 loop. Hence, this diversifying pattern of envelop protein in the Asia formulates the HIV-1 strains more pathogenic during the period of 17 years. These findings might help in understanding significant structural and functional constrains of the mutant viral strains and ultimately in vaccine development. Sobia Kanwal and Tariq Mahmood Copyright © 2012 Sobia Kanwal and Tariq Mahmood. All rights reserved. Viral Infection in Renal Transplant Recipients Wed, 02 May 2012 13:28:26 +0000 Viruses are among the most common causes of opportunistic infection after transplantation. The risk for viral infection is a function of the specific virus encountered, the intensity of immune suppression used to prevent graft rejection, and other host factors governing susceptibility. Although cytomegalovirus is the most common opportunistic pathogen seen in transplant recipients, numerous other viruses have also affected outcomes. In some cases, preventive measures such as pretransplant screening, prophylactic antiviral therapy, or posttransplant viral monitoring may limit the impact of these infections. Recent advances in laboratory monitoring and antiviral therapy have improved outcomes. Studies of viral latency, reactivation, and the cellular effects of viral infection will provide clues for future strategies in prevention and treatment of viral infections. This paper will summarize the major viral infections seen following transplant and discuss strategies for prevention and management of these potential pathogens. Jovana Cukuranovic, Sladjana Ugrenovic, Ivan Jovanovic, Milan Visnjic, and Vladisav Stefanovic Copyright © 2012 Jovana Cukuranovic et al. All rights reserved. A Real-Time PCR Assay for Bat SARS-Like Coronavirus Detection and Its Application to Italian Greater Horseshoe Bat Faecal Sample Surveys Sun, 29 Apr 2012 15:36:28 +0000 Bats are source of coronaviruses closely related to the severe acute respiratory syndrome (SARS) virus. Numerous studies have been carried out to identify new bat viruses related to SARS-coronavirus (bat-SARS-like CoVs) using a reverse-transcribed-polymerase chain reaction assay. However, a qualitative PCR could underestimate the prevalence of infection, affecting the epidemiological evaluation of bats in viral ecology. In this work an SYBR Green-real time PCR assay was developed for diagnosing infection with SARS-related coronaviruses from bat guano and was applied as screening tool in a survey carried out on 45 greater horseshoe bats (Rhinolophus ferrumequinum) sampled in Italy in 2009. The assay showed high sensitivity and reproducibility. Its application on bats screening resulted in a prevalence of 42%. This method could be suitable as screening tool in epidemiological surveys about the presence of bat-SARS-like CoVs, consequently to obtain a more realistic scenario of the viral prevalence in the population. Andrea Balboni, Laura Gallina, Alessandra Palladini, Santino Prosperi, and Mara Battilani Copyright © 2012 Andrea Balboni et al. All rights reserved. The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A Thu, 26 Apr 2012 15:23:05 +0000 Protein engineering provides an opportunity to generate new immunogens with desired features. Previously, we have demonstrated that hamster polyomavirus major capsid protein VP1-derived virus-like particles (VLPs) are highly immunogenic and can be employed for the insertion of foreign epitopes at certain surface-exposed positions. In the current study, we have designed pseudotype VLPs consisting of an intact VP1 protein and VP2 protein fused with the target antigen—cellular marker p16INK4A—at its N terminus. Both proteins coexpressed in yeast were self-assembled to pseudotype VLPs harbouring the inserted antigen on the surface. The pseudotype VLPs were used for generation of antibodies against p16INK4A that represents a potential biomarker for cells transformed by high-risk human papillomavirus (HPV). The pseudotype VLPs induced in immunized mice a strong immune response against the target antigen. The antisera raised against pseudotype VLPs showed specific immunostaining of p16INK4A protein in malignant cervical tissue. Spleen cells of the immunized mice were used to generate monoclonal antibodies against p16INK4A protein. The specificity of antibodies was proven by the immunostaining of HPV-transformed cells. In conclusion, the current study demonstrates the potential of pseudotype VLPs with inserted target antigen as a new type of immunogens to generate antibodies of high diagnostic value. Rita Lasickienė, Alma Gedvilaite, Milda Norkiene, Vaida Simanaviciene, Indre Sezaite, Dovile Dekaminaviciute, Evelina Shikova, and Aurelija Zvirbliene Copyright © 2012 Rita Lasickienė et al. All rights reserved. Characterization of the Immune Response Induced by a Commercially Available Inactivated Bluetongue Virus Serotype 1 Vaccine in Sheep Tue, 24 Apr 2012 11:52:57 +0000 The protective immune response generated by a commercial monovalent inactivated vaccine against bluetongue virus serotype 1 (BTV1) was studied. Five sheep were vaccinated, boost-vaccinated, and then challenged against BTV1 ALG/2006. RT-PCR did not detect viremia at any time during the experiment. Except a temperature increase observed after the initial and boost vaccinations, no clinical signs or lesions were observed. A specific and protective antibody response checked by ELISA was induced after vaccination and boost vaccination. This specific antibody response was associated with a significant increase in B lymphocytes confirmed by flow cytometry, while significant increases were not observed in T lymphocyte subpopulations (CD4+, CD8+, and WC1+), CD25+ regulatory cells, or CD14+ monocytes. After challenge with BTV1, the antibody response was much higher than during the boost vaccination period, and it was associated with a significant increase in B lymphocytes, CD14+ monocytes, CD25+ regulatory cells, and CD8+ cytotoxic T lymphocytes. Ana Cristina Pérez de Diego, Pedro José Sánchez-Cordón, Ana Isabel de las Heras, and José Manuel Sánchez-Vizcaíno Copyright © 2012 Ana Cristina Pérez de Diego et al. All rights reserved. Parvovirus B19 Infection and Severe Anemia in Renal Transplant Recipients Tue, 24 Apr 2012 11:34:19 +0000 Kidney transplant (KT) recipients can develop symptomatic Parvovirus (PV) B19 infections, frequently associated with persistent anemia. The aim of this study was to evaluate the prevalence and clinical significance of PV B19 infection in anemic and non-anemic KT patients. Overall, out of 64 patients monitored for the presence of PV B19 by real-time PCR, 2 (3.12%) had an active PV B19 infection, in absence of other viral coinfections. The 2 cases occurred in nonanemic kidney transplant patients group (2/50, 4%), while none of the anemic transplant patients (0/14) was found to suffer from this infection. Moreover, patients affected by active PV B19 infection showed viral loads not exceeding 1×105 genome copies/reaction. In conclusion, in this study, PV B19 infection was not common in renal transplant population and wasn’t associated with severe anemia. Antonio Carraturo, Valentina Catalani, Donatella Ottaviani, Patrizia Menichelli, Maurizio Rossini, Delia Terella, and Brunello Biondi Copyright © 2012 Antonio Carraturo et al. All rights reserved. Cytotoxic, Virucidal, and Antiviral Activity of South American Plant and Algae Extracts Tue, 24 Apr 2012 08:46:31 +0000 Herpes simplex virus type 1 (HSV-1) infection has a prevalence of 70% in the human population. Treatment is based on acyclovir, valacyclovir, and foscarnet, three drugs that share the same mechanism of action and of which resistant strains have been isolated from patients. In this aspect, innovative drug therapies are required. Natural products offer unlimited opportunities for the discovery of antiviral compounds. In this study, 28 extracts corresponding to 24 plant species and 4 alga species were assayed in vitro to detect antiviral activity against HSV-1. Six of the methanolic extracts inactivated viral particles by direct interaction and 14 presented antiviral activity when incubated with cells already infected. Most interesting antiviral activity values obtained are those of Limonium brasiliense, Psidium guajava, and Phyllanthus niruri, which inhibit HSV-1 replication in vitro with 50% effective concentration (EC50) values of 185, 118, and 60 μg/mL, respectively. For these extracts toxicity values were calculated and therefore selectivity indexes (SI) obtained. Further characterization of the bioactive components of antiviral plants will pave the way for the discovery of new compounds against HSV-1. Paula Faral-Tello, Santiago Mirazo, Carmelo Dutra, Andrés Pérez, Lucía Geis-Asteggiante, Sandra Frabasile, Elina Koncke, Danilo Davyt, Lucía Cavallaro, Horacio Heinzen, and Juan Arbiza Copyright © 2012 Paula Faral-Tello et al. All rights reserved. Human Papillomavirus Is Associated with Breast Cancer in the North Part of Iran Sun, 01 Apr 2012 08:51:41 +0000 We have analyzed the possible relevance of HPV infection for breast cancer risk among Iranian women from north part of Iran. Among women with breast cancer, 25.9% had positive test results for HPV DNA in breast tumor samples in contrast to 2.4% of women with noncancer status (𝑃=0.002). The infection of HPV has increased the risk of breast cancer (OR 14.247; 95% CI 1.558–130.284, 𝑃=0.019). The high-risk HPV genotypes (types 16 and 18) in samples of breast cancer patients were the predominant types (53.34%). Other genotypes detected in breast cancer were HPV-6, HPV-11, HPV-15, HPV-23, and HPV-124, and one isolate could not be genotyped compared to HPV reference sequences. While the sole detected HPV in control specimens was HPV-124. Our study reveals that HPV infection and age are the risk factors in breast cancer development in the north part of Iran. Afsaneh Sigaroodi, Seyed Alireza Nadji, Farshad Naghshvar, Rakhshandeh Nategh, Habib Emami, and Ali Akbar Velayati Copyright © 2012 Afsaneh Sigaroodi et al. All rights reserved. Microscopic Analysis of Severe Structural Rearrangements of the Plant Endoplasmic Reticulum and Golgi Caused by Overexpression of Poa semilatent virus Movement Protein Wed, 04 Jan 2012 10:56:00 +0000 Cell-to-cell transport of plant viruses is mediated by virus-encoded movement proteins and occurs through plasmodesmata interconnecting neighboring cells in plant tissues. Three movement proteins coded by the “triple gene block” (TGB) and named TGBp1, TGBp2 and TGBp3 have distinct functions in viral transport. TGBp1 binds viral genomic RNAs to form ribonucleoprotein complexes representing the transport form of viral genome, while TGBp2 and TGBp3 are necessary for intracellular delivery of such complexes to plasmodesmata. Recently, it was revealed that overexpression of Potato virus X TGBp3 triggers the unfolded protein response mitigating the endoplasmic reticulum (ER) stress leading to cell death if this protein reaches high levels in the ER. Here we report microscopic studies of the influence of the Poa semilatent hordeivirus TGBp3 overexpressed in Nicotiana benthamiana epidermal cells by particle bombardment on cell endomembranes and demonstrate that the protein C-terminal transmembrane segment contains a determinant responsible for vesiculation and coalescence of the endoplasmic reticulum and Golgi presumably accompanying the ER stress that can be induced upon high-level TGBp3 expression. Andrey G. Solovyev, Joachim Schiemann, and Sergey Y. Morozov Copyright © 2012 Andrey G. Solovyev et al. All rights reserved. AFRICAN SWINE FEVER VIRUS (ASFV) MULTIGENE FAMILIES 360 AND 530 GENES PROMOTE INFECTED MACROPHAGE SURVIVAL Mon, 01 Jan 1900 00:00:00 +0000 L. Zsak, J.H. Sur, T.G. Burrage, J.G. Neilan, and D.L. Rock Copyright © 2001 L. Zsak et al. All rights reserved. Ras Signalling Pathway: A Gateway for HSV-1 Infection Mon, 01 Jan 1900 00:00:00 +0000 Faris Farassati and Patrick W.K. Lee Copyright © 2003 Faris Farassati and Patrick W.K. Lee. All rights reserved. Apoptosis of Spleenocytes and Expression of HIV Gene Products in the HIV-1 Transgenic Rat Mon, 01 Jan 1900 00:00:00 +0000 D. Clark, F.J. Denaro, N. Hayes, O. Jones, M. McCready, H. Davis, W. Reid, and J. Bryant Copyright © 2001 D. Clark et al. All rights reserved. Regulation of Apoptosis in African Swine Fever Virus–Infected Macrophages Mon, 01 Jan 1900 00:00:00 +0000 A number of viruses have evolved antiapoptotic mechanisms to promote infected-cell survival, either to ensure efficient productive viral replication or to promote long-term survival of virus-infected cells. Recent studies identified critical African swine fever virus genes involved in the complex regulation of ASFV-host interactions. Here we review the present knowledge of the recently identified ASFV genes with special attention to those which affect viral virulence, host range, and pathogenesis by regulating viral-induced apoptotic mechanisms. Laslo Zsak and John G. Neilan Copyright © 2002 Laslo Zsak and John G. Neilan. All rights reserved. Far Out! Foot-and-Mouth, Ribosomes, and Other Tales Mon, 01 Jan 1900 00:00:00 +0000 The spreading scourge of foot-and-mouth disease heads Nature’s news this week. Science leads this week with a story about stunning new images of ribosomes. Copyright © 2001 TheScientificWorldJOURNAL All rights reserved. Structure of Influenza A Virus Promoter and its Implications for Viral RNA Synthesis Mon, 01 Jan 1900 00:00:00 +0000 Since the worst worldwide pandemic ever recorded — the 1918 Spanish influenza outbreak that killed more than 20 million people — we have achieved significant advances in understanding the influenza virus. However, the fear of such a pandemic remains strong. For example, in 1997, when a lethal influenza variant afflicted eight people in Hong Kong, contributing to the death of six, officials feared the next wave had begun. They managed to solve the problem quickly, however, by destroying all of the poultry in Hong Kong[1]. Sung-Hun Bae and Byong-Seok Choi Copyright © 2001 Sung-Hun Bae and Byong-Seok Choi. All rights reserved. POLYOMAVIRUS LARGE T ANTIGEN INTERACT WITH THE DISC AND PROTECT AGAINST FAS INDUCED APOPTOSIS Mon, 01 Jan 1900 00:00:00 +0000 Rodier F., Bertrand R., and A-M. Mes-Masson Copyright © 2001 Rodier F. et al. All rights reserved.