Search results for: newcastle disease virus
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4243

Search results for: newcastle disease virus

4063 The Origin, Diffusion and a Comparison of Ordinary Differential Equations Numerical Solutions Used by SIR Model in Order to Predict SARS-CoV-2 in Nordic Countries

Authors: Gleda Kutrolli, Maksi Kutrolli, Etjon Meco

Abstract:

SARS-CoV-2 virus is currently one of the most infectious pathogens for humans. It started in China at the end of 2019 and now it is spread in all over the world. The origin and diffusion of the SARS-CoV-2 epidemic, is analysed based on the discussion of viral phylogeny theory. With the aim of understanding the spread of infection in the affected countries, it is crucial to modelize the spread of the virus and simulate its activity. In this paper, the prediction of coronavirus outbreak is done by using SIR model without vital dynamics, applying different numerical technique solving ordinary differential equations (ODEs). We find out that ABM and MRT methods perform better than other techniques and that the activity of the virus will decrease in April but it never cease (for some time the activity will remain low) and the next cycle will start in the middle July 2020 for Norway and Denmark, and October 2020 for Sweden, and September for Finland.

Keywords: forecasting, ordinary differential equations, SARS-COV-2 epidemic, SIR model

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4062 Epstein, Barr Virus Alters ATM-Dependent DNA Damage Responses in Germinal Centre B-Cells during Early Infection

Authors: Esther N. Maina, Anna Skowronska, Sridhar Chaganti, Malcolm A. Taylor, Paul G. Murray, Tatjana Stankovic

Abstract:

Epstein-Barr virus (EBV) has been implicated in the pathogenesis of human tumours of B-cell origin. The demonstration that a proportion of Hodgkin lymphomas and all Burkitt’s lymphomas harbour EBV suggests that the virus contributes to the development of these malignancies. However, the mechanisms of lymphomagenesis remain largely unknown. To determine whether EBV causes DNA damage and alters DNA damage response in cells of EBV-driven lymphoma origin, Germinal Centre (GC) B cells were infected with EBV and DNA damage responses to gamma ionising radiation (IR) assessed at early time points (12hr – 72hr) after infection and prior to establishment of lymphoblastoid (LCL) cell lines. In the presence of EBV, we observed induction of spontaneous DNA DSBs and downregulation of ATM-dependent phosphorylation in response to IR. This downregulation coincided with reduced ability of infected cells to repair IR-induced DNA double-strand breaks, as measured by the kinetics of gamma H2AX, a marker of double-strand breaks, and by the tail moment of the comet assay. Furthermore, we found that alteration of DNA damage responses coincided with the expression of LMP-1 protein. The presence of the EBV virus did not affect the localization of the ATM-dependent DNA repair proteins to sites of damage but instead lead to an increased expression of PP5, a phosphatase that regulates ATM function. The impact of the virus on DNA repair was most prominent 24h after infection, suggesting that this time point is crucial for the viral establishment in B cells. Our results suggest that during an early infection EBV virus dampens crucial cellular responses to DNA double-strand breaks which facilitate successful viral infection, but at the same time might provide the mechanism for tumor development.

Keywords: EBV, ATM, DNA damage, germinal center cells

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4061 Anterior Uveitis Caused by Infection with Cytomegalovirus and Herpes Simplex Virus Type I at Cicendo Eye Hospital Bandung

Authors: Shinta Stri Ayuda Nur Setyaningsih

Abstract:

Anterior uveitis is often triggered by viral infections such as herpes simplex virus (HSV) and cytomegalovirus (CMV). This study aims to provide an overview of the demographic and clinical characteristics of patients with anterior uveitis caused by CMV and HSV infection at PMN Cicendo Eye Hospital Bandung. This study used a retrospective observational method. Data were collected from the medical records of patients who visited the PMN Infection and Immunology Polyclinic at Cicendo Eye Hospital between February and July 2023. The results showed that anterior uveitis associated with HSV and CMV viruses often occurs in the elderly and more in women. The most common clinical symptoms are red eyes and decreased visual acuity, with a gradual onset of symptoms. Complications that often arise are cataracts and glaucoma. This study provides a deeper understanding of anterior uveitis caused by infection with HSV and CMV viruses.

Keywords: uveitis anterior, cytomegavirus, herpes simplex virus type I ELISA

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4060 Detection of Elephant Endotheliotropic Herpes Virus in a Wild Asian Elephant Calf in Thailand by Using Real-Time PCR

Authors: Bopit Puyati, Anchittha Kaewchana, Nuntita Ruksachat

Abstract:

In January 2018, a male wild elephant, approximately 2 years old, was found dead in Phu Luang Wildlife Sanctuary, Loei province. The elephant was likely to die around 2 weeks earlier. The carcass was decayed without any signs of attack or bullet. No organs were removed. A deadly viral disease was suspected. Different organs including lung, liver, intestine and tongue were collected and submitted to the veterinary research and development center, Surin province for viral detection. The samples were then examined with real-time PCR for detecting U41 Major DNA binding protein (MDBP) gene and with conventional PCR for the presence of specific polymerase gene. We used tumor necrosis factor (TNF) gene as the internal control. In our real-time PCR, elephant endotheliotropic herpesvirus (EEHV) was recovered from lung, liver, and tongue whereas only tongue provided a positive result in the conventional PCR. All samples were positive with TNF gene detection. To our knowledge, this is the first report of EEHV detection in wild elephant in Thailand. EEHV surveillance in this wild population is strongly suggested. Linkage between EEHV in wild and domestic elephants should be further explored.

Keywords: elephant endotheliotropic herpes virus, PCR, Thailand, wild Asian elephant

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4059 A Small-Molecular Inhibitor of Influenza Virus via Disrupting the PA and PB1 Interaction of the Viral Polymerase

Authors: Shuofeng Yuan, Bojian Zheng

Abstract:

Assembly of the heterotrimeric polymerase complex of influenza virus from the individual subunits PB1, PA, and PB2 is a prerequisite for viral replication, in which the interaction between the N-terminal of PB1 (PB1N) and the C terminal of PA (PAC) may be a desired target for antiviral development. In this study, we first compared the feasibility of high throughput screening by enzyme-linked immunosorbent assay (ELISA) and fluorescence polarization (FP) assay. Among the two, ELISA was demonstrated to own broader dynamic range so that it was used for screening inhibitors, which blocked PA and PB1 interaction. Several binding inhibitors of PAC-PB1N were identified and subsequently tested for the antiviral efficacy. Apparently, 3-(2-chlorophenyl)-6-ethyl-7-methyl[1,2,4]triazolo[4,3-a]pyrimidin-5-ol, designated ANA-1, was found to be a strong inhibitor of PAC-PB1N interaction and act as a potent antiviral agent against the infections of multiple subtypes of influenza A virus, including H1N1, H3N2, H5N1, H7N7, H7N9 and H9N2 subtypes, in cell cultures. Intranasal administration of ANA-1 protected mice from lethal challenge and reduced lung viral loads in H1N1 virus infected BALB/c mice. Docking analyses predicted that ANA-1 bound to an allosteric site of PAC, which would cause conformational changes thereby disrupting the PAC-PB1N interaction. Overall, our study has identified a novel compound with potential to be developed as an anti-influenza drug.

Keywords: influenza, antiviral, viral polymerase, compounds

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4058 Detection of Bcl2 Polymorphism in Patient with Hepatocellular carcinoma

Authors: Mohamed Abdel-Hamid, Olfat Gamil Shaker, Doha El-Sayed Ellakwa, Eman Fathy Abdel-Maksoud

Abstract:

Introduction: Despite advances in the knowledge of the molecular virology of hepatitis C virus (HCV), the mechanisms of hepatocellular injury in HCV infection are not completely understood. Hepatitis C viral infection (HCV) influences the susceptibility to apoptosis. This could lead to insufficient antiviral immune response and persistent viral infection. Aim of this study: was to examine whether BCL-2 gene polymorphism at codon 43 (+127G/A or Ala43Thr) has an impact on development of hepatocellular carcinoma caused by chronic hepatitis C Egyptian patients. Subjects and Methods: The study included three groups; group 1: composing of 30 patients with hepatocellular carcinoma (HCC), group 2 composing of 30 patients with HCV, group 3 composing of 30 healthy subjects matching the same age and socioeconomic status were taken as a control group. Gene polymorphism of BCL2 (Ala43Thr) were evaluated by PCR-RFLP technique and measured for all patients and controls. Results: The summed 43Thr genotype was more frequent and statistically significant in HCC patients as compared to control group. This genotype of BCL2 gene may inhibit the programmed cell death which leads to disturbance in tissue and cells homeostasis and reduction in immune regulation. This result leads to viral replication and HCV persistence. Moreover, virus produces variety of mechanisms to block genes participated in apoptosis. This mechanism proves that HCV patients who have 43Thr genotype are more susceptible to HCC. Conclusion: The data suggest for the first time that the BCL2 polymorphism is associated with the susceptibility to HCC in Egyptian populations and might be used as molecular markers for evaluating HCC risk. This study clearly demonstrated that Chronic HCV exhibit a deregulation of apoptosis with the disease progression. This provides an insight into the pathogenesis of chronic HCV infection, and may contribute to the therapy.

Keywords: BCL2 gene, Hepatitis C Virus, Hepatocellular carcinoma, sensitivity, specificity, apoptosis

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4057 Detection of Respiratory Syncytial Virus (hRSV) by PCR Technique in Lower Respiratory Tract Infection (LRTI) in Babylon City

Authors: Amal Raqib Shameran, Ghanim Aboud Al-Mola

Abstract:

Respiratory syncytial virus (hRSV) is the major pathogens of respiratory tract infections (RTI) among infants and children in the world. They are classified in family Paramyxoviridae and sub-family Pneumovirinae. The current work aimed to detect the role of RSV in the lower respiratory tract infection (LRTI) in Hilla, Iraq. The samples were collected from 50 children who were admitted to hospital suffering from lower respiratory tract infections (LRTI). 50 nasal and pharyngeal swabs were taken from patients at the period from January 2010 till April 2011, hospitalized in Hilla Maternity and Children Hospital. The results showed that the proportion of children infected with hRSV accounted for 24% 12/50 with lower respiratory tract infections (LRTI) when they tested by polymerase chain reaction (RT-PCR).

Keywords: respiratory syncytial virus, respiratory tract infections, infants, polymerase chain reaction (PCR)

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4056 Facile Synthetic Process for Lamivudine and Emtricitabine

Authors: Devender Mandala, Paul Watts

Abstract:

Cis-Nucleosides mainly lamivudine (3TC) and emtricitabine (FTC) are an important tool in the treatment of Human immune deficiency virus (HIV), Hepatitis B virus (HBV) and Human T-Lymotropoic virus (HTLV). Lamivudine and emtricitabine are potent nucleoside analog reverse transcriptase inhibitors (nRTI). These two drugs are synthesized by a four-stage process from the starting materials: menthyl glyoxylate hydrate and 1,4-dithane-2,5-diol to produce the 5-hydroxy oxathiolane which upon acetylation with acetic anhydride to yield 5-acetoxy oxathiolane. Then glycosylation of this acetyl product with silyl protected nucleoside to produce the intermediate. The reduction of this intermediates can provide the final targets. Although there are several different methods reported for the synthesis of lamivudine and emtricitabine as a single enantiomer, we required an efficient route, which was suitable for large-scale synthesis to support the development of these compounds. In this process, we successfully prepared the intermediates of lamivudine and emtricitabine without using any solvents and catalyst, thus promoting the green synthesis. All the synthesized compound were confirmed by TLC, GC, Mass, NMR and 13C NMR spectroscopy.

Keywords: emtricitabine, green synthesis, lamivudine, nucleoside

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4055 A DNA-Based Nano-biosensor for the Rapid Detection of the Dengue Virus in Mosquito

Authors: Lilia M. Fernando, Matthew K. Vasher, Evangelyn C. Alocilja

Abstract:

This paper describes the development of a DNA-based nanobiosensor to detect the dengue virus in mosquito using electrically active magnetic (EAM) nanoparticles as the concentrator and electrochemical transducer. The biosensor detection encompasses two sets of oligonucleotide probes that are specific to the dengue virus: the detector probe labeled with the EAM nanoparticles and the biotinylated capture probe. The DNA targets are double hybridized to the detector and the capture probes and concentrated from nonspecific DNA fragments by applying a magnetic field. Subsequently, the DNA sandwiched targets (EAM-detector probe–DNA target–capture probe-biotin) are captured on streptavidin modified screen printed carbon electrodes through the biotinylated capture probes. Detection is achieved electrochemically by measuring the oxidation–reduction signal of the EAM nanoparticles. Results indicate that the biosensor is able to detect the redox signal of the EAM nanoparticles at dengue DNA concentrations as low as 10 ng/ul.

Keywords: dengue, magnetic nanoparticles, mosquito, nanobiosensor

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4054 Investigation of Clubroot Disease Occurrence under Chemical and Organic Soil Environment

Authors: Zakirul Islam, Yugo Kumokawa, Quoc Thinh Tran, Motoki Kubo

Abstract:

Clubroot is a disease of cruciferous plant caused by soil born pathogen Plasmodiophora brassicae and can significantly limit the production through rapid spreading. The present study was designed to investigate the effect of cultivation practices (chemical and organic soils) on clubroot disease development in Brassica rapa. Disease index and root bacterial composition were investigated for both chemical and organic soils. The bacterial biomass and diversity in organic soil were higher than those in chemical soil. Disease severity was distinct for two different cultivation methods. The number of endophytic bacteria decreased in the infected root for both soils. The increased number of endophytic bacterial number led to reduce the proliferation of pathogen spore inside the root and thus reduced the disease severity in organic plants.

Keywords: clubroot disease, bacterial biomass, root infection, disease index, chemical cultivation, organic cultivation

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4053 Viral Metagenomics Revealed a Novel Cardiovirus in Feces of Wild Rats

Authors: Asif Mahmood, Shama Shama, Hao Ni, Hao Wang, Yu Ling, Hui Xu, Shixing Yang, Qais Ahmad Naseer, Wen Zhang

Abstract:

Cardiovirus is a genus of viruses belonging to the family Picornaviridae. Here, we used viral metagenomic techniques to detect the viral nucleic acid in the fecal samples from wild rats in Zhenjiang city in China. Fecal samples were collected from 20 wild rats and pooled into four sample pools and then subjected to libraries construction which were then sequenced on Illumina MiSeq platform. The sequenced reads were analyzed using viral metagenomic analysis pipeline. A novel cardiovirus from feces of a wild rat was identified, named amzj-2018, of which the complete genome was acquired. Phylogenetic analysis based on the complete amino acid sequence of polyprotein revealed that amzj-2018 formed a separate branch located between clusters of Saffold virus and Rat Theilovirus 1 (RTV-1). Phylogenetic analysis based on different regions of the polyproteins, including P1, P2, P3, and P2+P3, respectively, showed discordant trees, where the tree based on P3 region indicated that amzj-2018 clustered separately between Theiler's murine encephalomyelitis virus and RTV-1. The complete genome of a cardiovirus was determined from the feces of wild rats which belonged to a novel type of cardiovirus based on phylogenetic analysis. Whether it is associated with disease needs further investigation.

Keywords: cardiovirus, viral metagenomics, genomic organization, phylogenetic analysis

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4052 Cardiovascular Disease Is Common among Patients with Systemic Lupus Erythematosus

Authors: Fathia Ehmouda Zaid, Reim Abudelnbi

Abstract:

Cardiovascular disease is a major cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE). Patients and method: Cross-section study (68) patients diagnosed as systemic lupus erythematosus (SLE), who visited the outpatient clinic of rheumatology, these patients were interviewed with a structured questionnaire about their past and current clinically for presence of Cardiovascular disease in systemic lupus and use SLEDAI, specific tests [ECG –ECHO –CXRAY] the data are analyzed statistically by Pearson's correlation coefficient was calculated and statistical significance was defined as P< 0.05,during period (2013-2014). Objective: Estimation Cardiovascular disease manifestation of systemic lupus erythematosus, correlation with disease activity, morbidity, and mortality. Result: (68) Patients diagnosed as systemic lupus erythematosus' age range from (18-48 years), M=(13±29Y), Sex were female 66/68 (97.1%), male 2/68 (2.9%),duration of disease range[1-15year], M =[7±8y], we found Cardiovascular disease manifestation of systemic lupus erythematosus 32/68 (47.1%), correlation with disease activity use SLEDAI,(r= 476** p=0.000),Morbidity,(r= .554**; p=0.000) and mortality (r=.181; p=.139), Cardiovascular disease manifestations of systemic lupus erythematosus are pericarditis 8/68 (11.8%), pericardial effusion 6/68 (8.8%), myocarditis 4/68 (5.9 %), valvular lesions (endocarditis) 1/68 (1.5%), pulmonary hypertension (PAH) 12/68 (17.6%), coronary artery disease 1/68 (1.5%), none of patients have conduction abnormalities involvement. Correlation with disease activity use SLEDAI, pericarditis (r= .210, p=.086), pericardial effusion (r= 0.079, p=.520), myocarditis (r= 272*, p=.027), valvular lesions (endocarditis) (r= .112, p= .362), pulmonary hypertension (PAH) (r= .257*, p=.035) and coronary artery disease (r=.075, p=.544) correlation between cardiovascular disease manifestations of systemic lupus erythematosus and specific organ involvement we found Mucocutaneous (r=.091 p= .459), musculoskeletal (MSK) (r=.110 p=.373), Renal disease (r=.278*, p=.022), neurologic disease (r=.085, p=.489) and Hematologic disease (r=-.264*, p=.030). Conclusion: Cardiovascular manifestation is more frequent symptoms with systemic lupus erythematosus (SLE) is 47 % correlation with disease activity and morbidity but not with mortality. Recommendations: Focus research to evaluation and an adequate assessment of cardiovascular complications on the morbidity and mortality of the patients with SLE are still required.

Keywords: cardiovascular disease, systemic lupus erythematosus, disease activity, mortality

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4051 The Four-Way Interactions among Host Plant-Whitefly-Virus-Endosymbionts in Insect and Disease Development

Authors: N. R. Prasannakumar, M. N. Maruthi

Abstract:

The whitefly, Bemisia tabaci (Gennadius) (Hemiptera; Aleyrodidae) is a highly polyphagous pest reported to infest over 600 plant hosts globally. About 42 genetic groups/cryptic species of B. tabaci exist in the world on different hosts. The species have variable behaviour with respect to feeding, development and transmission of viral diseases. Feeding on diverse host plants affect both whitefly development and the population of the endosymbionts harboured by the insects. Due to changes in the level of endosymbionts, the virus transmission efficiency by the vector also gets affected. We investigated these interactions on five host plants – egg plant, tomato, beans, okra and cotton - using a single whitefly species Asia 1 infected with three different bacteria Portiera, Wolbachia and Arsenophonus. The Asia 1 transmits the Tomato leaf curl Bangalore virus (ToLCBV) effectively and thus was used in the interaction studies. We found a significant impact of hosts on whitefly growth and development; eggplant was most favourable host, while okra and tomato were least favourable. Among the endosymbiotic bacteria, the titre of Wolbachia was significantly affected by feeding of B. tabaci on different host plants whereas Arsenophonus and Portiera were unaffected. When whitefly fed on ToLCBV-infected tomato plants, the Arsenophonus population was significantly increased, indicating its previously confirmed role in ToLCBV transmission. Further, screening of total proteins of B. tabaci Asia 1 genetic group interacting with ToLCBV coat protein was carried out using Y2H system. Some of the proteins found to be interacting with ToLCBV CP were HSPs 70kDa, GroEL, nucleoproteins, vitellogenins, apolipophorins, lachesins, enolase. The reported protein thus would be the potential targets for novel whitefly control strategies such as RNAi or novel insecticide target sites for sustainable whitefly management after confirmation of genuine proteins.

Keywords: cDNA, whitefly, ToLCBV, endosymbionts, Y2H

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4050 Evaluation of Disease Risk Variables in the Control of Bovine Tuberculosis

Authors: Berrin Şentürk

Abstract:

In this study, due to the recurrence of bovine tuberculosis, in the same areas, the risk factors for the disease were determined and evaluated at the local level. This study was carried out in 32 farms where the disease was detected in the district and center of Samsun province in 2014. Predetermined risk factors, such as farm, environmental and economic risks, were investigated with the survey method. It was predetermined that risks in the three groups are similar to the risk variables of the disease on the global scale. These risk factors that increase the susceptibility of the infection must be understood by the herd owners. The risk-based contagious disease management system approach should be applied for bovine tuberculosis by farmers, animal health professionals and public and private sector decision makers.

Keywords: bovine tuberculosis, disease management, control, outbreak, risk analysis

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4049 A Novel Small-Molecule Inhibitor of Influenza a Virus Acts by Suppressing PA Endonuclease Activity of the Viral Polymerase

Authors: Shuafeng Yuan, Bojian Zheng

Abstract:

The RNA-dependent RNA polymerase of influenza a virus comprises conserved and independently folded subdomains with defined functionalities. The N-terminal domain of the PA subunit (PAN) harbors the endonuclease function so that it can serve as a desired target for drug discovery. To identify a class of anti-influenza inhibitors that impedes PAN endonuclease activity, a screening approach that integrated the fluorescence resonance energy transfer based endonuclease inhibitor assay with the DNA gel-based endonuclease inhibitor assay was conducted, followed by the evaluation of antiviral efficacies and potential cytotoxicity of the primary hits in vitro and in vivo. A small-molecule compound ANA-0 was identified as a potent inhibitor against the replication of multiple subtypes of influenza A virus, including H1N1, H3N2, H5N1, H7N7, H7N9 and H9N2, in cell cultures. Combinational treatment of zanamivir and ANA-0 exerted synergistic anti-influenza effect in vitro. Intranasal administration of ANA-0 protected mice from lethal challenge and reduced lung viral loads in H1N1 virus infected BALB/c mice. Docking analyses predicted ANA-0 bound the endonuclease cavity of PAN by interacting with the metal-binding and catalytic residues. In summary, ANA-0 shows potential to be developed to novel anti-influenza agents.

Keywords: anti-influenza, novel compound, inhibition of endonuclease, PA

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4048 Neuroblastoma in Children and the Potential Involvement of Viruses in Its Pathogenesis

Authors: Ugo Rovigatti

Abstract:

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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4047 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

Abstract:

Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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4046 Effect of Time of Planting on Powdery Mildew Development on Cucumber

Authors: H. Parameshwar Naik, Shripad Kulkarni

Abstract:

Powdery mildew is a serious disease among the fungal in high humid areas with varied temperature conditions. In recent days disease becomes very severe due to uncertain weather conditions and unique character of the disease is, it produces white mycelia growth on upper and lower leaf surfaces and in severe conditions it leads to defoliation. Results of the experiment revealed that sowing of crop in the I fortnight (FN) of July recorded the minimum mean disease severity (7.96 %) followed by crop sown in II FN of July (13.19 %) as against the crop sown in II FN of August (41.44 %) and I FN of September (33.78 %) and the I fortnight of October (33.77 %). In the first date of sowing infection started at 45 DAS and progressed till 73 DAS and it was up to 14.66 Percent and in second date of sowing disease progressed up to 22.66 percent and in the third date of sowing, it was up to 59.35 percent. Afterward, the disease started earlier and progressed up to 66.15 percent and in sixth and seventh date of sowing disease progressed up to 43.15 percent and 59.85 percent respectively. Disease progress is very fast after 45 days after sowing and highest disease incidence was noticed at 73 DAS irrespective of dates of sowing. From the results of the present study, it is very clear that disease development will be very high if crop sown in between 1st fortnight of August and the 1st fortnight of September.

Keywords: cucumber, India, Karnataka, powdery mildew

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4045 Hanta Virus Infection in a Child and Sequelae

Authors: Vijay Samuel, Tina Thekkekkara, Shoma Ganguly

Abstract:

There is no reported Hanta Seoul virus infection in children in the UK so far, making it quite challenging for clinicians in diagnosing, predicting and prognosticating the outcome of the infection to patients and parents. We report a case of a ten-year-old girl who presented with pyrexia associated with headache, photophobia and abdominal pain. The family had recently acquired two pet rats six weeks ago. She appeared flushed with peri-oral pallor, coated the strawberry tongue, inflamed tonsils and bilateral cervical lymphadenopathy. Her liver and splenic edges were palpable. Investigations showed that she was thrombocytopenic with deranged renal and liver functions. An ultrasound abdomen demonstrated a mildly enlarged spleen, peripancreatic lymph node and an acalculous cholecystitis. In view of her clinical presentation, a diagnosis of leptospirosis was considered and she was commenced on intravenous benzylpenicillin. The following day she became oliguric, developed significant proteinuria and her renal function deteriorated. Following conservative management, her urine output gradually improved along with her renal function, proteinuria and thrombocytopaenia. Serology for leptospirosis and various other viruses were negative. Following discussion with the Rare and Imported Pathogens Laboratory at Porton hanta virus serology was requested and found to be strongly positive for Seoul hanta virus. Following discharge she developed palpitations, fatigue, severe headache and cognitive difficulties including memory loss and difficulties in spelling, reading and mathematics. Extensive investigations including ECG, MRI brain and CSF studies were performed and revealed no significant abnormalities. Since 2012, there have been six cases of acute kidney injury due to Hantavirus infection in the UK. Two cases were from the Humber region and were exposure to wild rats and the other four were exposed to specially bred pet fancy rats. Hanta virus infections can cause mild flu like symptoms but two clinical syndromes are associated with severe disease including haemorrhagic fever with renal syndrome, which may be associated with thrombocytopenia and Hantavirus cardiopulmonary syndrome. Neuropsychological impairments reported following hantavirus pulmonary syndrome and following Puumala virus infection have been reported. Minor white matter lesions were found in about half of the patients investigated with MRI brain. Seoul virus has a global distribution owing to the dispersal of its carrier host rats, through global trade. Several ports in the region could explain the possible establishment of Seoul virus in local populations of rats in the Yorkshire and Humber region. The risk of infection for occupationally exposed groups is 1-3% compared to 32.9% for specialist pet rat owners. The report highlight’s the importance of routinely asking about pets in the family. We hope to raise awareness of the emergence of hantavirus infection in the UK, particularly in the Yorkshire and Humber region. Clinicians should consider hantavirus infection as a potential cause of febrile illness causing renal impairment in children. Awareness of the possible neuro-cognitive sequele would help the clinicians offer appropriate information and support to children and their families. Contacting Rare and Imported Pathogens Laboratory at Porton is a useful resource for clinicians in UK when they consider unusual infections.

Keywords: Seoul hantavirus in child Porton, UK Acute kidney injury

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4044 Distribution of HLA-DQA1 and HLA-DQB1 Alleles in Thais: Genetics Database Insight for COVID-19 Severity

Authors: Jinu Phonamontham

Abstract:

Coronavirus, also referred to as COVID-19, is a virus caused by the SARS-Cov-2 virus. The pandemic has caused over 10 million cases and 500,000 deaths worldwide through the end of June 2020. In a previous study, HLA-DQA1*01:02 allele was associated with COVID-19 disease (p-value = 0.0121). Furthermore, there was a statistical significance between HLA- DQB1*06:02 and COVID-19 in the Italian population by Bonferroni’s correction (p-value = 0.0016). Nevertheless, there is no data describing the distribution of HLA alleles as a valid marker for prediction of COVID-19 in the Thai population. We want to investigate the prevalence of HLA-DQA1*01:02 and HLA-DQB1*06:02 alleles that are associated with severe COVID-19 in the Thai population. In this study, we recruited 200 healthy Thai individuals. Genomic DNA samples were isolated from EDTA blood using Genomic DNA Mini Kit. HLA genotyping was conducted using the Lifecodes HLA SSO typing kits (Immucor, West Avenue, Stamford, USA). The frequency of HLA-DQA1 alleles in Thai population, consisting of HLA-DQA1*01:01 (27.75%), HLA-DQA1*01:02 (24.50%), HLA-DQA1*03:03 (13.00%), HLA-DQA1*06:01 (10.25%) and HLA-DQA1*02:01 (6.75%). Furthermore, the distributions of HLA-DQB1 alleles were HLA-DQB1*05:02 (21.50%), HLA-DQB1*03:01 (15.75%), HLA-DQB1*05:01 (14.50%), HLA-DQB1*03:03 (11.00%) and HLA-DQB1*02:02 (8.25%). Particularly, HLA- DQA1*01:02 (29.00%) allele was the highest frequency in the NorthEast group, but there was not significant difference when compared with the other regions in Thais (p-value = 0.4202). HLA-DQB1*06:02 allele was similarly distributed in Thai population and there was no significant difference between Thais and China (3.8%) and South Korea (6.4%) and Japan (8.2%) with p-value > 0.05. Whereas, South Africa (15.7%) has a significance with Thais by p-value of 0.0013. This study supports the specific genotyping of the HLA-DQA1*01:02 and HLA-DQB1*06:02 alleles to screen severe COVID-19 in Thai and many populations.

Keywords: HLA-DQA1*01:02, HLA-DQB1*06:02, Asian, Thai population

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4043 Rapid Detection and Differentiation of Camel Pox, Contagious Ecthyma and Papilloma Viruses in Clinical Samples of Camels Using a Multiplex PCR

Authors: A. I. Khalafalla, K. A. Al-Busada, I. M. El-Sabagh

Abstract:

Pox and pox-like diseases of camels are a group of exanthematous skin conditions that have become increasingly important economically. They may be caused by three distinct viruses: camelpox virus (CMPV), camel contagious ecthyma virus (CCEV) and camel papillomavirus (CAPV). These diseases are difficult to differentiate based on clinical presentation in disease outbreaks. Molecular methods such as PCR targeting species-specific genes have been developed and used to identify CMPV and CCEV, but not simultaneously in a single tube. Recently, multiplex PCR has gained reputation as a convenient diagnostic method with cost- and time–saving benefits. In the present communication, we describe the development, optimization and validation a multiplex PCR assays able to detect simultaneously the genome of the three viruses in one single test allowing for rapid and efficient molecular diagnosis. The assay was developed based on the evaluation and combination of published and new primer sets, and was applied to the detection of 110 tissue samples. The method showed high sensitivity, and the specificity was confirmed by PCR-product sequencing. In conclusion, this rapid, sensitive and specific assay is considered a useful method for identifying three important viruses in specimens from camels and as part of a molecular diagnostic regime.

Keywords: multiplex PCR, diagnosis, pox and pox-like diseases, camels

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4042 Effects of Propolis on Immunomodulatory and Antibody Production in Broilers

Authors: Yu-Hsiang Yu

Abstract:

The immunomodulatory effect of propolis has been widely investigated in the past decade. However, the beneficial effects in broilers are still poorly understood. The aim of this study was to evaluate the effect of propolis added in drinking water on immunomodulatory and antibody production in broiler. Total of 48 chicks were randomly allocated into four groups with 12 broilers per group. All birds were intranasal inoculated with Newcastle Disease vaccine at 4 and 14 days old of age. Four groups, including control without any treatment, groups of A, B and F [3 days of anterior (A), 3 days of posterior (P) and 6 days of full (F)] were supplied the propolis at 300 ppm in drinking water when vaccination was performed, respectively. Our results showed that no significant difference was found in growth performance, antibody production and immune organ index among groups. However, propolis treatments in broilers significantly reduced IL-4 expression in spleen at 14 days-old of age and bursa at 28 days-old of age compared with control group. The expression of IFN-gamma in spleen (A, P and F group) and bursal (F group) were elevated compared with control group at 28 days-old of age. In conclusion, our results indicated that propolis-treated birds could bear the capability for immunomodulatory effects by change Th1 subset cytokine expression in vaccination.

Keywords: propolis, broiler, immunomodulatory, vaccination

Procedia PDF Downloads 329
4041 Exploring Emerging Viruses From a Protected Reserve

Authors: Nemat Sokhandan Bashir

Abstract:

Threats from viruses to agricultural crops could be even larger than the losses caused by the other pathogens because, in many cases, the viral infection is latent but crucial from an epidemic point of view. Wild vegetation can be a source of many viruses that eventually find their destiny in crop plants. Although often asymptomatic in wild plants due to adaptation, they can potentially cause serious losses in crops. Therefore, exploring viruses in wild vegetation is very important. Recently, omics have been quite useful for exploring plant viruses from various plant sources, especially wild vegetation. For instance, we have discovered viruses such as Ambrossia asymptomatic virus I (AAV-1) through the application of metagenomics from Oklahoma Prairie Reserve. Accordingly, extracts from randomly-sampled plants are subjected to high speed and ultracentrifugation to separated virus-like particles (VLP), then nucleic acids in the form of DNA or RNA are extracted from such VLPs by treatment with phenol—chloroform and subsequent precipitation by ethanol. The nucleic acid preparations are separately treated with RNAse or DNAse in order to determine the genome component of VLPs. In the case of RNAs, the complementary cDNAs are synthesized before submitting to DNA sequencing. However, for VLPs with DNA contents, the procedure would be relatively straightforward without making cDNA. Because the length of the nucleic acid content of VPLs can be different, various strategies are employed to achieve sequencing. Techniques similar to so-called "chromosome walking" may be used to achieve sequences of long segments. When the nucleotide sequence data were obtained, they were subjected to BLAST analysis to determine the most related previously reported virus sequences. In one case, we determined that the novel virus was AAV-l because the sequence comparison and analysis revealed that the reads were the closest to the Indian citrus ringspot virus (ICRSV). AAV—l had an RNA genome with 7408 nucleotides in length and contained six open reading frames (ORFs). Based on phylogenies inferred from the replicase and coat protein ORFs of the virus, it was placed in the genus Mandarivirus.

Keywords: wild, plant, novel, metagenomics

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4040 Addressing Factors Associated with Vertical HIV Transmission among Pregnant Women in Rwanda

Authors: Murorunkwere Marie Claire

Abstract:

Introduction: In Sub-Saharan Africa and specifically in Rwandan rural areas, mother-to-Child human immunodeficiency virus transmission remains a big challenge. This is mainly due to lack of awareness and ignorance among pregnant rural women, leading to neglect regular taking of prophylactic antiretroviral treatment and to persistently beliefs in traditional healers and home deliveries. This paper explores the factors associated with stagnant reduction in human immunodeficiency virus vertical transmission among pregnant rural women and provides solutions to tackle it. Methodology: The first phase of this research will be a qualitative survey was conducted to assess the knowledge, attitudes and practices towards vertical human immunodeficiency virus transmission among pregnant women in one rural district in Rwanda. The data generated from phase one of this research will be used to address the main factors revealed through community mobilization and motivation on attending required antenatal consultations and hospital deliveries, proper and regular antiretroviral treatment taking, and discouraging beliefs in traditional healers and home deliveries. Refresher training seminars will also be organized for healthcare providers qualified on conducting deliveries about current measures to maximize the reduction of chances that can lead to mother -child contamination (to avoid early rupture of membranes and to prevent any source of contamination). Results: This paper is expected to contribute in a significant reduction of the vertical human immunodeficiency virus transmission burden among pregnant rural women. Conclusion: Strong campaigns on prevention of mother- to-child human immunodeficiency virus transmission and community mobilization of pregnant rural women, and house to house education and continuous reminders as well as training seminars to health care personnel on updated measures is, key in addressing vertical human immunodeficiency virus transmission.

Keywords: attitudes transformation, community mobilisation, pregnant rural women, vertical HIV transmission

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4039 Level of Awareness of Genetic Counselling in Benue State Nigeria: Its Advocacy on the Inheritance of Sickle Cell Disease

Authors: Agi Sunday

Abstract:

A descriptive analysis of reported cases of sickle cell disease and the level of awareness about genetic counselling in 30 hospitals were carried out. Additionally, 150 individuals between ages 16-45 were randomly selected for evaluation of genetic counselling awareness. The main tools for this study were questionnaires which were taken to hospitals, and individuals completed the others. The numbers of reported cases of sickle cell disease recorded in private, public and teaching hospitals were 14 and 57; 143 and 89; 272 and 57 for the periods of 1995-2000 and 2001-2005, respectively. A general informal genetic counselling took place mostly in the hospitals visited. 122 (86%) individuals had the knowledge of genetic disease and only 43 (30.3%) individuals have been exposed to genetic counselling. 64% of individuals agreed that genetic counselling would help in the prevention of genetic disease.

Keywords: sickle disease, genetic counseling, genetic testing, advocacy

Procedia PDF Downloads 389
4038 Analyzing the Impact of Migration on HIV and AIDS Incidence Cases in Malaysia

Authors: Ofosuhene O. Apenteng, Noor Azina Ismail

Abstract:

The human immunodeficiency virus (HIV) that causes acquired immune deficiency syndrome (AIDS) remains a global cause of morbidity and mortality. It has caused panic since its emergence. Relationships between migration and HIV/AIDS have become complex. In the absence of prospectively designed studies, dynamic mathematical models that take into account the migration movement which will give very useful information. We have explored the utility of mathematical models in understanding transmission dynamics of HIV and AIDS and in assessing the magnitude of how migration has impact on the disease. The model was calibrated to HIV and AIDS incidence data from Malaysia Ministry of Health from the period of 1986 to 2011 using Bayesian analysis with combination of Markov chain Monte Carlo method (MCMC) approach to estimate the model parameters. From the estimated parameters, the estimated basic reproduction number was 22.5812. The rate at which the susceptible individual moved to HIV compartment has the highest sensitivity value which is more significant as compared to the remaining parameters. Thus, the disease becomes unstable. This is a big concern and not good indicator from the public health point of view since the aim is to stabilize the epidemic at the disease-free equilibrium. However, these results suggest that the government as a policy maker should make further efforts to curb illegal activities performed by migrants. It is shown that our models reflect considerably the dynamic behavior of the HIV/AIDS epidemic in Malaysia and eventually could be used strategically for other countries.

Keywords: epidemic model, reproduction number, HIV, MCMC, parameter estimation

Procedia PDF Downloads 366
4037 Role of DatScan in the Diagnosis of Parkinson's Disease

Authors: Shraddha Gopal, Jayam Lazarus

Abstract:

Aims: To study the referral practice and impact of DAT-scan in the diagnosis or exclusion of Parkinson’s disease. Settings and Designs: A retrospective study Materials and methods: A retrospective study of the results of 60 patients who were referred for a DAT scan over a period of 2 years from the Department of Neurology at Northern Lincolnshire and Goole NHS trust. The reason for DAT scan referral was noted under 5 categories against Parkinson’s disease; drug-induced Parkinson’s, essential tremors, diagnostic dilemma, not responding to Parkinson’s treatment, and others. We assessed the number of patients who were diagnosed with Parkinson’s disease against the number of patients in whom Parkinson’s disease was excluded or an alternative diagnosis was made. Statistical methods: Microsoft Excel was used for data collection and statistical analysis, Results: 30 of the 60 scans were performed to confirm the diagnosis of early Parkinson’s disease, 13 were done to differentiate essential tremors from Parkinsonism, 6 were performed to exclude drug-induced Parkinsonism, 5 were done to look for alternative diagnosis as the patients were not responding to anti-Parkinson medication and 6 indications were outside the recommended guidelines. 55% of cases were confirmed with a diagnosis of Parkinson’s disease. 43.33% had Parkinson’s disease excluded. 33 of the 60 scans showed bilateral abnormalities and confirmed the clinical diagnosis of Parkinson’s disease. Conclusion: DAT scan provides valuable information in confirming Parkinson’s disease in 55% of patients along with excluding the diagnosis in 43.33% of patients aiding an alternative diagnosis.

Keywords: DATSCAN, Parkinson's disease, diagnosis, essential tremors

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4036 Sensing Endocrine Disrupting Chemicals by Virus-Based Structural Colour Nanostructure

Authors: Lee Yujin, Han Jiye, Oh Jin-Woo

Abstract:

The adverse effects of endocrine disrupting chemicals (EDCs) has attracted considerable public interests. The benzene-like EDCs structure mimics the mechanisms of hormones naturally occurring in vivo, and alters physiological function of the endocrine system. Although, some of the most representative EDCs such as polychlorinated biphenyls (PCBs) and phthalates compounds already have been prohibited to produce and use in many countries, however, PCBs and phthalates in plastic products as flame retardant and plasticizer are still circulated nowadays. EDCs can be released from products while using and discarding, and it causes serious environmental and health issues. Here, we developed virus-based structurally coloured nanostructure that can detect minute EDCs concentration sensitively and selectively. These structurally coloured nanostructure exhibits characteristic angel-independent colors due to the regular virus bundle structure formation through simple pulling technique. The designed number of different colour bands can be formed through controlling concentration of virus solution and pulling speed. The virus, M-13 bacteriophage, was genetically engineered to react with specific ECDs, typically PCBs and phthalates. M-13 bacteriophage surface (pVIII major coat protein) was decorated with benzene derivative binding peptides (WHW) through phage library method. In the initial assessment, virus-based color sensor was exposed to several organic chemicals including benzene, toluene, phenol, chlorobenzene, and phthalic anhydride. Along with the selectivity evaluation of virus-based colour sensor, it also been tested for sensitivity. 10 to 300 ppm of phthalic anhydride and chlorobenzene were detected by colour sensor, and showed the significant sensitivity with about 90 of dissociation constant. Noteworthy, all measurements were analyzed through principal component analysis (PCA) and linear discrimination analysis (LDA), and exhibited clear discrimination ability upon exposure to 2 categories of EDCs (PCBs and phthalates). Because of its easy fabrication, high sensitivity, and the superior selectivity, M-13 bacteriophage-based color sensor could be a simple and reliable portable sensing system for environmental monitoring, healthcare, social security, and so on.

Keywords: M-13 bacteriophage, colour sensor, genetic engineering, EDCs

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4035 Biochemical and Antiviral Study of Peptides Isolated from Amaranthus hypochondriacus on Tomato Yellow Leaf Curl Virus Replication

Authors: José Silvestre Mendoza Figueroa, Anders Kvarnheden, Jesús Méndez Lozano, Edgar Antonio Rodríguez Negrete, Manuel Soriano García

Abstract:

Agroindustrial plants such as cereals and pseudo cereals offer a substantial source of biomacromolecules, as they contain large amounts per tissue-gram of proteins, polysaccharides and lipids in comparison with other plants. In particular, Amaranthus hypochondriacus seeds have high levels of proteins in comparison with other cereal and pseudo cereal species, which makes the plant a good source of bioactive molecules such as peptides. Geminiviruses are one principal class of pathogens that causes important economic losses in crops, affecting directly the development and production of the plant. One such virus is the Tomato yellow leaf curl virus (TYLCV), which affects mainly Solanacea family plants such as tomato species. The symptoms of the disease are curling of leaves, chlorosis, dwarfing and floral abortion. The aim of this work was to get peptides derived from enzymatic hydrolysis of globulins and albumins from amaranth seeds with specific recognition of the replication origin in the TYLCV genome, and to test the antiviral activity on host plants with the idea to generate a direct control of this viral infection. Globulins and albumins from amaranth were extracted, the fraction was enzymatically digested with papain, and the aromatic peptides fraction was selected for further purification. Six peptides were tested against the replication origin (OR) using affinity assays, surface resonance plasmon and fluorescent titration, and two of these peptides showed high affinity values to the replication origin of the virus, dissociation constant values were calculated and showed specific interaction between the peptide Ampep1 and the OR. An in vitro replication test of the total TYLCV DNA was performed, in which the peptide AmPep1 was added in different concentrations to the system reaction, which resulted in a decrease of viral DNA synthesis when the peptide concentration increased. Also, we showed that the peptide can decrease the complementary DNA chain of the virus in Nicotiana benthamiana leaves, confirming that the peptide binds to the OR and that its expected mechanism of action is to decrease the replication rate of the viral genome. In an infection assay, N. benthamiana plants were agroinfected with TYLCV-Israel and TYLCV-Guasave. After confirming systemic infection, the peptide was infiltrated in new infected leaves, and the plants treated with the peptide showed a decrease of virus symptoms and viral titer. In order to confirm the antiviral activity in a commercial crop, tomato plants were infected with TYLCV. After confirming systemic infection, plants were infiltrated with peptide solution as above, and the symptom development was monitored 21 days after treatment, showing that tomato plants treated with peptides had lower symptom rates and viral titer. The peptide was also tested against other begomovirus such as Pepper huasteco yellow vein virus (PHYVV-Guasave), showing a decrease of symptoms in N. benthamiana infected plants. The model of direct biochemical control of TYLCV infection shown in this work can be extrapolated to other begomovirus infections, and the methods reported here can be used for design of antiviral agrochemicals for other plant virus infections.

Keywords: agrochemical screening, antiviral, begomovirus, geminivirus, peptides, plasmon, TYLCV

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4034 Prolonged Synthesis of Chitin Polysaccharide from Chlorovirus System

Authors: Numfon Rakkhumkaew, Takeru Kawasaki, Makoto Fujie, Takashi Yamada

Abstract:

Chlorella viruses or chloroviruses contain a gene that encodes a function for chitin synthesis, which is expressed early in viral infection to produce chitin polysaccharide, a polymer of β-1, 4-linked GlcNAc, on the outside of Chlorella cell wall. Interestingly, chlorovirus system is an eco-friendly system which converses CO2 and solar energy from the environment into useful materials. However, infected Chlorella cells are lysed at the final stage of viral infection, and this phenomenon is caused the breaking down of polysaccharide. To postpone the lysing period and prolong the synthesis of chitin polysaccharide on cells, the slow growing virus incorporated with aphidicolin treatment, an inhibitor of DNA synthesis, was investigated. In this study, a total of 25 virus isolates from water samples in Japan region were analyzed for CHS (the gene for CH synthase) gene by PCR (polymerase chain reaction). The accumulation and appearance of chitin polysaccharide on infected cells were detected by biotinylated chitin-binding proteins WGA (wheat germ agglutinin)-biotin for chitin in conjunction with avidin-Cy 2 or Cy 3 and investigated by fluorescence microscopy, observed as green or yellow fluorescence over the cell surface. Among all chlorovirus isolates, cells infected with CNF1 revealed the accumulation of chitin over the cell surface within 30 min p.i. and continued to accumulate on cells until 4 h p.i. before cell lyses which was 1.6 times longer accumulation period than cells infected with CVK2 (prototype virus). Furthermore, addition of aphidicolin could extend the chitin accumulation on cells infected with CNF1 until 8 h p.i. before cell lyses. Whereas, CVK2-infected cells treated with aphidicolin could prolong the chitin synthesis only for 6 h p.i. before cell lyses. Therefore, chitin synthesis by Chlorella-virus system could be prolonged by using slow-growing viral isolates and with aphidicolin.

Keywords: chitin, chlorovirus, Chlorella virus, aphidicolin

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