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      Rapid identification of inhibitors that interfere with poliovirus replication using a cell-based assay

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          Abstract

          A small molecule library containing 480 known bioactive compounds was screened for antiviral activity against poliovirus (PV) using a cellular fluorescence resonance energy transfer (FRET) assay for viral protease activity. The infected reporter cells treated with the viral replication-suppressing compounds were examined via fluorescence microscope 7.5 h postinfection. Twelve molecules showed moderate to potent antiviral activity at concentrations less than 32 μM during the primary screening. Three compounds, anisomycin, linoleic acid, and lycorine, were chosen for validation. A dose-dependent cytotoxicity assay and a secondary screening using conventional plaque assay were conducted to confirm the results. The developed method can be used for rapid screening for molecules with antiviral activity.

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          FRET imaging.

          Elizabeth A Jares-Erijman, Thomas M. Jovin (2003)
          Förster (or Fluorescence) Resonance Energy Transfer (FRET) is unique in generating fluorescence signals sensitive to molecular conformation, association, and separation in the 1-10 nm range. We introduce a revised photophysical framework for the phenomenon and provide a systematic catalog of FRET techniques adapted to imaging systems, including new approaches proposed as suitable prospects for implementation. Applications extending from a single molecule to live cells will benefit from multidimensional microscopy techniques, particularly those adapted for optical sectioning and incorporating new algorithms for resolving the component contributions to images of complex molecular systems.
          Article 0 comments Cited 329 times – based on 0 reviews     Review now
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            Identification of natural compounds with antiviral activities against SARS-associated coronavirus

            Shi-you Li, Cong Chen, Hai-qing ZHANG … (2005)
            More than 200 Chinese medicinal herb extracts were screened for antiviral activities against Severe Acute Respiratory Syndrome-associated coronavirus (SARS-CoV) using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay for virus-induced cytopathic effect (CPE). Four of these extracts showed moderate to potent antiviral activities against SARS-CoV with 50% effective concentration (EC50) ranging from 2.4 ± 0.2 to 88.2 ± 7.7 μg/ml. Out of the four, Lycoris radiata was most potent. To identify the active component, L. radiata extract was subjected to further fractionation, purification, and CPE/MTS assays. This process led to the identification of a single substance lycorine as an anti-SARS-CoV component with an EC50 value of 15.7 ± 1.2 nM. This compound has a CC50 value of 14980.0 ± 912.0 nM in cytotoxicity assay and a selective index (SI) greater than 900. The results suggested that four herbal extracts and the compound lycorine are candidates for the development of new anti-SARS-CoV drugs in the treatment of SARS.
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              Hepatitis C virus RNA replication is regulated by host geranylgeranylation and fatty acids.

              F V Chisari, Sharookh B Kapadia (2005)
              Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Our laboratory has previously demonstrated that high-level HCV replication during acute infection of chimpanzees is associated with the modulation of multiple genes involved in lipid metabolism, and that drugs that regulate cholesterol and fatty acid biosynthesis regulate the replication of the subgenomic HCV replicon in Huh-7 cells. In this article, we demonstrate that Huh-7 cells harboring replicating, full-length HCV RNAs express elevated levels of ATP citrate lyase and acetyl-CoA synthetase genes, both of which are involved in cholesterol and fatty acid biosynthesis. Further, we confirm that the cholesterol-biosynthetic pathway controls HCV RNA replication by regulating the cellular levels of geranylgeranyl pyrophosphate, we demonstrate that the impact of geranylgeranylation depends on the fatty acid content of the cell, and we show that fatty acids can either stimulate or inhibit HCV replication, depending on their degree of saturation. These results illustrate a complex cellular-regulatory network that controls HCV RNA replication, presumably by modulating the trafficking and association of cellular and/or viral proteins with cellular membranes, suggesting that pharmacologic manipulation of these pathways may have a therapeutic effect in chronic HCV infection.
              Article 0 comments Cited 133 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Marylynn V. Yates
                Journal
                Journal ID (nlm-ta): Antiviral Res
                Journal ID (iso-abbrev): Antiviral Res
                Title: Antiviral Research
                Publisher: Elsevier B.V.
                ISSN (Print): 0166-3542
                ISSN (Electronic): 1872-9096
                Publication date PMC-release: 16 January 2008
                Publication date (Print): March 2008
                Publication date (Electronic): 16 January 2008
                Volume: 77
                Issue: 3
                Pages: 232-236
                Affiliations
                [a ]Department of Environmental Sciences, University of California, Riverside, CA 92521, United States
                [b ]Department of Chemical and Environmental Engineering, University of California, Riverside, CA, United States
                [c ]Department of Microbiology and Molecular Genetics, Harvard Medical School, MA, United States
                Author notes
                [* ]Corresponding author. Tel.: +1 951 827 2358; fax: +1 951 827 3993. marylynn.yates@ 123456ucr.edu
                Article
                Publisher ID: S0166-3542(07)00504-9
                DOI: 10.1016/j.antiviral.2007.12.009
                PMC ID: 7114228
                PubMed ID: 18243348
                SO-VID: df6e7519-a9d6-4094-bdce-3b2af2d44ad4
                Copyright statement: Copyright © 2007 Elsevier B.V. All rights reserved.
                License:

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                Date received : 3 August 2007
                Date accepted : 15 December 2007
                Categories
                Subject: Article

                ScienceOpen disciplines: Infectious disease & Microbiology
                Keywords: 2apro, 2a protease,fret, fluorescence resonance energy transfer,cellular sensor,protease,viral replication,fret
                Data availability:
                ScienceOpen disciplines: Infectious disease & Microbiology
                Keywords: 2apro, 2a protease, fret, fluorescence resonance energy transfer, cellular sensor, protease, viral replication, fret

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