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All Journal Medical Journal of Indonesia Universa Medicina
Sylvia Y. Muliawan
Department of Microbiology, Faculty of Medicine Universitas Indonesia, Jakarta 10320
Health Professions Immunology & microbiology Medicine & Pharmacology Public Health

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Journal : Universa Medicina

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Profile of nonstructural glycoprotein NS1 as a diagnostic marker in dengue type 2 virus infection Muliawan, Sylvia Y.
Universa Medicina Vol 26, No 2 (2007)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2007.v26.73-79

Abstract

BackgroundSeveral studies in dengue on cloned or cell culture passaged in wild and attenuated virus have been performed, but each report described different sites of nucleotide and amino acid changes possibly associated with virulence. The aim of this study was to investigate protein expression of dengue type 2 virus infected C6/36 cells through two-dimensional gel electrophoresis followed by N-terminal Edman degradation sequencing to identify target proteins.MethodsTwo type of samples; dengue type 2 virus-infected C6/36 cells and uninfected C6/36 cells were prepared. Immobilized non-linear pH gradient strips, pH 3-10 were used in isoelectric focusing, and 10% homogeneous gels were used in Sodium dodecyl sulphate - Polyacylamide gel electrophoresis. The target protein spots were subsequently transferred onto polyvinyldifluoride membrane by western blotting and visualized by coomassie brilliant blue for N-terminal sequencing purpose.ResultsThis study revealed four target protein spots in dengue type 2 virus-infected C6/36 cells that were not present in uninfected C6/36 cells. The N-terminal sequencing result was D-S-G-C-V-V-S-W-K-N-K which was identical to nonstructural glycoprotein NS1 (Swissprot-database) associated with the replication process of flaviviruses. ConclusionNonstructural NS1 might be used as a diagnostic marker and/or as a parameter to evaluate the effect of antiviral agents for dengue type 2 virus infection/replication.
Effect of Dillenia suffruticosa extract on dengue virus type 2 replication Muliawan, Sylvia Y.
Universa Medicina Vol 27, No 1 (2008)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2008.v27.1-5

Abstract

Drugs of plant origin can have easy acceptability as they are non-toxic and inexpensive. Dillenia suffruticosa (D.suffruticosa), also known as Simpoh air in Malaysia, has antibacterial and antifungal activity, but no reported antiviral activity. This led the investigation of this plant against dengue virus type 2 (DV2) replication. Two type of samples: DV2-infected C6/36 cells and uninfected C6/36 cells were prepared. D.suffruticosa extract was prepared in water. The plant extract, D.suffruticosa, was subjected to C6/36 cells with following concentrations, 0.025, 0.05, 0.1, 0.2, 0.4 mg/ml and incubated for 2 hours before infected by DV2. Immobilized non-linear pH gradient strips, pH 3-10 were used in isoelectric focusing, and 10% homogeneous gels were used in sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The results were stained by silver staining for visualization. It was found that NS1 protein expression on infected C6/36 cells was down regulated in intensity of concentrations 0.025, 0.05 and 0.1 mg/ml. However, when higher concentration of D.suffruticosa extract were used (0.2 and 0.4 mg/ml), the cells exhibited slightly reduced NS1 protein spots, when compared to the above concentrations (0.025, 0.05, 0.1 mg/ml). D.suffruticosa extract has an inhibitory effect on the replication of DV2. Non-structural NS1 could be used as a diagnostic marker and/or as a parameter to evaluate the effect of antiviral agents for dengue type 2 virus infection/replication.
Co-Authors L.H. Moehario Lucky H. Moehario P. Sudarmono