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Detection of Type 2 Dengue Virus by in situ Hybridization Radji, Maksum; Soebandrio, Amin; Sudiro, Mirawati; Sudarmono, Pratiwi
Jurnal Mikrobiologi Indonesia Vol 4, No 1 (1999): JURNAL MIKROBIOLOGI INDONESIA
Publisher : Jurnal Mikrobiologi Indonesia

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Abstract

Demonstration of dengue virus DV in infected cell would enable elaboration of pathogenesis and pathoplsysiology of dengueFever and dengue Haemorrhagic Fever. Molecular detection of DV would give high sensitivity as well as specifity. C6136 mosquitocell line artificially infected with DV was used as a model of DV infected cell. 290-bp eDNA of envelope region of DV type 2 DV-2labeled with digoxigenin-il-dUT? was used as probe. Hybridization was performed directly to infected cell fixed on to glass slide insitu. 10 ng/ul of the probe was able to detect as low as lOx TCJD 50 infecting DV-2. The signal produced was not found in negativecontrol and was clearly increasing in infecting viral dose dependent manner. There was no cross reactivity between DV-2 probe andDV-3 and vice versa. The DV-2 probe was sensitive yet specific in demonstrating the presence of Dv-2 in infected cell.
Optimization of Surfactin Production by Bacillus amyloliquefaciens MD4-12 using Response Surface Methodology WIBISANA, AHMAD; SUMARYONO, WAHONO; SUDIRO, MIRAWATI; SUDARMONO, PRATIWI PUDJILESTARI
Microbiology Indonesia Vol 9, No 3 (2015): September 2015
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5454/mi.9.3.4

Abstract

Surfactin is a lipopeptide biosurfactant that show potential biomedical application due to its activities such as antiviral, antibacterial, antifungi, anticancer, and antimycoplasma. Bacillus amyloliquefaciens MD4-12, isolated from oil-contaminated soil, produced promising yield of surfactin in McKeen medium. The production of surfactin was influenced by many fermentation process parameters such as carbon, nitrogen, minerals and also environmental conditions such as pH and agitation. Therefore, to obtain high yield of surfactin by Bacillus amyloliquefaciens MD4-12, optimization of process production was conducted in shake flask fermentation using response surface methodology. McKeen medium composition was used as basal medium.  Screening of the best carbon and nitrogen source were selected in preliminary experiments followed by selection of the influencing significant parameters on surfactin production using Plackett-Burman design. Selected parameters were optimized by central composite design and for the data analysis was used response surface methodology. The result showed that the optimum medium composition contained (g/L) 45.0 glucose, 6.33 urea, 1.0 monosodium glutamate, 1.85 MgSO4.7H2O, 0.4 KCl, 0.5 K2HPO4 and 0.5 mL trace elements. The surfactin yield at optimal condition was 1.25 g/L, increased 2.4 times compared to condition prior to optimization. 
Antiviral Effect of Pterocarpus indicus Willd Leaves Extract Against Replication of Dengue Virus (DENV) In Vitro Dewi, Beti Ernawati; Angelina, Marissa; meilawati, lia; Hartati, Sri; Dewijanti, Indah Dwiatmi; Santi, Mei Ria; Desti, Hidayati; Sudiro, Mirawati
Journal of Tropical Life Science Vol 8, No 1 (2018)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.08.01.10

Abstract

Dengue hemorrhagic fever (DHF) is major public health problem in tropical and subtropical areas of the world with lack of approved vaccines and effective antiviral therapies. With no current treatment for illness attributed to dengue virus (DENV) infection other than supportive care, therapeutic strategies that use natural extract was developed. Indonesia have many plants that potential for antiviral drµgs such as Pterocarpus indicus Willd (P. indicus). The objective of this study was to determine the effect of P. indicus to inhibit DENV replication. We used a well-differentiated hepatocytes-derived cellular carcinoma cell line (Huh-7 it-1 cells) to determine and select antiviral activity. The toxicity effects were determined by MTT assay. Then, the suppression of DENV replication was determined by Focus assay. Dengue infected cells with DMSO were used as control. We found that crude extract (Pi), hexane (Pi.1) and ethyl acetate (Pi.2) extract showed strong inhibition with high selectivity index (SI) of 1,392; 285.36 and 168.56 respectively.  Sub fraction of Pi.1 and Pi.2 still showed strong inhibition with high SI.  Further sub-sub fraction of Pi.2 such as Pi.2.12 and Pi.2.12.1 still showed inhibition of DENV replication but there was reduction of SI value. The mechanism experiment of Pi.2.12, we found that Pi 2.12 more profound to inhibit in the post infection stage that entry or pre-infection. We conclude that the sub-fraction of Pi.2.12 has potential antiviral activity against DV infection in vitro. Further studies are still needed to investigate the pure compound of Pi.2.12 that inhibit and have advantages in the future as alternative for treatment of DENV infection.