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Objective To determine anti-viral activities of three Artocarpus species: Artocarpus altilis , Artocarpus camansi , and Artocarpus heterophyllus ( A. heterophyllus ) against Hepatitis C Virus (HCV). Methods Antiviral activities of the crude extracts were examined by cell culture method using Huh7it-1 cells and HCV genotype 2a strain JFH1. The mode of action for anti-HCV activities was determined by time-of-addition experiments. The effect on HCV RNA replication and HCV accumulation in cells were analyzed by quantitative reverse transcription-PCR and western blotting, respectively. Results The dichloromethane (DCM) extract of A. heterophyllus exhibited strong anti-HCV activity with an inhibitory concentration (IC 50 ) value of (1.5 ± 0.6) μg/mL without obvious toxicity. The DCM extracts from Artocarpus altilis and Artocarpus camansi showed moderate anti-HCV activities with IC 50 values being (6.5 ± 0.3) μg/mL and (9.7 ± 1.1) μg/mL, respectively. A time-of-addition studies showed that DCM extract from A. heterophyllus inhibited viral entry process though a direct virucidal activity and targeting host cells. HCV RNA replication and HCV protein expression were slightly reduced by the DCM treatment at high concentration. Conclusions The DCM extract from A. heterophyllus is a good candidate to develop an antiviral agent to prevent HCV grant reinfection following liver transplantation. [ABSTRACT FROM AUTHOR]

Hepatitis C Virus (HCV) has infected approximately 2-3% (130-170 million) of the world's population. No vaccine is available to prevent HCV infection. Investigation of anti-HCV agent is thus deemed necessary. Various plants have been explored for their anti-HCV activity. A. serratus is a member of Sapindaceae family, which fruit and seed were traditionally used as insecticide. Anti-HCV activity tested on A.serratus leaves extract has been done. The result showed that leaves extract exhibited anti-HCV with IC 50 value of 14.9 μg/ml and 9.8 μg/ml against HCV J6/JFH1 and JFH1a, respectively. The cytotoxicity assay results showed that A.serratus leaves extract was not toxic and has CC 50 >100 μg/ml. Mode of action experiment results suggested that A.serratus extract inhibited HCV at the post-entry step. Further fractionation of leaves extract by open column chromatography resulted in 4 fractions. Only Fraction 1 (AP-5F.1) exhibited anti-HCV with IC 50 value of 1.2 μg/ml against HCV JFH1a. Separation of AP-5F.1 by open column chromatography resulted in 15 fractions. Fraction number 13 (AP-5F.1.13) exhibited anti-HCV with IC 50 value of 0.43 μg/ml against HCV JFH1a. Separation of AP-5F.1.13 by semi preparative-HPLC resulted in isolate identified by TLC and LC-MS method as chlorophyll derivate. There was a possibility that chlorophyll derivate has participated in performing the anti-HCV activity of fractions and extract besides the other compounds contained. In this study, we concluded that A. serratus leaves extract, AP-5F.1, and AP-5F.1.13 exhibited anti-HCV activity against JFH1a virus. [ABSTRACT FROM AUTHOR]

Hepatitis C Virus (HCV) is a major global disease which often leads to chronicity and is potential to liver failure. There is no anti-HCV vaccine and the high diversity of viral genotypes will probably make it very difficult to develop a vaccine. Therefore, the development of new drugs for HCV treatment is highly required. It is commonly known that numerous important modern drugs have been developed from molecules originally isolated from natural sources. In this study, we tested the leave extract and fractions of Ficus fistulosa for their anti-HCV activities by cell culture method using Huh7it cells and HCV JFH1a. The result showed that ethanol extract of Ficus fistulosa (FFL) inhibited HCV JFH1a with IC 50 value of 20.43±4.51 μg/ml. Toxicity test also indicated that FFL was not toxic with CC 50 value of >200 μg/ml. The extract was further fractionated using chloroform (FFLC) and butanol (FFLB) successively. FFLC showed anti-HCV activity with IC 50 value of 5.67±1.54 μg/ml and CC 50 value of >100 μg/ml (Selectivity index >17.65). Further separation of FFLC by open column chromatography resulted in 12 subfractions (FFLC1-C12). Two subfractions, FFLC10, and FFLC11 showed high selectivity index (>100) with IC 50 value of 0.60±0.30 μg/ml and 0.43±0.29 μg/ml, respectively. Therefore the leave extract (FFL) and fractions (FFL10, FFL11) of Ficus fistulosa would be a good candidate to develop antiviral against HCV. [ABSTRACT FROM AUTHOR]

Dengue virus has four serotypes that cause a public health problem in Indonesia. Currently, there is no preventative vaccine for this disease, but some model vaccines are in development. The envelop (E) protein genes from three isolates of dengue virus (DENV-1, -3 and -4) were isolated, cloned into Escherichia coli and then sub-cloned into a baculovirus vector before co-transfection into Sf9 cells. Recombinant E genes were inserted between the Sma l and Sacl sites of the plasmid, adjacent to the baculoviral structural gene, polyhedrin. The sequence of recombinant E gene was relatively stable with 97–98% homology, although there were amino acid substitutions in some regions. The recombinant protein was more antigenic when exposed to polyclonal sera from infected humans than sera from immunized mice, but its binding to monoclonal antibodies IgG1a and IgG2b was stronger than other isotopes, including IgM, IgG and Ig1b. Recombinant E protein induced cellular immune responses in immunized mice, as demonstrated by lymphocyte secretion of IL-3. This study indicates that recombinant E protein expressed in a baculovirus system can induce humoral and cellular immune responses. [ABSTRACT FROM AUTHOR]

Rotavirus A (RVA) is a major cause of acute gastroenteritis in humans and animals worldwide. As a result of the segmented nature of the rotavirus genome, genetic reassortment commonly occurs. This study aims to clarify the genetic characteristics of RVAs circulating in Indonesia. From June 2015 through August 2016, stool samples were collected from 134 children aged <5 years (71 male and 63 female) with acute gastroenteritis who were inpatients at a private hospital in Surabaya, Indonesia. All stool samples were screened for RVA antigen using immunochromatography. Forty-two samples (31.3%, 42/134) were RVA antigen-positive. All RVA positive samples tested showed the unusual combinations of G3P[8] ( n  = 36) and G3P[6] ( n  = 3) with a short RNA pattern by G/P typing and polyacrylamide gel electrophoresis (PAGE). Whole genome analysis by next-generation sequencing (NGS) was performed for 11 strains to determine the RVA genotypes. Eleven rotavirus strains were found to carry a DS-like genetic backbone; nine strains showed a G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genome constellation, which was recently reported in Australia, Hungary, Spain and Brazil; as well, two strains showed a G3-P[6]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genome constellation. The phylogenetic tree based on the VP7 gene showed that all 11 strains were classified as equine-like G3, which is genetically distinct and different in origin from typical human G3 strains. The phylogenetic tree based on the NSP4 gene showed that six strains were classified as bovine-like strain and the remaining five were classified as human strain. In conclusion, we identified the strains which are intergenogroup reassortants containing an equine-like G3 VP7, a P[8])/P[6] VP4, with a DS-1-like genetic backbone. These findings suggest that equine-like G3P[8] and P[6] RVA strains have been circulating in the Indonesian population for at least 1 year and probably longer, indicating a diversity of RVAs in this area. [ABSTRACT FROM AUTHOR]

Norovirus (NoV) is a major cause of nonbacterial acute gastroenteritis worldwide in all age groups, and asymptomatic individuals may contribute to NoV transmission as a reservoir. Nonetheless, little information is available regarding asymptomatic NoV infection in Indonesia. We performed an epidemiological analysis of NoV infection among asymptomatic healthy volunteers in the city of Surabaya, Indonesia (population ~ 2.75 million). A total of 512 stool samples from 18 individuals (age range 20–42 years) collected from July 2015 to June 2016 were examined. The detection of NoV and the genotype classification were carried out by a reverse transcription-polymerase chain reaction (RT-PCR) direct sequencing method. NoV was detected in 14 of the 512 stool samples (2.7%), with 7 individuals (38.9%) having at least 1 positive stool sample. All 14 of the NoV strains detected belonged to genogroup GII. The phylogenetic analysis indicated that 10 strains (71.4%) were grouped with GII.2, 2 (14.3%) were GII.17, 1 was GII.4 Sydney 2012, and 1 was GII.1. The circulation of GII.Pg/GII.1 and GII.Pe/GII.4 Sydney 2012 recombinant variants was detected among an asymptomatic population in Surabaya, Indonesia. Of the 7 positive individuals, 2 were repeatedly infected with the same strain and heterogenous strains. Taken together, our results suggest that the excretion of NoV from healthy individuals is one of the sources of NoV outbreak. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) infection is highly prevalent among global populations, with an estimated number of infected patients being 170 million. Approximately 70–80% of patients acutely infected with HCV will progress to chronic liver disease, such as liver cirrhosis and hepatocellular carcinoma, which is a substantial cause of morbidity and mortality worldwide. New therapies for HCV infection have been developed, however, the therapeutic efficacies still need to be improved. Medicinal plants are promising sources for antivirals against HCV. A variety of plants have been tested and proven to be beneficial as antiviral drug candidates against HCV. In this study, we examined extracts, their subfractions and isolated compounds of Ruta angustifolia leaves for antiviral activities against HCV in cell culture. We isolated six compounds, chalepin, scopoletin, γ-fagarine, arborinine, kokusaginine and pseudane IX. Among them, chalepin and pseudane IX showed strong anti-HCV activities with 50% inhibitory concentration (IC 50 ) of 1.7 ± 0.5 and 1.4 ± 0.2 μg/ml, respectively, without apparent cytotoxicity. Their anti-HCV activities were stronger than that of ribavirin (2.8 ± 0.4 μg/ml), which has been widely used for the treatment of HCV infection. Mode-of-action analyses revealed that chalepin and pseudane IX inhibited HCV at the post-entry step and decreased the levels of HCV RNA replication and viral protein synthesis. We also observed that arborinine, kokusaginine and γ-fagarine possessed moderate levels of anti-HCV activities with IC 50 values being 6.4 ± 0.7, 6.4 ± 1.6 and 20.4 ± 0.4 μg/ml, respectively, whereas scopoletin did not exert significant anti-HCV activities at 30 μg/ml. [ABSTRACT FROM AUTHOR]