Judul: Isolasi, Identifikasi dan Karakterisasi Molekular Virus Hepatitis B pada Owa Jawa (Hylobates moloch) di Indonesia
Abstrak: Hepatitis B virus (HBV) was reported not only able to infect human being, but also non-human primates, especially the great apes group such as orangutans, gorillas, chimpanzees, as well as the lesser apes from the family of Hylobatidae. Among other species within the genus, Hylobates moloch (owa jawa or silvery/ javan gibbons), which is considered as one of Indonesia’s endemic endangered species, has been reported to harbor HBV of their own strain. Analyses of HBV isolated from infected gibbons were done in this study to reveal their relatedness to the published data. Plasma samples from nine javan gibbons were obtained as part of diagnostic purpose. Those animals were previously tested positive for the presence of hepatisis B-surface antigen (HBsAg) by enzyme-linked immunosorbent assay (ELISA). DNA samples were extracted from these nine plasma samples to be used as templates for amplification of the Pre-S1 region by polymerase chain reactions (PCR) technique using semi degenerate primers that could amplify several strains of HBVs. The PCR products of 457 - 459 base pairs were then subject to restriction fragment length polymorphism (RFLP) analyses using Bst2UI enzyme to evaluate the pattern of cleaved fragments. The PCR products were sequenced for further homology analyses and to create phylogenetic tree of the sequenced data obtained from the study in comparison with published data. Two plasma samples were used as templates for whole region amplification using sets of 4 primer. The PCR products of 3192 base pairs were sequenced for further homology analyses and to create phylogenetic tree. Phylogenetic tree analyses for Pre-S1 nucleotide region and whole region showed that the javan gibbon isolates formed their own clusters, separate from the other nonhuman primate isolates.
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Judul: Study of Tupaia javanica Hepatocytes Culture as In Vitro Replication Media of Nonhuman Primate HBV
Abstrak: Virus hepatitis B (VHB) adalah virus DNA yang menginfeksi organ hati sebagai target utama. Infeksi ini menyebabkan hepatitis yang dapat bersifat kronis dan berlanjut menjadi sirosis, kanker hati bahkan kematian. Pengembangan model in vivo dan in vitro berkaitan dengan hepatitis B menjadi perhatian khusus bagi para peneliti. Kultur hepatosit banyak digunakan untuk mempelajari berbagai ha] terkait hepatitis B baik dari aspek virus, penyakit, pencegahan, dan pengobatan. Pengembangan hepatosit asal hewan kecil sepe1ii Tupaia sp. untuk model in vitro merupakan salah satu altematif dalam penyediaan kultur hepatosit. Tujuan penelitian ini adalah mengembangkan kultur primer dan lestari hepatosit T. javanica yang mampu mendukung replikasi VHB orangutan (OuHBV) dan VHB owa jawa (GiHBV), dan dapat menjawab kebutuhan akan model in vitro kultur hepatosit. Isolasi hepatosit T. javanica dilakukan dengan prosedur sebagai berikut: organ hati yang telah dicacah dalam cawan petri dengan larutan perfusi, diinkubasi pada 37 °C. Suspensi disentrifugasi 300-500 g sehingga didapatkan pelet sel hati. Pelet yang sudah dicuci kemudian diresuspensi dengan Hepatocyte Basal Media untuk menumbuhkan hepatosit dan selanjutnya hepatosit akan ditransformasi menggunakan perangkat Lenti-SV 40. Sodium taurocholate cotransporting polypeptide (NTCP) yang diindikasikan sebagai reseptor hepatosit untuk melekatnya VHB, nampak pada hepatosit baik pada kultur primer maupun kultur yang ditransformasi. Reseptor hepatosit spesifik ini terlihat sebagai pendaran hijau dengan uji imunofluoresensi. Kultur hepatosit primer T. javanica berhasil dikembangkan selama 14 hari untuk mencapai kepadatan 80%, dan inokulasi GiHBV dan OuHBV ke dalam kultur pada hari ke-15 menunjukkan adanya replikasi virus hingga hari ke-8 (berdasarkan hasil PCR). Kuantifikasi virus dengan real-time PCR memperlihatkan replikasi tertinggi pada hari kedua setelah inokulasi. Penjajaran sebagian protein permukaan setelah virus dinokulasikan memperlihatkan kesamaan asam amino dengan virus wild type sebesar 99.29% dan 95.71 % pada GiHBV dan OuHBV., Hepatitis B virus (HBV) is a DNA virus which infects liver as primary target organ. This i11fection causes hepatitis which could become chronic and progress to cirrhosis, liver cancer and even death. Development of in vitro and in vivo model related to hepatitis B is a concern for researchers. Hepatocyte cultures widely used to study a variety of aspects of hepatitis B such as the virus, disease, prevention and treatment. Development qf hepatocyte from small animal e.g. Tupaia sp. for the in vitro model is an alternative of hepatocyte cultures. The purpose of the study is to develop primary and immortal T. javanica hepatocytes cultures capable to replicate orangutan HBV (OuHBV) and javan gibbon HBV (GiHBV), and to answer the need for in vitro model ofhepatocytes. Isolation of hepatocytes from T. javanica was done as followed: mincedliver were transported into petri dish containing perfusion buffer solution to be incubated at 37 °C. The suspension was centrifuged at 300-500 g to obtain liver cell pellets. The washed-pellet was resuspended in Hepatocyte Basal Media to let specific hepatocytes grow and later be transformed by Lenti-SV40 kit. Sodium taurocholate cotransporting polypeptide (NTCP) has been ident(fied as hepatocyte receptor for HBV attachment, and was seen on hepatocytes from both primary and transformed cultures. This hepatocytes specific receptor was shown as green luminenscent by immunofluorescence assay. Primary T. javanica hepatocytes cultures was successfully maintained for
Keyword: Tupaia javanica, in vitro model, hepatocytes culture, hepatitis B virus, Liver cancer, Picornaviridae, Hepadanaviridae, Flaviviridae, Caliciviridae

Judul: Pertukaran dan Pengadukan Massa Air Kaitannya dengan Transfort Sedimen di Perairan Teluk Kendari
Abstrak: Sedimen dan transfort sedimen di perairan teluk dikontrol oleh tingkat pengadukan, pertukaran (sirkulasi) dan run off massa air.
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