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All Journal Jurnal Sain Veteriner JURNAL TEKNIK MESIN Indonesian Journal of Biotechnology Jurnal Medik Veteriner Jurnal AME (Aplikasi Mekanika dan Energi): Jurnal Ilmiah Teknik Mesin MOTIVECTION : Journal of Mechanical, Electrical and Industrial Engineering Indonesian Journal of Advanced Research (IJAR) Jurnal Kedokteran Hewan Jurnal Sosialita: Jurnal Kajian Sosial dan Pendidikan
Aris Haryanto
Bagian Klinik Hewan, Fakultas Kedokteran Hewan, Universitas Udayana, Bali
Humanities Automotive Engineering Biochemistry, Genetics & Molecular Biology Civil Engineering, Building, Construction & Architecture Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Education Energy Environmental Science Immunology & microbiology Industrial & Manufacturing Engineering Languange, Linguistic, Communication & Media Materials Science & Nanotechnology Mechanical Engineering Medicine & Pharmacology Social Sciences Veterinary

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Nuclear Import Analysis of Two Different Fluorescent Marker Proteins into Hepatocyte Cell Lines (HuH-7 Cell) Haryanto, Aris; Kann, Michael
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

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Abstract

The application of fluorescent proteins as expression markers and protein fusion partners has provedimmensely valuable for resolving the organization of biological events in living cells. EGFP and DsRed2 arecommonly fluorescent marker protein which is used for biotechnology and cell biology research. The presentstudy was designed to identify the expression vector that suitable to ligate with DNA encoding HBV coreprotein for intracellular localization study in hepatocyte cell, which were expressed as fusion proteins. We alsocompared and quantified the expressed fluorescent protein which predominantly localized in the cellcompartment. The results indicated that DsRed2 shown as less than ideal for intracellular localization study ofthan EGFP, because of its tetrameric structure of the fluorescent protein and when fused to a protein of interest,the fusion protein often forms aggregates in the living cells. In contrast, EGFP fluorescent protein shown a muchhigher proportion of cytoplasmic localization, thus being more suitable for analysis of intracellular localizationthan DsRed2 fluorescent protein. EGFP fluorescent protein is also capable to produce a strong green fluorescencewhen excited by blue light, without any exogenously added substrate or cofactor, events inside living cell canthus be visualized in a non-invasive way. Based on our present quantitative data and some reasons above shownthat EGFP is more suitable than DsRed2 as a fluorescent marker protein for intracellular localization study intoHuH-7 cell.Keywords: EGFP, DsRed2 fluorescent protein , HuH-7 cell, HBV, intracellular localization
Effect of Nuclear Export Inhibitor Leptomycin B on the Intracellular Localization of HBV Core Protein into Hepatocytes Cell Line Huh-7 and HepG2 Cells Haryanto, Aris; Wijayanti, Nastiti; Kann, Michael
Indonesian Journal of Biotechnology Vol 15, No 2 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

Leptomycin B (LMB) was originally discovered as a potent anti-fungal antibiotic from Streptomyces species. The cellular target of LMB has been identified as the nuclear export receptor CRM-1 or exportin-1, which is involved in nuclear trafficking of cellular RNAs or proteins containing the nuclear export sequence (NES). CRM-1 is the main mediator of nuclear export in many cell types including hepatocyte cell lines. The ability of LMB to inhibit nuclear export has made it a useful tool in the study of the intracellular localization of manyregulatory proteins. In this study, we evaluated the effect of nuclear export inhibitor LMB treatment on the intracellular localization of HBV core protein into the hepatocyte cell lines, Huh-7 and HepG2 cells. We also reported the quantification of the distribution of EGFP-Core fusion protein with redundant core NLS as well as SV-40 NLS into cell compartments. Results shown that in Huh-7 cells treatment of LMB caused retention of EGFP-Core fusion protein into the nucleus, so increased the nuclear localization of EGFP-Core and all variants.In HepG2 cells, although not significantly, treatment of LMB increased a number of nuclear localization in all EGFP-Core constructions, even the nuclear localization in HepG2 cells is not so high as in Huh-7 cells. Keywords: Leptomycin B, HBV, core protein, intracellular localization, NLS, Huh-7, HepG2 cell
Effect of the HBV Capsid Assembly Inhibitor Bayer 41-4109 on the Intracellular Localization of EGFP-Core Fusion Proteins Haryanto, Aris; Wijayanti, Nastiti; Kann, Michael
Indonesian Journal of Biotechnology Vol 12, No 2 (2007)
Publisher : Universitas Gadjah Mada

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Abstract

Bayer 41-4109 is heteroarylpyrimidine (HAP) which has been identified as potent of HBV capsid assemblyinhibitor. The present study was to study effect of Bayer 41-4109 treatment on the intracellular localization ofEGFP-Core fusion proteins into HepG2 cells. Three recombinant plasmids of pEGFP-Core with single, double andtriple NLS of HBV core (EGFP-Core 1C, 2C and 3C ) and two recombinant plasmids with single and triple NLS ofSV-40 (EGFP-Core 1 and 3 SV-40) were used in this work. After transient transfected into HepG2 cells and treatedwith Bayer 41-4109, the intracellular localization of expressed fusion proteins from all plasmid constructions weredetermined and quantified under confocal laser microscope. Results shown that Bayer 41-4109 treatment in HepG2cells inhibited the nuclear localization of EGFP-Core with single of triple HBV core NLS. As well as the constructionsof expressed fusion protein with single and triple SV-40 NLS (EGFP-Core 1 and 3 SV-40 NLS) showeddecreasing the nuclear localization after treated with Bayer 41-4109, even not as strong as EGFP-Core 1C and 3CNLS. Bayer 41-4109 has been identified as a potent inhibitors of HBV replication which has multiple effects on HBVcapsid assembly. It may inhibit virus replication by inducing assembly inappropriately and by misdirectingassembly decreasing the stability of normal capsids.Keywords: HBV capsid, Bayer 41-4109, EGFP-Core fusion protein, HepG2 cell
Molecular Genotyping of HBV by using Nested PCR-RFLP among Hepatitis B Patients in Daerah Istimewa Yogyakarta Province and Surrounding Area Haryanto, Aris; Mulyani, Nenny Sri; Widowati, Titis; Wijayanti, Nastiti; Hadi, Purnomo
Indonesian Journal of Biotechnology Vol 13, No 2 (2008)
Publisher : Universitas Gadjah Mada

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Abstract

Hepatitis B virus (HBV) can be classified into 8 genotypes, genotype A to H. Genotype of HBV is important for clinical and etiological investigations. Research for HBV genotyping, HBV transmission study using nested PCR and HBV genotyping based on RFLP using restriction enzymes have been reported. However, both of those methods have been not applied for HBV genotyping study among hepatitis B patients in endemic area, like Indonesia yet. Molecular genotyping of HBV will describe epidemiology, pathogenesis and clinical implication of HBV. Combination of nested PCR and RFLP (nested PCR-RFLP) method to determine HBV genotype in Indonesia is still less information. The objectives of study were to develop a system for HBV genotyping by nested PCR combined with RFLP (nested PCR-RFLP) method based on nucleotide sequence of surface protein encoding</div><div>gene (S gene) in HBV genome and to confirm HBV genotypes which predominantly found among hepatitis B patients in Daerah Istimewa Yogyakarta Province and surrounding area. Total of 149 sera from chronic hepatitis B patients from Daerah Istimewa Yogyakarta and surrounding areas were collected for in this work. Viral DNA were extracted from sera of hepatitis B patients and used as template for first round nested PCR amplification using outer primers set. Amplicons of first round PCR were used as template for second round amplification using inner primers set. Then, amplicons of second round nested PCR were restriction digested by Sty I and Bsr I enzymes. For HBV genotyping then the restriction products were analyzed by RFLP based on restriction pattern. Results showed that the first round nested PCR amplification generated DNA fragments of whole S gene in length 1.233 bp, and in second round nested PCR amplification using inner primer set generated DNA fragments 585 bp in length. Genotype analysis for all samples using nested PCR-RFLP methods by restriction digested of Sty I and Bsr I enzymes found only 2 HBV genotypes among hepatitis B patients, namely genotype B and C. Quantification</div><div>data showed that most of hepatitis B patients found infected by HBV genotype B (92,8%), genotype C (3,6%) and unidentified genotype (3,6%). Nested PCR-RFLP methods for HBV genotyping is simple and inexpensive for clinical diagnostic in large scale.
Effect of Staurosporine on the Intracellular Localization of Hepatitis B Virus Core Protein Haryanto, Aris; Wijayanti, Nastiti; Kann, Michael
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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Abstract

protein is also including in the HBV genome targeting into the nucleus through modulating carboxyl residues byphosphorylation. Nuclear localication Signal (NLS) in HBV core protein is inside the virion structure and it must beunmasked in order to function, perhaps by phosphorylation. Phosphorylation of of HBV core protein in turn couldbegin to alter capsid conformation. Staurosporine is a natural product originally isolated from bacteriumStreptomyces staurosporeus. Staurosporine was discovered to have biological activities ranging from anti-fungal toanti-hypertensive. The interest in these activities resulted in a large investigative effort in chemistry and biology andthe discovery of the potential for anti-cancer treatment. The main biological activity of Staurosporine is the inhibitionof protein kinases through the prevention of ATP binding to the kinase. In the present study, we have studied theintracellular localization of EGFP-Core fusion protein with triple HBV core and SV-40 nuclear localization signal atits carboxyl terminal in presence and absence of Staurosporine. We also to study the effect of Staurosporine treatmenton the intracellular localization of EGFP-Core fusion protein in the hepatocyte cells line of HepG2 cell. Resultsshowed that effect of Staurosporine is prevent the nuclear localization of EGFP-Core fusion protein into nucleusthrough an inhibition of the phosphorylation of core protein. Stauroporine also prevents cell division so that passivetrapping of core protein is inhibited.
Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR Wijayanti, Nastiti; Wibawa, Tri; Nirwati, Hera; Haryanto, Aris; S, Sutaryo1
Indonesian Journal of Biotechnology Vol 11, No 2 (2006)
Publisher : Universitas Gadjah Mada

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Abstract

world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detectingdengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primersthat conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirusand there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virususing multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assaycan be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiologyand also evolutionary studies.Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR
Expression Analysis and Nuclear Import Study of Full-length Isoforms Importin α as 6x Histidin-tagged Fusion Protein on the Intracellular Localization of Recombinant HBV Core Protein Haryanto, Aris
Indonesian Journal of Biotechnology Vol 10, No 1 (2005)
Publisher : Universitas Gadjah Mada

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Abstract

Isoform importin α molecules play a central role in the classical nuclear import pathway, that occurs throughthe nuclear pore complex (NPC) and typically requires a specific nuclear localization signal (NLS). In this study,it was investigated the role of isoforms importin α in the nuclear import of wild type recombinant hepatitis B viruscore protein (WT rHBc), phosphorylated recombinant HBV core (rHBc) and recombinant HBV core without NLSby co-immunoprecipitation. Four recombinant full-length isoforms importin α as 6x histidin-tagged fusion proteinwere expressed and analysed from expression plasmid vectors Rch1, pHM 1969, pHM 1967 and pHM 1965. Theresults indicated that importin α-1, importin α-3, importin α-4 and importin α-5 can be expressed and isolatedfrom E. coli transformed recombinant DNA plasmid as protein in size around 58-60 kDa. By the nuclear transportstudy shown that isoforms importin α are involved in the nuclear import of WT rHBc, phosphorylated rHBc andrHBc without NLS. It also indicated that they have an important role for nuclear transport of from cytoplasm intothe nucleus.Keywords: NPC, NLS, importin α, importin β, isoforms importin α as 6x histidin-tagged fusion protein, WTrHBc, SV40 Tag, co-immunoprecipitation, westernblotting.
Biokimia Haryanto, Aris
Jurnal Sain Veteriner Vol 34, No 1 (2016): Juni
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.22204

Abstract

Biokimia
Molecular Fish Sexing on Taisho Sanshoku Koi (Cyprinus carpio) Based on ArS.9-15 Gene Amplification using PCR Method Novindasari, Balqis Bahiya Milan; Nurrahmi, Isti Ananda; Andrian, Krisna Noli; Haryanto, Aris
Jurnal Medik Veteriner Vol. 7 No. 2 (2024): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jmv.vol7.iss2.2024.255-263

Abstract

Taisho Sanshoku is a variant of Koi fish (Cyprinus carpio) that has high demand due to its high economic value and relatively expensive price. This study aimed to determine the sex of the Taisho Sanshoku Koi fish by molecular sexing using the PCR method to amplify the ArS.9-15 gene. This study was initiated by rearing a 4–6 month-old of 10 Taisho Sanshoku Koi fish in a fish tank with a filter and oxygen aeration. The fish were fed with fish pellets for 1–3 days. The Koi fishes were then anesthetized using Koi anesthesia containing β-hydroxyethyl phenyl ether. Each fish's peripheral blood was collected as much as 0.5 mL per fish and then stored in tubes containing Ca-EDTA anticoagulant. The genomic DNA was extracted from peripheral blood samples and used as a template DNA for PCR amplification targeting ArS.9-15 gene. Agarose with 1.5% concentration and CybrSafe staining was used in electrophoresis for visualization of the PCR results then visualized in a dark chamber using a UV transilluminator. The Taisho Sanshoku Koi fish's sex was determined using descriptive analysis based on the electrophoresis results. According to the PCR results, the female Taisho Sanshoku Koi fish only produced one 800 bp DNA band, whereas the male fish produced two 800 bp and 1,100 bp DNA bands. The outcome of molecular fish sexing of the 10 Taisho Sanshoku Koi fish reported that 60% were male and 40% were female.
Analisis Dinamika Kendaraan Mobil Antawirya Pada Saat Cornering Haryanto, Aris; Setiyana, Budi; Kurdi, Ojo
JURNAL TEKNIK MESIN Vol 12, No 1 (2024): VOLUME 12, NOMOR 1, JANUARI 2024
Publisher : Jurusan Teknik Mesin, Fakultas Teknik, Universitas Diponegoro

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Abstract

Mobil saat ini menjadi alat transportasi yang sangat dibutuhkan masyarakat umum. Produsen mobil saat ini yang terus mengembangkan performa mesin dan desain agar selalu dalam kondisi aman dan nyaman pada saat dikendarai terutama didalam ruang kemudi agar minim terjadi kecelakaan. Salah satu faktor yang mempengaruhi terjadinya kecelakaan yaitu mengalami slip condition pada saat kendaraan melakukan cornering. Pentingnya faktor keamanan dan keselamatan kendaraan mobil. Multibody Dynamics (MBD) simulation merupakan salah satu metode untuk menguji kinematika dan dinamika dari rancangan. Pada penelitian ini dilakukan pengujian MBD dengan variasi radius putar dan kecepatan yaitu radius 10 m, 15 m, dan 20 m sedangkan variasi kecepatan 20 km/h, 30 km/h, 40km/h. Model kendaraan yang dijadikan referensi pada penelitian ini adalah mobil Antawirya Undip. Pengujian constant speed dan constant radius tertentu mendapatkan kondisi dinamik dari mobil antawirya yang terdiri dari turning radius, steering wheel angle, lateral acceleration, vehicle roll, vehicle yaw rate, vehicle path, vehicle slip. Dari pengujian didapatkan kecepatan aman maksimum dengan radius 10 m yaitu 30 km/h, radius 15 m adalah 30 km/h dan pada radius 20 m yaitu 40 km/h.
Co-Authors Ahmad, Nurrudin Andrian, Krisna Noli Budi Setiyana Charles Rangga Tabbu Dini Wahyu Yudianingtyas Duhita Andinita Fachry Abda El Rahman Hera Nirwati Hevi Wihadmadyatami Kajang, Elphan Augusta Karnati, Srikanth Kayanaveda, Yohanna Khaerudin Khaerudin Medania Purwaningrum Michael Kann Muhammad Isa Nastiti Wijayanti Nenny Sri Mulyani Novindasari, Balqis Bahiya Milan Nurrahmi, Isti Ananda Nurul Ilmi, Nurul Ojo Kurdi Purnomo Hadi Putra, I Nengah Putra, Rizky Dwiandra Rini Widayanti salamah salamah Sawangmake, Chenphop Setya Permana Sutisna Siahaan, Timbul Sri Handayani Irianingsih Sutaryo1 S, Sutaryo1 Sutoyo, Edi Teuku Zahrial Helmi Teuku Zahrial Helmy Titis Widowati, Titis Tri Wibawa Wayan Tunas Artama Widagdo Sri Nugroho