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All Journal HAYATI Journal of Biosciences Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Biogenesis: Jurnal Ilmiah Biologi Indonesian Journal of Biotechnology Biota Mutiara Medika: Jurnal Kedokteran dan Kesehatan Pharmacon Molecular and Cellular Biomedical Sciences (MCBS) Jurnal Biologi Tropis Jurnal Medik Veteriner Proceedings of the International Conference on Applied Science and Health Global Health Management Journal Biota Jurnal Biomedika JURNAL PERIKANAN TROPIS Narra J Makara Journal of Science Biological Environment and Pollution Rumphius Pattimura Biological Journal Berkala Ilmiah Biologi Kesmas: Jurnal Kesehatan Masyarakat Nasional (National Public Health Journal)
Nastiti Wijayanti
Universitas Gadjah Mada
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Dentistry Earth & Planetary Sciences Education Energy Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Nursing Public Health Veterinary

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Journal : Indonesian Journal of Biotechnology

 1 
Early Detection and Serotyping of Dengue Viruses Clinical Isolates Using Reverse Transcription Polymerase Chain Reaction (RT-PCR) 2 Primers Siregar, Abdul Rahman; Wibawa, Tri; Wijayanti, Nastiti
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

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Abstract

Recently several methods for confirming Dengue Virus have been developed involve virus isolation, detection of virus antigen, and nucleic acid using PCR. It has been reported that rapid detection method for confirming DHF by Multiplex RT-PCR had been successfully developed. It was more effective than the other methods with a high sensitivity and specivicity were 100% at the early phase (day 1-3). This study was designed to develop rapid detection and serotyping methods for Dengue Virus using RT-PCR 2 primers (Dcon and preM) with annealing temperature was 57oC. The whole blood samples were collected from suspected dengue fever patients that had been confirmed with NS1 kit from R.S. Persahabatan DKI Jakarta and R.S. Prof. Dr. Sardjito DI Yogyakarta during Februari-August 2009. The PCR products showed that in 12 samples, 100 % were postitive with different pattern among the serotypes especially for DEN1 and DEN2, but not for DEN3 and Den4.  This method was also able to confirm the double infection DEN2-DEN3, but not for the other ones because of the unspecific pattern. From the results, it indicated that the 2 primers can be a promising early detection and serotyping method of Dengue Virus which infected the DHF patients. Key words: Dengue Virus, DHF, early detection, serotyping, RT-PCR 2 primers.
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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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
Diversity of Nonribosomal Peptide Synthetase Genes in the AnticancerProducing Actinomycetes Isolated from Marine Sediment in Indonesia Camelia Herdini; Shinta Hartanto; Sofia Mubarika; Bambang Hariwiyanto; Nastiti Wijayanti; Akira Hosoyama; Atsushi Yamazoe; Hideaki Nojiri; Jaka Widada
Indonesian Journal of Biotechnology Vol 20, No 1 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (236.873 KB) | DOI: 10.22146/ijbiotech.15266

Abstract

Marine actinomycetes is a group of bacteria that is highly potential in producing novel bioactivecompound. It has unique characteristics and is different from other terrestrial ones. Extreme environmentalcondition is suspected to lead marine actinomycetes produce different types of bioactive compoundfound previously. The aim of this study was to explore the presence and diversity of NRPS genes in 14anticancer-producing actinomycetes isolated from marine sediment in Indonesia. PCR amplificationand restriction fragment analysis of NRPS genes with HaeIII from 14 marine actinomycetes were doneto assess the diversity of NRPS genes. Genome mining of one species of marine actinomycetes (strainGMY01) also was employed towards this goal. The result showed that NRPS gene sequence diversity in 14marine actinomycetes could be divided into 4 groups based on NRPS gene restriction patterns. Analysisof 16S rRNA gene sequences of representatives from each group showed that all isolates belong to genusof Streptomyces. Genome mining result showed that strain GMY01 harboring 10 different NRPS geneclusters that encode secondary metabolites, as pure NRPS or hybrid between NRPS and other compounds.These results indicated that marine actinomycetes having a high potential to be developed as source ofanticancer drugs development.
Limited evidence for white spot syndrome virus susceptibility associated with expression of PmVRP15 in local population of giant tiger shrimp (Penaeus monodon) Aushia Tanzih Al Haq; M. Murwantoko; T. Trijoko; Nastiti Wijayanti; Ch. Retna Handayani; Rarastoeti Pratiwi
Indonesian Journal of Biotechnology Vol 20, No 2 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (912.481 KB) | DOI: 10.22146/ijbiotech.24189

Abstract

White spot syndrome virus (WSSV) is a devastating viral disease in shrimp aquaculture. Infection ofWSSV in penaeid shrimps affects immune defense and changes gene expression. PmVRP15 has been reported as a part of the WSSV propagation pathway that is highly up-regulated in hemocytes at the acute phase of WSSV infection. This study analyzed the expression of PmVRP15 in local populations of giant tiger shrimp (Penaeus monodon) to be associated with susceptibility to WSSV. Tested populations consisted of an inbreeding population (G8) and outbreeding population (G8iA) from Jepara, Indonesia. Susceptibility was determined by cumulative mortality, median lethal time (LT50), and severity of infection at time of death. Though all populations were susceptible to WSSV, the frst mortality in G8 occurred at 18 hours post-infection (hpi) with mild infection, while frst mortality of G8iA occurred at 30 hpi with severe infection. The LT50 of G8 was signifcantly lower than that of G8iA, indicating that G8iA was less susceptible to WSSV than G8. Relative PmVRP15 transcripts of G8iA were insignifcantly down-regulated, whereas relative PmVRP15 transcripts of G8were insignifcantly upregulated. Although it’s still not conclusive, the results of this study suggest that PmVRP15 has weak potentialas a WSSV susceptibility marker in G8 and G8iA broodstock selection.
Evaluation of synthetic‐gene recombinant LipL32 antigen for IgM ELISA detection of Leptospira infection Widiastuti, Dyah; Paramita, Dewi Kartikawati; Murwanti, Retno; Kusrini, Ina; Wijayanti, Nastiti; Haryanto, Aris
Indonesian Journal of Biotechnology Vol 30, No 4 (2025)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.111102

Abstract

Leptospirosis presents with nonspecific clinical features and requires time‐consuming laboratory tests for gold standard diagnosis. This study aims to design and characterize the recombinant LipL32 from synthetic gene and assess its performance as an antigen for detecting leptospirosis. The antigen was developed by cloning the LipL32 gene conserved portion of Leptospira interrogans serovar Icterohaemorrhagiae strain Langkawi. The immunoinformatic was used to characterize the developed rLipL32. Western blot results using anti‐histidine revealed a band of rLipL32 protein at ~40 kDa. Subsequently, it was used to examine the IgM antibody on human sera by using ELISA. The IgM‐LipL32 ELISA was evaluated using 67‐positive and 25‐negative sera and compared with a commercial ELISA. With a cut‐off value of 0.8, it showed 85.7% sensitivity, 83.3% specificity, a 48% positive prediction value (PPV), and 97% negative prediction value (NPV), indicating modest performance compared to existing commercial kits. The rLipL32 is a potential antigen for detecting IgM using ELISA; however, for use in low incidence areas, a confirmation test is crucial.
Co-Authors . Sismindari Abdul Rahman Siregar, Abdul Rahman Abrory Agus Cahya Pramana Adelfiani, Adelfiani Adi Sofyan Ansori, Muhammad Aeniah, Siti Agustiningsih Agustiningsih Akira Hosoyama Allimi, Hayu Swari Amalia, Nur Rofika Ayu Shinta Andi Mahendra Andrian, Krisna Noli Anika Prastyowati, Anika Apriliyani, Tia Ardaning Nuriliani ARI SUSILOWATI Ariesti, Wiwin Arif Luthfi Nurul Huda Aris Haryanto Atsushi Yamazoe Aushia Tanzih Al Haq Bambang Hariwiyanto Bambang Retnoaji BUDI SAKSONO Camelia Herdini Ch. Retna Handayani Dafa, Maula Haqul Dafip, Muchamad Dewi Kartikawati Paramita Diani Mentari Dwi Liliek Kusindarta Dyah Widiastuti Empra, Desi Eka Putri ERLIN LISTIYANINGSIH Fajar Sofyantoro Hakim, Mohamad Saifudin Halim, Shuha Ma’muriyah Hanbal, Mayland Muhammad Hendry Tri Sakti Saragih Hera Nirwati Heriyanto, Didik S. Hevi Wihadmadyatami Hideaki Nojiri Ibnu Agus Ariyanto Illiyin, Sirayya Ina Kusrini Jajah Fachiroh, Jajah JAKA WIDADA Juliadmi, Dian Karlina, Ina Karnati, Srikanth Kartika W. Taroeno-Hariadi Kartika, Aprilia I. Lisna Hidayati Mayani, Olvita Mei Ria Santi Michael Kann Muhammad Adnan Mumtazah, Dzul Fithria Munawir Sazali Murwantoko . Naima, Mirtani Nenny Sri Mulyani Ni Wayan Erly Sintya Dewi Nisa, Mutia Khoirun Nuhamunada, Matin Oktaviana, Shintia Peni Indrayudha Pertiwi, Anggita Endar Pertiwi, Gian Aditya Podhi, Felisitas Moli Pradnya Paramita Pratitis, Visi Endah Primahesa, Alfian Prissandi, Anthera Al Firdaus Priyambada, Fajar Purnomo Hadi Pusparini, Nur Ainun Oktavia Puspitasari, Pinki Anggrahini Putra, Hendy Eka R.C. Hidayat Soesilohadi Rahayu, Sekar Ramadhani, Eka Rarastoeti Pratiwi Retno Murwanti Rinda Binugraheni, Rinda Rohmah, Zuliyati Rumansara, Papuani Samparisna Sadewo, Imran Saeed, Faisal Salamah, Rohmi Saragih, Hendry T. S. S. G. Sari, Ria Vinola Septhya Saribu, Ruth Liananda Citra Dolok Septriani, Nur Indah Septriani, Nur Indah Setiawibawa, Raden Aditya Aryandi Shinta Hartanto Simanungkalit, Eben Ezer Slamet Widiyanto Sofia M. Haryana Sofia Mubarika Haryana Subchan, Aditya Nur Sularsah, Sais Supraitno, Murtiadi Erlan Sutaryo1 S, Sutaryo1 T. Trijoko Tasania, Nadia Nisa Titis Widowati, Titis To'bungan, Nelsiani Tri Rini Nuringtyas Tri Wibawa Triska Desi Sundari Tsabitah, Khansa Wulansari, Riska Yundari, Yundari Zulfa, Amania Zulfadhli Zulfadhli Zumrotus Sholichah Zusrina, Laili Mufli ‘Aliyah, Siti Hamidatul