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All Journal Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Indonesian Journal of Biotechnology BERITA BIOLOGI Proceedings of the International Conference on Applied Science and Health Jurnal Dunia Kesmas BALABA (JURNAL LITBANG PENGENDALIAN PENYAKIT BERSUMBER BINATANG BANJARNEGARA)

Nastiti Wijayanti

Unknown Affiliation

Author-ID : 328644

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Education Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Nursing Public Health Veterinary

Published : 11 Documents Claim Missing Document

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Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR Nastiti Wijayanti; Hera Nirwati; Tri Wibawa; Aris Haryanto; S. Sutaryo
Indonesian Journal of Biotechnology Vol 11, No 2 (2006)
Publisher : Universitas Gadjah Mada

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

Effect of Staurosporine on the Intracellular Localization of Hepatitis B Virus Core Protein Aris Haryanto; Nastiti Wijayanti; Michael Kann
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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.

Effect of the HBV Capsid Assembly Inhibitor Bayer 41-4109 on the Intracellular Localization of EGFP-Core Fusion Proteins Aris Haryanto; Nastiti Wijayanti; Michael Kann
Indonesian Journal of Biotechnology Vol 12, No 2 (2007)
Publisher : Universitas Gadjah Mada

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 Aris Haryanto; Nenny Sri Mulyani; Titis Widowati; Nastiti Wijayanti; Purnomo Hadi
Indonesian Journal of Biotechnology Vol 13, No 2 (2008)
Publisher : Universitas Gadjah Mada

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 Nuclear Export Inhibitor Leptomycin B on the Intracellular Localization of HBV Core Protein into Hepatocytes Cell Line Huh-7 and HepG2 Cells Aris Haryanto; Nastiti Wijayanti; Michael Kann
Indonesian Journal of Biotechnology Vol 15, No 2 (2010)
Publisher : Universitas Gadjah Mada

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

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

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.

Aktivitas Sitotoksik dan Apoptosis Ekstrak Spons Spesies A Anggota Ordo Astroporida terhadap Sel HeLa (Cervical Cancer Cell Line) Ardaning Nuriliani; Ibnu Agus Ariyanto; Mei Ria Santi; Andi Mahendra; Ni Wayan Erly Sintya Dewi; Arif Luthfi Nurul Huda; Nastiti Wijayanti
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 18, No 1 (2013): February 2013
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v18i1.263

Spons merupakan fauna laut yang diketahui memiliki berbagai senyawa bioaktif. Senyawa tersebut berpotensi sebagai antibakteri, antivirus, dan antikanker. Penelitian ini bertujuan mempelajari aktivitas sitotoksik dan apoptosis ekstrak spons spesies A anggota ordo Astrophorida terhadap sel HeLa. Pada penelitian ini pengujian aktivitas sitotoksik ekstrak etanolik, metanolik, dan kloroform spons spesies A terhadap sel HeLa dilakukan menggunakan MTT assay dan uji apoptosis menggunakan double staining, yaitu etidium bromida-acridine orange. Deteksi golongan senyawa yang terkandung di dalam spons spesies A dilakukan menggunakan Kromatografi Lapis Tipis (KLT). Hasil uji sitotoksisitas menunjukkan bahwa ekstrak etanolik, metanolik, dan kloroform spons spesies A masing-masing memiliki nilai IC50 sebesar 18,25; 27,87; dan 13,87 Âµg/mL. Ekstrak etanolik, metanolik, dan kloroform spons spesies A pada konsentrasi 31,25 Âµg/mL dapat menginduksi kematian sel melalui apoptosis masing-masing sebesar 35,3 Â± 11,16%; 82,64 Â± 16,21%; dan 86,76 Â± 9,27%. Berdasarkan uji menggunakan KLT diketahui bahwa spons spesies A menggandung golongan senyawa alkaloid, flavonoid, fenol, dan terpenoid. Oleh karena itu, dapat disimpulkan bahwa ekstrak spons spesies A berpotensi untuk dikembangkan sebagai obat antikanker.Kata kunci: ekstrak spons spesies A, sitotoksik, apoptosis, sel HeLa

PEMERIKSAAN VIRUS DENGUE-3 PADA NYAMUK Aedes aegypti YANG DIINFEKSI SECARA INTRATHORAKAL DENGAN TEKNIK IMUNOSITOKIMIA MENGGUNAKAN ANTIBODI DSSE10 Dyah Widiastuti; Sitti Rahmah Umniyati; Nastiti Wijayanti
BALABA: JURNAL LITBANG PENGENDALIAN PENYAKIT BERSUMBER BINATANG BANJARNEGARA Volume 8 Nomor 1 Juni 2012
Publisher : Balai Penelitian dan Pengembangan Kesehatan Banjarnegara Badan Litbangkes Kemenkes RI

Dengue viruses, globally the most prevalent arboviruses, are transmitted to humans by persistently infected Aedes mosquitoes. The most important vector of Dengue virus is the mosquito Ae.aegypti, which should be the main target of surveillance and control activities. Virologic surveillance for dengue viruses in its vector has been used as an early warning system to predict outbreaks. Detection of Dengue virus antigen in mosquito head squash using immunocytochemical streptavidin biotin peroxidase complex (SBPC) assay is an alternative method for dengue vector surveillance. The study aimed to develope immunocytochemical SBPC assay to detect Dengue virus infection in head squash of Ae.aegypti. The study design was experimental. Artificially-infected adult Ae. aegypti mosquitoes of DENV 3 were used as infectious samples and non-infected adult Ae. aegypti mosquitoes were used as normal ones. The immunocytochemical SBPC assay using monoclonal antibody DSSE10 then was applied in mosquito head squash to detect Dengue virus antigen. The results were analyzed by descriptive analysis. The immunocytochemical SBPC assay can detect Dengue virus antigen in mosquito head squash at day 2 postinfection. There are some false positive results found in immunocytochemical SBPC assay.

POTENTIAL CHEMOPREVENTIVE AGENT: STUDY OF APOPTOSIS IN THE EXTRACTS OF SPONGE-ASSOCIATED FUNGI FROM YOGYAKARTA AGAINST CERVICAL CANCER HeLa CELL LINE Eka Ramadhani; Fajar Priyambada; Abrory Agus Cahya Pramana; Aditya Nur Subchan; Gian Aditya Pertiwi; Raden Aditya Aryandi Setiawibawa; Hendy Eka Putra; Nur Rofika Ayu Shinta Amalia; Nastiti Wijayanti
Proceedings of the International Conference on Applied Science and Health No. 1 (2017)
Publisher : Yayasan Aliansi Cendekiawan Indonesia Thailand (Indonesian Scholars' Alliance)

Show Abstract | Download Original | Original Source | Check in Google Scholar

Background: Cervical cancer is one of the leading-cancers affecting women. Cancer drugs that do not originate from natural ingredient, chemotherapy drugs, have side and resistant effects. Thus study about the natural products treating cancer cells is needed. Secondary metabolites isolated from sponge-associated fungi are expected to have a potency to fight cancer cells. In addition, the production of anticancer compounds from microorganisms has several advantages, including rapid growth and can be manipulated to increase productivity. The isolation and testing cytotoxicity against 3 fungal isolates from Yogyakarta have been done on the previous research. All three isolates have a potential candidate as anticancer drug. Aims: The purpose of this advanced study was studying bioactive compounds induced apoptosis pathway of sponge-associated fungi against cervical cancer HeLa cells. Methods: This study has been carried out for approximately 5 months. The method conducted in this research including the sponge cultivation (covers growth and isolation of secondary metabolites), the mycelium extraction of fungi, the cytotoxicity assay against HeLa cells using MTT Assay and Apoptosis Staining was to see the induction of apoptosis pathway. Results: Based on the research showed that ethyl acetate extract from mycelium is 0.22 grams. The cytotoxicity assay from mycelium extract showed IC50 value of 164 μg/mL against HeLa cell line. Conclusion: The findings is carrying to a possibility to develop the extracts of sponge-associated fungi as candidate of anti-cancer compound. By apoptosis staining, showed the cells coloured green are still alive, and cells undergoing apoptosis have nucleus that appears orange to red. We assuming that the apoptosis was caused by the possibility of peptide compounds that induce apoptosis through the mitochondrial pathway, by increasing the activity of the protein expression of apoptosis, which are Bcl-2 and Bcl-xl.

ANALISIS SEKUEN NUKLEOTIDA E/NS1 GENE JUNCTION VIRUS DENGUE SEROTIPE 2 ASAL DKI JAKARTA, INDONESIA Nurhaida Widiani; Tri Wibawa; Nastiti Wijayanti
JURNAL DUNIA KESMAS Vol 1, No 1 (2012): Volume 1 Nomor 1
Publisher : Fakultas Kesehatan Masyarakat Universitas Malahayati

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33024/jdk.v1i1.314

Demam dengue atau demam berdarah dengue merupakan salah satu masalahkesehatan di daerah tropis dan subtropis. Penyakit ini disebabkan oleh virus dengue(genus flavivirus, famili flaviviridae). Vektor pembawa virus dengue adalah nyamukAedes aegypti dan Aedes albopictus. Genom virus dengue tersusun atas tiga proteinstruktural (protein nukleokapsid, protein envelope, dan protein pre-membran) dan tujuhprotein nonstruktural (NS1, NS2a, NS2b, NS3, NS4a, NS4b, dan NS5). Usaha untukmengontrol penyakit ini tergantung pada pemahaman patogenesis virus dengue. Tetapipengetahuan mengenai patogenesis virus dengue masih belum banyak diketahui karenabelum adanya model yang cocok baik in vitro maupun in vivo. Sehingga dilakukananalisis sekuen nukleotida E/NS1 gene junction virus dengue tipe 2 asal DKI Jakarta,Indonesia untuk mengetahui hubungan filogeni virus dengue yang beredar saat ini.Sekuen nukleotida (240 bp) dari E/NS1 gene junction merupakan segmen yang biasadigunakan untuk analisis perbandingan sekuen. Hasil analisis menunjukkan strain virusyang diteliti berkerabat dengan virus DEN-2 asal Asia yaitu Myanmar, Pakistan, SriLanka, Brunei, dan Malaysia. Tetapi kelima virus ini berada dalam kelompok yangberbeda dengan virus asal Amerika latin (Brazil). Virus Dengue dari Asia diduga lebihvirulen dibandingkan virus dengue dari daerah lain. Sehingga kemungkinan terjadipeningkatan kasus DBD di Indonesia.Kata kunci: Virus Dengue, E/NS1 gene junction, serotype, filogeni, DEN-2

Co-Authors Abdul Rahman Siregar Abrory Agus Cahya Pramana Aditya Nur Subchan Andi Mahendra Arif Luthfi Nurul Huda Aris Haryanto Assyafiya Salwa Dyah Widiastuti Eka Ramadhani Fajar Priyambada Gian Aditya Pertiwi Hendy Eka Putra Hera Nirwati Ibnu Agus Ariyanto Lisna Hidayati Mei Ria Santi Michael Kann Nenny Sri Mulyani Ni Wayan Erly Sintya Dewi Nur Rofika Ayu Shinta Amalia Nurhaida Widiani Purnomo Hadi Raden Aditya Aryandi Setiawibawa S. Sutaryo Sitti Rahmah Umniyati Titis Widowati Tri Rini Nuringtyas Tri Wibawa Yundari, Yundari

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