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ANALISIS GEN HAEMAGGLUTININ PADA VIRUS CAMPAK LIAR Subangkit, Subangkit; Mursinah, Mursinah; Bela, Budiman; Ibrahim, Fera
Media Penelitian dan Pengembangan Kesehatan Vol 25, No 1 Mar (2015)
Publisher : Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (360.115 KB)

Abstract

AbstrakPenyakit Campak disebabkan oleh virus campak yang termasuk genus Morbilivirus dan Family Paramyxoviridae. Penyakit campak masih menjadi masalah kesehatan karena masih ditemukan Kejadian Luar Biasa (KLB) di Indonesia. Salah satu penyebab terjadinya KLB tersebut diduga sebagaiakibat perbedaan antigenesitas antara strain vaksin yang digunakan dengan strain virus campak liar yang beredar di Indonesia. Penelitian ini bertujuan mendapatkan gambaran tentang karakteristik genetik gen Haemagglutinin virus campak liar yang ada di Indonesia. Spesimen yang digunakan sebanyak 27 isolat virus penyebab KLB dari 17 propinsi selama periode tahun 2003-2010. Isolat virus dilakukan pemeriksaan secara RT-PCR dan sekuensing dengan metode Sanger. Hasil sekuensing dianalisis dengan menggunakan perangkat lunak Bioedit 7.0 dan MEGA 4.0. Hasil penelitian didapatkan perbedaan 10 asam amino antara virus campak strain vaksin CAM-70 dan virus campak liar pada posisi D416N; K424T; V451M; N455T; V466I; I473T; F476L; Y481S atau Y481N; H495N; G505D. Kesimpulan penelitian ini adalah terdapat perbedaan karakteristik genetik antara virus campak liar di Indonesia berbeda dengan strain virus vaksin CAM-70.Kata kunci : Campak, Analisis Molekuler, Hemagglutinin, CD46AbstractMeasles is caused by virus belonging to the genus Morbilivirus and Family Paramyxoviridae. Measles is still a public health problem because outbreak of measles still found in Indonesia. Outbreak is suspected as a result of differences in antigenicity between vaccine strains used with wild-type measles virus strains circulating in Indonesia. This study aims to get genetic characteristics of wild-type measles virus haemagglutinin gene in Indonesia. The specimens were used 27 viral isolates from 17 provinces period 2003-2010. Viral isolates examined by RT-PCR and sequencing with Sanger method. Sequencing analysis were conducted using Bioedit 7.0 and MEGA 4.0 software. The results showed 10 amino acid differences between the vaccine strain measles virus CAM-70 and wild-type measles virus in position D416N; K424T; V451M; N455T; V466I; I473T; F476L; Y481S or Y481N; H495N; G505D. Conclusion: There is a difference between the genetic characteristics of wild-type measles virus in Indonesia and vaccine strain CAM-70.Keywords : Measles, Molecular Analisys, Haemagglutinin, CD46
In vitro transcription of HIV-1 RNA for standard RNA Yasmon, Andi; Bela, Budiman; Ibrahim, Fera; Syahruddin, Elisna
Medical Journal of Indonesia Vol 20, No 3 (2011): August
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (119.131 KB) | DOI: 10.13181/mji.v20i3.447

Abstract

Background: The quantitative assays are important tests in the management of patients with HIV-1/AIDS. The important step in developing the assay is the availability of the standard HIV-1 RNA. For this purpose, we optimized in vitro HIV-1 RNA transcription to produce the standard HIV-1 RNA.Methods: The HIV-1 DNA was amplified from pNL43 by PCR using a primer pair that was specific for conserved region of HIV-1 Gag gene. The PCR product was further cloned into pBluescript II KS. The recombinant plasmid was restricted with EcoRI enzyme. Then, the linearized plasmid was used as template for RNA transcription. RT-PCR and PCR were performed simultaneously for confirmation of synthesized RNA fragment.Results: A 115 bp DNA of HIV-1 Gag gene has been cloned into pBluescript II SK with the exact true orientation. The reaction of the RNA transcription was also successfully performed. The RNA transcripts have been confirmed and showed the accuracy of the transcripts.Conclusion: We successfuly constructed the recombinant plasmid containing a conserved region of HIV-1 Gag gene, and the HIV-1 RNA has been transcribed in vitro as well. (Med J Indones. 2011; 20:185-9)Keywords: HIV-1/AIDS, Quantitative assay, RNA transcription
A second generation of RT-PCR assay for detection of human immunodeficiency virus type 1 (HIV-1) infection Yasmon, Andi; Fatmawati, Ni N.D.; Ibrahim, Fera; Bela, Budiman
Medical Journal of Indonesia Vol 19, No 3 (2010): August
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (120.308 KB) | DOI: 10.13181/mji.v19i3.397

Abstract

Aim A spesific and rapid diagnosis such as RT-PCR assay is the most needed to minimize transmission of HIV-1 infection. Therefore, in this study we developed the RT-PCR assay that was spesific against the gag gene of HIV-1.Methods The developed RT-PCR assay was evaluated against 46 specimens that were obtained from voluntary counseling and testing for HIV (VCT) in Rumah Sakit Umum Pemerintah (RSUP) Sanglah, Bali. To get the sensitivity and specificity of RT-PCR assay, the results of assays were compared with the results of commercially serologic teststhat were commonly used in Indonesia.Results The RT-PCR assay could detect 21 of 26 serologic test-positive specimens and showed 19 negative results of 20 serologic test-negative specimens. There was one specimen that was positive in RT-PCR but negative in serologic assay, which might depict a true yield at particular condition when the serologic assay was unable to detect. Five serologic positive-test specimens were negative by RT-PCR that was possibly caused by low detection level of the assay.Conclusion The RT-PCR assay is potential to be used for the detection of HIV-1 infection with a sensitivity and specificity of 80.8% and 95.0% respectively. (Med J Indones 2010;19:154-7)Key words: AIDS, diagnosis, PoL, sensitivity, specificity
Optimizations of expression and purification of recombinant HIV-1 CRF01_AE p24 protein in Escherichia coli for development of immunodiagnostic assay Simaremare, Ade P.R.; Bela, Budiman; Yasmon, Andi; Ibrahim, Fera
Medical Journal of Indonesia Vol 24, No 1 (2015): March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1028.215 KB) | DOI: 10.13181/mji.v24i1.1166

Abstract

Background: Conventional method for confirmation of HIV infection is Western blot. However, it has limitations because of contamination by human cellular antigen and genetic diversity among the HIV-1 subtypes that show indeterminate result and inaccuracy for the diagnosis of different strains. Most of Western blot developed are based on HIV-1 B subtype. In Indonesia HIV-1 CRF01_AE subtype is dominantly circulated. Therefore, we optimized the expression, purification of the recombinant HIV-1 CRF01_AE p24 protein for development of immunodiagnostic assay.Methods: Optimization of protein expression in Escherichia coli strain BL21CP was performed including induction time, isopropyl-1-thio-d-galactopyranoside (IPTG) and immidazole consentrations, and induction temperature. Purification of the recombinant p24 protein was used by using Ni-NTA (Qiagen) purification system in native condition. Results: Expression and purification of HIV-1 CRF01_AE p24 protein have been performed. Confirmation of the recombinant protein by Western blot showed the expression and purification of recombinant p24 protein has been optimized well and reactive with sera of patients with HIV-1 CRF01_AE subtype positive.Conclusion: The recombinant HIV-1 CRF01_AE p24 protein has been expressed and purified successfully, and it is potential to be used as antigen for immunodiagnostic assay.
Simultaneous detection of Legionella species and Legionella pneumophila by duplex PCR (dPCR) assay in cooling tower water samples from Jakarta, Indonesia Yasmon, Andi; Yusmaniar, Yusmaniar; Anis, Anis; Bela, Budiman
Medical Journal of Indonesia Vol 19, No 4 (2010): November
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (158.177 KB) | DOI: 10.13181/mji.v19i4.408

Abstract

Aim: Since culture method is time-consuming and has low  sensitivity, we developed a duplex PCR (dPCR) assay for the detection of Legionella sp. and L. pneumophila in cooling tower samples. We used culture method as a gold standard.Methods: Optimization of dPCR method was performed to obtain an assay with high sensitivity and specifi city. The optimized method was used to detect Legionella sp. dan L. pneumophila in 9 samples obtained from 9 buildings in Jakarta. For culture method, the bacteria were grown or isolated on selective growth factor supplemented-buffered charcoal yeast extract (BCYE) media.Results: Of 9 samples tested by dPCR assay, 6 were positive for Legionella species,1 was positive for L. pneumophila, and 2 showed negative results. For the same samples, no Legionella sp. was detected by the culture method.Conclusion: dPCR assay was much more sensitive than the culture method and was potentially used as a rapid, specifi c and sensitive test for routine detection of Legionella sp. dan for L. pneumophila in water samples. (Med J Indones 2010; 19:223-7)Keywords: BCYE media, mip gene, 16S-rRNA gene
Teknik reverse transcription – polymerase chain reaction (RT-PCR) dan hibridisasi dot blot dengan pelacak DNA untuk deteksi human immunodeficiency virus (HIV) dalam serum darah Rosilawati, Maria Lina; Bela, Budiman
Universa Medicina Vol 26, No 3 (2007)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2007.v26.111-119

Abstract

LATAR BELAKANGTeknik biologi molekuler seperti teknik reverse transcription – polymerase chain reaction (RT-PCR) dot blot hybridization dengan pelacak DNA berlabel biotin dapat mendeteksi human immunodeficiency virus (HIV) dalam serum darah. Teknik ini selanjutnya dapat diterapkan untuk skrining HIV donor jaringan biologi terutama dari Bank Jaringan Riset Badan Tenaga Nuklir Nasional (BATAN), seperti amnion, allograft steril radiasi, melalui darahnya. Penelitian ini bertujuan untuk membandingkan sensitivitas metode RT-PCR hibridisasi dot blot dengan pelacak DNA berlabel biotin dan RT-PCR elektroforesis gel agarosa untuk deteksi HIV.METODEPenelitian ini menggunakan serum darah dari Rumah Sakit Ketergantungan Obat (RSKO) Fatmawati. Jumlah serum yang dipakai sebanyak 55 sampel terdiri dari 5 sampel negatif HIV hasil uji serologi dengan rapid test dan 50 sampel dengan enzyme linked immunoassay (ELISA). Ekstraksi RNA HIV sampel darah dilaksanakan menggunakan kit RNA viral extraction sedangkan teknik one step RT-PCR digunakan untuk amplifikasi DNA. HASILHasil penelitian menunjukkan pada 55 sampel yang diuji baik dengan teknik RT-PCR elektroforesis gel agarosa maupun RT-PCR hibridisasi dot blot, 43 sampel positif mengandung HIV. Hasil RT-PCR hibridisasi dot blot jauh lebih jelas dibanding dengan RT-PCR-elektroforesis gel agarosa. Hal ini terlihat munculnya dot hitam tebal pada film sedangkan pada gel agarosa pita DNA tampak tipis untuk beberapa sampel positif HIV yang sama KESIMPULANTeknik RT-PCR hibridsasi dot blot dengan pelacak DNA berlabel biotin lebih sensitif dibanding dengan RT-PCR elektroforesis gel agarosa untuk mendeteksi HIV.
Antiproliferative activity and caspase enhancement properties of Annona muricata leaves extract against colorectal cancer cells Indrawati, Lili; Ascobat, Purwantyastuti; Bela, Budiman; Abdullah, Murdani; Surono, Ingrid S.; Pramono, Suwijiyo
Medical Journal of Indonesia Vol 25, No 3 (2016): September
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (498.684 KB) | DOI: 10.13181/mji.v25i3.1449

Abstract

Background: The prevalence of colorectal cancer is rising in Asia including Indonesia. Annona muricata tea leaves, that is traditionally used for maintaining health, and lately being used by cancer patients. The objectives of this study is to investigate its effects in human colorectal cancer cell in vitro and ex vivo.Methods: Thirty patients with colorectal cancer (CRC) were enrolled in a randomized double-blind placebo-controlled trial. They were equally divided into two groups: those treated with 300 mg A. muricata leaf extract and placebo daily for 8 weeks. Serum from supplemented CRC patients of both groups was compared for caspase 9 and caspase 8 enhancement activity. Antiproliferative effect of water extract of A. muricata leaves and its fractions were evaluated against colorectal cancer cell line (DLD-1 and COLO 205) compared with 5-fluorouracil and placebo, the dose range was 62.5-2,000 µg/mL. Method used was 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data were analyzed by Mann-Whitney U test. The p value was set at 0.05.Results: Ethanol-soluble fraction of A. muricata leaves extract water extract (ESFAM) leaves extract had cytotoxicity effects on DLD-1 as well as COLO 205 cell line, as shown by the lower IC50 compared to 5-fluorouracil and placebo, 20.59 μg/mL and 654.9μg/mL, respectively. Serum of subjects supplemented with extract significantly induced caspase 9 (p=0.001) activity of DLD-1 colorectal cancer cell line, but not for caspase 8 activity (p=0.372).Conclusion: The study's results suggest the cytotoxicity potential of  A. muricata  leaves extract  in in vitro and ex vivo studies.
DETEKSI MUTASI GEN KATG MYCOBACTERIUM TUBERCULOSIS DENGAN METODE PCR (POLYMERASE CHAIN REACTION) - HIBRIDISASI DOT BLOT MENGGUNAKAN PELACAK OLIGONUKLEOTIDA BERTANDA 32 Maria Lina R.; Budiman Bela; Andi Yasmon
Jurnal Ilmiah Aplikasi Isotop dan Radiasi Vol 5, No 1 (2009): Juni 2009
Publisher : BATAN

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17146/jair.2009.5.1.525

Abstract

Penanganan dan pengendalian penyakit tuberkulosis (TB), penyakit yang disebabkan kuman M. tuberculosis, menjadi semakin sulit dengan meningkatnya kasus resistensi kuman penyebab terhadap obat anti tuberkulosis (oat) sepertiisoniazid. Resistensi dapat terjadi karena penggunaan obat yang tidak tepat dan tidak teratur, sehingga menimbulkan mutasi pada gen yang mengkode/menyandi target oatseperti gen katG (katalase peroxidase G) untuk isoniazid. Teknik biologi molekuler seperti PCR dilanjutkan dengan hibridisasi dot blot dengan pelacak oligonukleotida berlabel radioisotop merupakan teknik deteksi cepat adanya mutasi pada gen tersebut. Percobaan ini bertujuan menerapkan teknik PCR-hibridisasi dot blot dengan pelacak oligonukleotida bertanda radioisotop (32P) untuk mendeteksi adanya mutasigen katG pada kodon 315 yang menyebabkan M. tuberculosis resisten terhadap isoniazid. Dalam penelitian ini digunakan 89 contoh sputum dan kuman standar M. tuberculosis H37Rv. Ekstraksi DNA dilakukan dengan menggunakan metode BOOMuntuk contoh sputum dan metode fenol kloroform untuk kuman standar. Primer oligonukleotida yang dipakai untuk proses PCR adalah Pt8 & Pt9 untuk mendeteksi Mycobacterium penyebab tuberkulosis dan RTB 59 & RTB36 untuk mendeteksi keberadaan gen katG, yang masing-masing dirancang dari daerah pada sekuens sisipan IS6110 dan gen katG M. tuberculosis. Hasil PCR dideteksi dengan teknik elektroforesis gel agarosa. Metode hibridisasi dot blot dengan pelacak oligonukleotida 315 mu bertanda radioisotop 32P, dilaksanakan untuk mengetahui adanya mutasi gen katG dari hasil PCR dengan primer RTB59 & RTB36. Proses PCR dengan primer Pt8dan Pt9 menunjukkan hasil positif untuk semua contoh sputum yang menyatakan sputum mengandung Mycobacterium penyebab tuberkulosis. Hasil proses hibridisasi dot blot dengan pelacak oligonukleotida 315 mu bertanda 32P, memperlihatkan, 11 strain dari 89 strain M. tuberculosis pada contoh sputum, mengalami mutasi pada kodon 315 dari gen katG. Berdasarkan hasil tersebut, dapat dinyatakan bahwa teknik PCR-hibridisasi dot blot dengan pelacak oligonukleotida bertanda 32P adalah metode yang cepat, spesifik, dan sensitif untuk mendeteksi adanya mutasi gen katG M. tuberculosis yang berkaitan dengan resistensinya terhadap isoniazid.
Five Unique Amino Acid Residues of Hemagglutinin (HA) Proteins of Swine Influenza A (H1N1) Detected in 2009 in Jakarta, Indonesia ANDI YASMON; YULIANTY MUHAYAR; VIVI SETIAWATY; BETI ERNAWATI DEWI; BUDIMAN BELA; FERA IBRAHIM
Microbiology Indonesia Vol. 6 No. 2 (2012): June 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (669.535 KB) | DOI: 10.5454/mi.6.2.3

Abstract

Nine HA genes of influenza A (H1N1) viruses originating from swine which were detected in 2009 in Jakarta, Indonesia, were characterized in this study. Nasopharyngeal and/or pharyngeal samples were extracted to obtain viral RNA genomes. Amplification of the HA segment was performed by using the reverse transcription-polymerase chain reaction (RT-PCR), and followed by nested PCR in cases of RT-PCR negative. DNA sequencing was performed by using eight overlapping primers. All the Jakarta strains were closely related to vaccine strain A/California/07/2009. Nine amino acid changes were found in the Jakarta strains, and 5 (P100S, S220T, G239D, R240Q, and I338V) of those were unique to all Jakarta strains with respect to strain A/California/07/2009 used to produce vaccine. An I338V substitution was detected in a cleavage site of HA and no amino acid changes were detected in potential sites for N-linked glycosylation. For seven sites (positions 131, 158, 160, 183, 187, 222, and 227) playing an important role in viral attachment to host receptor, none of the expected amino acid changes was detected; however, a S220T substitution close to amino acid 222 was found in all the Jakarta strains. All amino acid changes potentially affect the pathogenicity of the viruses and the efficacy of strain A/California/07/2009 in neutralizing the Jakarta strains.
Konstruksi Plasmid Pengekspresi Antigen Rekombinan HCV Berbasis Multiepitop untuk Deteksi Antibodi Anti-HCV Dian Amirulloh; Silvia Tri Widyaningtyas; Budiman Bela
Media Penelitian dan Pengembangan Kesehatan Vol 28 No 3 (2018)
Publisher : Sekretariat Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/mpk.v28i3.39

Abstract

AbstractHepatitis C virus (HCV) infection can cause chronic liver disease that develops into cirrhosis and liver cancer. It is estimated that are more than 170 million of th world’s population suffering from HCV. Accurate diagnosis is needed to provide appropriate early treatmen, including preventing further transmission of the virus. The purpose of this study was to construct plasmid expression of recombinant antigen for detection of anti-HCV antibodies. The antigen coding gene is designed so that is composed to epitopes that are immunodominant, sustainable and and represent HCV subtypes circulating in Indonesia and globally. Furthermore, the gene was made by synthetic DNA techniques by DNA synthesis service providers and accepted by the researchers in the form of blinding on the PUC57 plasmid to pQE80L plasmid with BamHI and HindIII cloning sites. Subcloned recombinant plasmids were then propagated on Top10 Escherichia coli cells and verified by PCR colony tecnique, restriction, and sequencing analysis. HCV recombinant antigen coding gene is 1200 bp. Cloning of these gene on the PUC57 vector produced a plasmid pUC57-HCV_ME (3910 bp) and subcloned in the pQE80L vector producing pQE80L-HCV_ME plasmid (5909bp). Based on verification results of pQE80L-HCV_ME plasmid the expression of recombinant antigen for detection of anti-HCV antibodies has been successfully constructed. AbstrakInfeksi hepatitis C virus (HCV) dapat menyebabkan penyakit hati kronis yang berkembang menjadi sirosis dan kanker hati. Diperkirakan terdapat lebih dari 170 juta penduduk dunia menderita HCV. Diagnosis yang akurat diperlukan untuk memberikan penanganan tepat secara dini, termasuk mencegah penularan virus tersebut lebih lanjut. Tujuan penelitian ini adalah mengonstruksi plasmid pengekspresi antigen rekombinan untuk deteksi antibodi anti-HCV. Gen pengode antigen tersebut dirancang sedemikian rupa sehingga tersusun atas epitop yang bersifat imunodominan, lestari, serta mewakili subtipe HCV yang bersirkulasi di Indonesia maupun global. Selanjutnya gen tersebut dibuat dengan teknik DNA sintetik oleh perusahaan penyedia jasa sintesis DNA dan diterima oleh peneliti dalam bentuk terklona pada plasmid pUC57. Untuk ekspresi pada sel Escherichia coli, gen penyandi antigen rekombinan disubklona dari plasmid pUC57 ke plasmid pQE80L dengan situs pengklonaan BamHI dan HindIII. Plasmid rekombinan hasil subklona kemudian dipropagasi pada sel Escherichia coli Top10 dan diverifikasi dengan teknik PCR koloni, analisis dengan enzim restriksi dan sekuensing. Gen penyandi antigen rekombinan HCV berbasis epitop multipel (HCV_ME) berukuran 1200 pb. Pengklonaan gen tersebut pada vektor pUC57 menghasilkan plasmid pUC57-HCV_ME (3910 pb) dan subklona pada vektor pQE80L menghasilkan plasmid pQE80L-HCV_ME (5909 pb). Berdasarkan pada hasil verifikasi plasmid pQE80L-HCV_ME pengekspresi antigen rekombinan untuk deteksi antibodi anti-HCV telah berhasil dikonstruksi.
Co-Authors Ade P.R. Simaremare Andi Yasmon Andrijono Andrijono Anggraini Alam Anis Anis Ardiana Kusumaningrum Armi Susandi Aroem Naroeni Aroem Naroeni Ascobat, Purwantyastuti ASRI SULFIANTI Beti Ernawati Dewi Cahyani, Hesti Pramesti Christina, Diana Cintera Rahmagiarti Diah Iskandriati Dian Amirulloh Dimas Seto Prasetyo Dondin Sajuthi Elisna Syahruddin Evy Yunihastuti FERA IBIRAHIM Fera Ibrahim Firdaus Sirait Ganjar Noviar Gede Eko Darmono Geraldine, Vanessa Hanifi, Muhammad Hartiyowidi Yuliawuri Hartono, Soni Herna Herna Herna, Herna Hertanto, Robby Ibnu Agus Ariyanto Iin Maemunah Ingrid S. Surono Jeanne Adiwinata Pawitan Jeanne Elvia Christian Kamila, Etri Dian Ketut Tuti Parwati Lili Indrawati Maria Lina R. Maria Lina Rosilawati Melinda Remelia Muhayya, Syarifah Raisha Murdani Abdullah Mursinah Mursinah Mursinah Mursinah Ni Ken Ritchie Ni Nengah Dwi Fatmawati Nia Kurniati Nuzli Fahdia Mazfufah Pamungkas, Joko Pauline Phoebe Halim Pramita Gayatri Pratiwi, Ekawati Betty Radiana Dhewayani Antarianto SAHLAN, MUHAMAD Sari, Siska Yuliana Satyapertiwi, Dwiantari Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Tri Widyaningtyas Silvia Triwidyaningtyas Sofy Meilany Subangkit Subangkit Subiantistha, Tanaya Sugiarta, Gede Yuda SUWIJIYO PRAMONO Utami, Mardhah Sastri VIVI SETIAWATY Widianingtyas, Silvia Widyaningtyas, Silvia Tri Wiseso, Anggoro Yohda, Masafumi YULIANTY MUHAYAR Yuliar Budi Hartanto Yusmaniar Yusmaniar Yuyun SM Soedarmono Yuyun Soedarmono Zakiudin Munasir