DIAH ISKANDRIATI
Primate Research Center Bogor Agricultural University, Bogor, Indonesia

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Biocompatibility of various hydoxyapatite scaffolds evaluated by proliferation of rat’s bone marrow mesenchymal stem cells: an in vitro study Kamal, Achmad F.; Iskandriati, Diah; Dilogo, Ismail H.; Siregar, Nurjati C.; Hutagalung, Errol U.; Susworo, R.; Yusuf, Achmad A.; Bachtiar, Adang
Medical Journal of Indonesia Vol 22, No 4 (2013): November
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (516.004 KB) | DOI: 10.13181/mji.v22i4.600

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Background: Scaffold (biomaterial) biocompatibility test should be performed in vitro prior to in vivo stem cell application in animal or clinical trial. These test consists of direct and indirect toxicity test (MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]). Those tests were used to identify cell morphological changes, cell-substrate adhesion impairment, and reduction in cell proliferation activity.Methods: The tested scaffolds were hydroxyapatite-calcium sulphate (HA-CaSO4) (scaffold I), nano-particular HA paste (scaffold II), synthetic HA granule (scaffold III), bovine HA granule (scaffold IV), and morsellized bovine xenograft (scaffold V). Direct contact toxicity test and MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] were performed on those groups. In direct contact toxicity test, we put granules of various scaffolds within plates and incubated together with mesenchymal stem cells (MSCs). In MTT assay we included phenol 20 mg/mL and 100 mg/mL group as positive control. Morphology, cell adhesion impairment, and cell growth were monitored daily until day-7. Cells counting in the direct contact toxicity test was conducted on day-7.Results: There were no changes on 24 hours observation after direct contact. On day-7, an impairment of cell adhesion to plastic substrates, changes in cell morphology, and cell death were observed, especially in scaffold I, scaffold II, and scaffold V. In MTT assay, only scaffold I, phenol 20 mg/mL, and phenol 100 mg/mL showed more than 50% inhibition at 24-hour and 7-day-observation. Extracts from scaffold II, III, IV, and V did not affect the viability and proliferation of bone marrow MSCs (inhibition value < 50%). Scaffold II, III, IV and V were proven non-cytotoxic and have good biocompatibility in vitro,  no statistical significant differences were observed among the scaffold groups (p > 0.05).Conclusion: We understand which scaffold was nontoxic or the least toxic to MSCs in vitro. Scaffold IV (bovine HA granule) showed the least toxic effect to rat’s bone marrow MSCs on direct contact test and MTT assay. (Med J Indones. 2013;22:202-8. doi: 10.13181/mji.v22i4.600)Keywords: Biocompatibility test, direct contact test, hydroxyapatite, MTT assay, scaffold
MAMMARY GLAND CELL CULTURE OF MACACA FASCICULARIS AS A RESERVOIR FOR STEM CELLS Mariya, Silmi; Dewi, Fitriya Nur Annisa; Suparto, Irma Herawati; Wilkerson, Gregory K.; Cline, J. Mark; Permanawati, .; Iskandriati, Diah; Budiarsa, I Nengah; Sajuthi, Dondin
HAYATI Journal of Biosciences Vol. 24 No. 3 (2017): July 2017
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1422.555 KB) | DOI: 10.4308/hjb.24.3.136

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The mammary gland contains adult stem cells that are capable of self-renewal and are likely target for neoplastic transformation leading to breast cancer. In this study, we developed a cell culture derived from the mammary glands of cynomolgus monkeys (Macaca fascicularis)(MfMC) and furthermore identified the expression of markers for stemness and estrogenreceptor-associated activities. We found that the primary culture can be successfully subcultured to at least 3 passages, primarily epithelial-like in morphology, the cultured cells remained heterogenous in phenotype as they expressed epithelial cell markers CD24, CK18, and marker for fibroblast S1004A. Importantly, the cell population also consistently expressed the markers of mammary stem cells (ITGB1 or CD29 and ITGA6 or CD49f), mesenchymal stem cells (CD73 and CD105) and pluripotency (NANOG, OCT4, SOX2). In addition to this, the cells were also positive for Estrogen Receptor (ER), and ER-activated marker Trefoil Factor 1, suggesting an estrogen responsiveness of the culture model. These results indicate that our cell culture model is a reliable model for acquiring a population of cells with mammary stem cell properties and that these cultures may also serve as a reservoir from which more purified populations of stem cell populations can be isolated in the future.
ISOLATION AND CHARACTERIZATION OF C-C CHEMOKINE LIGAND 7 (CCL7) IN CYNOMOLGUS MACAQUES Mariya, Sela S.; Dewi, Fitriya N.; Villiandra, Villiandra; Paramastri, Yasmina A.; Iskandriati, Diah; Saepuloh, Uus; Hayes, Eric; Pamungkas, Joko; Sajuthi, Dondin
HAYATI Journal of Biosciences Vol. 26 No. 3 (2019): July 2019
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (261.18 KB) | DOI: 10.4308/hjb.26.3.%x

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Cynomolgus macaques (Macaca fascicularis) are an established animal model of asthma, which exhibit different responses to allergen exposure that are clinically relevant. The chemokine ligand gene (CCL7) encodes Monocyte Chemotactic Protein-3, which has an important role in asthma pathogenesis. While CCL7 polymorphism in humans is associated with asthma phenotype, very little is known about CCL7 in nonhuman primate models of respiratory disease. The objective of this study was to isolate and characterize CCL7 gene in cynomolgus macaques of Indonesian origin. In this study, we used sequencing and bioinformatics technique for gene isolation, characterization, and protein 3D structure prediction. We isolated a 2253 base-pair (bp) sequence of CCL7 in cynomolgus macaques, which exhibited 95% similarity in coding sequence to human CCL7. The amino acid sequence was more closely clustered with human CCL7 than with that of rodents. Importantly, the predictive protein structure of CCL7 was similar to that in humans. These similarities in CCL7 suggests the potential of cynomolgus macaque as a translational model to study asthma, particularly in the context of genetics and role of chemokines such as CCL7.
ANALYSIS OF INTESTINAL MUCOSAL IMMUNOGLOBULIN A IN SPRAGUE DAWLEY RATS SUPPLEMENTED WITH TEMPEH SOKA, SUSAN; SUWANTO, ANTONIUS; RUSMANA, IMAN; SAJUTHI, DONDIN; ISKANDRIATI, DIAH; JESSICA, KATHARINA
HAYATI Journal of Biosciences Vol. 22 No. 1 (2015): January 2015
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1507.883 KB) | DOI: 10.4308/hjb.22.1.48

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Tempeh is a well-known Indonesian fermented food made from soybean. During the fermentation process, microorganisms play an important role in the flavor, texture, and nutritional quality of tempeh. Tempeh has been show to have immuno-modulatory and immune-stimulating properties that may also be caused by the microorganisms in tempeh as they interact between the microbial population in the intestinal tract. The objective of this study was to quantify IgA gene expression at both the transcription and translation levels in Sprague Dawley (SD) rats supplemented with tempeh. A total of 6 female SD rats were divided into 3 groups of 2 rats. The first group was the control and was fed a standard diet without tempeh. The second- and third group were fed with a standard diet supplemented with raw and cooked tempeh, respectively. Ileum tissue samples were collected after tempeh supplementation for 28 days. RNA was extracted from ileum samples, and measurement of IgA gene expression was further analyzed using semi quantitative real-time PCR. The concentration of IgA protein was quantified from ileum lysate using the half sandwich ELISA method. IgA gene expressions in rats supplemented with raw, and with cooked tempeh, were 1.18 and 1.17 fold higher, respectively, compared to the control group. Moreover, IgA protein secretion levels also increased 2.46 and 2.08 fold, respectively, compared to the control group. The result of this study indicates that both raw and cooked tempeh may stimulate IgA secretion, and also that both viable and non-viable microorganisms might stimulate IgA gene expression.
Identifikasi Molekuler Virus Papilloma Genital Pada Dua Spesies Primata di Fasilitas Penangkaran Pusat Studi Satwa Primata-Institut Pertanian Bogor Sari, Isti Kartika; Suparto, Irma H; Iskandriati, Diah
JURNAL BIOLOGI INDONESIA Vol 10, No 1 (2014): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v10i1.339

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Antiproliferative Effect of Electric Fields on Breast Tumor Cells In Vitro and In Vivo Alamsyah, Firman; Ajrina, Izzatun Niswah; Dewi, Fitriya Nur Annisa; Iskandriati, Diah; Prabandari, Silvia Arin; Taruno, Warsito P.
Indonesian Journal of Cancer Chemoprevention Vol 6, No 3 (2015)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev6iss3pp71-77

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Our research focused on the antiproliferative effect of low intensity (18 Vpp) and intermediate frequency (100 KHz) electrostatic wave between two capacitive electrodes on breast tumor cells in vitro and in vivo. In vitro study has been conducted by using MCF-7 cell lines treated with external electrostatic for 24, 48, and 72 hours of treatment and the cells number were calculated during treatment by using hemocytometer and presented as Growth Inhibition (GI)% efficacy. For in vivo, we used female mice (Mus musculus) strain C3H as animal model. The mice were injected with either MCF-7 cells, mammary tumor cells from C3H donor, or NaCl 0.9% (placebo) subcutaneously into the axilla area and exposed by external electrostatic in each cage for 12 hours in 2 weeks before necropsied. The adjacent and breast tissue were collected and stained with Hematoxylin – Eosin then analyzed for histopathological profile. In vitro study revealed the number of exposed cells decreased with lower proliferation rate than the non-exposed cells. Moreover, the external electrostatic caused 28-39% growth inhibition efficacy of MCF-7 cells. After 2 weeks of exposure, placebo mice were physically normal, whereas the tumor undergone significant shrinkage of more than 67% in size. Histopathological analysis of the mammary glands indicated infiltration of macrophages into the tumor area through the blood vessel. No abnormality was found in the skin layer and mammary glands of the breast tissue of placebo mice. Here, we present new knowledge of electro-capacitive cancer therapy (ECCT) as a novel treatment modality.Keywords : ECCT, tumor, in vitro, in vivo, breast cancer cells, antiproliferative
Bat Coronavirus of Pteropus alecto from Gorontalo Province, Indonesia Febriani, Wenty Dwi; Saepuloh, Uus; Ayuningsih, Ellis Dwi; Saputra, R. Suryo; Purbatrapsila, Azhari; Nangoy, Meis Jacinta; Ransaleh, Tiltje Andretha; Wahyuni, Indyah; Dako, Safriyanto; Noviana, Rachmitasari; Iskandriati, Diah; Tumbelaka, Ligaya ITA; Pamungkas, Joko
The International Journal of Tropical Veterinary and Biomedical Research Vol 3, No 2 (2018): Vol. 3 (2) November 2018
Publisher : The Faculty of Veterinary Medicine of Syiah Kuala University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (249.276 KB) | DOI: 10.21157/ijtvbr.v3i2.12359

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Bats have been known as natural reservoirs for potential emerging infectious viruses, such as Lyssaviruses, Coronaviruses, Ebola viruses, Nipah virus, and many others. Because of their abudance in population, wide distribution and mobility, bats have a greater risk as source for zoonotic transmission than other animals. Despite the facts of their role as reservoirs for many pathogens, not until an epidemic of Severe Acute Respiratory Coronavirus (SARS-CoV) in 2003 and Middle-East Respiratory Syndrome Coronavirus (MERS-CoV) in 2012, that people pay much attention about coronavirus in bats. SARS-like virus also found in bats with a higher prevalence rate. This study aims to detect the coronavirus of bats in Gorontalo province Indonesia, characterization at the molecular level of the coronavirus genome and determining the level of kinship (through trees filogenetic). This study was conducted as part of bigger PREDICT Indonesia project, in particular to examine coronavirus in bats from Gorontalo province, Indonesia.  As many as  95 rectal swab samples collected from flying foxes (Pteropus alecto) were analyzed in the laboratory using Consensus Polymerase Chain Reaction (PCR) technique to amplify the target sequence from RNA-dependent RNA Polymerase (RdRp) gene with 434 basepair product, resulted 24 samples determined as presumptive positive. Eight out of 24 presumptive positive samples by PCR were analyzed further by nucleotide sequencing and confirmed coronavirus positive. Phylogenetic tree analyses to the eight coronavirus confirmed-sequences were constructed with MEGA-6.0 . The conclusion was 24 out of 95 samples suggested as presumptive positive to Bat CoV. Eight out of 24 samples were analyzed further by nucleotide sequencing and have similarities in the kinship. Three samples had the 98% nucleotide identity to BatCoV from Indonesia and five samples were 85-88% nucleotide identity to BatCoV from Thailand.
EKSPRESI ENZIM REKOMBINAN REVERSE TRANSCRIPTASE (RTRNASE H) SIMIAN BETARETROVIRUS SEROTIPE-2 ASAL MACACA FASCICULARIS INDONESIA DALAM SISTEM EKSPRESI ESCHERICIA COLI Saepuloh, Uus; Iskandriati, Diah; Pamungkas, Joko; Sajuthi, Dondin
Jurnal Ilmu Pertanian Indonesia Vol. 18 No. 1 (2013): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (295.513 KB)

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Reverse transcriptase (RT) enzyme is a key tool in molecular biology for the synthesis of complementary DNA (cDNA) from messenger RNA (mRNA). Combining RT activity with PCR amplification has been a gold standard as the first step in cloning the coding region of any gene of interest. Evidently, RTs have been critical in advancing molecular biology, genetics and medicine to their current stage. In this study, we were developing the RTDRNase H recombinant enzyme isolated from serotype-2 simian betaretrovirus-2 (SRV-2) infected Indonesian Macaca fascicularis using Escherichia coli expression system. The study was conducted using RT SRV-2 gene expression using E. coli expression system, proteins purification, and application to RT PCR technique. The SDS PAGE expression analysis showed a specific band size of 32.7 kDa assumed as RT protein enzyme. Application of RT SRV-2 enzyme generated to the RT PCR technique of ?-globin CDV and SRV-2 env gene target showed that the RT SRV-2 enzyme was capable to reverse transcribed mRNA into cDNA as indicated by the presence of specific DNA band compared with commercial RT enzymes. This RT SRV-2 enzyme showed its activity similar to that of commercial one, although the activity was lower.
IDENTIFIKASI MOLEKULER VIRUS PAPILLOMA GENITAL PADA DUA SPESIES PRIMATA DI FASILITAS PENANGKARAN PUSAT STUDI SATWA PRIMATA-INSTITUT PERTANIAN BOGOR Sari, Isti Kartika; Suparto, Irma H; Iskandriati, Diah
JURNAL BIOLOGI INDONESIA Vol 10, No 1 (2014): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v10i1.339

Abstract

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