Medania Purwaningrum
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Penerapan Metode Diagnosis Cepat Virus Avian Influenza H5N1 dengan Metode Single Step Multiplex RT-PCR Aris Haryanto; Ratna Ermawati; Medania Purwaningrum; Dini Wahyu Yudianingtyas; Michael Haryadi Wibowo; Charles Rangga Tabbu
Jurnal Veteriner Vol 11 No 4 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (162.68 KB)

Abstract

Avian influenza (AI) virus is a segmented single stranded (ss) RNA virus with negative polarity andbelong to the Orthomyxoviridae family. Diagnose of AI virus can be performed using conventional methodsbut it has low sensitivity and specificity. The objective of the research was to apply rapid, precise, andaccurate diagnostic method for AI virus and also to determine its type and subtype based on the SingleStep Multiplex Reverse Transcriptase-Polymerase Chain Reaction targeting M, H5, and N1 genes. In thismethod M, H5 and NI genes were simultaneously amplified in one PCR tube. The steps of this researchconsist of collecting viral RNAs from 10 different AI samples originated from Maros Disease InvestigationCenter during 2007. DNA Amplification was conducted by Simplex RT-PCR using M primer set. Then, bysingle step multiplex RT-PCR were conducted simultaneously using M, H5 and N1 primers set. The RTPCRproducts were then separated on 1.5% agarose gel, stained by ethidum bromide and visualized underUV transilluminator. Results showed that 8 of 10 RNA virus samples could be amplified by Simplex RTPCRfor M gene which generating a DNA fragment of 276 bp. Amplification using multiplex RT-PCRmethod showed two of 10 samples were AI positive using multiplex RT-PCR, three DNA fragments weregenerated consisting of 276 bp for M gene, 189 bp for H5 gene, and 131 bp for N1. In this study, rapid andeffective diagnosis method for AI virus can be conducted by using simultaneous Single Step Multiplex RTPCR.By this technique type and subtype of AI virus, can also be determined, especially H5N1.
Osteogenic Differentiation Potential of Equine Dental Pulp vs. Periodontal Ligament Stem Cells: A Comparative In Vitro Study Purwaningrum, Medania; Haryanto, Aris; Kayanaveda, Yohanna; Sawangmake, Chenphop
Jurnal Medik Veteriner Vol. 8 No. 2 (2025): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jmv.vol8.iss2.2025.262-272

Abstract

Equine mesenchymal stem cells (MSCs) are promising for bone tissue engineering (BTE) because of their capabilities of differentiating into osteoblasts. Cell therapy using equine MSCs has been introduced. Recently, dental-derived MSCs have gained significant attention due to their capabilities and ease of collection with minimally invasive collection methods. Dental stem cells show high plasticity, accessibility, and applicability for regenerative medicine and are thus considered alternative sources of MSCs. This study evaluated the characterization, osteogenic differentiation potential, and migration assay of equine dental pulp stem cells (eDPSCs) as compared with equine periodontal ligament stem cells (ePDLSCs). Equine dental stem cells from eDPSCs and ePDLSCs (n = 4) were isolated and expanded to passage 3. The morphology, colony-forming capability, cell proliferation assay, stemness and surface markers, trilineage differentiation potential, and migration assay were investigated in vitro. Both eDPSCs and ePDLSCs exhibited a fibroblast-like morphology; showed a colony-forming capability; were able to proliferate based on the results of the cell proliferation assay; expressed stemness and surface markers (NANOG, CD29, CD44, CD90, CD18); maintained the ability to differentiate into osteocytes, chondrocytes, and adipocytes and demonstrated migration capacity based on the migration assay. Surprisingly, ePDLSCs showed significant differences in matrix mineralization, quantification of Alizarin Red staining by cetylpyridinium chloride, and mRNA expression of the osteogenic marker RUNX2. ePDLSCs and eDPSCs may be better alternative MSCs than dental stem cells for the further design of therapeutic regimens for BTE and wound-healing therapy.