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"Indonesian National Committee for human immunodeficiency virus (HIV) and acquired immune deficiency syndrome(AIDS) has reported the significant increase of HIV infected individual in Indonesia. A sensitive accurate diagnosticsare urgently needed to prevent the dissemination of HIV and also to provide a suitable therapy. For this reason, we havedeveloped HIV diagnostic method based on PCR to elucidate this problem. For this research, samples were collectedfrom hospitals and Indonesian Red Cross that consist of samples possesing HIV serological test positive andindeterminate. Ribonucleic Acid (RNA) were isolated from blood plasma. These RNA then were amplified afterReverse transcriptase reaction by using primers which have been designed using env (C2V3C3), Long TerminalRepeats (LTR) (U3) and Capsid gag (p24) as target regions. The obtained results shown 26/34 (76.5%) positive in LTR,10/33 (36.4%) positive in Env and 11/33 (33.3%) positive in p24. These results showed that LTR primers detect HIVmore than other primers. It suggests that LTR could be used as detection target as complement of env and p24 Theseresults need to be explored further by using sequencing method."

"Beberapa hasil uji serologi HIV indeterminate pada tes skrining darah ditemukan di Indonesia. Prosedur skrining darahyang dilakukan saat ini sesuai ketentuan yang ditetapkan oleh WHO untuk skrining darah, yaitu 3 tes uji serologi HIVselama pemeriksaan darah. Ketidaksesuaian hasil yang satu dengan yang lain didefinisikan sebagai hasil indeterminate.Penelitian ini bertujuan untuk mengidentifikasi galur-galur HIV yang sulit teridentifikasi dari darah dengan uji serologiHIV intermediate dan mengevaluasi apakah galur HIV yang beredar di Indonesia mempunyai kemungkinan lolos darisistem pendeteksian yang ada. Deteksi RT-PCR dilakukan pada 40 sampel RNA HIV dari donor darah yangmempunyai hasil uji serologi indeterminate dengan sebelumnya melakukan uji konfirmasi dengan menggunakanwestern blot. Deteksi RT-PCR menunjukkan bahwa sebanyak 24/32 (75%) sampel positif LTR, 4/31 (13%) positif poldan 3/5 (60%) positif env. Amplifikasi pada daerah p24, pita-pita yang ditemukan pada sampel selalu lebih rendah dariyang diharapkan. Sekuensing dilakukan untuk mengkonfirmasi hasil amplifikasi menunjukkan bahwa perlu analisislebih lanjut untuk mengetahui apakah perubahan ini yang menyebabkan hasil indeterminate.Indeterminate results ofserological HIV test have been found in Indonesia. The screening procedure is following the prescribed by WHO forscreening of blood donors which is based on 3 different serological HIV test during screening of blood donors.Discordant results are interpreted as indeterminate. This research aims to identify GIV strains that previously difficult todetermine, and to evaluate whether the HIV strains present in Indonesia could pass the existing screening system. RTPCRdetection test of HIV RNA were conducted for 40 blood donors samples with indeterminate serological HIV-testafter a confirmatory test using western blot. Preliminary results showed that 24/32 (75%) of the samples are positiveLTR, 4/31 (12%) positive pol and 1/3 (33%) positive env. Amplification in p24 region showed that bands found havelower size than expected. Sequencing performed to confirm these findings show that further analysis is needed todetermine whether this change is what behind the indeterminate results."

"Penggunaan agen biologi di laboratorium selain memiliki dampak negatif juga memiliki dampak positif. Dampak negatif yang muncul dalam penggunaan agen biologi di laboratorium adalah timbulnya risiko infeksi pada pekerja akibat bahan-bahan biologi berbahaya. Selain itu, dampak negatif lainnya adalah munculnya risiko penyalahgunaan agen biologi oleh pihak-pihak tertentu. Universitas Indonesia yang memiliki tujuan untuk menjadi universitas riset, harus juga memperhatikan hal tersebut. Dampak-dampak negatif yang dapat muncul dari penggunaan bahan biologi tersebut dapat ditanggulangi dengan menerapkan biorisk management yang dapat melindungi pekerja maupun lingkungan sekitar. Tujuan dari penelitian ini adalah untuk mengetahui implementasi dari biorisk management yang terdapat di laboratorium COE IBR-GS FMIPA UI yang dianalisis dengan menggunakan standar WHO, CDC, dan Laboratory Biosafety Guideline yang dikeluarkan pemerintah Kanada. Dalam laboratorium COE IBRGS, sudah terdapat beberapa sistem yang mendukung keselamatan dan kesehatan bagi pekerja. Namun, terdapat beberapa komponen dari biorisk management yang belum dilaksanakan, tidak sesuai, dan harus diperbaiki.

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The use of biological agents in the laboratory in addition to having positive impact also has a a negative impact. Negative impact that arise in the use of biological agents in the laboratory is the emergence of an infection risk to workers due to hazardous biological materials. In addition, other negative impact is the emergence of the risk of misuse of biological agents by certain parties. University of Indonesia, which has a goal to become a research university, must also pay attention to it. Negative impacts that may arise from the use of biological materials can be overcome by applying biorisk management that can protect workers and the environment.

The purpose of this study is to know the implementation of the biorisk management in the laboratory COE IBR-GS FMIPA UI which analyzed using the standard WHO, CDC, and the Laboratory Biosafety Guidelines from Canadian government. In the laboratory COE IBR-GS, there have been some systems that support the safety and health for workers. However, there are several components of the biorisk management have not been implemented, not appropriate, and should be improved."

"ABSTRAKPenggunaan sel punca sebagai anti fibrosis hati cukup menjanjikan. Sel punca CD34 asal darah tali pusat sudah banyak digunakan dalam studi anti fibrosis hati. Penelitian ini menjelaskan efek ko-kultur antara sel stelata hepatik HSC LX-2 dan sel punca CD34 asal darah tali pusat dalam morfologi sel dan ekspresi TGF-?, tenascin-C dan kolagen tipe 1A1. Metode : Sel CD34 diisolasi dari sel darah tali pusat manusia yang dikriopreservasi menggunakan separasi magnet. Sel HSC LX-2 dikultur sebagai kontrol monokultur. Sebagian dipanen dan dihitung untuk dilakukan ko-kultur dengan sel CD34 dalam rasio 1:1. Ko-kultur CD34 dan LX-2 dilakukan dengan metode kultur konvensional 2D dan 3D hanging drop. Hasil monokultur dan ko-kultur dipanen pada hari ke1, 2 dan 3 dan dilakukan pewarnaan imunositokimia tenascin-C ekstraksi RNA untuk analisis kuantitatif dengan real time PCR ekspresi TGF-? dan kolagen tipe 1A1.Hasil : Hasil menunjukkan perbedaan morfologi ko-kultur 2D dan 3D hanging drop dibandingkan kontrol monokultur. Pada ko-kultur 2D terdapat mikromassa, sedangkan pada monokultur 2D tidak ada mikromassa yang terbentuk. Pada ko-kultur 3D hanging drop, terdapat spheroid yang lebih kecil hambatan pembentukan spheroid dibandingkan monokultur 3D hanging drop. Sel CD34 memiliki efek direk terhadap aktivitas sinyal sel stelata hepatik dengan adanya kecenderungan penurunan ekspresi TGF-?. Analisis imunositokimia tenascin-C dalam mikromassa dan spheroid masih perlu dioptimasi. Ko-kultur 2D dan 3D hanging drop method sel punca CD34 asal darah tali pusat dan sel stelata hepatik memiliki efek terhadap penurunan ekspresi kolagen tipe 1A1.Kesimpulan : Sel punca CD34 asal darah tali pusat memiliki efek direk terhadap morfologi sel, inhibisi aktivitas sel stelata hepatik LX-2 yang ditandai dengan penurunan ekspresi TGF-beta dan inhibisi deposisi matriks ekstrasel yang ditandai penurunan ekspresi kolagen tipe 1A1.Kata kunci: sel punca asal darah tali pusat CD34 , sel stelata hepatik, liver fibrosis, TGF-beta, tenascin-C, kolagen 1A1.

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ABSTRACTBackground The development of stem cell therapy antifibrotik placing as one of the promising therapy. Umbilical cord blood CD34 stem cells has been widely used in the study antifibrosis. This study describes the effect of co culture between hepatic stellate cells HSC LX 2 and umbilical cord blood CD34 stem cells on cell morphology and expression of TGF , tenascin C and collagen type 1A1.Method CD34 cells were isolated from thawed cryopreserved human umbilical cord blood cells using magnetic separation. LX 2 cells culture were harvested and counted. CD34 and LX 2 cells were mixed in suspension with 1 1 ratio v v . Cell suspension divided into 2 sets 2D co culture plated in standard well plate and 3D co culture as hanging drops. LX 2 monoculture, CD34 dan LX 2 coculture were harvested on day 1, 2 and 3 as sample for further analysis. Tenascin C expression was analysed by imunocytochemistry techniques. TGF Beta and collagen type 1A1 expression was analysed by qPCR.Result The result showed different morphology between co culture and monoculture on 2D and 3D hanging drop. The 2D co culture showed micromass formation, instead of no micromass formation on monoculture. The 3D hanging drop showed smaller spheroid formation spheroid formation inhibition compared with monoculture. CD34 cells showed direct effect on hepatic stellate cell signalling activity represented by the decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in Collagen type 1A1 expression.Conclusion UCB CD34 cells showed direct effect on cell morphology, inhibition of hepatic stellate cell LX 2 activity represented by a decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in collagen type 1A1 expression. "

"Diferensiasi osteogenik dari Sel punca mesenkim (MSC) menjadi osteoblas memiliki signifikansi klinis yang sangat penting untuk mengobati cedera tulang. Berbagai penelitian telah dilakukan untuk meneliti faktor-faktor yang dapat meningkatkan diferensiasi osteogenik, termasuk pengembangan perancah untuk kultur MSC. Perancah Polivinil Alkohol (PVA)/ Fibroblast-derived Matrix (hFDM) asal manusia menjadi salah satu kandidat perancah yang diduga dapat mendukung diferensiasi osteogenik MSC. Keberadaan matriks ekstraseluler (ECM) pada perancah dapat meregulasi berbagai aktivitas seluler melalui komponen protein matriks yang terdapat pada ECM. Protein matriks berperan sebagai sekuesterasi berbagai faktor pertumbuhan. Faktor pertumbuhan seperti BMP2 dan chordin diketahui dapat meregulasi diferensiasi osteogenik. Proses terjadinya diferensiasi osteogenik dapat diamati melalui akumulasi mineral kalsium yang terdeposit pada matriks ekstraseluler. Tujuan dari penelitian ini adalah untuk mengetahui metode optimum dalam pembuatan perancah PVA/hFDM, danmengetahui peran perancah PVA/hFDM dalam mempengaruhi diferensiasi osteogenik MSC dengan mengukur ekspresi gen BMP2, dan chordin, serta ekspresi kadar kalsium relatif pada matriks ekstraseluler. Optimasi pembuatan perancah PVA hFDM dimulai dengan optimasi medium kultur, waktu kultur, preparasi, dan teknik deselulerisasi. hFDM dikarakterisasi menggunakan pewarnaan Hematoxylin, Masson Trichrome, dan Imunohistokimia untuk mengetahui keberadaan protein matriks. MSC dikultur pada perancah PVA/hFDM untuk uji diferensiasi osteogenik selama 21 hari. Sampel RNA diisolasi pada hari ke-7,14, dan 21. Ekspresi gen BMP2 dan chordin dianalisis menggunakan metode qRT-PCR. Adapun ekspresi kadar kalsium relatif dianalisis dengan uji kualitatif dan kuantitatif pewarnaan Alizarin Red. Hasil penelitian ini menunjukkan protokol pembuatan perancah PVA/hFDM telah dioptimasi, dan karakterisasi hFDM memperlihatkan keberadaan protein matriks berupa kolagen dan biglycan. Ekspresi gen BMP2 menurun pada kelompok MSC yang dikultur pada perancah PVA/hFDM baik di hari ke-7, 14, dan 21. Sedangkan ekspresi gen chordin meningkat pada kelompok MSC yang dikultur pada perancah PVA/hFDM di hari ke 7, dan 14, kembali menurun di hari ke-21. Ekspresi kadar kalsium relatif cenderung meningkat pada kelompok MSC yang dikultur pada perancah PVA/hFDM dengan gambaran mikroskopis berupa bercak merah pada permukaan perancah. Kesimpulan dari penelitian ini adalah perancah PVA/hFDM cenderung dapat mendukung diferensiasi osteogenik MSC. Hasil penelitian menunjukkan bahwa penggunaan perancah PVA/hFDM dapat menurunkan ekspresi gen BMP2, dan meningkatkan ekspresi gen chordin, serta cenderung meningkatkan ekspresi kadar kalsium relatif yang terdeposit pada matriks ekstraseluler.

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Osteogenic differentiation from Mesenchymal Stem Cell (MSC) to osteoblasts has great clinical significance for treating bone injury. Various studies have been conducted to investigate factors that can enhance osteogenic differentiation, including scaffold development for MSC culture. Scaffold Polyvinyl Alcohol (PVA) / human Fibroblast-derived Matrix (hFDM) is a scaffold candidate assumed to support osteogenic differentiation of MSCs. The extracellular matrix (ECM) presence on the scaffold can regulate various cellular activities through the matrix protein components contained in the ECM. Matrix protein plays a role in sequestering multiple growth factors. Growth factors such as BMP-2 and chordin are to regulate osteogenic differentiation. The process of osteogenic differentiation can be observed by accumulating calcium minerals in the extracellular matrix. The purpose of this study was to determine the optimal method for making PVA / hFDM scaffold and to determine the role of the PVA / hFDM scaffold in affecting MSC osteogenic differentiation by measuring the expression of BMP2 and chordin genes, as well as the expression of relative calcium levels in the extracellular matrix. Optimization of making hFDM PVA scaffold begins with the optimization of culture medium, culture time, preparation, and decellularization techniques. hFDM was characterized using Hematoxylin, Masson Trichrome, and Immunohistochemical staining to determine matrix proteins' presence. MSCs were cultured on the PVA / hFDM scaffold for osteogenic differentiation assay for 21 days. RNA samples were isolated on day 7,14 and 21. Expression of BMP2 and chordin genes were analyzed using the qRT-PCR method. The expression of relative calcium levels was analyzed by qualitative and quantitative tests of Alizarin Red staining. The results of this study indicate that the PVA / hFDM scaffold preparation protocol has been optimized, and the hFDM characterization shows the presence of matrix proteins in the form of collagen and biglycan. BMP-2 gene expression decreased in the MSC group cultured on the PVA / hFDM scaffold on days 7, 14, and 21. In contrast, the chordin gene expression increased in the MSC group cultured on the PVA / hFDM scaffold on days 7, and 14, back down on day 21. The expression of relative calcium levels tended to increase in the MSC group cultured on the PVA / hFDM scaffold with a microscopic appearance of red spots on the scaffold surface. This study concludes that Scaffold PVA / hFDM tends to support osteogenic differentiation of MSCs. The results showed that the use of the PVA / hFDM scaffold could decrease the expression of the BMP2 gene, and increase the expression of the chordin gene, and tended to increase the expression of the relative calcium levels deposited in the extracellular matrix.

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