Heru Susetya

Human umbilical mesenchymal stem cells conditioned medium promote primary wound healing regeneration.

Aim: This research was conducted to clarify the capability of human umbilical mesenchymal stem cells conditioned medium (HU-MSCM) to promote regenerations of primary wound healing on the incision skin injury. Materials and Methods: In this study, two approaches in vitro and in vivo already done. On in vitro analysis, tube formation was performed using HU vein endothelial cells in the presence of HU-MSCM, in some experiments cells line was incubated prior the presence of lipopolysaccharide and HU-MSCM then apoptosis assay was performed. Furthermore, in vivo experiments 12 female rats (Rattus norvegicus) were used after rats anesthetized, 7 mm wound was made by incision on the left side of the body. The wound was treated with HU-MSCM containing cream, povidone iodine was run as a control. Wound healing regenerations on the skin samples were visualized by hematoxylin-eosin staining. Results: In vitro models elucidate HU-MSCM may decreasing inflammation at the beginning of wound healing, promote cell migration and angiogenesis. In addition in vivo models show that the incision length on the skin is decreasing and more smaller, HE staining describe decreasing of inflammation phase, increasing of angiogenesis, accelerate fibroplasia, and maturation phase. Conclusions: Taken together our observation indicates that HU-MSCM could promote the acceleration of skin tissue regenerations in primary wound healing process.

The structural and functional recovery of pancreatic β-cells in type 1 diabetes mellitus induced mesenchymal stem cell-conditioned medium

Aim: Various studies have shown that secreted factors alone in culture medium without stem cell are capable of repairing tissues by itself in various conditions involving damaged tissue/organ. Therefore, this study was aimed to investigate the role of human umbilical cord mesenchymal stem cell-derived conditioned medium (CM) on the recovery of pancreatic β-cells in Wistar rats (Rattus norvegicus) with type 1 diabetes mellitus. Materials and Methods: The 0.05 ml CM induction was applied to the diabetic group of rats in weeks 1, 2, 3, and 4. 1 week after each CM induction, insulin concentration was analyzed using ELISA. The pancreas was divided into 3 regions, processed by paraffin method, stained with hematoxylin-eosin, and immunohistochemical method for insulin. Results: This study indicated the decrease in the total number of islets and insulin concentration after the injection of single dose of alloxan. The exocrine acini were also damaged. Microscopic observation detected the presence of small islets in the diabetic group 1 week after the first 0.05 ml CM induction. The number and size of the islets increased in line with the CM doses and time of inductions. Immunohistochemically, the presence of low intensity of insulin-positive cells could be recognized at the splenic and duodenal regions of the pancreas, but not gastric region, 1 week after the first and second 0.05 ml CM induction. The intensity of staining and the number of insulin-positive cells increased dramatically in 1 week after the third and fourth 0.05 ml of CM induction in all regions of the pancreas. The data of insulin blood concentration showed clear differences between the second and the fourth induction of 0.05 ml CM induction. Conclusions: This study showed very strong evidence on the role of human umbilical cord mesenchymal stem cell-derived CM in recovering the pancreatic β-cells damage in Wistar rats (R. norvegicus) with type 1 diabetes mellitus, structurally and functionally.

Phylogeography of the current rabies viruses in Indonesia

Rabies is a major fatal zoonotic disease in Indonesia. This study was conducted to determine the recent dynamics of rabies virus (RABV) in various areas and animal species throughout Indonesia. A total of 27 brain samples collected from rabid animals of various species in Bali, Sumatra, Kalimantan, Sulawesi, Java, and Flores in 2008 to 2010 were investigated. The cDNA of the nucleoprotein gene from each sample was generated and amplified by one-step reverse transcription-PCR, after which the products were sequenced and analyzed. The symmetric substitution model of a Bayesian stochastic search variable selection extension of the discrete phylogeographic model of the social network was applied in BEAST ver. 1.7.5 software. The spatial dispersal was visualized in Cartographica using Spatial Phylogenetic Reconstruction of Evolutionary Dynamics. We demonstrated inter-island introduction and reintroduction, and dog was found to be the only source of infection of other animals. Ancestors of Indonesian RABVs originated in Java and its descendants were transmitted to Kalimantan, then further to Sumatra, Flores, and Bali. The Flores descendent was subsequently transmitted to Sulawesi and back to Kalimantan. The viruses found in various animal species were transmitted by the dog.

Detection of Trenbolone Acetate in Beef Muscle and Liver Using High-Performance Liquid Chromatography Method

The use of trenbolone acetate (TBA) as growth promotor was legal in some regions, and it has to be considered since there are no rules yet in Indonesia committed to TBA application in beef. As a step for preconditioning the policy of TBA used as growth promotor in beef, the validated method of hormone detection is needed to be established. This research was aimed to validate the high-performance liquid chromatography (HPLC) method for TBA detection as a tool to control the hormone residue in meat. The imported beef muscles and liver were obtained from a supermarket in Yogyakarta. All samples (n = 10) were kept in the freezer (−18 °C) before analysis. The HPLC method is developed from sample extraction, cleanup with solid-phase extraction (SPE) cartridge, and the hormone concentration analysis using standard curve of TBA. The method has validated and gave the values of LOD (limit of detection): 1 × 10−5 μg · L−1 (muscle) and 1 × 10−4 μg · L−1 (liver). The LOQ (limit of quantification): 1 × 10−4 μg · L−1 (muscle) and 1 × 10−3 μg · L−1 (liver) and R2 = 0.99 (muscle and liver). The analysis results were three positive TBA in muscles (0.730 μg · L−1, 0.074 μg · L−1 and 0.116 μg · L−1) and two positive in livers (0.080 μg · L−1 and 0.130 μg · L−1). All TBA levels in tissues were not exceeding the maximum residue limits of TBA according to the Codex Alimentarius Commissions. This HPLC analysis procedure could be done as a detection method for TBA in beef tissues.

Potential risk of viral transmission from flying foxes to domestic animals and humans on the southern coast of West Java, Indonesia

Flying foxes have been considered to be involved in the transmission of serious infectious diseases to humans. Using questionnaires, we aimed to determine the direct and/or indirect contacts of flying foxes in an Indonesian nature conservation area with domestic animals and humans living in the surrounding area. We surveyed 150 residents of 10 villages in West Java. Villages were classified into 3 groups: inside and/or within 1 km from the outer border of the conservation area and 1–5 km or 5–10 km away from the reserve’s outer border. Data were collected by direct interview using a structured questionnaire consisting of the respondent characteristics (age, sex and occupation); histories of contacts between flying foxes and humans, dogs and other domestic animals; and knowledge about infectious diseases, mainly rabies, in flying foxes. We found that flying foxes from the nature conservation area often enter residential areas at night to look for food, especially during the fruit season. In these residential areas, flying foxes had direct contacts with humans and a few contacts with domestic animals, especially dogs. People who encounter flying foxes seldom used personal protective equipment, such as leather gloves, goggles and caps. The residents living around the conservation area mostly had poor knowledge about flying foxes and disease transmission. This situation shows that the population in this region is at a quite high risk for contracting infectious diseases from flying foxes.

STATUS VAKSINASI RABIES PADA ANJING DI KOTA MAKASSAR RABIES VACCINATION STATUS OF DOGS IN MAKASSAR

Telah dilakukan suatu analisis cakupan vaksinasi rabies pada anjing di kota Makassar. Tujuan penelitian ini adalah untuk mengetahui cakupan vaksinasi anjing bertuan, serta asosiasi antara status vaksinasi dan faktorfaktor manajemen pemeliharaan anjing. Sebanyak 246 sampel anjing dipilih untuk mengetahui status vaksinasi rabies. Analisis status vaksinasi dan faktor pemeliharaan anjing dilakukan secara deskriptif. Uji Chi square (c2) dan odds ratio (OR) digunakan mengetahui asosiasi antara faktor-faktor yang diteliti dan status vaksinasi. Hasil penelitian menunjukkan bahwa cakupan vaksinasi pada anjing bertuan di kota Makassar sangat rendah (21 %). Rendahnya cakupan vaksinasi tersebut diperkirakan berhubungan dengan cara pemeliharaan (P=0,002; OR=4,3), tujuan pemeliharaan (P=0,009; OR=2,3), pengetahuan pemilik tentang rabies (P=0,001; OR=3,0), penghasilan di atas Rp 2.000.000,00 (p=0,000; OR=5,0), dan lokasi pemeliharaan urban area (P=O,OOOI; OR=4, 1). Kata kunci: Cakupanvaksinasi, rabies, faktor risiko