Widagdo Sri Nugroho

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.

Frequency and risk-factors analysis of Escherichia coli O157:H7 in Bali-cattle

Cattle are known as the main reservoir of zoonotic agents verocytotoxin-producing Escherichia coli. These bacteria are usually isolated from calves with diarrhea and/or mucus and blood. Tolerance of these agents to the environmental conditions will strengthen of their transmission among livestock. A total of 238 cattle fecal samples from four sub-districts in Badung, Bali were used in this study. Epidemiological data observed include cattle age, sex, cattle rearing system, the source of drinking water, weather, altitude, and type of cage floor, the cleanliness of cage floor, the slope of cage floor, and the level of cattle cleanliness. The study was initiated by culturing of samples onto eosin methylene blue agar, then Gram stained, and tested for indole, methyl-red, voges proskauer, and citrate, Potential E.coli isolates were then cultured onto sorbitol MacConkey agar, and further tested using O157 latex agglutination test and H7 antisera. Molecular identification was performed by analysis of the 16S rRNA gene, and epidemiological data was analyzed using STATA 12.0 software. The results showed, the prevalence of E. coli O157:H7 in cattle at Badung regency was 6.30% (15/238) covering four sub districts i.e. Petang, Abiansemal, Mengwi, and Kuta which their prevalence was 8.62%(5/58), 10%(6/60), 3.33%(2/60), and 3.33(2/60)%, respectively. The analysis of 16S rRNA gene confirmed of isolates as an E. coli O157:H7 strain with 99% similarities. Furthermore, the risk factors analysis showed that the slope of the cage floor has a highly significant effect (P<0.05) to the distribution of infection. Consequently, implementing this factor must be concerned in order to decrease of infection.

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.

Isolasi and Identification of Bacteria from the Urine Fluid Organic Fertilizer (POC)Ettawa Crossbred (PE) in The Sleman Regency

Urine is a by product from Ettawa crossbred (PE) goat farm in Sleman regency. In addition, the manure urine goats can be used for fluid organic fertilizer (POC). Recently, organic farming is more interesting for people because it does not use any chemical fertilizers. Salmonella sp and E. coli O157:H7 are pathogenic bacteria that can contaminate agricultural products and dangerous for public health. One of sources of contamination in agricultural products is due to the organic fertilizer. Therefore, the aim of the present study was to determine pathogenic bacteria from POC urine PE goats in Sleman regency. A total of 8 POC samples were collected from PE goat urine in Sleman regency. These samples were analyzed for Salmonella sp , E. coli O157:H7 and other pathogenic bacteria by using biochemical reactions. All samples are negative for Salmonella sp. and E. coli O157:H7 as well. Whereas, 85% of 8 samples are positive for Pseudomonas sp . The present study showed that POC from PE goats farm in Sleman regency are negative for pathogenic bacteria.

Confirmation test of suspected Mycobacterium avium subspecies paratuberculosis (MAP) isolated using PCR F57

Seropositive and isolate suspected as Mycobacterium avium subspecies paratuberculosis (MAP) was detected at dairy cows in West Java. This bacteria causes Johne’s disease (JD) and potentially becomes a new emerging disease for Indonesian dairy cows. The aim of this study was to confirm the suspected local isolate as a MAP distinctively by PCR. Reculture of MAP reference isolate, suspected local isolate done by resuspending bacteria in PBS 0.5% and inoculating it in Herrold’s egg yolk medium with mycobactin J (HEYM) and than inoculating it in 37 o C for 16 weeks. The cultures grew in various time, Mycobacterium avium subspecies avium was detected in 3 rd week, MAP reference was detected in 7 th week, and local isolate was detected in 14 th week. The confirmation test was carried out by PCR with primer F57. The PCR F57 result showed that MAP suspected isolate was not a Mycobacterium avium subspecies paratuberculosis . Key Words: Local Isolate, Mycobacterium avium Subspecies Paratuberulosis , PCR F57