Wayan Tunas Artama

Piloting the One Health Systems Mapping and Analysis Resource Toolkit in Indonesia

As a global network, countries are being asked to meet goals set forth in the Global Health Security Agenda (GHSA) for a workforce capable of effective and efficient prevention, detection and response to infectious disease threats. There is great need for a cross-sectoral workforce that can innovate and problem-solve. To achieve GHSA goals, countries need a way to visualize their existing system, identify opportunities for improvement, and achieve improved cross-sectoral interactions. The One Health Systems Mapping and Analysis Resource Toolkit (OH-SMART) was successfully piloted in West Sumatra, Indonesia, and was used to enhance multi-agency collaboration around infectious disease outbreaks and proved to be an adaptable, scalable process requiring minimal resources. The authors present OH-SMART as a potential tool to help countries analyze their existing health system and create relevant action steps to improve cross-sectoral collaborations.

Pelacakan Protein Wingless-Type MMTV Integration Site Family Member-4 pada Uterus Mencit (DETECTION WINGLESS-TYPE MMTV INTEGRATION SITE FAMILY MEMBER-4 PROTEIN OF MOUSE UTERUS)

The aims of this study was to detect the expression of Wingless-type MMTV integration site familymember 4 (Wnt4) protein in the uterus of Swiss Webster mice. Laboratory animals that were used areadult Swiss Webster mice weighing 25-30 grams. Mice were kept and mated in a controlled laboratoryconditions. Pregnancy was determined by the presence of vaginal plug in female mice after breeding.Protein was isolated from the uterus at seven days of gestation. Immunoblotting was performed usingChemiluminescent Western Blot kit. The primary antibody used was anti Wnt-4 antibody with a 1: 1000dilution. The results showed protein bands with molecular weights ranging from 40 kDa showed a positivereaction to the primary antibody that were used so that it can be concluded that the protein is Wnt4protein.

Regulatory elements of stx2 gene and the expression level of Shiga-like toxin 2 in Escherichia coli O157:H7

BACKGROUND/PURPOSE Shiga-like toxin (Stx) is an important factor in the pathogenesis of Escherichia coli O157:H7 infection and is responsible for some severe complications. Stx2 is usually associated with hemolytic uremic syndrome in humans. Its expression is regulated by elements located upstream of the stx2 gene, including stx2-promoter sequence, ribosome binding site, and the antiterminator q gene. The present study aimed to find the correlation between regulatory elements and the expression level of Stx2 in two local isolates of E. coli O157:H7. METHODS Two local E. coli O157:H7 strains SM-25(1) and KL-48(2), originating from human and cattle feces, respectively, and an E. coli reference strain, ATCC 43894, were investigated. The complete stx2 gene covering the sequences of promoter, ribosome binding site, and open reading frame and q gene of each strain was analyzed. The magnitude of Stx2 production was detected with a reverse passive latex agglutination method and Stx mediated cellular damage was determined with the Vero cell assay. RESULTS A comparison of the complete stx2 gene contained stx2-promoter, ribosome binding site, and q genes of two local strains KL-48(2) and SM25(1), and the E. coli ATCC 43894 showed that the amino acid sequences were identical. Both local isolates were Stx negative in the reverse passive latex agglutination test and nontoxic in the Vero cell assay. CONCLUSION The expression level of Shiga-like toxin of the two local isolates of E. coli O157:H7 did not only depend on the regulatory elements of the stx2 gene.

Cloning and Clone Analysis of GRA1 Gene from Local Isolate Toxoplasma gondii Tachyzoite

The GRA1 gene of Toxoplasma gondii encoding protein called GRA1 protein. GRA1 protein known to be immunogenic and essentialy involved in modification of parasitophorus vacoule which has role in immune evasion and virulency of organism. The local isolate of T. gondii is successfuly isolated and known as highly pathogenic isolate similarly as its RH strain. Unfortunately, the homology sequence of GRA1 gene between those isolate still unknown. The purpose of the research are to clone the GRA1 gene and to analyze the homology from pathogenic T. gondii isolate and RH strain. Tachyzoite of T. gondii was grown in mice peritoneum by intraperitoneal injection. Then, total mRNA was isolated and purified. cDNA was synthesized from mRNA and then amplified using F1 dan R1 primers to get clone of GRA1 from local isolate. Homology analysis was perform using several bioinformatic softwares. The result showed that cDNA of GRA1 from local isolate has 84% homologs with RH strain of T.gondii . However, when subsequently editing performed to parts of suspected non coding sequence of cDNA GRA1 to get CDS of GRA1, the homology was increase to 100% compare to CDS of GRA1 of RH strain. Key words: GRA1, Toxoplasma gondii , Clonning, Expression