The Indonesian Biomedical Journal

139-50

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: The average lifespan of humans is increasing, and with it the percentage of people entering the 65 and older age group is growing rapidly and will continue to do so in the next 20 years. Within this age group, cardiovascular disease will remain the leading cause of death, and the cost associated with treatment will continue to increase. Aging is an inevitable part of life and unfortunately poses the largest risk factor for cardiovascular disease. CONTENT: We provide an overview of some of the molecular mechanisms involved in regulating lifespan and health, including mitochondria, telomeres, stem cells, sirtuins, Adenosine Monophosphate-activated Protein Kinase, Mammalian Target of Rapamycin and Insulin-like Growth Factor 1. We also provide future perspectives of lifespan and health, which are intimately linked fields. SUMMARY: Aging remains the biggest non-modifiable risk factor for cardiovascular disease. The biological, structural and mechanical changes in senescent cardiovascular system are thought to contribute in increasing incidence of cardiovascular disease in aging. Understanding the mechanisms contributing to such changes is therefore crucial for both prevention and development of treatment for cardiovascular diseases. KEYWORDS: cardiovascular aging, mitochondria, telomeres, sirtuin, stem cells

161-8

Authors: Yuliana Sambara, Mansyur Arif, Uleng Bahrun

BACKGROUND: Hyperglycemia in diabetic patients cause both chronic inflammation and extracellular matrix accumulation that can lead to progressive renal damage. Albumin, Gammaglutamytransferase (GGT) and clusterin in urine are markers to detect damage in glomerulus, cell of the tubules and proximal tubules, respectively. METHODS: This study aimed to evaluate the pathomechanism of haemoglobin A1c (HbA1c), albumin, GGT, clusterin, type IV collagen in urine, and high sensitivity C-reactive protein (hsCRP) in type 2 diabetes mellitus (DM) patients. The study was a cross sectional study involving 82 subjects consisting of 36 males and 46 females, 35-65 years old, divided into 3 groups: uncontrolled DM, controlled DM and non DM. Data were obtained from interviews, physical examinations (weight, height, blood pressure) and laboratory examinations (HbA1c, serum glutamic oxaloacetic (SGOT), serum glutamic pyruvic (SGPT), creatinine, hsCRP, urinary albumin, urinary GGT, urinary clusterin, and urinary type IV collagen). Statistical analysis was performed for correlation, difference and cross tabulation tests. RESULTS: The study results showed there were significant differences (p<0.05) between uncontrolled DM group compared with controlled DM and non DM groups in HbA1c, ratio of urinary type IV collagen and ratio of urinary albumin. However, there were no significant differences between controlled DM and non DM groups. There were positive significant correlations between HbA1c with hsCRP (r=0.223, p<0.05), HbA1c with ratio of urinary type IV collagen/creatinine (r=0.563, p<0.001), HbA1c with ratio of urinary albumin/creatinine (r=0.263, p<0.05), and ratio of urinary type IV collagen/creatinine with ratio urinary albumin/creatinine (r=0.613, p<0.001). CONCLUSION: Results of this study indicated that albumin and type IV collagen in urine play a role in renal damage caused by uncontrolled glucose level in subjects with type 2 DM. The increased concentration of both HbA1c and urinary type IV collagen indicates increased risk factor of glomerulus damage. Urinary type IV collagen is likely to be future potential marker for early detection of renal damage.

173-8

Authors: Ferry Sandra, Toshio Kukita, Tatsushi Muta, Tadahiko Iijima

BACKGROUND: Caffeic acid was reported in our previous study to have potential in inhibiting osteoclastogenesis through inhibition of nuclear factor κB (NFκB). Here in our current study, we would like to investigate further the caffeic acid-affected signaling pathway leading to NFκB inhibition. Since tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) plays important role in osteoclastogenesis, we applied TRAF6- transfected RAW264 cells D-Clone (RAW-D) cells as model in this study. METHODS: Caffeic acid in various concentrations was added to in vitro osteoclastogenesis of receptor activator nuclear factor κB ligand (RANKL)-TNFα-induced TRAF6-transfected RAW-D cells. Cells were collected, lysed and immunoblotted to detect TRAF6 expression. To detect tartrate resistant acid phosphatase (TRAP)+ polynucleated cells (PNCs), TRAP staining was performed. Meanwhile, to measure NFκB Activity, cells were transfected with pNFκB-TA-Luc and subjected to Dual Luciferase Reporter Assay System. RESULTS: Caffeic acid did not influence TRAF6 expression of RANKL-TNFα-induced TRAF6-transfected RAW-D cells. Caffeic acid diminished NFκB activity of RANKL-TNFα-induced TRAF6-transfected RAW-D cells in a concentration dependent manner. Significant NFκB activity inhibitions were seen under treatment of 1 and 10 μg/ml caffeic acid. By adding 10 μg/ml caffeic acid in RANKL-TNFα-induced TRAF6-transfected RAW-D cells, TRAP+ PNCs number was significantly suppressed. CONCLUSION: Caffeic acid inhibited RANKL-TNFα-TRAF6-induced osteoclastogenesis pathway. Since caffeic acid did not influence TRAF6 expression, TRAF6-RANK interactions and/or TRAF6 downstream signaling pathway should be further pursued to disclose inhibition mechanism of caffeic acid.

185-92

Authors: Winny Xie, Yusmiati Yusmiati

BACKGROUND: According to a report from WHO, cases of rubella infection in Indonesia has increased up to 10-fold from 2007 to 2011. Despite no data of congenital rubella syndrome in the report, there are approximately 45,000 cases of babies born with heart failure and 0.1-0.3% live births with congenital deafness in Indonesia. Allegedly, rubella infection during pregnancy may play a role in this condition. This study aimed to optimize and validate a real-time reverse transcriptase polymerase chain reaction (RT-qPCR) method to detect rubella virus RNA as an aid for the diagnosis of congenital rubella infection. METHODS: Method optimization was conducted using nucleic acids extracted from Trimovax Merieux vaccine with the High Pure Viral Nucleic Acid Kit. One step RT-qPCR was performed with Quantifast Multiplex RTPCR+R Kit. Target synthetic DNA was designed and used to determine the sensitivity of the method. RNA internal control was synthesized to control the process of extraction and amplification. RESULTS: The analytical sensitivity of this method was as low as 5 copies target synthetic DNA/μl. The mean Coefficient of Variation (CV) % of the critical threshold (Ct) obtained were 2.71%, 1.20%, 1.62%, and 1.59% for within run, between run, between kit lots, and between operators, respectively. Recovery of the target synthetic DNA from amniotic fluid was 100.51% (by the log copies/μl) at the concentration of 1,000,000 copies/μl. CONCLUSION: RT-qPCR is successfully used for the detection of rubella virus RNA in vaccine and synthetic nucleic acid. With its high sensitivity, good precision and recovery, this method offers a means to improve the diagnosis of congenital rubella infection in developing countries like Indonesia.