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Jakarta, Indonesia

Sandra F.,Prodia Clinical Laboratory | Sandra F.,Trisakti University | Oktaviono Y.H.,Airlangga University | Oktaviono Y.H.,Brawijaya University | And 4 more authors.
Molecular and Cellular Biochemistry

Endothelial progenitor cells (EPCs) clinical applications have been well reported. However, due to low number of EPCs that could be isolated, EPCs expansion study became one of the main focuses. Some optimized mediums to culture EPCs were currently available. However, the proliferation signaling pathway is not clearly disclosed yet. Peripheral blood was collected from eight healthy subjects, followed by mononuclear cells (MNCs) isolation. MNCs were then prepared and cultured for 2 days. After that, non-adherent cells were harvested and further cultured for 3 days. Resulted colony-forming unit (CFU)-Hill colonies were documented and enumerated under an inverted light microscope. To detect membrane markers, immunofluorescence was performed to detect CD34, VEGFR-2, and CD133. Cell documentation was conducted under a fluorescence microscope. To check cell proliferation, XTT Cell Proliferation Assay Kit was used according to kit insert. To detect possible activation of p44/42 MAPK, western blot was performed to detect p44/42 MAPK and phosphorylated p44/42 MAPK. All visualized bands were captured and quantified. Our results showed that EPCs markers (CD34, CD133 and VEGFR-2) were detected in 3 days culture. From XTT cell proliferation assay and CFU enumeration results, we found that EPCs proliferated significantly (p = 0.012) with addition of supplement. Phosphorylated-p42 MAPK expression of EPCs treated with supplement was significantly higher than the one of EPCs without treatment. Significant inhibition of p42 MAPK phosphorylation by U0126 was observed (p = 0.012). By pretreatment of U0126, number of viable cells and CFUs treated with supplement was significantly decreased (p = 0.012). Our results showed that MEK-dependent p42 MAPK pathway might play an important role in EPCs proliferation. © 2014, Springer Science+Business Media New York. Source

Yoo S.J.,Inje University | Wang L.L.,University of Sichuan | Ning H.-C.,Chang Gung Memorial Hospital | Tao C.M.,University of Sichuan | And 11 more authors.
Journal of Clinical Virology

Background: Early diagnosis of hepatitis C virus (HCV) infection is essential to allow appropriate treatment and prevent transmission. Objectives: To evaluate the Elecsys® Anti-HCV II assay as a routine screening assay in Asia using a large number of samples from different Asian Pacific populations and compare its performance with other HCV assays routinely used in the region. Study design: The sensitivity and specificity of the Elecsys® Anti-HCV II assay were determined using routine hospital samples and compared with at least one of the following comparator assays at nine independent centers: ARCHITECT™ Anti-HCV; Serodia®-HCV Particle Agglutination; Vitros® ECi Anti-HCV; Elecsys® Anti-HCV; ADVIA Centaur® HCV; InTec® HCV EIA; or Livzon® Anti-HCV. Commercially available seroconversion panels were used to assess sensitivity for early detection of infection. Results: The Elecsys® Anti-HCV II assay was more sensitive in recognizing early infection and detected acute HCV infection earlier on average than the comparator assays for all six panels tested. 7,726 routine samples were tested and 322 identified as HCV positive. Elecsys® Anti-HCV II had a sensitivity of 100% and a specificity of 99.66%, both of which were comparable or superior to the results obtained for competitor assays, which ranged from 87.5-100% and 98.98-100%, respectively. Conclusions: The Elecsys® Anti-HCV II assay has the sensitivity and specificity to support its use as a routine screening method in the Asia Pacific region. Furthermore, this assay shortens the diagnostic window between infection and the detection of antibodies compared with established methods. © 2015 The Authors. Source

Ichihara K.,Yamaguchi University | Ceriotti F.,San Raffaele Scientific Institute | Tam T.H.,Center for Standardization | Sueyoshi S.,Chiba Cardiovascular Center | And 13 more authors.
Clinical Chemistry and Laboratory Medicine

Background: A multicenter study conducted in Southeast Asia to derive reference intervals (RIs) for 72 commonly measured analytes (general chemistry, inflammatory markers, hormones, etc.) featured centralized measurement to clearly detect regionality in test results. The results of 31 standardized analytes are reported, with the remaining analytes presented in the next report. Method: The study included 63 clinical laboratories from South Korea, China, Vietnam, Malaysia, Indonesia, and seven areas in Japan. A total of 3541 healthy individuals aged 20-65 years (Japan 2082, others 1459) were recruited mostly from hospital workers using a well-defined common protocol. All serum specimens were transported to Tokyo at -80 C and collectively measured using reagents from four manufacturers. Three-level nested ANOVA was used to quantitate variation (SD) of test results due to region, sex, and age. A ratio of SD for a given factor over residual SD (representing net between-individual variations) (SDR) exceeding 0.3 was considered significant. Traceability of RIs was ensured by recalibration using value-assigned reference materials. RIs were derived parametrically. Results: SDRs for sex and age were significant for 19 and 16 analytes, respectively. Regional difference was significant for 11 analytes, including high density lipoprotein (HDL)-cholesterol and inflammatory markers. However, when the data were limited to those from Japan, regionality was not observed in any of the analytes. Accordingly, RIs were derived with or without partition by sex and region. Conclusions: RIs applicable to a wide area in Asia were established for the majority of analytes with traceability to reference measuring systems, whereas regional partitioning was required for RIs of the other analytes. © 2013 by Walter de Gruyter Berlin Boston 2013. Source

Mutakin,Gunma University | Mutakin,Padjadjaran University | Meiliana A.,Prodia Clinical Laboratory | Wijaya A.,Prodia Clinical Laboratory | And 5 more authors.
Journal of Trace Elements in Medicine and Biology

Background and aim: Previous evidence has suggested an association between selenium and cardiovascular disease, which is main outcome of metabolic syndrome. The aim of this study was to examine possible correlation between selenium nutritional status and metabolic risk factors in men with visceral obesity. Methods: Plasma samples were collected from 123 Indonesian men with visceral obesity. Their metabolic risk factors and selenium nutritional status were analyzed. The eligible subjects (n=78) were stratified according to the International Diabetes Federation: obese, obese plus one component, and obese plus two components or more. Obese plus two components or more were diagnostic criteria of metabolic syndrome. Pearson's correlation was performed to examine the correlation in each group. Results: In the obese group, selenium positively correlated with high-density lipoprotein (HDL) cholesterol (r=0.390, P<0.05) and with fatty acid binding protein-4 (FABP4) (r=0.474, P<0.05); glutathione peroxidase-3 (GPx3) activity was inversely correlated with FABP4 (r=-467, P<0.05). In the obese plus one component group, GPx3 activity positively correlated with HDL cholesterol (r=0.413, P<0.05). In the metabolic syndrome group, selenium negatively correlated with monocytes chemoattractant protein (MCP)-1 (r=-0.429, P<0.05). Conclusions: These results show that the association between selenium nutritional status and metabolic risk factors is limited to particular group of obese men with or without metabolic syndrome. © 2012. Source

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