Neonatal hyperbilirubinemia (NH) is a common finding in newborn babies in Indonesia. Common and rare variants of UGT1A1 have been known to contribute to NH etiology. This study aims to identify UGT1A1 genetic variation and haplotype associated with NH in Indonesian population. DNA was isolated from 116 cases and 115 controls and a targeted-deep sequencing approach was performed on the promoter, UTRs, and exonic regions of UGT1A1. Determining association of common variants and haplotype analysis were performed using PLINK and Haploview. Ten and 4 rare variants were identified in cases and controls, respectively. The UGT1A1 rare variants frequency in cases (5.17%) was higher than that in controls (1.7%). Four of those rare variants in cases (p.Ala61Thr, p.His300Arg, p.Lys407Asn, and p.Tyr514Asn) and three in controls (p.Tyr79X, p.Ala346Val, and p.Thr412Ser) are novel variants. The frequencies of p.Gly71Arg, p.Pro229Gln, and TA7 common variants were not significantly different between cases and controls. A haplotype, consisting of 3 major alleles of 3′ UTRs common variants (rs8330C>G, rs10929303C>T, and rs1042640C>G), was associated with NH incidence (p = 0.025) in this population. Using targeted-deep sequencing and haplotype analysis, we identified novel UGT1A1 rare variants and disease-associated haplotype in NH in Indonesian population.

BackgroundNeonatal jaundice is a common finding in newborns in Asia, including Indonesia. In some cases, the serum total bilirubin levels exceeds the 95th percentile for hours of life (neonatal hyperbilirubinemia). Severe neonatal hyperbilirubinemia (NH) could lead to kernicterus and neonatal death. Glucose-6-Phosphage Dehydrogenase (G6PD) genetic variations and deficiency have been reported in several studies to be associated with NH. This study aimed to analyze the G6PD genetic variations and its activity in neonates with and without hyperbilirubinemia in the Deutromalay Indonesian population.MethodsDeoxyribose Nucleic Acid (DNA) was isolated from peripheral blood of 116 and 115 healthy term neonates with and without hyperbilirubinemia. All infants underwent the following laboratory examinations: routine hematologic evaluation, Coombs test, G6PD activity measurement using the Randox kit method, and serum total bilirubin level. All exons of the G6PD gene were targeted for deep sequencing using MiSeq (Illumina). An association study of G6PD polymorphisms with NH was performed using PLINK.ResultsThe prevalence of G6PD deficiency in neonates with and without hyperbilirubinemia in Indonesian Deutromalay population were 1.72% (95% Confidence Interval (CI): 0.6–4.1%) and 1.74% (95% CI: 0.7–4.1%), respectively. The most common G6PD polymorphisms, i.e. rs1050757/c.* + 357A > G, rs2230037/c.1311C > T, and rs2071429/c.1365-13 T/IVS11, were identified. However, none of those polymorphisms and their haplotype were associated with NH (p > 0.05, Odds Ratio (OR) ~1.00). The prevalence of G6PD mutations in neonates with and without hyperbilirubinemia were 6.8% (95% CI: 2.3–11.5%) and 6.9% (95% CI: 2.3–11.6%), respectively. The most frequently identified G6PD mutation was the Viangchan variant (p.V291 M), which was followed by the Canton (p.R459L) and Vanua Lava (p.L128P) variants. Two novel mutations were identified both in case (p.V369A, p.I167F) and control (p.L474=, p.I36T) groups.ConclusionThe prevalence of G6PD deficiency is low in neonates with or without hyperbilirubinemia in Deutromalay Indonesian population. The majority of G6PD mutations identified among Indonesian Deutromalay population in this study are Viangchan, Canton and Vanua Lava variants.

Macrovascular diabetes complications are generally caused by a process called atherosclerosis. Evidences suggest that to initiate atherosclerosis, oxidated low-density lipoprotein (oxLDL) has to promote the expression of adhesion molecule. Several studies have evidenced the relevance of oxidative stress and atherosclerosis. However, the protective effect of alpha-lipoic acid (ALA) at atherosclerosis still needs to be explored. This study is aimed at investigating the concentration of plasma oxLDL and the expression of adhesion molecule of type 2 diabetes mellitus (DM) using rat model. Eighteen male rats were segregated into three groups labeled as control group, DM group and DM+ALA group. Type 2 diabetes was induced by intraperitoneal injection of streptozotocin (50 mg/kg) followed by nicotinamide (110 mg/kg). ALA was administered at a dose of 60 mg/kg body weight/day throughout the feeding period of 3 weeks. Plasma oxLDL concentration was measured by enzyme-linked immunosorbent assays and expression of vascular cell adhesion molecule-1 (VCAM-1) was measured by immunohistochemistry. Expression of abdominal aortic adhesion molecule was assessed by calculation with Adobe Photoshop CS3. Analysis of variance test was used to compare the concentration of plasma oxLDL and expression of adhesion molecule. A P -value of 0.05 was considered statistically significant. Plasma oxLDL was lower in diabetic rat+ALA compared with the diabetic rat. Percentage of area VCAM-1 in DM+ALA group was lower than DM group. There were no significant differences between groups in intensity of VCAM-1. In conclusion, ALA showed protective effects against early atherosclerosis in diabetic rats.

BACKGROUND: Diabetes mellitus (DM) is associated with an accelerated atherosclerotic macrovascular disease affecting medium-sized arteries. Several evidences support the role of oxidative stress in atherogenesis. However, the role of alpha lipoic acid (ALA) to prevent atherosclerosis is still debatable. This study was conducted to determine the effect of 60 mg/kg/day ALA for 21 days toward the expression of intercellular adhesion molecule-1 (ICAM-1) in rat model.METHODS: Eighteen male rats were divided into three groups labelled as control group, type 2 DM (T2DM) group, and T2DM+ALA group. The T2DM rat models were created by intraperitoneally injecting 50 mg/kg streptozotocin, followed by 110 mg/kg nicotinamide. Immunohistochemistry was used to evaluate the ICAM- 1 expression in rats. Quantitative image analysis of immunohistochemical stains was done on the abdominal aorta using Adobe Photoshop CS3 to find the area percentage and intensity. Kruskal-Wallis and Mann-Whitney tests were used to compare the mean value of area percentage and intensity.RESULTS: There was an increase in area percentage and intensity of ICAM-1 expression. The highest area percentage of ICAM-1 expression was found in the DM group, while the lowest was found in the control group. There were significant differences in the area percentage and intensity between DM+ALA group and DM group, where the area percentage and intensity of ICAM-I in DM group was higher than the DM+ALA group.CONCLUSION: In conclusion, our results demonstrate that ALA inhibits the expression of ICAM-1 in T2DM rat models.KEYWORDS: atherosclerosis, ICAM-1, alpha lipoic acid

Problem-based Learning (PBL) is an effective approach to promote student-centered learning. PBL tutorials were the key step in PBL process. Students' participation in PBL tutorials can be influenced by many factors. The aim of this study was to identify the factors that influence students' participation in PBL tutorials among students in Faculty of Medicine University of Riau. The research was a cross sectional study using self report questionnaires obtained from PBL students, conducted in April -October 2009. The students' participation was assessed using Visschers-Pleijers questionnaires. The result revealed the characteristics of parcipants which are female (79.5%), 19 years old (40.2%), had GPA 2.50 -3.00 (56.7%), had past experience in organization (89.0%). Among 44.1% of the participants showed active participation in tutorial processs. Male students more likely active than female students (P = 0.045). Another factors including age, GPA, past experience in organization, role of tutors, and quality of scenario were not influenced the participation of subject. Students' participation in PBL tutorials among students in Faculty of Medicine University of Riau was influenced by sex.