Rani Sauriasari

Oxidative stress biomarkers and lifestyles in Japanese healthy people

The urinary concentrations of 8-isoprostane and 8-hydroxy-2’-deoxyguanosine (8-OHdG), which are biomarkers of oxidative stress, were measured in 677 Japanese people without any diseases, and their correlations with lifestyle facotrs, lifestyle-related blood biochemical parameters, and dietary intake of antioxidative vitamins were investigated. The mean urinary concentration of 8-isoprostane and 8-OHdG was 0.58 ng/mg creatinine and 8.43 ng/mg creatinine, respectively. Mean urinary 8-isoprostane was significantly different in terms of age, gender, smoking and alcohol consumption but not different in terms of body mass index (BMI) and exercise. By multiple regression analysis, urinary 8-isoprostane was significantly influenced by smoking and age. On the other hand, mean urinary 8-OHdG showed differences only by age group. Multiple regression analysis revealed that urinary 8-OHdG was significantly influenced by age, smoking, body weight, levels of high-sensitivity C-reactive protein (Hs-CRP) and low density lipoprotein-cholesterol in females, although it was significantly influenced by body weight in males. The present study shows that urinary 8-isoprostane is associated with lipid peroxidation related-lifestyles such as smoking, and urinary 8-OHdG is associated with arteriosclerosis related-factors such as Hs-CRP. Our findings suggest that 8-isoprostane and 8-OHdG appear to be prospective biomarkers for early prediction of lifestyle related-disease risk at the population level.

Plasma 3-nitrotyrosine, urinary 8-isoprostane and 8-OHdG among healthy Japanese people

To examine the relation between lifestyle factors and oxidative stress biomarkers, this study analysed plasma 3-nitrotyrosine (3-NT), urinary 8-isoprostane and 8-hydroxy-2’- deoxyguanosine (8-OHdG) of 323 healthy Japanese without any disease. Plasma 3-NT was significantly increased by excessive exercise (p=0.010), but it was not significantly different in terms of sex, age (< 40, ≧40), BMI (<18.5, 18.5–24.9, ≧25.0), smoking (non-smokers, smokers) and alcohol drinking per week (non-drinkers, <10 units, ≧10 units). Urinary 8-isoprostane was significantly associated with alcohol drinking (p <0.01) and sex (p <0.01), although it had no significant relevance to age and exercise. Moreover, urinary 8-OHdG was positively associated with age (p <0.05) and negatively associated with BMI (p <0.05) and fasting insulin (p <0.001). However, it was not related with sex, smoking, alcohol drinking and exercise. In conclusion, the present results suggest that 3-NT, 8-isoprostane and 8-OHdG seem to be useful biomarkers for early prediction of lifestyle-related disease risk at the population level.

C-reactive protein is associated with cigarette smoking-induced hyperfiltration and proteinuria in an apparently healthy population

Although cigarette smoking is known to be an important risk factor for renal disease, the mechanism by which smoking induces progressive renal disease in a healthy population has not been established. We hypothesized that oxidative stress (measured as 8-iso-prostaglandin F2α, 8-iso-PGF2a), inflammation (highly sensitive C-reactive protein (CRP), hs-CRP) and nitric oxide may be associated with an alteration in the estimated glomerular filtration rate (eGFR) and proteinuria in otherwise healthy smokers. A total of 649 eligible subjects were classified according to their smoking status. Plasma NOx was measured using ozone-based chemiluminescence, urinary 8-iso-PGF2a was measured using enzyme immunoassay and serum hs-CRP was measured using a latex aggregation nephelometry method. The levels of 8-iso-PGF2a and hs-CRP increased in current smokers (P=0.001 and P=0.029, respectively), although there was not an increase in the NOx level. The prevalence of a high eGFR increased in light smokers (odds ratio (OR) 1.15 (95% confidence interval (CI), 0.61–2.17)) and heavy smokers (OR 2.33 (95% CI, 1.06–5.10)) when compared with non- and past smokers (P for trend=0.024). The multivariable-adjusted mean values of the eGFR in current smokers, reported from the lowest to the highest quintiles of hs-CRP levels, were 82.1, 85.1, 86.4 and 88.5 ml per min per 1.73 m2 (P for trend=0.027). The mean values of proteinuria were 28.6, 34.6, 37.2 and 39.5 mg g−1 creatinine (P for trend=0.003). The correlation coefficient between hs-CRP and eGFR was increased significantly (P=0.03) across non- (r=0.03), past (r=−0.17), light (r=0.13) and heavy smokers (r=0.31). In conclusion, cigarette smoking is a risk factor for renal function alteration in healthy smokers and is characterized by a high eGFR and a high urinary protein associated with an increase in the hs-CRP. This finding suggests that hs-CRP may help mediate the alteration of renal function in smokers.

The Correlation between Urinary 8-iso-Prostaglandin F2α and Hydrogen Peroxide toward Renal Function in T2DM Patients Consuming Sulfonylurea and Combination of Metformin-Sulfonylurea.

BACKGROUND Renal dysfunction is a common complication in type 2 diabetes mellitus patients associated with oxidative damage which could be characterized by 8-iso-prostaglandin F2α and hydrogen peroxide level as oxidative stress markers. OBJECTIVE The aim of our study is to determine if there is a difference in 8-iso-prostaglandin F2α and hydrogen peroxide levels between sulfonylurea and combination of metformin-sulfonylurea in diabetic patients. We also wanted to determine if these oxidative stress markers correlate with the estimated Glomerular Filtration Rate (eGFR). SUBJECTS AND METHODS We conducted a cross-sectional study with inclusion of 55 patients with type 2 diabetes mellitus in Dr. Sitanala Tangerang Hospital, Indonesia with purposive sampling. The value of eGFR was obtained by serum creatinine levels, while the level of 8-iso-prostaglandin F2α was measured by ELISA and urinary hydrogen peroxide using FOX-1 (Ferrous Ion Oxidation Xylenol Orange 1). RESULTS There was no difference in 8-iso-prostaglandin F2α and hydrogen peroxide level between the two groups (p=0.088 and p=0.848). Moreover, there was no difference in eGFR values between the two groups, measured by Cockroft-Gault, MDRD, and CKD-EPI. 8-iso-prostaglandin F2α (n=55) was positively correlated with eGFR based on Cockroft-Gault (r=0.382; p=0.009), whereas urinary hydrogen peroxide (n=47) also generate significant positive correlation with eGFR based on the MDRD equation (r=0.326; p=0.021). Linear regression analysis showed that 8-iso-prostaglandin F2α is the most predictive factor and the only significant factor for eGFR in Cockroft-Gault, MDRD and also CKDEPI, even after controlled by gender, age, BMI, HbA1c, systole, and H2O2. CONCLUSION The two treatments did not have any significant differences in antioxidant activity. However, an increase of urinary 8-iso-prostaglandin F2. and hydrogen peroxide which correlates with eGFR in the total sample may play a significant role in the pathophysiology of diabetic nephropathy.

Marker of lipid peroxidation related to diabetic nephropathy in Indonesian type 2 diabetes mellitus patients

OBJECTIVE Even though diabetes patients exhibit an increased oxidative stress, its correlation with diabetic nephropathy is not fully understood. The purpose of this study was to determine whether lipid peroxidation marker correlates well with eGFR and UACR in type 2 diabetes mellitus patients. METHODS We collected urine and serum samples of Indonesian type 2 diabetes mellitus outpatients with normo- and microalbuminuria at a Local Government Clinic (from ages: 39-74 years). Urinary 8-iso-PGF2α was measured by ELISA, the serum malondialdehyde by TBARS assay, and urinary albumin by BCG albumin assay. eGFR was calculated using the corrected-Cockcroft-Gault (CG), MDRD, and CKD-EPI equation. Other necessary data were obtained through questionnaires. RESULTS The results showed that the increasing level of malondialdehyde was mildly correlated with the decline in eGFR (MDRD). In contrary, there was a significant positive correlation between 8-iso-PGF2α concentration and eGFR based on the corrected-CG, MDRD study, and CKD-EPI equation (r=0.457, p<0.001; r=0.424, p<0.001; r=0.443, p<0.001). This relationship still persisted in the normoalbuminuric subjects (n=43) (r=0.491, p=0.001; r=0.461, p=0.002; r=0.455, p=0.002). The multivariate analysis showed that 8-iso-PGF2α together with fasting plasma glucose was the most predictive factor for the high 2-quantile eGFR (adjusted OR 1.001, (95% CI, 1.000-1.001)). However, there was no significant correlation between UACR with malondialdehyde (r=0.268, p=0.050) and 8-iso-PGF2α(r=-0.030, p=0.808). UACR itself was inversely correlated with eGFR based on the corrected-CG, the MDRD, and CKD-EPI (r=-0.232, p<0.05; r=-0.228, p<0.05; r=-0.232, p<0.05). CONCLUSIONS Increased 8-iso-PGF2α and malondialdehyde in type 2 diabetes mellitus patients may play a role in the pathophysiologic significance of diabetic nephropathy, even while considering the effect of potential confounders.

Arginase Inhibitory, Antioxidant Activity and Pharmacognosy Study of Sterculia macrophylla Vent. Leaves

Objective: The purpose of this study was to investigate the arginase inhibitory activity, antioxidant activity, and also pharmacognostical study of Sterculia macrophylla leaves. The main component of genus Sterculia was flavonoid that was well known to demonstrate arginase inhibitory activity. Methods: Sample was extracted gradually using n-hexane, ethyl acetate, and methanol solvents, subsequently. The n-hexane, ethyl acetate, and methanol extract were determined for their arginase inhibitory activity. The most active extract was methanol extract. This extract was determined for its antioxidant activity, arginase inhibitory activity, identification of chemical compound, chromatogram profile and determined the content of total flavonoid. The leaves and powder of Sterculia macrophylla were identified with microscopic and macroscopic evaluation. Results: The most active extract was methanol extract with IC50 114,659 μg/mL for arginase inhibitory activity and IC50 78.47 μg/mL for DPPH scavenging activity. The secondary metabolite of methanol extract presence compound of alkaloid, flavonoid, tannin, terpene, and glycoside. The total flavonoid content was 141.10 mg/gram extract. The star-shape trichoma was identified as a specific fragment. Conclusion: The methanol extract of Sterculia macrophylla showed activity as arginase inhibitor and antioxidant.

Antioxidant Activity of Fractions from Garcinia hombroniana Pierre Leaves Extracts

Introduction: Radicals were compounds that generated in normal metabolism and create cell damage. A significant increase of free radical and decreased radical elimination can lead to oxidative stress. Oxidative stress plays an important role in the development of many diseases. Enhanced supply of antioxidants will help prevent the morbidity of many diseases. Garcinia hombroniana Pierre has potency as an antioxidant, but study to evaluate the active fractions as an antioxidant has not been done. Objective: The objective of the study was to evaluate antioxidant activity of fractions separated from ethyl acetate (EtOAc) and methanol (MeOH) extract of Garcinia hombroniana leaves and to obtain active fractions to facilitate finding a pure antioxidant compound. Methods: The extract was fractionated using column chromatography, while antioxidant activity assay was conducted in vitro using spectrophotometric methods with DPPH and FRAP method. Results: EtOAc extract of G. hombroniana leaves yielded EA-8 with radical scavenging percentage 32.67% (10 ppm, with DPPH method) and EA-11 with antioxidant activity percentage 25.73% (10 ppm, with FRAP method) as the most active fraction from EtOAc extract, while MeOH extract yielded M-3 with radical scavenging percentage 37.42% (10 ppm, with DPPH method) and 26.70% (10 ppm, with FRAP method) as the most active fraction from MeOH extract Conclusion: Most active fractions has good antioxidant activity, worthy for further study to isolate antioxidant compound which is responsible for antioxidant activity. However, the percentage of radical scavenging or antioxidant activity of all active fractions were smaller than quercetin as a positive control.

Cost-effectiveness Analysis of Deferiprone and Deferasirox on Thalassemia Major Patients in Tangerang District Hospital, Indonesia

Background: Blood transfusion are needed in improving the quality of life of thalassemia major patients. However since it can lead to excess iron, the iron chelation therapy is needed. Deferiprone and deferasirox are the most often used therapy in Indonesia. Objective: The aim of this study was to compare the cost-effectiveness of deferasirox with deferiprone with costeffectiveness analysis (CEA). Methods: Data were taken retrospectively and sampling was done using total sampling based on medical records and hospital information system. Serum ferritin levels of patients consuming deferasirox (n=27) and deferiprone (n=33) were measured to observe the mean changes of serum ferritin levels as effectiveness’ parameter. The cost was median of the total direct medical cost, summed from the cost of drugs, medical devices, hospitalization, administration, physician, laboratories and blood bags. Results: Based on the results of this study, the effectiveness of deferasirox (1164 ng/mL) was greater than deferiprone (692 ng/mL). Median total cost of deferasirox was more expensive (Rp 76,610,618.69) than deferiprone (Rp 51,869,965.64). Cost-effectiveness ratio of deferasirox (CER: Rp 65,816.68/effectiveness) was lower than deferiprone (CER: Rp 74,956.60/ effectiveness). None of both medications was dominant and therefore we could not determine which medication was the most cost-effective therapy. Changing of medication from deferiprone to deferasirox requires an extra cost of Rp 52,416.64 per one incremental unit of effectivity. Conclusion : The policy maker in healthcare facility need to consider the budget and whether the incremental cost of deferasirox is proportional to its increased effectiveness. Key words: Cost-effectiveness analysis, deferiprone, deferasirox, iron chelation therapy, thalassemia major.

Fractionation and α-glucosidase Inhibitory Activity of Fractions from Garcinia hombroniana Pierre Leaves Extracts

Background: Diabetes mellitus become one of the biggest global health problems of the 21st century. Type 2 diabetes play role for the majority of cases of diabetes worldwide which is characterized by the increase of postprandial blood glucose level. Maintaining postprandial glucose level through inhibition of α-glucosidase is one of the essential strategies in the treatment of diabetes. Inhibitory effect of α-glucosidase was commonly used to identify active compounds potentially to treat diabetes. Natural resources have potency as antidiabetic that can be used in diabetes treatment. Objective: The objective of the study is to separate active fraction in the crude extract of Garcinia hombroniana leaves to facilitate obtaining a pure biologically active compound as the α-glucosidase inhibitor. Methods: Fractionation to separate active fraction was performed using column and thin layer chromatography methods while α-glucosidase inhibitory activity assay was performed in vitro using spectrophotometric methods at λ 400 nm. Results: Ethyl acetate and methanol extract of G. hombroniana yielded 14 and 12 fractions, respectively. Two fractions with the higher percent inhibition compared to other factions are fraction 8 from ethyl acetate extract (FEA8) and fraction 3 from methanol extract (FM3). The IC50 values of FEA8, FM3 and acarbose are 16.370 μg/mL, 59.042 μg/mL, and 39.534 μg/mL respectively. Conclusion: Fraction 8 from ethyl acetate extract of G. hombroniana leaves (FEA8) was separated and known in this study as the most bioactive α-glucosidase inhibitor agent compared with another extract, fractions, and acarbose.

α-Glucosidase inhibitory activity from ethyl acetate extract of Antidesma bunius (L.) Spreng stem bark containing triterpenoids

Background: Buni (Antidesma bunius [L.] Spreng) has been used as a traditional antidiabetic agent in Asia. Objective: The mechanism of antidiabetic properties was studied in this study by determine its α-glucosidase inhibitory activity. Method: Inhibition of α-glucosidase was performed in all fraction of Buni stem bark with acarbose and miglitol as standards. The half maximal inhibitory concentration (IC50) value of acarbose and miglitol was 5.75 and 59.76 μg/mL respectively while ethyl acetate (EtOAc) fraction was the most active fraction with IC50of 19.33 μg/mL. Three isolates (B1, B2, and B3) were found in the EtOAc fraction and elucidated by infrared, 1hydrogen-nuclear magnetic resonance, 13carbon-nuclear magnetic resonance, and two-dimensional nuclear magnetic resonance. Result: The chemical structures of the isolates were identified by the spectrum then compared with literature which concluded that B1 is friedelin, B2 is β-sitosterol, and B3 is betulinic acid. Inhibition of the α-glucosidase assay showed IC50values of B1, B2, and B3 were 19.51, 49.85, and 18.49 μg/mL, respectively. Abbreviations used: IC50: Half maximal inhibitory concentration; H-NMR: Hydrogen-nuclear magnetic resonance; C-NMR: Carbon nuclear magnetic resonance; 2D-NMR: Two dimensional-nuclear magnetic resonance; EtOH: Ethanol; EtOAc: Ethyl acetate; MeOH: Methanol; CHCl3: Chloroform; DMSO: Dimethyl sulfoxide; EtF: Ethyl acetate fraction; Na2CO3: Sodium carbonate; IR: Infrared; TGR5: Transmembrane G protein-coupled receptor 5; EC50: Half maximal effective concentration