Pramudji Hastuti, Pramudji
Department ofr Biochemistry, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta

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-429 T/C and -374 T/A Polymorphisms in Receptor Advanced Glycation Endproducts (RAGE) gene in Type 2 Diabetic Patients with Diabetic Retinopathy at the Dr. Sardjito General Hospital Yogyakarta Djuma, Agustina Welhelmina; ., Sunarti; Hastuti, Pramudji
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Receptor of advanced glycation endproduct (RAGE) plays an important role in the pathogenesis of diabetic vascular complications, such as diabetic retinopathy. The interaction between the RAGE and advanced glycation end product (AGE) leads to oxidative stress and could result in cellular activation and infl ammation. The production of AGE occurs normally during aging but it increases in hyperglycemia condition. The objective of this research was to investigate the association between -429 T/C and -374 T/A polymorphisms in RAGE gene with the risk of diabetic retinopathy (DR) of type 2 diabetic patients in Javanese population. This was a case control study which consisted of 40 type 2 diabetic patients with DR as case subjects and 40 type 2 diabetic patients without DR (NDR) as control subjects. Genotyping of polymorphism was performed by PCR-RFLP. Chi-square test and odds ratio models were used to evaluate the association of both polymorphisms and DR risk and to examine 2-SNP haplotype of -429 T/C and -374 T/A polymorphisms in RAGE gene on DR. The genotype frequencies of -429 T/C polymorphism in RAGE gene in DR subjects were TT = 72.5% and TC/CC = 27.5%; while in NDR subjects were TT = 80% and TC/ CC = 20%, with p = 0.431. The allele frequencies of -429 T/C polymorphism in DR subjects were T = 83.7% and C= 16.3%, while in NDR subjects were T = 87.5% and C = 12.5%, with p = 0.499. The genotype frequencies of -374T/A polymorphism in RAGE gene in DR subjects were TT = 67.5%, TA = 32.5% while in NDR subjects were TT =82.5%, TA = 17.5%, with p = 0.121. In DR subjects, the frequencies of T and A were 83.7% and16.3%, while in NDR subjects the frequencies of T and A were 91.2 % and 8.8%, with p = 0.151. Odds ratios of -429 T/C polymorphism were 1.52 (95% CI = 0.54 – 4.29) for TC/CC genotype and 1.358 (95% CI = 0.56 – 3.31) for C allele. Odds ratios of -374 T/A polymorphism were 2.27 (95% CI = 0.79 – 6.49) for TA genotype and 2.02 (95% CI = 0.76 – 5.37) for A allele. χ2-value for 2-SNP haplotype was p = 0.127. The -374 T/A polymorphism in RAGE gene was a stronger risk factor of DR than -429 T/C polymorphism in RAGE gene. There were not signifi cantly different of frequencies of genotypes, allele, and two-SNP haplotype of -429 T/C and -374 T/A polymorphisms in RAGE gene between DR subjects and NDR subjects.
Genetic Variation of Apolipoprotein E (ApoE) in Surabaya, Palu and Alor Populations of Indonesia Hastuti, Pramudji; Sofro, Abdul Salam Mudzakir; Asdie, Ahmad Husain; Sadewa, Ahmad Hamim
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

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AbstractApolipoprotein E (ApoE) has been considered to play an important role in cardiovascular disorders.Several studies reported that genetic variation in ApoE locus influence plasma lipoprotein level. The objectivesof this study was to compare the frequency of ApoE genotypes and alleles in some populations of Indonesia.One hundred and ninety five voluntarily unrelated apparently healthy individuals were recruited fromSurabaya, Palu and Alor representing the western, middle and eastern populations of Indonesia, respectively.Blood samples were collected from each subject for DNA extraction. The common allelic variants of ApoE werescreened using polymerase chain reaction (PCR) and restriction fragment length polymorphism. Three allelesi.e. ε2, ε3 and ε4 were identified and five genotypes i.e. ApoE ε2/ε2, ApoE ε2/ε3, ApoE ε3/ε3, ApoE ε2/ε4, ApoE ε3/ε4 were found in three populations studied, while ApoE ε4/ε4 was absent in Surabaya, representing the westernpopulations of Indonesia. The frequency of ε2, ε3 and ε4 alleles in the western population were 0.208, 0.701and 0.092 respectively; in the middle population were 0.242, 0.618 and 0.140 respectively and in the easternpopulation of Indonesia were 0.267, 0.466 and 0.267 respectively. The highest frequency of ε2 and ε4 allelewas found in the eastern population of Indonesia. The distribution of ε2 allele were not significantly differentamong all Indonesian populations, but significantly different were found in ε3 and ε4 allele in the easternpopulation compared to those in the western and middle populations of Indonesian. It can be concluded thatthe frequency of three ApoE alleles in the western and middle populations of Indonesia was not significantlydifferent however, significantly different was observed in the frequency of ApoE ε3 and ε4 alleles from theeastern compared to those in the western and middle populations of Indonesia.Keywords : Apolipoprotein E; genotypes; allele frequency; populations of Indonesia
Production and Optimization of Oleic Acid Ethyl Ester Synthesis Using Lipase From Rice Bran (Oryza sativa L.) and Germinated Jatropha Seeds (Jatropha curcas L.) by Response Surface Methodology Prastowo, Indro; Hidayat, Chusnul; Hastuti, Pramudji
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Recently, the fatty acid ethyl ester has been synthesized in place of fatty acid methyl ester since ethanol has been more renewable. In this research, oleic acid ethyl ester (OAEE) was synthesized using germinated jatropha seeds (Jatropha curcas.L) and rice bran (Oryza sativa) as source of lipase. The objective of the research was to optimize the synthesis conditions using Response Surface Methodology. Factors, such as crude enzyme concentration, molar ratio of oleic acid to ethanol, and the reaction time, were evaluated. The results show that lipase from germinated jatropha seeds had the hydrolitic and esterifi cation activity about 6.73 U/g and 298.07 U/g, respectively. Lipase from rice bran had the hydrolitic and esterifi cation activity about 10.57 U/g and 324.03 U/g, respectively. The optimum conditions of esterifi cation reaction using germinated jatropha seed lipase as biocatalyst were crude enzyme concentration of 0.31 g/ml, molar ratio of oleic acid to ethanol of 1 : 1.81, and reaction time of 50.9 min. The optimum conditions of esterifi cation reaction using rice bran lipase were crude enzyme concentration of 0.29 g/ml, molar ratio of oleic acid to ethanol of 1 : 2.05, and reaction time of 58.61 min. The obtained amounts of OAEE were 810.77 μmole and 626.92 μmole for lipases from rice bran and germinated jatropha seed, respectively.
T-786c Polymorphism in nitric oxide synthase 3 gene and Nitrit Oxide Level of Diabetic Retinopathy in Javanese Population Welkriana, Putri Widelia; -, Sunarti; Hastuti, Pramudji
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

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AbstractComplication of retinopathy in type 2 DM is caused of lower level of NO. Nitric oxide level is synthesizedfrom L-arginin in reaction that catalyze Nitric oxide synthase (NOS) 3. The T-786C mutation in NOS 3 genedecreases the expression of nitric oxide synthase (NOS) 3 so decreases NO synthesis. To investigate theassociation between T-786C polymorphism in NOS 3 gene with NO level of diabetic retinopathy patients. Thisstudy was a case control study, consist of 40 patient of type 2 diabetic with DR (case group) and 40 patient oftype 2 diabetic without DR (control group) of Javanese ethnic. The genotyping of T-786C polymorphism wasperformed by PCR-RLFP. Level of NO was measured by spectrophotometry. Chi square test and odd ratiowere used to analyze the association of the T-786C polymorphism in NOS 3 gene with DR. Differences ofNO level between TT and TC genotypes were analyzed using independent t test. The distribution of T-786Cpolymorphism in NOS 3 gene of DR subjects showed that frequency of TT genotype was 22.5% and TC genotypewas 77.5%. Non DR subjects showed the frequency of TT genotype was 50% and TC genotype was 50%, (p=0.011). Frequency of T allele in DR group was 61.25% and C allele was 38.75%, and frequency of T allele in nonDR group was 75% and C allele was 25%, (p= 0.62). Odd ratio of TC genotype was 3.444(CI; 95% : 0.964-3.735)and C allele was 1.898 (CI; 95% : 1.310-9.058). The NO level of TC genotype was 1.43+0.126 and TT genotypewas 11.27+5.87 (p=0.000). Level of NO between RD and non RD showed not different significantly (p=0.160)for retinopathy. The T-786C polymorphism of NOS 3 gene is risk factor for retinopathy in type 2 DiabetesMellitus. Individual with TC genotype of NOS 3 gene has lower level of NO than TT genotype.Keywords : Diabetic Retinopathy, Polymorphism, Nitric Oxide, Nitric Oxide Synthase.
Apolipoprotein E as Risk Factor for Coronary Heart Disease Hastuti, Pramudji; M Sofro, Abdul Salam; Asdie, Ahmad Husain; Sadewa, Ahmad Hamim
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

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Allelic variation of apolipoprotein E (apo E) has been shown to infl uence the concentrations of total cholesterol and low density lipoprotein cholesterol (LDL-C) and considered to play a role as one of risk factors for coronary heart disease (CHD). The aim of this study was to examine the relationship between Apo E polymorphism and the risk of CHD. Blood samples were collected from 33 CHD patients in Dr. Sardjito Hospital Yogyakarta, and 38 apparently healthy control individuals in a cross sectional study. The common allelic variants of ApoE were screened employing polymerase chain reaction and restriction fragment length polymorphism. The results obtained were analyzed by t-test and signifi cantly different if p <0.05 and risk factor was calculated by odd ratio. Frequency of ApoE ε2, ε2 and ε4 alleles in CHD patients were 12.1%, 69.7% and 18.2% while in controls were 18.4%, 72.4% and 9.2% respectively. Dyslipidemia condition was a strongrisk factor for CHD. By controlling lipid profi le and applying multifactorial statistic analysis, it was shown that ε4 gene carrier was the risk factor for CHD, but not in triglyceride level, whereas ε2 carrier gene was not the risk factor for CHD. Dislipidemia was the risk factor for CHD and ApoE ε4 gene carrier was the risk factor for CHD.Key words: apolipoprotein E, ApoE ε4 gene carrier, coronary heart disease, dyslipidemia.
Obesitas dan hubungannya dengan polimorfisme gena promoter 5-HT2A, tekanan darah, profil lipid, kadar glukosa, dan malondialdehid Hastuti, Pramudji; Anisah, Nur; Siagian, Johan Iswara; Binuko, Kautsar Prastudia Eko; Adhla, Aidil
Jurnal Gizi Klinik Indonesia Vol 8, No 2 (2011): Oktober
Publisher : Minat S2 Gizi dan Kesehatan, Prodi S2 IKM, FK-KMK UGM

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (448.084 KB) | DOI: 10.22146/ijcn.17772

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Background: Obesity among adults has risen significantly in the world-cutting across all ages, racial and ethnic groups and gender. The 5-HT 2A receptor is crucially involved in regulation of body weight and the appetite. Disturbances in the distribution and or gene regulation of the postsynaptic 5-HT2A receptor are implicated in the pathophysiology of conditions such as obesity, coronary heart disease, high blood pressure, diabetes, stroke dan some cancers.Objective: To test the association between obesity with polymorphism of promoter -1438G→A 5-HT2A receptors gene, blood pressure, lipid profiles, levels of blood glucose and malondialdehyde (MDA).Method: This cross-sectional study included thirty six unrelated obese young people (BMI ≥ 30) recruited from populations in Yogyakarta and 36 controls with age matched with BMI ≤ 25. Statistical differences between blood pressure, lipid profiles, glucose and MDA levels were assessed by t-test and genotypes by Chi square test.Results: There were no significant difference in blood pressure lipid profile, level of glucose and MDA in obese group compared with control (p>0,05). Allele A and G frequency in obese group 25% and 75% respectively, and controls 22.2% and 77.8% respectively, and no significant difference in all two groups, but G allele had higher risk to obese than A allele.Conclusion: These data indicated G allele was mild risk factor for obesity.
RETN rs3745368 polymorphism and resistin level in Javanese ethnic Indonesian obese: a case control study Utami, Rizki Fajar; Hastuti, Pramudji; Sadewa, Ahmad Hamim
Jurnal Teknologi Laboratorium Vol 8 No 1 (2019): 2019 (1)
Publisher : POLTEKKES KEMENKES YOGYAKARTA

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (329.825 KB) | DOI: 10.29238/teknolabjournal.v8i1.164

Abstract

Obesity has become a global public health problem. It occurs due to a positive energy balance leading to adipose tissue expansion. White adipose tissue was an endocrine organ which secreted resistin. Resistin also produced by immune cells due to low chronic level inflammation might cause higher resistin level in obese people. Polymorphism +62G>A RETN gene was reported has a relationship with low resistin level and A allele as a protective allele. This study aimed to determine genotype and allele frequency distribution concerning resistin level. Another objective aimed to know the correlation between resistin level with body mass index. The design of the research was a case-control study with 122 people (18-40 y.o.), divided equally in the case group (BMI ≥ 27 kg/m2) and control group (BMI 18.5-24.9 kg/m2) without diabetes mellitus. Blood was taken after fasting a minimal 8 hours. Plasma was used to measure the resistin level. DNA genotyping was analyzed using PCR-RFLP. Genotyping result showed three genotypes of RETN gene +62G>A polymorphism (GG, GA, AA). There was no significant difference in genotype and allele frequency distribution related to obesity status (p=0.680; p=1) and resistin level (p=0.537) between case and control group. There was no significant difference in resistin level between case and control group (p=0.770). Resistin level was correlated with BMI in obese group (p= 0.05; r= -0.25). The present study concludes that there is no significant difference in genotype and allele frequency distribution related to obesity status and resistin level. Resistin level has a negative correlation with BMI.
Vitamin D Status in Madura Pregnant Women with Hypertension: A Case Control Study Setiarsih, Dini; Hastuti, Pramudji; Nurdiati, Detty Siti; Putri, Lidia Aditama
Journal of Maternal and Child Health Vol. 7 No. 1 (2022)
Publisher : Masters Program in Public Health, Universitas Sebelas Maret, Indonesia

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Background: Vitamin D deficiency is inversely related to blood pressure and may contribute to the genesis and maintenance of hypertension. However, in many clinical studies the relationship between vitamin D status and blood pressure has not shown consistent results. This study aims to analyze differences in vitamin D status in Madurese ethnic pregnant women with hypertension and non-hypertension.Subjects and Method: This research is an analytical observational design with a case control study. The target population in this study were pregnant women who came from the Madurese ethnicity. The affordable population is pregnant women who come from the Madurese ethnicity in the Work Area of the Bangkalan District Health Office. The study was conducted in the period from May 2018 to February 2020. The sampling technique used was consecutive sampling. The sample size was 105 pregnant women with hypertension and 105 controls. The independent variables included consumption of food sources of vitamin D and exposure to ultraviolet B rays. The dependent variable was vitamin D levels. Data on consumption of foods that were sources of vitamin D was obtained using the Food Frequency Questionnaire (FFQ). Data on exposure to ultraviolet B (UV B) rays were obtained by questionnaire. Data on vitamin D levels were obtained from the collection of blood serum which was then analyzed by ELISA Kit. Independent Sample T and chi square test were used to analyze the differences in variables between the two groups.Results: The mean value of vitamin D levels in the case and control groups were 37.87 ng/mL and 37.89 ng/mL, respectively. Subjects with vitamin D insufficient status were more in the case group (19.00%) than the control group (16.20%). The results showed that there was no significant difference between the two groups in vitamin D levels (p= 0.984) and vitamin D status (p= 0.587). Insufficient exposure to UV B provides a 3.95 times greater risk of experiencing insufficient vitamin D.Conclusion: The conclusion of this study is that the role of vitamin D in the incidence of hypertension in pregnancy has not been determined.Keywords: hypertension pregnancy, pregnancy, UV B exposure, consumption patterns, vitamin DJournal of Maternal and Child Health (2022), 07(01): 1-8https://doi.org/10.26911/thejmch.2022.07.01.01