ASSOCIATION BETWEEN SMOKING BEHAVIOUR AND GLYCOHEMOGLOBINE LEVELS AMONG ADULT JAVANESE INDONESIAN SMOKERS

Nicotine, the active compound in cigarettes, can cause impaired glucose metabolism byincreasing insulin resistance as well as decreasing insulin secretion in ? cell pancreas. Thiscondition can increase the risk of type 2 diabetes in human. This study aims to evaluate the effectof smoking behaviour, determined by Cigarette per Day (CPD) and smoking duration, onglychohemoglobine (HbA1c) levels of Javanese Indonesian smokers. 30 smokers were studiedconsisting of 7 smokers with 10 CPD, 19 smokers with 11-20 CPD and 4 smokers with 21-30CPD. They had been smoking for more than 10 years. The whole blood sample was used toexamine the HbA1c levels. The HbA1c levels were tested at Bethesda Hospital's cliniclaboratories using Architect 600 instrument. The results showed that CPD and smoking durationsignificantly influenced HbA1c, in which F count was F table (370.541 3.354) withsignificance 0.05 (2.35. 10-20 0.05) and multiple correlation coefficient (R) of 0.982.Therefore, based on this research finding, it was concluded that longer smoking duration andhigher CPD caused higher smokers HbA1c level

CYP2A6*4 allele gene high frequency associated with low-density lipoprotein cholesterol (LDL-C) among Javanese Indonesian smokers

The CYP2A6 gene, which codes the CYP2A6 enzyme, has known to have ahigh polymorphism. This polymorphism could decrease, increase, or eliminate the CYP2A6 enzyme activity. CYP2A6*4, an inactive allele, decreased the CYP2A6 enzyme activity. One of the CYP2A6 enzyme-specific substrates is nicotine. This inactive allele could decrease nicotine metabolism that causes high nicotine levels in the blood. In addition, it caused the increasing levels of Low-Density Lipoprotein Cholesterol (LDL-C) by expanding the lipolysis process. The purpose of this research was to evaluate the effect of the CYP2A6*4 allele gene on LDL-C levels. Respondents in this study were 31 male Javanese smokers. This research is an analytic observational study with a cross-sectional design. Polymerase chain reaction (PCR) methods use to identification the CYP2A6*4 allele gene. This study shows that a high-frequency CYP2A6*4 alleles gene among the subject was detected, with an allele frequency is 93.55%. Furthermore, this CYP2A6*4 allele gene did not impact LDL-C levels, with the Odd Ratio value was 1.636 (P-Value = 0.737). In conclusion, the CYP2A6*4 allele gene does not significantly affect the LDL-C levels in Javanese Indonesian smokers

Distribution of cytochrome P450*4 (CYP2A6*4) allele gene among Javanese Indonesian T2DM patients

Smoking had been increasing the risk factor of type 2 diabetes mellitus (T2DM), both active and passive smokers, which is caused by nicotine contained in cigarettes. Nicotine has metabolized by cytochrome p450 2a6 (CYP2A6) enzyme coded by the CYP2A6 gene. This gene was a high polymorphism that is the CYP2A6*4 allele gene was inactive. Thus, our objective was to describe the CYP2A6*4 allele gene among active and passive Javanese smokers with T2DM. From this cross-sectional study, we identified this allele gene among 46 of the adults with T2DM, which were consist of 23 active smokers and 23 passive smokers. The CYP2A6*4 allele gene identification has done using polymerase chain reaction (PCR) methods. The CYP2A6*4 allele frequency was analyzed to describe the distribution of this allele among the participants. This study supports the hypothesis that smoking, including cigarette smoke, was an environmentally modifiable risk factor for developing T2DM. Based on our result, the allele frequency among the participants was 42.39%. A high frequency of the CYP2A4 allele gene among the participants was indicating that the CYP2A4 allele gene was also the other risk factor in developing T2DM

POLYMORPHISM OF CYTOCHROME P450 2A6 (CYP2A6*1 AND CYP2A6*4) AMONG JAVANESE INDONESIAN SMOKER AND NON SMOKER

Cytochrome P450 2A6 (CYP2A6) is the principal enzyme involved in the metabolic activation of tobacco-specific nitrosamines to their ultimate carcinogenic forms and metabolism of nicotine. The present study was developed to investigate the genetic polymorphism of CYP2A6 in Javanese Indonesian subjects carrying the CYP2A6*1 allele and the CYP2A6*4. The whole gene deletion of CYP2A6 (CYP2A6*4) may inhibit smokers from giving up smoking, but appears to function as a protective factor against to some cancer. However, the investigation of these allele, a major functional polymorphisms common in Asian populations, have not been reported among Javanese Indonesian population. A single polymerase chain reaction-restriction fragment length polymorphism was used to resolve the genotypes into CYP2A6*1 (wild type) and CYP2A6*4 (CYP2A6del). The sample studied consisted of 100 healthy subject that consist of 50 non smokers and 50 smoker from Javanese Indonesian population. The allele frequencies of *1 (wild type) and *4, were 47.5 and 52.5%, respectively. When the two allel were considered simultaneously, among the non-smokers, 45% were genotyped for CYP2A6*1/*4 and 5% were genotyped for CYP2A6*4/*4; on the other hand all of the smoker were genotyped for CYP2A6*1/*4 and there was no homozygote of wild type. Based on the data collected, it could be concluded that the polymorphism of CYP2A6 were detected in among Javanese population sample study and the allele frequencies of CYP2A6*4 were high.Key word: Polymorphism, CYP2A6*1, CYP2A6*4, Javanese Indonesia

PEMERIKSAAN PROFIL LIPID PADA JEMAAT LANSIA SALAH SATU GEREJA KRISTEN DI YOGYAKARTA

Lipid profile examination including total cholesterol level, triglycerides level and high-density lipoprotein cholesterol reduction is essential to be done as a routine activity quinquennially to prevent someone from the risk of cardiovascular diseases, diabetes and obesity. This kind of examination is mandatory for the elderly people to maintain their quality of life. In order to support the government program on the prevention of the cardiovascular diseases, a team of social service of Faculty of Pharmacy Universitas Sanata Dharma (FFUSD) was organizing an event of lipid profile examination in one of Christian church located in Kalasan, Yogyakarta. As many as 35 elderly people were enthusiastically participating on the event. A nurse from Poliklinik Sanata Dharma was appointed to take the blood samples, which were further taken to the Biochemistry Laboratory in FFUSD to get analysed. The results were sent to the participants in private. The overall blood lipid profiles of most participants showed good level of quality, indicating that they have been trying to manage their life healthily. However, as much as 62. 8% of participants showed a low level of the high-density lipoprotein cholesterol.  This was believed due to the food intake which was still rich in oil and fat.  The event was very succesful and the responses were very positive. The participants felt assisted with a fast and accurate results sent

Distribution of cytochrome P450*4 (CYP2A6*4) allele gene among Javanese Indonesian T2DM patients

Smoking had been increasing the risk factor of type 2 diabetes mellitus (T2DM), both active and passive smokers, which is caused by nicotine contained in cigarettes. Nicotine has metabolized by cytochrome p450 2a6 (CYP2A6) enzyme coded by the CYP2A6 gene. This gene was a high polymorphism that is the CYP2A6*4 allele gene was inactive. Thus, our objective was to describe the CYP2A6*4 allele gene among active and passive Javanese smokers with T2DM. From this cross sectional study, we identified this allele gene among 46 of the adults with T2DM, which were consist of 23 active smokers and 23 passive smokers. The CYP2A6*4 allele gene identification has done using polymerase chain reaction (PCR) methods. The CYP2A6*4 allele frequency was analyzed to describe the distribution of this allele among the participants. This study supports the hypothesis that smoking, including cigarette smoke, was an environmentally modifiable risk factor for developing T2DM. Based on our result, the allele frequency among the participants was 42.39%. A high frequency of the CYP2A4 allele gene among the participants was indicating that the CYP2A4 allele gene was also the other risk factor in developing T2DM

Uji toksisitas zat warna azo turunan odianisidin terhadap saccharomyces cerrevisae sebagai model uji ekotoksikologis = Toxicity test of O-dianisidine Azo Dyes to Saccharomyces cerrevisiae as A Model Ecotoxikological Test

O-dianisidine azo dyes are used in coloring processes of Indonesian batik industries. The toxicity of azo compuond is defined by its degradation, i.e. amine aromatic compound, nevertheless the toxicity of its parent compound is still unknown. There are posibilities to describe ecotoxicological interactions of xenobiotic using QSAR-model. The aim of this study is having more information of cause-effect relationship of xenobiotic o-dianisidine azo dyes toward Saccharomyces cerrevisae, as an eucaryotic model. The experiment begin with lipophilicity parameter determination (log k octanol/water, and log k paraffine/ water) followed by ecotoxicological parameter (EC 50). The determination of lipophilicity was carried out in two waysfirst, it using calculation based on the concept of additivity group contribution, developed by Leo-Hansch, to obtain its log k octanolsecond determination was carried in RPTLC (reversed phase thin Fayer chromatography) using paraffine as stationary phase and mixtures of aseton-water as mobile phase, to obtain log k octanol/water . EC 50 was obtained by measuring and calculating 50% Saccharomyces cerrevisae growth rate inhibition. Meanwhile, the possibilities of azo dyes degradation was studied with or without Saccharomyces cerrevisae. The calculated log k octanol/water of o-dianisidine azo dyes studied was 13 to 14, therefore it is considered as superhydrophobic compound and has difficulties to pass the cell wall of Saccharomycess cerrevisaenevertheless, it has EC 50 of 140 to 150 ppm. No degradation in media without Saccharomycess cerrevisae was observed, but there is azo dyes concentration decrease in the media containing Saccharomycess cerrevisae. The study on the cause of the decreasing concentration revealed that physically adsorption of azo dyes on the cell of Saccharomycess cerrevisae occured, according to Langmuir-Hinshelwood adsorption kinetics with k, = 5.67e3 to 6.79e3 mol/sec and Eads = 19.23 to 19.75 kj/mol. The evidence of physical adsorption was supported by the small CF value = 0.46 to 0.78. The azo dyes prefer to dissolved in organic solven, showed by log k paraffine/water between 3-4. Therefore, there is a linier correlation between log k paraffine/water with EC50(Y = -17.84X + 220.55). This correlation can be used to explain the dermatitis reaction on skin too. Keywords: Ecotoxicological, EC50, adsorption, azo dye, QSAR, lipophilicit

Metode Kromatografi Cair Kinerja Tinggi Menggunakan Kolom Oktil Silika Fully Endcapped Residual Silanol pada Pemisahan Kotinin dan 3-Hidroksikotinin dalam Sampel Urin

Cotinine (COT) and 3-hydroxycotinine (3-HCOT) are nicotine metabolite excreted in urine. Mediated by the enzyme cytochrome P450 2A6 (CYP 2A6), nicotine will be metabolized to COT and 3-HCOT. The activity of CYP 2A6 can be predicted from the ratio 3-HCOT to the COT, therefore the ratio of 3-HCOT and COT can be used as phenotyping and polymorphism studies of the enzyme. In this study, isolation COT and 3-HCOT of urine samples was carried out by liquid-liquid back extraction. Simultaneous analysis of COT and 3-HCOT using High Performance liquid Chromatography (HPLC) was performed by a reversed-phase octyl silica column (C8; Shimadzu 250 × 4.6 mm, 5 μm) fully endcapped residual silanol. The internal standard solution (SI) was acetanilide. The mobile phase which separate COT, 3-HCOT and SI was methanol : ammonium acetate 5 mM (50:50) at a flow rate 0.8 mL/min. Retention time (tR) of the three compounds was less than 10 minutes, with peak tailing factor (tf) was less than 2. The resolution (Rs) 3-HCOT to COT was 2.67, while the Rs COT to SI was 8.836.Kotinin (COT) dan 3-hidroksikotinin (3-HCOT) merupakan metabolit nikotin yang dikeluarkan melalui urin. Perubahan nikotin menjadi COT dan 3-HCOT diperantarai oleh enzim sitokrom P450 2A6 (CYP 2A6). Akitivitas enzim ini dapat diprediksi dari nilai rasio kadar 3-HCOT terhadap COT. Oleh karena itu nilai rasio 3-HCOT terhadap COT banyak digunakan pada studi fenotipe dan polimorfisme enzim tersebut. Pada penelitian ini, isolasi COT dan 3-HCOT dari sampel urin dilakukan dengan teknik isolasi cair-cair secara bertingkat. Analisis simultan COT dan 3-HCOT dilakukan dengan metode Kromatografi Cair Kinerja Tinggi (KCKT) fase terbalik menggunakan kolom oktil silika (C8; Shimadzu 250×4,6 mm, 5μm) fully endcapped residual silanol. Larutan standar internal (SI) yang digunakan adalah asetanilid. Sistem fase gerak yang dapat memisahkan COT, 3-HCOT dan SI adalah campuran metanol : amonium asetat 5mM (50:50) pada kecepatan alir 0,8 mL/menit. Nilai waktu retensi (tR) ketiga senyawa kurang dari 10 menit, dengan nilai peak tailing factor (tf) kurang dari 2. Nilai resolusi (Rs) 3-HCOT terhadap COT adalah 2,67, sedangkan nilai Rs COT terhadap SI adalah 8,8