13th balkan biochemical biophysical days & meeting on metabolic disorders’ programme & abstracts

Dokuz Eylül University School of Medicine, Departments of Biochemistry¹ and Pathology², Ege University School of Medicine, Department of Gastroenterology, Izmir, Turkey. e-mail: filiz.kuralay@deu.edu.tr

Induction of colitis by acetic acid(AA) in the rat is widely used experimental model of inflammatory bowel disease(IBD) and ulcerations. AA as an irritant induces colitis involving infiltration of colonic mucosa with neutrophils and increased production of inflammatory mediators, such as hydrogen peroxide(H₂O₂), nitric oxide(NO), myeloperoxidase activity(MPO), tumor necrosis factor(TNF-) levels. Trimetazidine(TMZ), an antianginal compound, was administered to investigate if its cytoprotective features in cardiac tissue are also effective in AA-colitis where ischemic injury contributes to colitis.

Administration of TMZ via IP improved the macroscopic and microscopic score alterations produced by AA. AA administration significantly elevated colonic MPO activities, however treatment with TMZ significantly lowered this enzyme activity compared to AA. AA administration significantly enhanced SOD activities, except for AA+TMZ-IR. TMZ treatment significantly lowered nitrate levels; but increased these levels. As for the TNF- levels, AA administration markedly lowered TNF- levels, but TMZ treatment elevated these levels to control.

This result supports the findings that overproduction of NO may be involved in the immunosuppression observed during acute AA-induced rat colitis. In conclusion, TMZ treatment was more effective via the IP compared to IR route, and may be beneficial in therapy of colitis.

MOLECULAR PATHOLOGY OF CYP1B1 GENE IN TURKISH PATIENTS

Sefayet BAGİYEVA¹, Rıza Köksal ÖZGÜL¹, Sinan M. SARICAOĞLU ², Cihan ÖNER¹ and Ay ÖĞÜŞ¹

sefaet@hacettepe.edu.tr

1 Hacettepe University, Department of Molecular Biology, Beytepe- Ankara,Turkey

2 Numune Research and Training Hospital, Department of Ophthalmology, Ankara,Turkey

Primary Congenital Glaucoma (PCG) or Buphthalmos (GLC3) is an autosomal recessive disorder, associated with unknown developmental defect(s) in the anterior chamber and manifests itself in early childhood, usually within the first year of life. The responsible gene for PCG phenotype is CYP1B1, the only known member of cytochrome P450 I subfamily of CYP. This gene has been reported to be responsible from 85% of cases in buphthalmos. In this study we investigated CYP1B1 gene mutations in the first locus (GLC3A), mapped to chromosome 2p21 in Turkish patients.

DNA samples were isolated from total of 18 PCG subjects. CYP1B1 gene was amplified by PCR. Nucleotide sequence of patients who revealed abnormal pattern in SSCP, were screened by DNA Sequence Analysis.

Two different mutations were detected in CYP1B1 gene in buphthalmos patients. The mutations are; 3987 G→A (G61E) in exon 2 and 8242 C→T (R469W) in exon 3. The frequencies of these mutations in Turkish patients are 11%. We also detected five different polymorphisms in different combinations (3947 cgg/ggg R48G; 4160 gcc/tcc A119S, 8131 gtg/ctg V432L; 8195 aac/agc N453S; 8184 gat/gac silent 449) in screened individuals.

The detection of the mutations in CYP1B1 gene will be helpful in early diagnosis of the disease, further understanding of its genetic base and the role of CYP1B1 gene in development and differentiation.

ATHEROSCLEROTIC POLYMORPHISMS IN POSTMENOPAUSAL WOMEN WITH ESTABLISHED CORONARY DISEASE

Lale AFRASYAP¹, Ibrahim BARIS<sup>2

1</sup>Mugla University, Health High School, Department of Biochemistry, Mugla ;

² Bogazici University, Department of Molecular Biology and Genetic, Istanbul/ TURKEY

laleafrasyap@hotmail.com

The incidence of coronary disease risk due to atherosclerosis is higher in men and postmenopausal women than in premenopausal women. Although the polymorphisms of the MTHFR (C677T and A1298C) and eNOS (G894T) genes were investigated in different population groups with coronary disease, very few studies have addressed about the association between these polymorphisms and coronary disease in postmenopausal women. The aim of study is to investigate if genetic mutations increase the risk of coronary disease in postmenopausal women. The study was organized for 40 postmenopausal women with an intact uterus. They were divided into two groups, according to angiography results. 1- 25 women with >50% stenosis affecting at least one artery were included in group with coronary heart disease (patients) 2-15 women with < 20 % stenosis were enrolled in group without disease (controls). Mean ages of patients and controls were 64,06±8,65 and 66,12±6,80, respectively. After DNA was extracted from whole blood samples with salting-out method, genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Statistical analyses were computed by SPSS 11,5 version, using nonparametric tests. Although the prevalences of 1298CC and 1298CC/AC were higher in patients with respect to controls (p=0.009; p=0,016, respectively),the significant difference was not observed in the prevalences of the other genotypes between the groups. There was the positive correlation between coronary disease and the frequency of 1298CC ( r=0,447 p=0,017) The odds ratio was 1,71 (p=0,038, 95% CI, 1,00 to 2,92) in the patients with 1298CC mutation with respect to without. It was also 1,71 ( p=0,026, 95% CI, 1,00 to 2,66) for 1298 CC as compared with 1298 AA/ AC combination. The high prevalence of the 1298 CC genotype might be effective on the genesis of the disease itself and an important risk factor in the occurrence of coronary disease in postmenopausal women.