Cox-2 gene polymorphism and IL-6 levels in coronary artery disease.

Coronary artery disease is one of the leading causes of mortality and diabetes mellitus is one of its main risk factors due to microvascular and macrovascular complications, such as atherosclerosis. Atherosclerosis is now known to be an inflammatory process mediated by prostaglandins and several interleukins. As both are important in inflammatory processes, we examined Cox-2 (-765G > C) polymorphism and interleukin-6 levels in coronary artery disease patients compared to healthy controls. We also divided the patients into diabetic and non-diabetic groups to check the effects of diabetes mellitus separately. We found that the GG allele frequency was significantly higher in the patient group. Patients with the GG genotype had an approximately 2.78-fold higher risk of coronary artery disease. We also found that the Cox-2 (-765G > C) polymorphism is associated with lower interleukin-6 levels, which decreased in the order: GG > GC > CC.

DNA repair genes in endometriosis.

Several polymorphisms in the DNA repair gene are thought to have significant effects on cancer risk. We investigated the association of polymorphisms in the DNA repair genes XRCC1 Arg399Gln, XRCC3 Thr241Met, XPD Lys751Gln, XPG Asp1104His, APE1 Asp148Glu, and HOGG1 Ser326Cys with endometriosis risk. Genotypes were determined by PCR-RFLP assays in 52 patients with endometriosis and 101 age-matched healthy controls. Although there were no significant (P > 0.05) differences in the frequencies of genotypes or alleles of APE1, XRCC1, XPD, XPG, and HOGG1 genes between patients and controls, the frequency of the XRCC3 Thr/Thr genotype was significantly greater in endometriosis patients compared with controls (P = 0.005). XRCC3 Thr/Met genotypes (P = 0.022), and the Met allele (P = 0.005) seem to have a protective role against endometriosis. The distributions of genotypes and alleles of the genes APE1, XRCC1, XRCC3, XPD, XPG, and HOGG1 were not significantly associated with the different stages of endometriosis (P > 0.05). We conclude that the XRCC3 Thr/Thr genotype is associated with endometriosis in Turkish women.

CCR2-64I is a risk factor for development of bladder cancer.

Chemokines are potent proinflammatory cytokines that are implicated in numerous inflammatory diseases. Proinflammatory gene polymorphisms lead to variations in the production and concentration of inflammatory proteins. We investigated a possible association between polymorphisms in chemokine and chemokine receptor genes (MCP-1 A-2518G and CCR2-V64I) and bladder cancer risk. Genotypes were determined by PCR-RFLP assays in 72 bladder cancer patients and 76 unrelated age-matched healthy controls. There were significant differences in the frequencies of the MCP-1 A-2518G (P = 0.012) and CCR2-V64I genotypes (P = 0.004) between the controls and patients. The MCP-1 A-2518G GG genotype frequencies for controls and cases were 0.039 and 0.11, respectively; individuals who had the GG genotype had a 3-fold increased risk of bladder cancer (P = 0.08). The CCR2-64I/64I genotype frequencies for controls and cases were 0.02 and 0.13, respectively; subjects carrying the 64I/64I genotype had a 5.9-fold increased risk of bladder cancer compared to the other genotypes. Individuals carrying the CCR2-V64I heterozygote or homozygous variant genotype (64I/64I + wt/64I) had a 2.9-fold increased risk of bladder cancer compared with the wild-type genotype (wt/wt). CCR2-V64I heterozygote or homozygous wild-type genotype (wt/wt + wt/64I) frequencies were significantly decreased in the patient group compared with controls. We conclude that CCR2-64I is a new risk factor for bladder cancer.

Paraoxonase (PON1) 55 and 192 polymorphism and its effects to oxidant-antioxidant system in turkish patients with type 2 diabetes mellitus.

Paraoxonase (PON1) is a serum enzyme with an antioxidant function, protecting the low density lipoproteins (LDL) from oxidative modifications. Because diabetic patients are at greater risk of oxidative stress, we investigated the effect of PON1 55 methione (M)/leucine (L) and PON1 192 glutamine (A)/arginine (B) polymorphisms on oxidant-antioxidant system in 213 individuals with type 2 diabetes mellitus and 116 non-diabetic control subjects from Turkish population were included in the study. Polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and agarose gel electrophoresis techniques were used to determine the PON1 genotypes. Thiobarbituric acid reactive substances (TBARS), conjugated dienes levels in the serum and glutathione (GSH) levels in whole blood were measured spectrophotometrically. In both groups PON1 192 AA and PON1 55 MM genotypes had higher TBARS, conjugated dienes levels and lower GSH levels, whereas PON1 192 BB and PON1 55 LL genotypes had lower TBARS, conjugated diene levels and higher GSH level than other genotypes. We thus conclude that PON1 192 BB and PON1 55 LL alleles have protective effect against oxidative stress.

The effects of Brassica oleraceae var capitata on epidermal glutathione and lipid peroxides in DMBA-initiated-TPA-promoted mice.

BACKGROUND: The objective of the present study was to determine if modulation of GSH-dependent antioxidant protective system by Brassica oleraceae var capitata might inhibit the molecular mechanism of skin tumor promotion. MATERIALS AND METHODS: In a two stages skin carcinogenesis model, the protocol used included a single topical application of 200 nmol of the initiator 7,12-dimethyl-benz(a) anthracene (DMBA) to the backs of mice, followed 1 week later by promotion with 10 nmol of 12-O-tetradecanoyl-phorbol-13 acetate (TPA) twice weekly for 30 weeks. In addition to this regimen, 0.1 g/L brassica extract was added water week prior to the initiating dose of DMBA in the treatment group. Tissue glutathione (GSH) contents and levels of lipid peroxidation products (measured as thiobarbituric-acid (TBA)-reactive substances) were quantitated in the skin tumors generated by the initiation-promotion protocol. RESULTS: It was observed that the tumor incidence and tumor multiplicity in the treatment group was highly significantly low compared to the first group of mice (p < 0.001 and p < 0.001, respectively). In the treatment group, GSH content in the papillomas was higher than in the non-involved skin surrounding the papillomas. CONCLUSIONS: We suggest that the anticarcinogenicity of Brassica may be linked to its ability to facilitate or enhance the activity of the natural GSH-dependent antioxidant protective system of the epidermal cells during the later stages of skin tumor promotion.

Is there additional effect of MTHFR C677T mutation on lipid abnormalities in renal allograft recipients?

BACKGROUND: The MTHFR C677T mutation and elevated atherogenic lipoprotein levels are known as cardiovascular risk factors in patients with renal transplantation treated with cyclosporine (CsA). The aim of the present study was to eveluate the contribution of MTHFR C677T mutation to the risk of dyslipidemia in renal transplant recipients. We also studied the effect of the MTHFR-C677 T genotype on transplant survival. METHODS: The study included 29 nondiabetic renal transplant recipients and 27 healthy controls. MTHFR C677T genotypes were determined by PCR and RFLP techniques. Biochemical parameters were measured in a computerized autoanalyzer. RESULTS: In the patient group, the distribution of the CC, CT, and TT genotypes was 44.8% (n = 13), 37.9% (n = 11), and 17.2% (n = 5), respectively. The frequencies of the C and T alleles were 0.64 and 0.36, respectively. Subjects with the T allele had the highest levels of TC (P <.05) and LDL-C (P <.05); subjects with the CC genotype had the lowest. CONCLUSIONS: We observed that the MTHFR T allele has an unfavorable effect on serum lipid profile, leading to a rise in the total and LDL cholesterol concentrations. Thus, we believe that MTHFR C allele has a protective effect and MTHFR T allele has a detrimental effect on the serum lipid profile.

Paraoxonase 192 polymorphism and its relationship to serum lipids in Turkish renal transplant recipients.

BACKGROUND: Cardiovascular disorders are the leading cause of death in patients with chronic renal insufficiency. Paraoxonase (PON1) gene variants have been identified as risk factors for cardiovascular disease (CVD). METHODS: We investigated the effect of PON1 192 polymorphisms on serum lipid profiles in 29 renal transplant recipients and 26 control subjects. Distribution of the PON1 192-gene polymorphism was determined by polymerase chain reaction-based restriction fragment length polymorphism. Serum lipid levels were measured enzymatically. RESULTS: Frequencies of PON1 192 AA, BB, and AB genotypes among the renal transplant recipients were 38.5%, 26.9%, and 34.6%, and among control subjects they were 35.75%, 17.9%, 46.4%, respectively. The genotype distribution for the PON1 192 polymorphism was not different between the two groups (P = NS, chi-square test). The PON1 192 polymorphisms failed to consistently influence the serum lipid profiles in renal transplant recipients. CONCLUSION: We have shown that the 192 polymorphism of the PON1 gene is not associated with increased cardiovascular risk in renal transplant recipients.

Interaction between apolipoprotein-E and angiotensin-converting enzyme genotype in Alzheimer's disease.

Both apolipoprotein-E (apo-E) epsilon 4 allele and angiotensin-converting enzyme (ACE) deletion (D) polymorphism have been associated with a high risk for coronary heart disease. Increased frequency of the epsilon 4 allele has also been reported in patients with late-onset of familial and sporadic Alzheimer's disease (AD). The primary aim of this study is to examine the possible relationship between the ACE gene polymorphism and AD. The second aim of this study is to explore the relation of the ACE and apo-E genotypes with AD. Polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and agarose gel electrophoresis techniques were used to determine the apo-E and ACE genotypes. The frequencies of ACE D and ACE insertion (I) allele among AD patients and controls were 55.7 percent versus 44.2 percent and 51.7 versus 48.2 percent, respectively. Apo-E allele frequencies in the AD group for epsilon 2, epsilon 3 and epsilon 4 were, 1.7 percent, 96.5 percent, and 1.7 percent, respectively. The apo-E allele frequencies of healthy groups for epsilon 2, epsilon 3 and epsilon 4 were 1 percent, 56 percent, and 1.7 percent, respectively. In conclusion ACE D and apo epsilon 4 allele were found to be more frequent in patients with Alzheimer's disease than in the control group.

Apolipoprotein-E gene polymorphism and lipid profiles in Alzheimer's disease.

In this study, the relationship between lipid profiles of sera and apolipoprotein E (apo E) gene polymorphism was investigated in 35 patients with Alzheimer's disease (AD) and 29 healthy people. Apo E genotypes and allele frequencies of the AD patient group were: apo E2/3, 2 (5.7 percent); apo E2/4, 1 (2.9 percent); apo E3/3, 26 (74.3 percent); apo E3/4, 5 (14.3 percent); apo E4/4, 1 (2.9 percent); epsilon 2, 3(4.2 percent); epsilon 3, 59 (84.2 percent); epsilon 4, 8 (11.4 percent). The healthy group's apo E genotypes and allele frequencies were: apo E2/3, 1 (3.4 percent); apo E3/3, 27 (93.1 percent); apo E3/4, 1 (3.4 percent); epsilon 2, 1 (1.7 percent); epsilon 3, 56 (96.5 percent); epsilon 4, 1 (1.7 percent). In Alzheimer's cases, epsilon 4 allele frequencies increased significantly as compared to the healthy group (p < 0.05). When the effects of the apo E isoforms on lipid profiles were evaluated, a relationship between apo E epsilon 4 allele and high total levels of serum cholesterol was found, whereas of apo E epsilon 2 allele was associated with the low total cholesterol of serum, although the difference was not statistically significant (p > 0.05). This study confirms the association of apo E epsilon 4 allele with lipid profiles in AD patients.

Cholesterol ester transfer protein, apolipoprotein E and lipoprotein lipase genotypes in patients with coronary artery disease in the Turkish population.

The aim of this study was to compare patients with coronary artery disease (CAD) to healthy objects, in order to explore a possible association between CAD and the variants in the gene encoding cholesterol ester transfer protein (CETP), apolipoprotein E (Apo E) and lipoprotein lipase (LPL). The relationship between CETP MspI, apo E and LPL PvuII gene polymorphisms and serum lipids were investigated in 173 patients with CAD and 111 healthy controls. The frequency of Apo epsilon4 (p < 0.05) and CETP M1 (p < 0.01) alleles were higher in the CAD group than in the control group. In the CAD group, those with the Msp M1 allele had higher levels of total cholesterol (TC) (p = 0026) and low-density lipoprotein cholesterol (LDL-C) than those with the Msp M2 allele. Subjects with an epsilon2 allele had the lowest levels of TC and LDL-C, while subjects with the epsilon4 allele had the highest. In the control group, CETP, the Msp M2 allele was associated with a higher level of high-density lipoprotein cholesterol (HDL-C) (p = 0.012) than the Msp M1 allele. The distributions of LPL genotype and allele did not differ between the CAD and control groups. The present study demonstrates that the CETP Msp1 and Apo E gene polymorphisms are associated with variations in lipids in patients with CAD and healthy controls in Turkish population.

Is epsilon4 allele of apolipoprotein E associated with more severe end-organ damage in essential hypertension?

The aim of the present study comparing patients with mild to moderate hypertension with controls, was to explore a possible association between hypertension-related target organ damage and evaluation found in the gene encoding apolipoprotein E (apo E) genotype. Detailed medical history was recorded and physical examination was performed for all patients in the study (88 hypertensives, 63 normotensive controls). PCR (Polymerase Chain Reaction), RFLP (Restriction Fragment Length Polymorphism), and agarose gel electrophoresis techniques were used to determine the apo E genotypes. The frequencies of apo epsilon2, apo epsilon3, and apo epsilon4 alleles were 3.97, 88.06, and 9.95%, respectively in the hypertensive group. The frequencies of apo epsilon2, apo epsilon3, and apo epsilon4 alleles were 5.5, 92.0, and 2.38%, respectively in the control group. There were about twice as many individuals in the heterozygote hypertensive group who had apo E3/4 as compared to the control group (7.30 vs. 2.38%) (p = 0.07). The hypertensive patients who were carriers of the apo epsilon4 had significantly higher organ damage (left ventricular hypertrophy (p < 0.001). dilated left atrium (p < 0.05), retinopathy (p < 0.05)) as compared to those who were not carriers of apo epsilon4. These results showed a trend for the epsilon4 allele to be associated with a higher prevalence of target organ damage in patients with mild to moderate hypertension.

Increased plasma levels of interleukin-6 and interleukin-8 in beta-thalassaemia major.

Several immunological defects can be found in patients with beta-thalassaemia, among which the impairment of neurophil and macrophage phagocytic and killing functions and the production of some cytokines are the most important. It is known that interleukin-6 (IL-6) and interleukin-8 (IL-8) are important components of the pro-inflammatory response. The plasma levels of these cytokines may be relevant in the pathophysiology of beta-thalassaemia. To assess this hypothesis, the plasma IL-6 and IL-8 concentrations in patients with beta-thalassaemia, were investigated. Fourteen patients with thalassaemia major were studied by evaluating body iron status, iron supply for erythropoiesis, and plasma IL-6 and IL-8 levels, together with 12 age-matched healthy controls. The plasma levels of IL-6 and IL-8 were determined by enzyme-linked immunosorbent assay (ELISA). Patients with beta-thalassaemia were found to have higher IL-8 concentrations than normal controls (p < 0.001) and plasma IL-6 concentrations increased significantly in the beta-thalassaemic patients compared with control subjects (p = 0.01). Serum ferritin levels of beta-thalassaemic patients were significantly higher than those of control groups (p < 0.05). IL-8 levels correlated with ferritin levels (r = 0.694; p < 0.05) and the total number of transfusions (r = 0.64; p < 0.05). Plasma IL-6 levels in beta-thalassaemic patients did not correlate with any clinical, haematological or biochemical parameters. It was also found that plasma IL-8 levels in the patients who had blood transfusions over 100 times were significantly higher than those of under 100 times (p < 0.05), whereas there was no statistical difference for IL-6. Markedly increased plasma IL-6 and IL-8 levels were documented in patients with beta-thalassaemia. Increased production of IL-6 and IL-8 might have contributed to abnormalities in iron metabolism and it is probably due to overstimulation of macrophages. Before a clinical value can be ascribed to these changes in plasma cytokine levels in beta-thalassaemia, the follow-up samples of larger series of patients with 8-thalassaemia should be evaluated.

Age-related changes in GM1, GD1a, GT1b components of gangliosides in Wistar albino rats.

In this study, age-related changes of GM1, GD1a, GT1b fractions of gangliosides were investigated in whole brain of male Wistar albino rats. Insignificant increases were detected in GM1 values from the third to the 24th month, whereas GD1a and GT1b concentrations of ganglioside in 24-month-old rats decreased significantly as compared to 6-month-old rats. Although there were no significant differences in the GD1a/GT1b ratio of any groups, GM1/GD1a and GM1/GT1b ratios were significantly increased as compared to 6-month-old rats. The increase in the ratios of gangliosides are not due to an increase of GM1 fractions; they result from a decrease of GD1a and GT1b fractions of gangliosides. In conclusion, the concentration of ganglioside decreased with ageing.

Association between angiotensin-converting enzyme gene polymorphism and coronary artery disease.

An insertion/deletion (I/D) polymorphism in the gene for angiotensin-converting enzyme (ACE) is associated with myocardial infarction and other cardiac pathology. There is evidence for a role of the renin-angiotensin system in cell growth and in the repair of damaged arterial walls, so the ACE gene is postulated to be a candidate gene affecting the important clinical problem of coronary artery disease (CAD). In view of the clinical importance of the ACE as a major marker of cardiovascular diseases, we investigated the I/D polymorphism of the ACE gene in Turkish CAD patients in comparison with control subjects to evaluate a possible association between CAD and the gene encoding ACE. Polymerase chain reaction, restriction fragment length polymorphism, and agarose gel electrophoresis techniques were used to determine the ACE genotype in 58 subjects. The frequencies of ACE D and ACE I allele among the patients with CAD were 62.26% and 37.73 % and in the control subjects were 49.3% and 50.76%, respectively. The greater frequency of deletion allele (D) was in the CAD group than in the control subjects was significant (P < 0.01).

Is there a role of angiotensin-converting enzyme gene polymorphism in the failure of arteriovenous femoral shunts for hemodialysis?

In humans, thrombosis and neointimal hyperplasia are the major factors responsible for prosthetic graft occlusion. Previous studies suggest that the renin-angiotensin system is one of the key enzymes in the vascular system and has been implicated in the pathogenesis of thrombosis and neointimal hyperplasia. We conducted a case-control study to determine the frequency of the different angiotensin-converting enzyme (ACE) genotypes among the patients who had PTFE graft implantation for hemodialysis access. Between 1997 and 1999, 30 graft implantations were performed. Twelve individuals (40%) developed thrombotic complications, 8 of the 12 patients had ACE ID polymorphism, and 2 patients had DD and 2 patients had II polymorphism. The ID polymorphism was significantly more frequent in the thrombosed arteriovenous (A-V) grafts than in nonthrombosed A-V grafts (chi2 = 7.57 and p = 0.02). Overall, the frequency of the D and I alleles was 66.6 and 33.3%, respectively. In conclusion, ID polymorphism of the ACE gene plays an important role in the pathogenesis of vascular access thrombosis in subjects undergoing hemodialysis for chronic renal failure.