Nihan Erginel Ünaltuna

HEREDITARY THROMBOPHILIC RISK FACTORS AND VENOUS THROMBOEMBOLISM IN ISTANBUL, TURKEY: THE ROLE IN DIFFERENT CLINICAL MANIFESTATIONS OF VENOUS THROMBOEMBOLISM

The aim of this study was to investigate the hereditary thrombophilic risk factors in patients with venous thromboembolism (VTE) and whether these risk factors play a different role in patients with isolated pulmonary embolism (PE) as compared with patients with deep vein thrombosis (DVT) and patients with PE + DVT. The protein C (PC), protein S, antithrombin activities, homocysteine levels, and factor V Leiden (FVL) G1691A and prothrombin G20210A mutations were evaluated in 191 patients with VTE and 191 controls. The prevalence of FVL and PC deficiency were higher in patients (P = .003 and P = .02, respectively). There was no significant difference for the other risk factors. The combination of thrombophilic risk factors was significantly higher in patients with DVT + PE as compared with patients with isolated PE or DVT (P = .04). In conclusion, the most important hereditary risk factors for VTE in this study were the FVL mutation and PC deficiency.

Gender specific association of ABCA1 gene R219K variant in coronary disease risk through interactions with serum triglyceride elevation in Turkish adults.

OBJECTIVE ATP binding cassette transporter A1 (ABCA1) controls the reverse cholesterol transport. Some ABCA1 variants are correlated with serum high-density lipoprotein cholesterol (HDL-C) and other lipid concentrations. We aimed to explore the relationship of ABCA1 gene with both the lipid profile and coronary heart disease (CHD) risk. METHODS Selected 627 individuals of the Turkish Adult Risk Factor Study were genotyped for ABCA1 R219K polymorphism using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method. Students t-test, one-way ANOVA, Chi-square test, linear and logistic regression was used for statistical analysis. RESULTS We demonstrated a gender-specific effect of the R219K polymorphism on plasma lipids and CHD. In men, while homozygosity of the K allele was associated with increased plasma low-density lipoprotein cholesterol (LDL-C) (p<0.05) and total cholesterol concentrations (p<0.05), carriage of this allele was associated with higher HDL-C concentrations (p<0.05) after adjustment for associated risk factors, but not with CHD. In women, however, without being related to HDL-C levels, each 219K allele was associated with 10% higher triglycerides (TG) concentrations (p<0.05). R219K heterozygosity in women independently doubled (95% CI 1.00; 3.80) the odds ratio for CHD risk in regression models, after adjustment for several variables. Interaction of TG elevation (>140 mg/dL) with CHD was demonstrated in female 219RK genotype carriers. CONCLUSION R219 allele of the ABCA1 gene independently confers CHD risk in heterozygote Turkish women, not via reduced HDL-C, but interacting with elevated TG expressed by the 219K allele, but not in men.

The variations of BOP gene in hypertrophic cardiomyopathy

OBJECTIVE The observation that Bop null allele mice show underdeveloped right ventricle and excessive development of left ventricle, suggests the possible relationship between human BOP gene and hypertrophic cardiomyopathy (HCMP). In our study, we investigated this possible relationship between BOP gene variations and QT dispersion, a noninvasive arrhythmic risk marker for HCMP. METHODS This cross-sectional study consisted of 50 patients clinically diagnosed with HCMP and 60 healthy subjects. Exonic regions of BOP gene were amplified by polymerase chain reaction and amplified exonic regions were analyzed by Single-Strand Conformation Polymorphisms (SSCP). The samples with different migration patterns were sequenced through an automated sequencing system. Continuous variables were compared by unpaired t-test for independent samples or Mann-Whitney U test. Genotype-disease relationship was tested by Chi-square test. RESULTS The nucleotide substitutions G275<or=A and C965<or=A in exon 2 and 7 were determined only in HCMP group. The G707<or=C, C710<or=T, T761<or=C, T1217<or=C SNPs in exon 6 and 9 are also found in the control group. Significant differences were found between two groups (p=0.002 and p=0.001). It was found that SNPs in exon 6 constitute a haplotype and that QT dispersion and corrected QT dispersion in the rare homozygote (707C/710T/761C) type carriers of HCMP patients for this haplotype were significantly lower than other genotypes (p=0.032 and p=0.030, respectively). CONCLUSION The human BOP gene was analyzed for the first time in HCMP to investigate possible association. The result that homozygosity of 707C/710T/761C haplotype is associated with lower QT dispersion and corrected QT dispersion supports the modifier role of BOP gene in HCMP.

Factor V leiden is a risk factor for myocardial infarction in young Turkish men.

Objective — Factor V Leiden is the most common known hereditary abnormality of the clotting system which leads to a reduced anticoagulant effect of activated protein C (APC resistance). Factor V Leiden has been shown to be the most frequent inherited thrombophilic disorder in patients with idiopathic venous thromboembolism. The relationship between this genetic abnormality and myocardial infarction is still unresolved. The aim of this study was to investigate whether factor V Leiden is a risk factor for myocardial infarction in young Turkish men or not. Methods and results — We compared 42 patients who had a diagnosis of acute MI and were younger than 40 years (35.6 ± 4.8 years) with 66 healthy, age and sex-matched control subjects. Blood samples from the patients and the controls were analysed for the factor V Leiden mutation by DNA analysis, using polimerase chain reaction. Factor V Leiden mutation was found in 10 of 42 (23.8%) patients with myocardial infarction and 6 of 66 (9%) control subjects (p < 0.001).The odds ratio for MI was 3.1. (CI 95% 1.0-8.9) Conclusion — The results of this study suggest that the presence of factor V Leiden increases the risk of MI in young Turkish men.

Platelet glycoprotein Ia 807C/T and 873G/A polymorphisms in patients with venous thromboembolism.

Two silent polymorphisms (807C/T and 873G/A) within glycoprotein Ia (GPIa) gene have been implicated in increased risk of developing thrombosis and myocardial infarction in affected individuals. The aim of this study was to investigate the GPIa gene polymorphism in patients with venous thromboembolism (VTE). A multiplexed allele specific-polymerase chain reaction (AS-PCR)–based method was used to determine the GPIa 807T/873A allele frequency in 77 patients with VTE and 106 healthy controls. The allelic frequency for 807T/873A was 33% in the patient group and 38% in the control group. The allelic frequency for 807C/873G was 66% in the patient group and 62% in the control group. The genotypic frequencies were 8% for 807TT/873AA, 42% for 807CC/GG, and 50% for 807CT/GA in the patient group. In the control group, the frequencies were 12% for 807TT/873AA, 35% for 807CC/873GG, and 52% for 807CT/873GA. As a result, the glycoprotein Ia 807C/T and 873G/A dimorphisms were not shown as risk factors for VTE.

Preheparin serum lipoprotein lipase mass interacts with gender, gene polymorphism and, positively, with smoking.

Abstract Background: Correlates of preheparin serum lipoprotein lipase (LPL) mass and its associations with the likelihood of metabolic syndrome (MS) and coronary heart disease (CHD) were investigated. Methods: A cross-sectional study was carried out in a population sample (n=352, median age 55). MS was defined according to modified Adult Treatment Panel III criteria. Results: Age-adjusted geometric mean preheparin LPL concentrations were 58.6±1.04 ng/mL in men and 66.9±1.03 ng/mL in women (p<0.004). A positive interaction with both the LPL X447 allele (p<0.034) and age-adjusted smoking status (p=0.026 in men and p=0.11 women) was observed. LPL mass was significantly correlated in both genders with high-density lipoprotein (HDL)-cholesterol and inversely with triacylglycerol levels and HOMA index. In multiple linear regression analysis, LPL mass was significantly associated with genotype, gender, age, adiponectin, smoking status and HDL-cholesterol, and in women with C-reactive protein after adjustment for body mass index, triacylglycerol and insulin. Significantly low sex- and age-adjusted serum LPL mass was observed in cases of MS, hypertension and CHD. Logistic regression analysis after adjustment for age, sex, adiponectin and S447X polymorphism demonstrated that LPL mass was inversely associated with CHD in men and both genders (p=0.02), with hypertension confined to women (p=0.04) and with MS likelihood in both genders combined and women [odds ratio 1.51 (95% CI 1.14–2.00) for halving the likelihood]. Conclusions: LPL X447 genotype, female gender and smoking habit interact in increasing preheparin serum LPL mass in Turkish adults. Serum LPL mass is inversely associated with MS and CHD, independent of confounders, and probably reflects insulin sensitivity. Clin Chem Lab Med 2009;47:208–15.

Expression of MMP-15 and MMP-24 in atherosclerotic and nonatherosclerotic coronary arteries

License. The full terms of the License are available at http://creativecommons.org/licenses/by-nc/3.0/. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. Permissions beyond the scope of the License are administered by Dove Medical Press Limited. Information on how to request permission may be found at: http://www.dovepress.com/permissions.php Metalloproteinases In Medicine 2014:1 15–20 Metalloproteinases In Medicine Dovepress

Role of simvastatin and RORα activity in the macrophage apoptotic pathway

Objective: Atherosclerosis is a chronic inflammatory condition and is one of the main causes of death worldwide. Macrophages play important roles in the formation of atherosclerotic plaques. Apoptosis is progressively observed while plaques develop, although the precise mechanisms and outcomes of apoptosis in atherosclerosis development and progression are still contradictory. This study was conducted to explore the effects of simvastatin and retinoic acid receptor-related orphan receptor alpha (RORa) ligands on apoptosis in human acute monocytic leukemia (THP-1) macrophage cells. Methods: Briefly, the occupancy of RORa in the promoter regions of apoptotic pathway genes was demonstrated in THP-1 cell lines using chromatin immunoprecipitation (ChIP) analysis. In order to modulate RORa activity, THP-1 macrophage cells were treated with specific ligands (CPG52608 and SR1001) and then viability as well as count of THP-1 macrophage cells were analyzed. Results: We observed that simvastatin and both RORa ligands had a tendency to decrease THP-1 macrophage cell viability in culture. When compared with non-treated controls, simvastatin significantly decreased cell viability (p=0.04) and cell count (p=0.03). However, this negative effect of simvastatin seemed to be partly prevented by RORa ligands. In addition, bioinformatics analysis of ChIP-on-chip data demonstrated that several genes that are involved in the apoptotic pathway were likely RORa target genes. These genes were involved in the regulation of apoptosis through various pathways. Conclusion: In summary, our study suggest that simvastatin-mediated macrophage apoptosis might be modulated by SR1001 administration. However, involvement of RORa in this modulation through potential apoptotic target genes remains elusive.OBJECTIVE Atherosclerosis is a chronic inflammatory condition and is one of the main causes of death worldwide. Macrophages play important roles in the formation of atherosclerotic plaques. Apoptosis is progressively observed while plaques develop, although the precise mechanisms and outcomes of apoptosis in atherosclerosis development and progression are still contradictory. This study was conducted to explore the effects of simvastatin and retinoic acid receptor-related orphan receptor alpha (RORα) ligands on apoptosis in human acute monocytic leukemia (THP-1) macrophage cells. METHODS Briefly, the occupancy of RORα in the promoter regions of apoptotic pathway genes was demonstrated in THP-1 cell lines using chromatin immunoprecipitation (ChIP) analysis. In order to modulate RORα activity, THP-1 macrophage cells were treated with specific ligands (CPG52608 and SR1001) and then viability as well as count of THP-1 macrophage cells were analyzed. RESULTS We observed that simvastatin and both RORα ligands had a tendency to decrease THP-1 macrophage cell viability in culture. When compared with non-treated controls, simvastatin significantly decreased cell viability (p=0.04) and cell count (p=0.03). However, this negative effect of simvastatin seemed to be partly prevented by RORα ligands. In addition, bioinformatics analysis of ChIP-on-chip data demonstrated that several genes that are involved in the apoptotic pathway were likely RORα target genes. These genes were involved in the regulation of apoptosis through various pathways. CONCLUSION In summary, our study suggest that simvastatin-mediated macrophage apoptosis might be modulated by SR1001 administration. However, involvement of RORα in this modulation through potential apoptotic target genes remains elusive.

Investigating the role of ceramide metabolism-associated CERS5 (LASS5) gene in atherosclerosis pathogenesis in endothelial cells

OBJECTIVE Ceramide, the backbone of sphingolipids, is the key component affecting atherosclerotic changes through its important second-messenger role. Previous studies have demonstrated protective role of AMP-activated protein kinase (AMPK) genes in regulating atherosclerosis and hypertension. Ceramide synthase 5 (LASS5 or CERS5) gene has function in de novo synthesis of ceramide, and has indirect effect on AMPK gene. Aim of the present study was to identify role of LASS5 gene in atherosclerosis. METHODS LASS5 gene-specific small interfering RNA (siRNA)-mediated gene silencing was performed in human umbilical vein endothelial cells (HUVEC) and differential expression of LASS5, AMPK-alpha and AMPK-alpha target genes were analyzed. HUVEC cells were then treated with AMPK activator in order to examine relationship of change in gene expression levels to AMPK activity. RESULTS Novel physiological function of LASS5 was identified. Downregulation of LASS5 was found to attenuate ceramide production and increase expression of some AMPK target genes in HUVEC. CONCLUSION This is the first study to demonstrate that LASS5 was involved in negative regulation of atherosclerosis-related genes, such as AMPK-alpha. These preliminary findings provide insight into molecular mechanism of atherosclerosis and are important for development of potential therapeutic agents in the treatment of atherosclerosis.

ACSL4 gen polimorfizminin (rs7886473) metabolik sendrom ve lipid düzeyleri üzerine etkisi

Amac: Lipid metabolizmasi uzerinden obezite ve metabolik sendromla iliskisi olabilecegini dusundugumuz ACSL4 geninin sik gorulen rs7886473 A>G polimorfizminin Turk toplumunda metabolik sendrom ve lipid duzeyleri uzerindeki etkisini arastirmayi amacladik. Gerec ve Yontem: Calismamiza, Turkiye genelinde takip edilen ve modifiye edilmis NCEP ATPIII Metabolik Sendrom tani kriterlerine gore 556 metabolik sendrom olan ve 520 metabolik sendrom olmayan eriskin birey dahil edildi. Metabolik sendrom olan ve olmayan bireylerin ACSL4 gen polimorfizmi Roche Light Cycler 480 Real-Time PCR ile genotiplendi ve karsilastirildi. Bulgular: Metabolik sendrom olan ve metabolik sendrom olmayan bireyler arasinda ACSL4 rs7886473 genotip dagilimlari arasinda anlamli bir farklilik bulunmadi. Tum erkeklerin ACSL4 rs7886473 polimorfizmine gore serum total kolesterol, HDL, LDL ve trigliserid duzeyleri karsilastirildiginda; GG ve AA genotipleri arasinda anlamli bir farklilik bulunmadi. Benzer sekilde tum kadinlarda da anlamli farklilik bulunmadi. Sonuc: Bu calisma, ACSL4 geni acisindan incelemis oldugumuz rs7886473 gen polimorfizminin metabolik sendrom ve serum lipid duzeyleri uzerine etkisi olmadigini gosterdi. Ancak calismamizda incelenmis olan ACSL4 geninde yaygin gorulen yalnizca bir polimorfizmdir. Bu durum ACSL4 geninin lipid metabolizmasi ve/veya metabolik sendrom uzerine etkili olmadigini degerlendirmek icin tek basina yeterli degildir.