Gene-gene interaction of ACE I/D, endothelial nitric oxide synthase 4 a/b and ApoE does not affect coronary artery disease severity.

OBJECTIVES: Previous studies have shown the impact of angiotensin converting enzyme (ACE) insertion/deletion (I/D), endothelial nitric oxide synthase (eNOS) polymorphisms and ApoE genotypes on coronary artery disease (CAD). The aim of this study is to investigate the relationship between the genetic polymorphisms and the severity of CAD and to evaluate their potential interactions. MATERIAL AND METHODS: All patients underwent coronary angiography; coronary score (CS) and severity score (SS) were calculated for them. ACE I/D, eNOS and ApoE polymorphisms were detected by polymerase chain reaction (PCR). RESULTS: Neither CS nor SS showed a direct relationship with eNOS and ApoE genotypes. CS and SS were found to be high in patients carrying the ACE DD allele (p = 0.034 and p = 0.009). In the gene interactions, there was an increase in the SS only in patients with coexisting eNOS b/b genotype and ACE DD allele (p = 0.043). CONCLUSIONS: The interactions of the gene polymorphisms investigated don't play an important role in determining an individual's risk for the severity of CAD.

Association of eNOS gene intron 4 a/b VNTR polymorphisms in children with nephrotic syndrome.

To investigate the association of endothelial nitric oxide synthase gene intron 4 (eNOS4) polymorphisms with nephrotic syndrome, the eNOS4 genotypes were assessed in 161 children with nephrotic syndrome in comparison with 78 healthy subjects. We classified the children with nephritic syndrome into 2 groups: as steroid-sensitive nephrotic syndrome (SSNS) (n=125) and steroid-resistant nephrotic syndrome (SRNS) (n=36). The eNOS4 polymorphisms were analyzed by polymerase chain reaction. The frequencies of eNOS4 aa, ab and bb genotypes were 3%, 31%, and 66% in all the nephrotic syndrome groups, and 1%, 23%, and 76% in the control group (x(2)=2.87, p>0.05). In addition, the frequencies of eNOS4 aa, ab and bb genotypes were 2%, 33%, and 65% in SSNS group, and 5%, 28%, and 67% in the SRNS group (x(2)=1.13, p=0.567). The present study is the first to investigate eNOS4 gene polymorphisms in children with SSNS and SRNS. Our data show that the eNOS4 gene polymorphisms were not associated with the development, frequent relapse and response to steroid in nephritic syndrome.

Ambulatory blood pressure monitoring and serum nitric oxide concentration in type 1 diabetic children.

Blood pressure can be determined more precisely with the use of 24 hours ambulatory measurement in type 1 diabetics. Nitric oxide (NO) has been suggested to be responsible for the vascular changes described in early diabetic nephropathy. We aimed to investigate serum NO concentration along with ambulatory blood pressure monitoring (ABPM) parameters in type 1 diabetic patients and to find out whether there are correlation between serum NO level and ABPM parameters. Forty type 1 diabetic subjects and 35 controls were enrolled. Diabetic subjects were grouped as microalbuminuric (n=16) and normalbuminuric (n=24). Casual and ambulatory blood pressure parameters and serum NO concentrations were measured in all study population. Microalbuminuric subjects had higher nighttime systolic blood pressure (SBP), 24 hours diastolic blood pressure (DBP) and 24 hours mean arterial pressure (MAP) than controls. Both microalbuminuric and normalbuminuric subjects had also significantly higher nighttime DBP and nighttime MAP than controls. Serum NO concentrations were higher in normalbuminuric and microalbuminuric subjects than controls. Serum NO concentrations were positively correlated with daytime DBP and MAP, nighttime SBP, DBP and MAP, and 24 hours DBP and MAP in microalbuminuric subjects. Serum NO concentrations were also positively correlated with nighttime DBP in normalbuminuric subjects. Multiple linear regression analysis revealed that serum NO(2)- + NO(3)- concentrations and 24 hours DBP were independently associated with the development of microalbuminuria. Albuminuria seems to be closely associated with serum NO concentrations and ABPM parameters in type 1 DM patients. A prospective follow-up study on diabetic patients with normo- and micro- albuminuria is needed to confirm the predictive values of increased NO concentrations and ABPM parameters on the development of albuminuria.

Genetic dilemma: eNOS gene intron 4a/b VNTR polymorphism in sepsis and its clinical features in Turkish children.

The role of endothelial nitric oxide synthase gene intron 4 a/b (eNOS4a/b) variable number of tandem repeats (VNTR) polymorphism in various diseases was investigated. We investigated whether this polymorphism is associated with susceptibility to sepsis and its clinical features such as acute respiratory distress syndrome (ARDS), multiorgan dysfunction syndrome (MODS) and shock. eNOS4a/b VNTR polymorphism was determined by the polymerase chain reaction in 100 children with sepsis and in 134 healthy controls. The genotype distribution of eNOS4 was not different between the patients and controls (p=0.44). There was no statistically significant association between genotypes/allele frequency and outcomes like mortality, MODS, ARDS, and shock (p>0.05). This is the first study that evaluates the effect of eNOS4a/b polymorphism in sepsis. We were unable to show a relationship between eNOS gene intron 4 a/b VNTR polymorphism and MODS, ARDS, mortality and shock. Larger studies that do research on the interaction of such genes are needed to clarify the association between eNOS4a/b polymorphism and sepsis.

Procollagen type I carboxy-terminal peptide shows left ventricular hypertrophy and diastolic dysfunction in hypertensive patients.

BACKGROUND: An excess of myocardial collagens in hypertension is a result of increased collagen synthesis and unchanged or decreased collagen degradation. Increased collagen content, which is shown by the procollagen type I carboxy-terminal peptide (PIP), promotes cardiac remodeling and function abnormalities. OBJECTIVES: The objectives of this study were to assess PIP levels as a marker of myocardial collagen synthesis and to investigate the relationship between PIP levels and left ventricular mass index (LVMI) as well as diastolic function in patients with mild-to-moderate essential hypertension. METHODS: The study subjects were divided into three groups: healthy subjects (Group I, n=30); hypertensive patients without left ventricular hypertrophy (Group II, n=30); and patients with left ventricular hypertrophy (Group III, n=30). Left ventricular diastolic function was assessed by standard echocardiography and tissue Doppler imaging. Serum PIP was measured by radioimmunoassay. RESULTS: The serum concentration of PIP was higher in Group III than in Groups I and II (P<.001). A positive correlation was found between serum PIP and LVMI in hypertensive patients (r=.57, P<.001). Patients with diastolic dysfunction (DD) had significantly higher PIP levels as compared with patients without DD (177.3+/-52.25 vs. 138.8+/-38.0 microg/L, P<.001). The cutoff values of PIP to predict left ventricular hypertrophy and DD were 155.0 microg/L (sensitivity, 84%; specificity, 73%) and 150.2 microg/L (sensitivity, 71%; specificity, 70%), respectively. CONCLUSION: An elevated serum concentration of PIP shows left ventricular hypertrophy and DD in the course of hypertension and may be used to follow up on the efficacy of the antihypertensive treatment used.

Allele frequency distributions of Apo B VNTR locus in Cukurova, Turkey.

The highly polymorphic minisatellites contain a variable number of tandemly repeated (VNTR) DNA sequences. They are extremely useful and informative markers to study genetic variation among human populations. We have analysed the allele frequency distribution at the highly polymorphic apolipoprotein B (Apo B) VNTR locus in order to obtain the population data for the Cukurova region in Turkey by using the polymerase chain reaction and polyacrylamide gel electrophoresis. We observed 10 different alleles and 21 genotypes in a sample of 100 unrelated individuals. The allele frequencies ranged from 0.01 to 0.4, with an expected heterozygosity of 0.69 for the Apo B locus. Alleles 37 (frequency = 0.4) and 35 (frequency = 0.17) were the most common in the Cukurova population. There was a significant deviation from the Hardy-Weinberg equilibrium (HWE) for genotype frequencies (chi2 = 29.12; df = 1; p = 0.000). This study possesses novelty as it is the first DNA polymorphism study conducted at the Cukurova population using an Apo B minisatellite locus.

Endothelial nitric oxide synthase gene intron 4 a/b VNTR polymorphism in children with APSGN.

The role of endothelial nitric oxide synthase gene intron 4 a/b (eNOS4a/b) variable number of tandem repeats (VNTR) polymorphism in various renal diseases was investigated. We investigated whether the eNOS4a/b VNTR polymorphism is associated with susceptibility to acute poststreptococcal glomerulonephritis (APSGN) and its clinical features. Endothelial NOS4a/b VNTR polymorphism is determined by the polymerase chain reaction in 60 children with APSGN, and 66 healthy controls. The genotype distribution of eNOS4 does not differ between the patients and the controls (X(2)=5.1, p=0.079). However, the frequency of eNOS4a (eNOS4a/a and eNOS4a/b) genotype is higher in the patients than in the controls (X(2)=4.5, p=0.046). In the APSGN group we performed renal biopsy on eight patients because of nephrotic syndrome accompanies acute nephritic syndrome or glomerular filtration rate (GFR) is lower than 50% of normal, and found that to carry a/a and a/b genotypes were a significant risk factor for this type presentation (OR=17.3, 95% CI:1.95-152.67, p=0.03). Mean serum creatinine values are found statistically significantly higher in a/a and a/b genotypes when compared with b/b genotypes (p=0.022). Children carrying the "aa" and "ab" genotype or "a" allele of eNOS4 have a greater tendency to develop and clinical presentation of APSGN.

ACE gene polymorphism in Turkish children with nephrotic syndrome.

Since 1990, the role of angiotensin converting enzyme (ACE) gene polymorphism in various renal and cardiac diseases is still debated. This study comprised 71 pediatric patients with nephrotic syndrome, 47 males (66%) and 24 females (34%) with a mean age of 57.4 +/- 37.6 months, and a control group of 83 healthy males (59%) and 57 healthy females (41%) with a mean age of 505 +/- 160.5 months. The distribution of the ACE genotype in the control group was II, 11%; ID, 53%; and DD, 36%, and the nephrotic syndrome was II, 4%; ID, 78%; and DD, 18%. Angiotensin-converting enzyme genotypes were significantly different between patients and control groups (p<0.05). The study groups consisted of 52 (73%) with steroid-sensitive nephrotic syndrome (SNSS) and 19 (27%) with steroid-resistant nephrotic syndrome (SRNS). The distribution of the ACE genotype was II, 6%; ID, 75%; and DD, 19% in the SSNS population and ID, 84% and DD, 16% in the SRNS population. No statistically significant difference was found between steroid sensitivity and ACE genotypes (p=0.5). The results show that ACE I/D polymorphism does not contribute to the steroid resistance, even though this study indicates that the presence of the I/D genotype has a much higher risk--approximately 2.8 times--of having nephrotic syndrome. Further studies with a larger number of patients are needed.

Insertion/deletion polymorphism of the angiotensin converting enzyme gene in coronary artery disease in southern Turkey.

Genetic factors are important in the pathogenesis of coronary artery disease (CAD). Angiotensin converting enzyme (ACE) gene insertion(I)/deletion(D) polymorphism is one of the genetic factor found to be related with CAD. We investigated the association between I/D polymorphism of the ACE gene and the presence of CAD. Three hundred and seven patients (187 males and 120 females, aged between 35-80, mean 54.3 +/-9.8 years) who underwent diagnostic coronary angiography were included in the study. ACE I/D polymorphism was detected by polymerase chain reaction. Of the 307, 176 had CAD. The most frequently observed genotype in all subjects was ID (47.9 %). However, in patients with CAD the frequency of II genotype was lower whereas DD genotype was higher compared to the controls (p < 0.05). The number of D allele carrying subjects were also higher (p < 0.05) in CAD patients. The logistic regression analysis indicated that the ACE D allele is an independent risk factor (odds ratio = 1.48, 95 % CI = 1.01-2.18, p < 0.05). In conclusion, the I/D polymorphism of ACE gene (carrying D allele) is an independent risk factor for CAD in the studied Turkish population.

Biochemical indicators and cardiac function tests in chronic alcohol abusers.

AIM: To determine the effect of chronic alcohol abuse on cardiac function, antioxidant system, trace elements, and liver function tests. METHODS: Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma glutamyl transferase (GGT), superoxide dismutase (SOD), malondialdehyde (MDA), as well as zinc, magnesium, and copper were assayed in 25 chronic alcoholic patients and their 25 healthy relatives matched in age and gender. Echocardiographic parameters were evaluated for subjects. RESULTS: Mean corpuscular volume (96.7 fL vs 92.4 fL) and mean corpuscular hemoglobin levels (31.4 pg vs 30.5 pg) were found to be significantly increased in the patient group (P=0.002 and P=0.048, respectively). The results of the SOD and MDA assays showed no significant differences between the two groups. AST (38.7 U/L vs 22.1 U/L) and GGT (104.2 U/L vs 34.2 U/L) levels were found to be significantly increased in the patient group compared with controls (P=0.005 and P<0.001, respectively). Magnesium (1.6 mmol/L vs 1.8 mmol/L) and zinc levels (14.9 micromol/L vs 19.2 micromol/L) were significantly decreased, whereas copper levels (19.3 micromol/L vs 17.9 micromol/L) were increased in alcoholics (P=0.042, P<0.001 and P=0.003, respectively). Echocardiographic examination showed a significant decrease in mitral and tricuspid ratio of peak early and atrial flow velocity (E/A ratio) in alcoholics. CONCLUSION: Decrease in mitral and tricuspid E/A ratios accompanied with low levels of magnesium and zinc, and increased levels of copper indicate that alcoholics already have heart muscle disease even chronic alcohol exposure.

eNOS gene intron 4 a/b VNTR polymorphism is a risk factor for coronary artery disease in Southern Turkey.

BACKGROUND: Nitric oxide (NO) plays a major role in the regulation of vascular tone Associations between NO genotypes, coronary artery disease (CAD) and other risk factors have been described by many authors. The aim of this study was to investigate the role of endothelial nitric oxide synthase (eNOS) gene intron 4 a/b variable number of tandem repeats (VNTR) polymorphism and other risk factors in the development of CAD in subjects living in Southern Turkey. METHODS: Two-hundred and sixty-six patients (154 males and 112 females, aged between 30 and 80 years, mean 52.4+/-10.3) whose coronary arteries were evaluated by means of coronary angiography were enrolled in the study. Of the total, 133 had CAD (Group I) and the remaining had normal coronary arteries (Group II). The eNOS gene intron 4 a/b VNTR polymorphism was analyzed by polymerase chain reaction. The plasma lipid levels and other risk factors were also determined in all subjects. RESULTS: The a allele frequencies and genotypes carrying a allele were significantly higher in Group I. Plasma lipids, except HDL-C, were also increased in this group. We found that hypertension (HT), diabetes mellitus (DM), male gender, age and smoking were the independent predictors of CAD. CONCLUSION: a allele of eNOS intron 4 a/b VNTR polymorphism is not an independent predictor of CAD. eNOS intron 4 a/b polymorphism (presence of a allele) is a risk factor in addition to HT, DM, male gender, age and smoking for the development of CAD in Southern Turkey.

Prenatal diagnosis of sickle cell anemia in twin pregnancies and identification by VNTRs.

BACKGROUND: Sickle cell anemia (SCA) is an inherited disorder of hemoglobin synthesis that is characterized by life-long severe hemolytic anemia, attacks of pain crisis, and chronic organ system damage. In this study, prenatal diagnosis was performed to three couples (families A, B, and C) with twin pregnancies who were at risk for SCA. METHODS: The SCA carrier state of the couples were confirmed at molecular level. Chorionic villus samples (CVS) of twins were obtained at 10-12 weeks of gestation. Amplification refractory mutation system (ARMS) and restriction fragment length polymorphism (RFLP) techniques were applied to determine and confirm the presence of the sickle cell anemia of the fetuses. Identification of each twin, confirmation of parentage and elimination of maternal contamination of chorionic villus samples were ruled out by variable number of tandem repeats (VNTR) analysis of four different loci [D1S80 (pMCT118), ApoB, IgJH, D4S95]. RESULTS: We found that one of the fetuses was heterozygous for SCA and the other was normal in family A; in family B both fetuses were heterozygous and in family C both fetuses were normal. CONCLUSION: Prenatal diagnosis is the major way of prevention of the genetic disorders including sickle cell anemia. Prenatal diagnosis of twins contains additional risks such as confusion in differentiation leading to false molecular diagnosis. VNTR loci analysis is a useful tool and can be safely used for the elimination of all problems mentioned above.

Relation between age and gender differences in plasma triglyceride concentrations and coronary artery disease in Southern Turkey.

BACKGROUND: Coronary artery disease (CAD) is still a leading cause of morbidity and mortality in both genders in Turkey as well as other countries. The role of plasma triglycerides (TG) as a CAD risk factor has been controversial. We investigated the relation between age and gender differences in plasma TG and CAD in patients with angiographically proven CAD. METHODS: The study population consisted of 937 patients (605 males and 332 females, aged between 32 and 82 years). The patients were divided into three age groups as < 45, 45-59 and > or = 60 years. Risk factors were considered as hypertension, hyperlipidemia, diabetes mellitus, cigarette smoking and family history. Hypertriglyceridemia was defined as TG concentration > 150 mg/dl (> 1.69 mmol/l). RESULTS: Of the total, 608 had CAD. TG was an independent risk factor for CAD only in women (p < 0.05). There was also a positive correlation between TG concentrations and number of the diseased vessels only in women (p = 0.001, r = 0.2). CONCLUSION: High TG concentrations increase CAD risk more in women than in men in Southern Turkey. Triglycerides should be taken into consideration as an important risk factor especially in women > 45 years of age.

The effects of methyl methacrylate on nasal cavity, lung, and antioxidant system (an experimental inhalation study).

Methyl methacrylate (MMA) is a monomer, commonly used in neurosurgery, orthopedic surgery, and in dental clinics. The adverse effects of this monomer are well described in the literature. This study was designed to evaluate the effects of MMA on nasal cavity, lung, and antioxidant status. For this purpose, two experimental groups of rats were exposed to MMA (at 1,000 ppm, 6 h/day, 5 days/week for 4 weeks) by inhalation under poor (group A, n = 12) and normal ventilation (group B, n = 11) conditions. A control group (group C, n = 10) received normal air. Degeneration of olfactory epithelium, bronchopneumonia, interstitial pneumonia, hemorrhage, atelectasis, edema, emphysema, and bronchial epithelial hyperplasia were observed in groups A and B. Emphysema was the most common lesion. Bronchopneumonia with abscesses was only observed in group A. Glutathione levels were significantly decreased and malondialdehyde levels were significantly increased in group A. No significant difference was observed in superoxide dismutase levels between the groups. The data presented indicate that before using MMA, adequate protection systems should be in place to prevent occupationally related MMA respiratory-tract injuries.

Apolipoprotein E polymorphism in childhood nephrotic syndrome.

Recent clinical reports have demonstrated that the progression and prognosis of renal diseases are possibly influenced by apolipoprotein E (apoE) genotypes and alleles. In this study we investigated whether apoE genotypes and alleles can be a prognostic criterion for the steroid responsiveness in childhood nephrotic syndrome. One hundred and seven pediatric patients with primary idiopathic nephrotic syndrome and 83 healthy volunteers were enrolled in the study. Eighty-seven of the patients had steroid-sensitive nephrotic syndrome (SSNS) and 20 had steroid-resistant nephrotic syndrome (SRNS). The varepsilon2 allele frequency and varepsilon2/3 genotype frequency of the SNRS group were statistically higher when compared with SSNS and control groups ( P<0.05). The higher frequency of the varepsilon2 allele in steroid resistant nephrotic patients suggests that the varepsilon2 allele gives a possible genetic predisposition to steroid resistance in our population, but further studies are needed to clarify this subject.