Nitric Oxide Synthase Polymorphisms Research Articles

Influence of endothelial nitric oxide synthase haplotypes on nitric oxide and peroxynitrite productions

The impact of four clinically significant genetic variants of endothelial nitric oxide synthase (eNOS) polymorphisms on the concentrations of nitric oxide [NO] and peroxynitrite [ONOO–] has been given scant consideration. This study utilized a [NO]/[ONOO–] ratio to determine the extent of endothelial dysfunction caused by these variations in the eNOS gene. The single nucleotide polymorphisms (T-786C, C-665T, and Glu298Asp) and a variable number of tandem repeats (intron 4 a/b/c) were genotyped in human umbilical vein endothelial cells (HUVEC), using sanger sequencing and DNA electrophoresis, respectively. Nanosensors were used to determine the maximal [NO] and [ONOO–], while traditional and low-temperature SDS-PAGE were used to evaluate the expression of eNOS and the eNOS dimer-to-monomer ratio, respectively. The study results indicate that the eNOS haplotype H3 (G T/C C 4a/c allele) may have a protective effect against cardiovascular disease (CVD) with the [NO]/[ONOO–] ratio higher than 2. However, the eNOS haplotypes H2 (G T/C C 4a/b) and H5 (T T/C C 4b) increase the susceptibility to CVD with [NO]/[ONOO–] ratio lower than 1. The results suggest that certain eNOS genetic variants may influence susceptibility to cardiovascular disease (CVD) while other variants may have a protective effect.

The Effect of Selected Nitric Oxide Synthase Polymorphisms on the Risk of Developing Diabetic Nephropathy.

Nitric oxide synthase (NOS) is an enzyme that catalyzes the formation of nitric oxide (NO), the altered production of which is characteristic of diabetic nephropathy. NOS exists in three isoforms: NOS1, NOS2, and NOS3. Moreover, there are reports about the potential role of NOS3 polymorphisms in the development of diabetes complications. The aim of this study was to assess the role of selected NOS polymorphisms-rs3782218 (NOS1), rs1137933 (NOS2), rs1799983, rs2070744, and rs61722009 (NOS3)-in the risk of developing diabetic nephropathy and in the likelihood of renal replacement therapy. The studied polymorphisms were analyzed in a group of 232 patients divided into three groups. Four polymorphisms (rs3782218, rs1137933, rs1799983, rs2070744) were genotyped using the PCR-RFLP, while the rs61722009 polymorphism was genotyped using the PCR. The C/C genotype and the C allele of the rs3782218 polymorphism (NOS1) were associated with an increased risk of developing diabetic nephropathy and an increased likelihood of renal replacement therapy. In turn, the G allele of the rs1137933 polymorphism (NOS2) reduces the likelihood of renal replacement therapy. The specific genotypes or alleles of the rs3782218 (NOS1) and rs1137933 (NOS2) polymorphisms seem to be potential risk factors for diabetic nephropathy and renal replacement therapy.

Involvement of Total Antioxidant Activity and eNOS Gene rs1799983/ rs2070744 Polymorphisms in Breast Carcinogenesis

Globally, breast cancer is the common malignancy affecting women and understanding its associated molecular events could help in disease prevention and management strategies. The present study was set to investigate an association between total antioxidant capacity (TAC) and endothelial nitric oxide synthase (eNOS) polymorphisms with breast cancer. For this purpose, 100 subjects were participated in this work, including 50 female patients diagnosed with breast cancer recruited from Oncology hospital, Baghdad - Iraq and 50 healthy women as a control group. The concentration of antioxidants was measured in the serums collected from blood samples of breast cancer patients and healthy controls. While eNOS SNPs (rs1799983, G894T and rs2070744, T 786C) were assessed using TaqMan SNP genotyping and utilising genomic DNA extracted from the participants. The results showed that the antioxidants levels were significantly (P˂0.0001) reduced in blood samples of breast cancer patients in comparison to that of that healthy controls (0.144± 0.097 and 0.587±0.239 respectively). Additionally, the homozygous GG genotype G894T (rs1799983) could retain beneficial impact for the protection from breast cancer potential. While SNP genotyping results showed that both of the homozygous CC and heterozygous TC genotypes (rs2070744 T >C SNP) seem to contribute to the susceptibility of breast cancer development in the investigated set of patients. Overall, the present study findings suggest an association between reduced antioxidant capacity and eNOS gene polymorphisms in breast carcinogenesis.

Functional polymorphisms of NOS3 and GUCY1A3 affect both nitric oxide formation and association with hypertensive disorders of pregnancy.

Impaired nitric oxide (NO) formation may be associated with endothelial dysfunction and increased cardiovascular disease risk in preeclampsia (PE). Functional single-nucleotide polymorphisms (SNPs) of nitric oxide synthase 3 (NOS3) (rs3918226) and guanylate cyclase 1, soluble, alpha 3 (GUCY1A3) (rs7692387) increase susceptibility to the adverse consequences due to inadequate generation of NO by the endothelium. However, no previous study has examined whether these SNPs affect NO formation in healthy pregnancy and in gestational hypertension (GH) and PE. Here, we compared the alleles and genotypes of NOS3 (rs3918226) and GUCY1A3 (rs7692387) SNPs in normotensive pregnant women (NP, n = 153), in GH (n = 96) and PE (n = 163), and examined whether these SNPs affect plasma nitrite concentrations (a marker of NO formation) in these groups. We further examined whether the interaction among SNP genotypes is associated with GH and PE. Genotypes were determined using TaqMan allele discrimination assays, and plasma nitrite concentrations were determined by an ozone-based chemiluminescence assay. Multifactor dimensionality reduction was used to examine the interactions among SNP genotypes. Regarding NOS3 rs3918226, the CT genotype (p = 0.046) and T allele (p = 0.020) were more frequent in NP than in GH, and GH patients carrying the CT+TT genotypes showed lower nitrite concentrations than NP carrying the CT+TT genotypes (p < 0.05). Regarding GUCY1A3 rs7692387, the GA genotype (p = 0.013) and A allele (p = 0.016) were more frequent in PE than in NP, and NP women carrying the GG genotype showed higher nitrite concentrations than GH or PE patients carrying the GG genotype (p < 0.05). However, we found no significant interactions among genotypes for these functional SNPs to be associated with GH or PE. Our novel findings suggest that NOS3 rs3918226 and GUCY1A3 rs7692387 may affect NO formation and association with hypertensive disorders of pregnancy.

Risk association of the nitric oxide synthase VNTR intron 4 a/b variant with diabetic nephropathy – a pilot study

Diabetic nephropathy (DN) is known to be a leading complication of type 2 diabetes mellitus (T2D). This study evaluated whether the VNTR intron 4 a/b and rs1799983 polymorphisms of endothelial-derived nitric oxide synthase (eNOS) gene modulated the risk of developing DN in Asian Indian patients. The eNOS variants were genotyped in 200 patients, 100 with DN and 100 without DN. A significant risk association was observed for the VNTR intron 4 a/b (p < 0.05). Haplotype analysis revealed that the allele combination of rs1799983894 G/Intron 4b and rs1799983894 T/Intron 4b had a statistically significant inverse association with DN.

Genetic Polymorphisms of Endothelial Nitric Oxide Synthase Associated with Hypertension and Blood Homocysteine Levels.

Endothelial dysfunction is a key mechanism in the development of hypertension and is closely linked to impairment of endothelial nitric oxide synthase (eNOS) and hyperhomocysteinemia. Genetic polymorphisms of eNOS (rs1799983 and rs2070744) are strongly associated with the risk of hypertension in individuals of Asian ethnicities. This study aimed to investigate the relationship between these polymorphisms and the risk of hypertension associated with homocysteine levels. For this cross-sectional study, we enrolled 370 Thai men aged 40-60 years from the Electricity Generating Authority of Thailand cohort study for both variants genotyping by TaqMan allelic discrimination analysis. Clinical, anthropometric, and biochemical parameters were also analyzed. In the high blood pressure group (n = 267), systolic and diastolic blood pressure and triglyceride levels were higher in those with homocysteine levels ≥ 15 µmol/L than in those with homocysteine levels < 15 µmol/L (p < 0.05). Significant risk of hypertension was found in GG and GT of rs1799983 (G894T), and in TT and TC of rs2070744 (T-786C), with higher ORs in heterozygous genotypes (all p values < 0.05). Further evaluation of the interactions between SNPs and HCY revealed that individuals with the GT or TC genotype, together with hyperhomocysteinemia, had an increased risk of hypertension (all p<0.05). eNOS variants rs1799983 and rs2070744 may be risk factors for hypertension linked to hyperhomocysteinemia. These findings provide potentially useful healthcare strategies for the management of hypertension.

ВЛИЯНИЕ ОДНОНУКЛЕОТИДНЫХ ПОЛИМОРФИЗМОВ ГЕНОВ IL1β, EDN1 И NOS3 НА ИНДИВИДУАЛЬНЫЙ ГЕНЕТИЧЕСКИЙ ПРОФИЛЬ ПАЦИЕНТОВ С ИШЕМИЧЕСКОЙ БОЛЕЗНЬЮ СЕРДЦА В РЕСПУБЛИКЕ КРЫМ

Abstracts. Introduction. Coronary heart disease prevalence rate grows, which determines the importance of researchers’ inquiries aiming to improve the prevention, diagnosis, and treatment of this pathology. Aim. This study was aimed at identifying the associations of polymorphisms of genes interleukin 1-beta, endothelin-1, and endothelial nitric oxide synthase in patients with coronary heart disease. Materials and Methods. We studied polymorphic markers of endothelin-1 (rs5370), endothelial nitric oxide synthase (rs2070244), and interleukin 1-beta genes (rs1143634 and rs16944) in patients with ischemic heart disease (n=229) and in controls (n=56) by multiplex allele-specific polymerase chain reaction with electrophoretic detection. Odds ratio (OR) was used to state the association of genotypes with the disease; 95% confidence interval limits were calculated to assess the significance of OR. Results and Discussion. Analysis of the T-511C polymorphism in the interleukin 1-beta gene demonstrated a trend towards an increased risk of coronary heart disease associated with the TT genotype in the study group (Odds ratio=2.2, 95%. Confidence interval: 1.3-3.6). At the same time, the CC genotype can be associated with a protective effect (Odds Ratio=0.3, 95%. Confidence Interval: 0.2-0.6). When analyzing the C3953T polymorphism in the interleukin 1-beta gene, the TT genotype showed a significantly increased disease development risk within the pathology under study (Odds Ratio=6.4, 95%. Confidence Interval: 1.7-23.4). Analysis of the polymorphic marker C786T in the NOS3 gene showed a correlation between the TT genotype and susceptibility to coronary heart disease (Odds ratio=2.07, 95%. Confidence interval: 1.3-3.2), while the CC genotype showed a protective effect (Odds ratio=0.3, 95%. Confidence interval: 0.2-0.5). A similar result was obtained in an experiment with the Lys198Asn polymorphism in the endothelin-1 gene. TT genotype showed an association with the increased risk of coronary heart disease (Odds ratio=6.5, 95%. Confidence Interval: 3.4-12.4). Odds ratio analysis of the effect of heterozygotes showed no risk of coronary heart disease (Odds ratio=0.3, 95%. Confidence Interval: 0.1-0.5). Conclusions. This study allowed us to find an association between the polymorphic markers of genes endothelin-1 (rs5370), eNOS (rs2070244), and interleukin-1β (rs114634 and rs16944) and the coronary heart disease risk in the population, which may be useful for both stratification and selection of further therapeutic strategies.

Single nucleotide polymorphisms in inducible nitric oxide synthase gene are not associated with persistent apical periodontitis.

The aim of this study was to investigate whether there is an association between inducible in single nucleotide polymorphisms in nitric oxide synthase (rs2297518 and rs2779249) and persistent apical periodontitis. A total of 291 Brazilian subjects were included: 125 with signs/symptoms of persistent apical periodontitis and 166 with root canal-treated teeth exhibiting healthy perirradicular tissues. Endodontically treated patients were followed up after 1 year. The two single nucleotide polymorphisms in nitric oxide synthase were analysed using real-time polymerase chain reaction. Chi-square test and odds ratio with 95% confidence intervals were performed to compare genotype distributions between 'healed' and 'persistent apical periodontitis' groups (p < 0.05). Logistic regression analysis was used to evaluate SNP-SNP interactions. The allele and genotype distributions for the polymorphisms between the persistent apical periodontitis and healed groups were not statistically significant (p > 0.05). In the logistic regression analysis, the polymorphisms were not associated with persistent apical periodontitis and SNP-SNP interactions.

Endothelial Nitric Oxide Synthase T-786C and G-894T Gene Polymorphisms: A Risk Assessment of Coronary Heart Disease.

The polymorphism of the endothelial nitric oxide synthase (eNOS) gene is thought to enhance the risk of coronary heart disease (CHD). The most protruding reason for endothelial dysfunction is the diminished production of NO. In the eNOS encoding gene, a number of mutations were associated with decreased NO effect promoting the presence of CHD. This study aims to look at the role of polymorphisms in the eNOS genes G-894T and T-786C in young South Indians. From January 2022 to May 2022, 91 angiographically proven CHD subjects attending the Department of Cardiology and Medicine and 91 controls from a master health checkup in the age group of 45 years participated in this observational cross-sectional study at SRM Medical College Hospital and Research Centre in Chennai, Tamil Nadu, India. Overnight fasting plasma samples were taken for analysis of the lipid profile as well as NO by Griess reaction utilizing an enzyme-linked immunosorbent assay (ELISA) technique. Polymerase chain reaction (PCR) and restricted fragment length polymorphism (RFLP) were used to amplify the T-786C and G-894T eNOS genes. When compared to controls, the mean level of serum NO in CHD patients was considerably lower. For eNOS T-786C polymorphism, the distribution of TC genotype (p = 0.017), odds (OD) ratio = 2.1, CC genotype (p = 0.011), OD ratio = 3.75, and minor C allele frequency (p = 0.001). And for eNOS G-894T polymorphism, the distribution of GT genotype (p = 0.01), TT genotype (p =0.02) with OD ratio and minor T allele frequency (p = 0.002) with OD ratio = 1.07 and 0.83. Our study concludes that the polymorphism of eNOS T-786C and G-894T genes may be the main causative association for the presence of CHD. How to cite this article: Jaishankar T, Shivasekar M, Vinodhini VM. Endothelial Nitric Oxide Synthase T-786C and G-894T Gene Polymorphisms: A Risk Assessment of Coronary Heart Disease. J Assoc Physicians India 2023;71(9):45-50.

Glu298Asp polymorphism of the endothelial nitric oxide synthase (eNOS) gene (rs1799983) is associated with dysmenorrhea

BackgroundWith a huge impact on the quality of life and productivity of young women, dysmenorrhea remains a major gynecological disorder. Although abnormal myometrial activity and vasoconstriction are believed to play a major role, the pathophysiology of dysmenorrhea is not fully understood. Our aim was therefore to examine a possible relationship between the Glu298Asp polymorphism (rs1799983) of the endothelial nitric oxide synthase (eNOS) gene and dysmenorrhea. MethodsWe sought the participation of women of reproductive age (n = 239). Each participant completed a questionnaire to obtain data on their menstrual cycles and a Numeric Pain Rating Scale (NPRC) to rate their pain experience during menstrual periods. Genotyping was done by PCR-RFLP method while a possible association between the presence and severity of menstrual pain and genotypes was determined. ResultsAmong the participants, pain was reported by 167 (89.9%) while 72 (30.1%) reported no pain. Seventy-one (29.7%) and 93 (38.9%) participants reported moderate and severe pains respectively while others reported no or mild pain. The frequencies of G and T alleles were 316 and 162 while frequencies of GG, GT, and TT were 119, 78, and 42 respectively. The population deviated from Hardy-Weinberg equilibrium (χ2 = 17.6, df = 1, p < 0.01). Participants with a T allele were more than three times as likely as those without a T allele to report pain (OR = 3.3, p < 0.01 95%CI = 1.8–5.9). ConclusionCarrying a T (Asp298) allele of Glu298Asp polymorphism of the eNOS gene may predispose to dysmenorrhea indicating that nitric oxide might play a role in dysmenorrhea.

Exploring nitric oxide as a crucial prognostic biomarker of coronary artery disease

PurposeThe study aimed to examine if the polymorphism of the endothelial nitric oxide synthase (eNOS) gene variable number of tandem repeats (VNTR) and the serum NO levels are associated with CAD. Materials/methodsCase-control study, 70 CAD and 30 control subjects were enrolled. The eNOS gene polymorphism was measured by polymerase chain reaction–agarose gel electrophoresis and the serum NO was assessed by using an ELISA plate and reader covering 540 nm. ResultsUncovering the area under curve (AUC) for serum NO, which was (0.6821), indicating that NO seemed to be a critical prognostic biomarker of CAD; also, glucose, serum creatinine and total bilirubin proved to be significant predictors of CAD with AUC (0.6793, 0.6717 and 0.6662) respectively. Furthermore, higher serum NO levels were associated with the eNOS (ab) genotype. Revealing the intron (a) allele was protective against CAD. Moreover, diminished levels of serum NO in CAD groups compared to controls (P < 0.05). Additionally, Multiple logistic regression analysis shows a significantly high Odds ratio associated with CAD in the Duhok population. ConclusionsThe eNOS (ab) variant seems to be a protective CAD factor for patients. Low serum NO levels are another risk factor for the advancement of CAD, suggesting their involvement in atherosclerosis. The (a) allele's protective effect is mediated through changes in eNOS promoter activity and higher NO levels.

Genetic polymorphism of prolactin and nitric oxide synthase in Holstein cattle.

Bacterial and viral infections affect the welfare of animals and lead to large economic losses in dairy cattle breeding due to decreased productive indicators and increased culling rates. In modern dairy farming, farmers are looking for effective solutions to prevent and minimize infectious disease risks. To this end, the most relevant study field is the search for gene sites that impact production and health. This study aimed to determine the nature of the distribution of the relative frequencies of alleles and genotypes of polymorphic prolactin (PRL) and nitric oxide synthase (NOS2) in Holstein cows and identify the relationship of these genes with resistance to mastitis and bovine leukemia. For this study, we chose cows because infectious diseases affect the amount of lactation and milk quality. Holstein cattle with mastitis and bovine leukemia were selected. Animal genotypes were determined by restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR) products. The results were analyzed using a nonparametric statistical method using Microsoft Excel 2010 and Statistica 6.0. In healthy animals, 94 genotypes were identified for both genes under study. For bPRL, bPRL-RsaIAA (72) was the most common genotype and bPRL-RsaIBB (4) the least; for NOS2, bNOS2 -HinfIAB (47) was the most common genotype and bNOS2 -HinfIAA the least (21). In animals with leukemia, 34 genotypes were identified. For PRL, bPRL-RsaIAA (25) was the most common genotype and bPRL-RsaIBB (2) the least; for NOS2, bNOS2 -HinfIBB (17) was the most common genotype and bNOS2 -HinfIAA (3) the least. In animals with mastitis, 67 genotypes were identified. For PRL, bPRL-RsaIAA (43) was the most common genotype and bPRL-RsaIBB (6) the least; for NOS2, bNOS2 -HinfIBB (31) was the most common genotype and bNOS2-HinfIAA (7) the least. The distribution of genotypes of polymorphic bPRL and bNOS2 generally coincides, and bPRL-RsaIBB is the most common genotype. In groups of sick animals, the number of bNOS2 -HinfIAA homozygotes was lower than that of the control group. In particular, the proportion of animals with the bNOS2 -HinfIAA genotype with bovine leukemia was 8.7% and with mastitis was 10.3% compared with 22.4% in healthy animals. These data support the possible association of the bNOS2 -HinfIAA genotype with resistance to infection. The frequency of the bPRL-RsaIB allele was higher in groups of sick animals. This allele is associated with increased milk productivity, suggesting that highly productive animals are less resistant to the incidence of viral bovine leukemia and mastitis of bacterial etiology. DNA amplification of Holstein cattle for the polymorphic regions of PRL and NOS2 using the PCR-RFLP method revealed a possible connection between the distribution of relative allele frequencies of bPRL and bNOS2 and resistance to viral and bacterial infections. Thus, in groups of sick animals, the frequency of bPRL-RsaIBB, associated with increased milk production compared with the theoretically calculated equilibrium value was higher and the number of homozygotes bNOS2 -HinfIAA was lower than in the control group. In conclusion, animals with increased milk production were more prone to diseases, such as mastitis and bovine leukemia.

Роль генетических факторов в развитии гнойно-септических осложнений в акушерстве

The purpose to study the role of polymorphisms of endothelial nitric oxide synthase (eNOS3 G894T), transforming growth factor beta TGF-β (CA rs1800470) and plasminogen activator inhibitor (SERPINE-1 5G (-675)4G) in full-term pregnant women in the development of purulent-septic complications after cesarean section in the lower uterine segment. Material and methods. The authors analyzed obstetric complications after cesarean section in the lower uterine segment. Prospectively, 81 maternity patients after single-fetus delivery by caesarean section in the lower uterine segment were monitored and treated for emergency indications in order to diagnose purulent-septic complications and perform genetic analysis. The Statistica 10.0 and Eviews 12.0 software packages were used for statistical processing of the obtained results. Results. A direct correlation was established between the presence of a homozygous mutation of the transforming growth factor-β1 gene (TGF-β (CA rs1800470)), as well as the presence of a homozygous mutation of the endothelial nitric oxide synthase eNOS3 (G894T) gene, with significant risk factors for purulent-septic complications after cesarean section in full-term pregnancy: obesity 3-grade II (rs = 0.467, p &lt; 0.001; rs = 0.461, p = 0.005) and pathological overall weight gain during pregnancy (rs = 0.660, p &lt; 0.001; rs = 0.385, p = 0.024). Conclusion. The genetic study data allow concluding that homozygous polymorphisms of the studied genes of transforming growth factor-β1 (TGF-β (CA rs1800470)) and endothelial nitric oxide synthase eNOS3 (G894T) are associated both with the presence of purulent-septic diseases in the postpartum period after cesarean section in the lower uterine segment, and with metabolic disorders such as obesity of the 3rd degree and pathological overall weight gain during pregnancy. It is the combination of these factors that significantly increases the risks of obstetric purulent-septic complications in women with this genotype.

Lack of association between the eNOS rs1800779 (A/G) polymorphism and the myocardial infarction incidence among the Iraqi Kurdish population

ObjectivesThe genetic polymorphisms of the endothelial nitric oxide synthase (eNOS) gene are strongly associated with several cardiovascular diseases (CVDs) in various populations. The current study aimed to investigate the association of the eNOS rs1800779 (A/G) polymorphism with the progress of myocardial infarction (MI). MethodsEighty-five healthy subjects and 80 patients with MI admitted to the Erbil Cardiac Centre in the Kurdistan Region of Iraq were enrolled in the study. All participants were Kurdish from the same ethnic group. The amplification refractory mutation system polymerase chain reaction (ARMS-PCR) was used to determine the rs1800779 (A/G) polymorphism of eNOS, and the nitric oxide (NO) serum level was detected by spectrophotometer. ResultsThe genotypic frequencies of the eNOS rs1800779 AA (wild type), AG, and GG were 58.75%, 33.75%, and 7.50%, respectively, in the MI patients, and 49.41%, 43.53%, and 7.06%, respectively, for the control group. The frequencies of the A and the G alleles were 75.6% and 24.4%, respectively, in the MI group, and 71.2% and 28.8%, respectively, in the control subjects. The results revealed a lack of association of the rs1800779 genotype distribution with the level of NO serum and increased risk of MI. ConclusionThe study concluded that there is a lack of association between the genotypes and alleles of the rs1800779 eNOS and susceptibility to MI in the studied population.

Association of Nitric Oxide Synthase Polymorphism and Coagulopathy in Patients with Osteonecrosis of the Femoral Head.

Genetic polymorphism of nitric oxide synthase (NOS) can cause reduction of nitric oxide (NO) levels and may be associated with osteonecrosis of the femoral head (ONFH). However, the association of coagulopathy and NOS polymorphism in ONFH patients has not been confirmed. Between November 2005 and October 2013, 155 patients with ONFH were recruited in the study of serum coagulation profiles and NOS polymorphism. Another 43 patients who had dysplasia, osteoarthritis, or trauma of hip joints were included as controls. PCR genotyping for the analysis of NOS 27-bp polymorphism in intron 4 was performed. The analysis of coagulation profiles included fibrinogen, fibrinogen degradation product (FDP), protein S, protein C, and anti-thrombin III. The results showed that 27-bp repeat polymorphism was significantly associated with ONFH (OR 4.32). ONFH patients had significantly higher fibrinogen, FDP, protein S, and anti-thrombin III levels than that of the controls. The incidence of coagulopathy was significantly higher in ONFH patients (73.2%), and the odds ratio increased from 2.38 to 7.33 when they had 27-bp repeat polymorphism. Patients with hyperfibrinogenemia, elevated FDP levels, and with the risk factor of alcohol or steroid use had significantly higher risks of bilateral hip involvement. This study demonstrated the presence of NOS polymorphism, and a resultant reduction in NO production was associated with coagulopathy, which in turn might contribute to higher risks of bilateral ONFH. Our data suggests that checking NOS polymorphism and coagulopathy may provide a new avenue in managing ONFH.

Inducible Nitric Oxide Synthase iNOS-954-G>C and Ex16+14-C>T Gene Polymorphisms and Susceptibility to Vitiligo in the Saudi Population.

ObjectiveVitiligo is an acquired pigmentary skin disorder with regional disappearance of melanocytes. Multigenic inheritance has been proposed in the pathogenesis of vitiligo. The present study aimed to investigate the possible association of inducible nitric oxide synthase polymorphisms iNOS-954-G/C (rs1800482 G>C) and iNOS-Ex16+14-C/T (rs2297518 C>T) with vitiligo in the Saudi population, if any.MethodsWe included 120 vitiligo cases and an equal number of age matched healthy controls. Polymerase chain reaction with restriction fragment length polymorphism method was used for the analysis of genetic polymorphisms.ResultsThe heterozygous (GC), (GC + CC) combined genotype and variant allele; C allele of rs1800482 G>C were associated significantly (p < 0.005, after Bonferroni correction) with increased risk of vitiligo (OR = 3.46, 95% CI = 1.99–6.01, p = 0.001), (OR = 3.30, 95% CI= 1.93–5.65, p = 0.001) and (OR = 1.94, 95% CI = 1.31–2.87, p = 0.001) respectively. When GC genotype of rs1800482 G>C was co-inherited with common genotype (CC) and heterozygous genotype (CT) of rs2297518 C>T, the risk of vitiligo was significantly increased ((OR = 4.51, 95% CI = 2.18–9.33, p = 0.001) and (OR = 3.60, 95% CI = 1.61–8.01, p = 0.001)) respectively. None of the rs1800482 G>C and rs2297518 C>T genotypes and alleles have been associated with non-segmental vitiligo in terms of gender, age of onset, and types of vitiligo.ConclusionThe heterozygous (GC), (GC+CC) combined genotype and variants allele; C allele of rs1800482 G>C, may cause overproduction of NO, which has been linked to melanocyte loss by increasing oxidative stress and decreasing melanocyte adhesion to the extracellular matrix components, and thus could be an associative risk factor for vitiligo.

Effects of Nitric Oxide Synthase 3 Gene Polymorphisms on Cardiovascular Events in a General Japanese Population - The Yamagata (Takahata) Study.

Background: Single nucleotide polymorphisms (SNPs) in nitric oxide synthase 3 (NOS3) are associated with cardiovascular risk factors. However, it is not clear whether the NOS3 SNP is a genetic risk factor for cardiovascular diseases.Methods and Results: This prospective cohort study included 2,726 subjects aged ≥40 years who participated in a community-based health checkup. We genotyped 639 SNPs, including 2 NOS3 SNPs (rs1799983 and rs1808593). All subjects were monitored prospectively over a median follow-up period of 16.0 years, with the endpoint being cardiovascular events, including cardiovascular death and/or non-fatal myocardial infarction. Kaplan-Meier analysis demonstrated that both rs1799983 GT/TT and rs1808593 GG carriers had a higher risk of the endpoint than non-carriers. Univariate and multivariate Cox proportional hazard regression analyses revealed that both rs1799983 GT/TT and rs1808593 GG were independently associated with cardiovascular events after adjusting for confounding risk factors. The net reclassification index and integrated discrimination index were significantly improved by the addition of NOS3 SNPs as cardiovascular risk factors.Conclusions:NOS3 gene polymorphisms could be genetic risk factors for cardiovascular events in the general Japanese population, and could be used to facilitate the early identification of individuals at high risk of cardiovascular events.

Prognostic significance of three endothelial nitric oxide synthase (eNOS) polymorphisms and metabolic syndrome (MetS) in patients with colorectal cancer.

Polymorphisms of endothelial nitric oxide synthases (eNOS) have been associated with cancer susceptibility. Also, metabolic syndrome is associated with cancer malignancy. However, the effect of eNOS polymorphisms and metabolic syndrome on colorectal cancer (CRC) prognosis remains unclear. To investigated whether three genetic polymorphisms (-786T > C rs2070744, 4a4b rs869109213, and 894G > T rs1799983) in the eNOS and metabolic syndrome (MetS) were associated with CRC patient survival. We genotyped three polymorphisms of eNOS (-786T > C, 4a4b, and 894G > T) in 312 CRC cases from the Korean population by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Although the three eNOS polymorphisms were not causative of MetS, the TT genotype of the 894G > T polymorphism was associated with a worse survival rate compared with the GG genotype in the CRC group with MetS than in the CRC group without MetS (5-years survival; adjusted HR = 54.777; 95% CI 5.073-591.487 and RFS; adjusted HR = 14.909; 95% CI 1.571-141.528). The eNOS polymorphisms were not associated with metabolic syndrome prevalence in CRC patients. However, our findings suggest that the eNOS 894G > T polymorphism with MetS was associated with poor clinical outcomes.

Are P-glycoprotein (ABCB1/MDR1) and endothelial nitric oxide synthase (eNOS) polymorphisms related to severity of the coronary artery disease?

Background/Aim: Atherosclerotic cardiovascular disease is one of the most common causes of morbidity and mortality in developed countries. Genetic and environmental factors are associated with atherosclerosis development. Some single nucleotide polymorphisms have also been directly related to atherosclerosis, for example, polymorphisms that reduce nitric oxide (NO) levels and/or activity have been linked to atherosclerotic diseases. However, the multidrug resistance gene 1 (MDR-1) polymorphism is related to repeated cardiovascular events. This study aimed to investigate the relationship between MDR-1 and endothelial NO synthase (e-NOS) polymorphism and severe coronary artery disease (CAD). Methods: In total, 90 patients presenting with acute coronary syndrome were included in this cross-sectional study. Patients with at least > 70% stenosis in ≥ 2 coronary vessels were defined as severe CAD (Group 1), while those with this same level of involvement in < 2 vessels were diagnosed with single-vessel disease (Group 2). MDR 3435C-T and eNOS T-786C were determined by polymerase chain reaction (PCR) amplification. Comparison of parametric and nonparametric values between the two groups was performed using the Student’s t- or Mann–Whitney U test. Categorical variables were analyzed using the χ2 test. Risk estimations for the association of severe CAD with the polymorphisms were calculated using odds ratio (OR) and 95% confidence intervals by comparing the genotypic combinations. Results: Baseline demographic parameters were similar in both groups, except for the presence of DM and glucose level. T allele of MDR was 42% and 40% in groups 1 and 2, respectively (OR = 1.12). The C allele of eNOS was 34% and 30% in groups 1 and 2, respectively (OR = 1.16). Fourteen and 15 patients (40% and 27%, respectively) had both T and C alleles in patients in groups 1 and 2, respectively (OR = 1.77). All P-values were > 0.05. Conclusion: This study is the first one that shows that MDR1 and e-NOS polymorphisms are frequent in patients with ≥ 2 vessel disease and may be associated with severe CAD.

Effect of endothelial nitric oxide synthase gene polymorphism on cardiovascular death and nonfatal myocardial infarction in Japanese general population

Abstract Introduction Single nucleotide polymorphisms (SNP) of endothelial nitric oxide synthase (NOS3) have been reported to be associated with diabetes mellitus and myocardial infarction. However, few reports have prospectively investigated the effects of NOS3 SNP on cardiovascular death and nonfatal myocardial infarction. Purpose The purpose of this study was to investigate the impact of NOS3 SNP on cardiovascular death and the development of nonfatal myocardial infarction. Methods This prospective cohort study included 2,752 subjects (aged ≥40) who participated in a community based health checkup. We genotyped two SNPs within NOS3 (rs1808593, rs1799983). All subjects were prospectively followed during the median follow-up period of 15.4 years with the end point of cardiovascular death and nonfatal myocardial infarction. Results The homozygous G-allele (GG), heterozygous (GT), and homozygous T-allele (TT) carriers of rs1808593 were identified in 60 (2%), 706 (26%), and 1,986 (72%) subjects, respectively. Kaplan-Meier analysis demonstrated that homozygous G-allele carriers of rs1808593 had the greater risk than those without. Multivariate Cox proportional hazard regression analysis revealed that the homozygous G allele of rs1808593 was associated with cardiovascular death and the development of nonfatal myocardial infarction after adjusting for confounding risk factors. Conclusions NOS3 gene polymorphism could be a genetic risk factor for cardiovascular death and nonfatal myocardial infarction in the Japanese general population. Funding Acknowledgement Type of funding sources: None. Figure 1