Investigation of endothelin-1 type A receptor gene polymorphism (−231 G > A) in preeclampsia susceptibility

Objective. Preeclampsia is considered as a multifactorial disorder with a genetic predisposition. Alterations in the endothelin-1 (ET-1) system are considered to take part in triggering the vasoconstriction seen in preeclampsia.

Methods. In order to investigate the possible association of the ?231 G > A polymorphism in the endothelin-1 type A receptor gene (EDNRA), previously shown to be associated with other conditions characterized by vasospasm, we examined 77 Caucasian preeclamptic women and 67 matched controls including normotensive subjects without history of thromboembolic event, abnormalities in blood pressure, proteinuria, edema and preeclampsia. The genotype was assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) on genomic DNA extracted from blood samples. Case vs. control allele frequencies and genotype distributions were compared.

Results. No significant differences were found when considering both genotype (χ² = 0.58, p = 0.75) and allelic frequencies (χ² = 0.08, p = 0.77). Furthermore, no significant genotype-related difference was found in relation to clinical features, such as gestational age at onset, systolic and diastolic blood pressure, proteinuria on admission and delivery week.

Conclusions. No association between the ?231 G > A polymorphism in the EDNRA gene and preeclampsia as well as any correlation with the main clinical features of the disorder were found, thus excluding a role for this polymorphism in susceptibility to preeclampsia.     

Association between the CLOCK gene 3111 T > C polymorphism and an irregular menstrual cycle in Korean adolescents

Abstract

The menstrual cycle is an example of a human infradian rhythm, but an altered sleep–wake cycle or a disrupted circadian rhythm can change the regularity of the menstrual cycle. In this study, we investigated whether an irregular menstrual cycle is associated with polymorphisms in the CLOCK (3111T?>?C) and/or PER3 (variable number tandem repeat, VNTR) genes, which are known to have an impact on the circadian rhythm. One hundred ninety-seven postmenarchal, adolescent girls from two girls’ high schools in Seoul, Korea, were studied. All participants were requested to complete the Perceived Stress Scale (PSS), the State-Trait Anxiety Inventory (STAI), and the Beck Depression Inventory (BDI) to assess the emotional distress that might cause menstrual irregularity. Every participant donated a blood sample from which DNA was extracted and genotyped for the CLOCK 3111T?>?C and PER3 VNTR polymorphisms. A significant association was found between the CLOCK 3111T?>?C genotype and irregular menstrual cycles. Subjects with the 3111T?>?C genotype had a high risk of an irregular menstrual cycle compared with 3111T/T homozygous subjects (odds ratio [OR]?=?2.88; 95% confidence interval [CI]: 1.26–6.55). When multivariate logistic regression analysis was performed to adjust for age, PSS, STAI, BDI and BMI, subjects with the 3111T?>?C polymorphism showed a significantly increased OR for irregular menstrual cycles (OR?=?3.09; 95% CI: 1.32–7.21). There was no significant association between the PER3 VNTR polymorphism and the irregularity of the menstrual cycle (p?>?0.05). The results of this study suggest that the CLOCK 3111T?>?C polymorphism could be an independent risk factor for irregular menstrual cycles, irrespective of psychological distress and endocrine or metabolic conditions, and could be used as a molecular marker for gynecological studies on this aspect.     

The −2549 insertion/deletion polymorphism in the promoter region of the VEGFA gene in couples with idiopathic recurrent spontaneous abortion

PurposeThe vascular endothelial growth factor A (VEGFA) is crucial for normal vasculogenesis and angiogenesis during pregnancy, and alterations in the VEGFA gene expression were detected in women with idiopathic recurrent spontaneous abortion (IRSA) and spontaneously aborted conceptuses. Our aim was to evaluate whether there is an association between the functional −2549 insertion/deletion (I/D) polymorphism in the promoter region of the VEGFA gene and IRSA in reproductive couples.MethodsWe performed a case-control study involving 149 women and their 140 partners with three or more IRSA and 149 control women and men. Allele-specific polymerase chain reaction was used for genotyping.ResultsWe found no association of the −2549 I/D polymorphism with IRSA in women. However, men with the DD genotype have a 1.75-fold increased risk of IRSA compared with men carrying the ID and II genotypes (95 % confidence interval (CI) = 1.05–2.93, P = 0.032). In addition, the D allele in men contributes to a 1.42-fold increased risk of IRSA (95 % CI = 1.02–1.97, P = 0.036) compared to men carrying the I allele.ConclusionsOur results indicate that the −2549 I/D polymorphism in the VEGFA gene in men might be associated with IRSA. Additional genetic association studies including both partners, as well as expression studies, are needed to elucidate the role of this polymorphism in IRSA.     

Association between the growth arrest-specific 6 (Gas6) gene polymorphism c.834 + 7G>A and preeclampsia

Objective: Preeclampsia (PE) is a hypertensive disease of pregnancy complicating 2–8% of all pregnancies. The exact pathophysiology still remains unknown. Growth arrest-specific 6 (Gas6) is a member of the vitamin K-dependent protein family and it has been suggested as a novel atherothrombotic risk factor with anti-angiogenic and pro-atherogenic properties. The goal of the our study was to investigate the relationships between the c.834?+?7G?>?A polymorphism of GAS6, plasma Gas6 levels and PE.

Methods: A total of 150 women, including 82 preeclamptic pregnant women and 68 normotensive pregnant (NP) women, were recruited in the current study. Blood samples were taken from all participitants. Plasma Gas6 levels measured by an enzyme-linked immunosorbent assay. GAS6 polymorphism was determined using a PCR-RFLP method.

Results: The plasma Gas6 levels of preeclamptic patients were significantly lower than those of NP women (8.65?±?3.70?ng/ml and 10.89?±?4.23?ng/ml respectively, p?<?0.001). The GG genotype was the most prevalent, and the risk of PE was 3.5-fold higher in pregnant women with GG genotype compared to woman with AA genotype (p?<?0.01). The A allele was less frequent in preeclamptic patients than in control subjects (OR?=?2.118, 95% CI?=?1.330–3.371, p?<?0.001).

Conclusions: Our results suggest that GAS6 c.834?+?7G?>?A polymorphism may have a pivotal role in the pathogenesis of PE suggesting that the A allele has a protective role for PE.     

The vascular endothelial growth factor (VEGF) +405 G/C polymorphism and its relationship with recurrent implantation failure in women in an IVF programme with ICSI

Purpose

Successful embryo implantation depends on trophoblast proliferation, migration and, lastly, invasion of the endometrium (to anchor the trophoblast to the uterus). This invasion is mediated by locally produced soluble factors. Of these, vascular endothelial growth factor (VEGF) is the best characterized regulator of angiogenesis. Here, we investigate the association between the VEGF + 405 C/G genotype and the recurrence of embryo implantation failure in women undergoing in vitro fertilization (IVF) program with intracytoplasmic sperm injection (ICSI).

Methods

Forty women with recurrent implantation failure defined by absence of pregnancy after transfer of more than 10 embryos and 131 women control, with at least one live birth after the transfer of fewer than 10 embryos were included. Genomic DNA was analysed with an allele-specific polymerase chain reaction and a Chi-2 test was used to compare the respective VEGF + 405 C/G genotype frequencies in cases and controls.

Results

The frequency of the VEGF +405C/C genotype was higher in women with recurrent implantation failure after ICSI-embryo transfer than in controls (17.5 % and 5.3 %, respectively, p = 0.01).

Conclusion

The VEGF +405 G/C polymorphism may influence embryo implantation and VEGF + 405 C/C genotype may predispose to recurrent implantation failure after ICSI-ET.     

Involvement of <Emphasis Type="Italic">17β-hydroxysteroid dehydrogenase type</Emphasis> gene <Emphasis Type="Italic">1</Emphasis> 937 A&gt;G polymorphism in infertility in Polish Caucasian women with endometriosis

Purpose

Endometriosis is considered to be an estrogen-related chronic inflammatory disease. The 17β-hydroxysteroid dehydrogenase 1 (HSD17B1) converts estrone to 17β estradiol. The role of HSD17B1 937 A>G (rs605059) single nucleotide polymorphism (SNP) in development of endometriosis is still disputable. This study evaluated the association of the HSD17B1 937 A>G (rs605059) SNP with infertile women affected by endometriosis from Polish Caucasian population.

Methods

The genotyping of cases (n = 290) and fertile women (n = 410) was conducted by high-resolution melting curve analysis.

Results

Statistical analysis demonstrated that the HSD17B1 937 A>G SNP is associated with endometriosis in stages I and II. The p _trend and p _allelic values calculated for the HSD17B1 937 A>G polymorphism were statistically significant and were equal to 0.001 and 0.0009, respectively. There was a significant association for the dominant model: (AG + GG vs AA) OR = 1.973 (95% CI = 1.178–3.304), p = 0.009, and for the recessive model: (GG vs AG + AA) OR = 1.806 (95% CI = 1.178–2.770), p = 0.006. However, we did not find statistical association of HSD17B1 937 A>G polymorphism with all infertile women with endometriosis or infertile women with endometriosis in stages III and IV.

Conclusion

Our genetic study demonstrated HSD17B1 937 G variant as a risk factor for infertility in women with stage I and II endometriosis in Polish Caucasian patients.

     

The significance of serum anti-Müllerian hormone (AMH) levels in patients over age 40 in first IVF treatment

Purpose

Although studies of serum anti-Müllerian hormone (AMH) in predicting ovarian reserve are numerous, many studies utilized patients under age 40. However, the assessment of ovarian reserve is especially critical in older infertile women. This study evaluates the significance of AMH level in patients over age 40 at the time of their first in vitro fertilization (IVF) treatment.

Methods

Forty-nine women over age 40 were studied. Although serum samples were taken prior to their IVF treatments, the data of serum AMH of patients were not taken into consideration to determine the therapy strategy, including follicle induction in which clomiphene citrate and human menopausal gonadotropin were used.

Result(s)

Twelve out of 49 patients achieved a clinical pregnancy (24.4 %). There was a positive correlation between serum AMH levels and the number of oocytes retrieved (P < 0.0001). The ROC curve analysis for prediction of poor ovarian response, ≤3 retrieved oocytes, showed that the optimum cut-off level was < 1.0 ng/mL for AMH. The lower AMH group (AMH < 1.0 ng/ml) showed less chance of undergoing embryo transfer than the higher AMH group (AMH ≥1.0 ng/ml). There was no difference in pregnancy rate between the two groups. Five out of 12 pregnant women exhibited AMH levels of less than 0.4 ng/ml.

Conclusion(s)

Assessment of serum AMH concentration in older patients is useful for the prediction of oocytes numbers which may be obtained in IVF. A cut-off level of 1.0 ng/ml AMH can be used to predict poor ovarian response. This cut-off level of AMH of 1.0 ng/ml might be useful to predict whether patients could have an embryo transfer, but had no power to predict achieving pregnancy. On the other hand, our data also showed that patients over age 40 with extreme low levels of AMH still had a chance of pregnancy.     

The mechanism of anticancer activity of the new synthesized compound - 6,7-Methylenedioxy-4-(2,4-dimethoxyphenyl)quinolin −2(1H)-one(12e) in human ovarian cancer cell lines

ObjectiveOvarian cancer is the most lethal of the gynecologic malignancies. Most women have advanced disease at diagnosis and require extensive debulking surgery and aggressive chemotherapy. Induction of apoptosis in cancer cells has been used as an important approach for cancer therapy. We examined the anticancer effect of 6,7-methylenedioxy-4-(2,4-dimethoxyphenyl)quinolin-2(1H)-one (12e) in human ovarian cancer cell line.Materials and methodsThe 6,7-methylenedioxy-4- (2,4-dimethoxyphenyl) quinolin-2 (1H) -one (12e) was synthesized and provided by Dr. Li-Jiau Huang of China Medical University. Cell viability analysis showed that 12e inhibited cell growth and induced cell death in time- and dose-dependent manners. In order to study the underlying cell death mechanism, 2774 and SKOV3 cells treated with 12e were studied by morphology, DAPI/TUNEL double staining, DNA gel electrophoresis. To search the mechanisms of anti-proliferative effect of 12e, cell cycle analysis was performed. Changes in proteins related to cell death were analyzed by Western blot.Results12e significantly induced apoptosis evidenced by morphological changes, TUNEL-DAPI double-staining and DNA fragmentation. Western blot analysis demonstrated that the protein level of Bcl-2 was decreased after treatment with 12e, while the level of p53 and Bax was increased. 12e treatment resulted in G2/M arrest through down modulation of cyclin B1 and cdk1.ConclusionThese results suggested that 12e -induced growth inhibition was associated with cell cycle arrest and apoptosis.     

The significance of an early (premature) rise of plasma progesterone in in vitro fertilization cycles induced by a “long protocol” of gonadotropin releasing hormone analogue and human menopausal gonadotropins

Objective: Our purpose was to assess and clarify the mechanism of whether an early progesterone rise in cycles with gonadotropin-releasing hormone agonist (GnRH-a) is associated with an impairment of IVF outcome Methods: Seven hundred eighty-six cycles were induced with GnRH-a and human menopausal gonodotropin (hMG) (long protocol). Plasma progesterone (PP) levels on the day of human chorionic gonadotropin (hCG) administration were divided into three groups: <0.9 ng/ml (Group A), 1–2 ng/ml (Group B), and >2 ng/ml (Group C). We also analyzed the pregnancies achieved in our egg donation protocol in relation to the PP levels of each donor on the day of hCG administration. Results: Group A involved 525 cycles, Group B had 223, and Group C had 38. The overall pregnancy rate per egg transfer was 19.2%, with the highest for Group A (22.3%), declining for Groups B (14.3%) and C (7.9%) (A = B = C; P<0.005). The embryo implantation rate was found to be negatively correlated with the PP levels on the day of hCG administration. In contrast, there was an opposite trend between PP levels and the chance of conception in 30 pregnancies achieved by egg donation. Conclusions: Since premature luteinization is very unlikely to occur under the conditions of this study, our findings suggest that an early PP rise has a negative impact on endometrial receptivity but not on egg and embryo quality.