APOE-E4 allele as a potential marker for implantation failure: A comparison between fertile women, ART success and RIF patients.

OBJECTIVE: Apolipoprotein E (APOE) is the most important precursor for the production of steroid hormones and is also involved in regulating the function of steroid hormones, hence playing a significant role in reproductive processes. So, APOE gene expression may be correlated with the implantation process. This study tries to make a better clarification of the correlation between APOE gene polymorphisms and recurrent implantation failure (RIF), where we compared the frequency of APOE polymorphisms in RIF patients, assisted reproductive treatment (ART) success cases and fertile women. METHOD: In all, 100 women with successful ART who got pregnant (fetal heart rate positive) in their first or second cycle of in vitro fertilization or intracytoplasmic sperm injection, 100 infertile RIF cases, and 100 normal fertile control cases with at least one live birth were included in present study. Following DNA extraction, genotypes were determined through polymerase chain reaction-restriction fragment length polymorphism method using HhaI restriction enzyme. Finally, statistical analysis was performed by chi-squared (χ2) test in SPSS software (P < 0.05). RESULTS: The RIF group showed significantly higher frequency for E3/E4 genotype (29%) compared with the other two control groups (fertile = 15%, ART success [ART+] = 13%) (P = 0.007). There was also a significantly higher frequency of the E4 allele in the RIF group (14.5%) compared with both of the control groups (fertile = 7.5%, ART+ = 6.5%) (P = 0.018). CONCLUSION: APOE4 is correlated with recurrent failure in the process of embryo implantation and, accordingly, it may potentially be considered a possible risk factor to the implantation process. The presence of E4 can be proposed as a predictive indicator in determining the results of assisted reproductive techniques.

The association between thrombophilic genes alterations and poor ovarian response in infertile women: a retrospective case-control study.

This research aimed to retrospectively investigate the possible association between poor ovarian stimulation and selected thrombophilia markers in Iranian women with infertility. For this study 100 Iranian infertile women, with a history of at least three Assisted Reproduction Technology (ART) failures (50 with a poor ovarian response and 50 with a normal response), referred to Royan Institute were selected. Targeted genetic variation evaluation for Factor V G1691A, F II Prothrombin G20210A, MTHFR C677T, MTHFR A1298C was performed by PCR-RFLP followed by Sanger Sequencing. The association between these variants and the ovarian response was examined. The results showed an association between Factor V G1691A mutation and poor ovarian response. The heterozygosity rate of the FVL was significantly different between poor responders compared with the normal response group (p-value ≤ 0.05). In conclusion screening of this polymorphism can be used as a genetic determinant of ovarian response functioning through a vascular mechanism. A larger study with bigger sample size is recommended.Impact statementWhat is already known on this subject? Thrombophilia is a multi-genetic disease that is associated with changes in homeostatic mechanisms. Some studies have suggested that thrombophilia has no relationship with poor ovarian response and reduced ovarian reserve in general infertile population undergoing ART.What do the results of this study add? Our results showed a significant association between the FVL heterozygote mutation and poor ovarian response.What are the implications of these findings for clinical practice and/or further research? Screening of FVL polymorphism may be suggested as a predictive test for ovarian stimulation response in infertile women undergoing ART. Further prospective studies with bigger sample size evaluating other thrombophilia markers and ovarian response, as well as further in-vitro studies may help clarify the biological mechanisms behind the effect of the FVL polymorphism on ovarian response, oocyte quality and embryo quality.

Oxidative stress-related miRNAs in spermatozoa may reveal the severity of damage in grade III varicocele.

Varicocele is associated with excessive production of reactive oxygen species (ROS). Although the harmful effects of ROS on sperm DNA, proteins and lipids are well documented, its impact on the expression of miRNAs in spermatozoa has not been fully understood. In this study, the expression patterns of microRNAs (miRNAs), miR-21, miR-34a and miR-122a as well as the level of ROS in the fertile control (FC; proven fertility without varicocele, n = 15) and grade III varicocele patients with normal (VN; n = 15) and abnormal (VA; n = 15) spermogram were investigated. The real-time PCR was performed to analyse the expression of the miRNAs, while oxidative stress was evaluated by measuring the concentrations of MDA. Our results showed that the expression levels of miR-21 (p = .001), miR-34a, (p = .007) and miR-122a (p < .001) were significantly decreased in spermatozoa of VN and VA patients in comparison with the fertile group. Also, increased levels of oxidative stress were detected in semen samples of varicocele patients compared with the fertile control (p < .0001). Overall, these findings demonstrate oxidative stress changes the expression pattern of some miRNAs, and these alterations could be a valuable diagnostic marker for the diagnosis and prognosis of varicocele-induced oxidative stress to retain the male fertility during the spermatogenesis process.

Birth of a healthy boy following preimplantation genetic diagnosis for congenital adrenal hyperplasia.

Classical 3ß-HSD deficiency due to mutations in the HSD3B2 gene is responsible for a rare form of congenital adrenal hyperplasia (CAH) and is identified by varying degrees of salt wasting. Preimplantation genetic diagnosis (PGD) was performed in a couple carrying mutation c.690 G>A in the HSD3B2 gene. Four polymorphic short tandem repeat markers closely linked to the HSD3B2 gene (D1S185, D1S453, D1S514, D1S540) for linkage analysis in conjunction with the direct mutation analysis were used in embryo genotyping. Two CODIS STRs (VWA and THO1) were also used to confirm embryo zygosity and rule out possible contaminations. Finally, SRY and AMYLOGENIN markers were used for embryo sex determination. PGD was performed by fluorescent multiplex seminested polymerase chain reaction and sequencing. Six embryos were tested and one male carrier embryo was transferred, resulting in the birth of a healthy boy.