Association of Follicular Fluid Antioxidants Activity with Aging and In Vitro Fertilization Outcome: A Cross-Sectional Study.

BACKGROUND: This research was aimed at assessing the relationship between the follicular fluid (FF) antioxidants activity, aging and in vitro fertilization (IVF) outcome. MATERIALS AND METHODS: The present cross-sectional study was carried out on 65 women undergoing IVF/intracytoplasmic sperm injection (IVF/ICSI) cycles due to unexplained infertility. Ovarian stimulation was performed using the long gonadotropin-releasing hormone (GnRH) agonist protocol. After ovum pickup, FF was collected and processed to measure the level of superoxide dismutase (SOD), catalase (CAT) activity, total antioxidant capacity (TAC) and glutathione (GSH). Day 3 after ICSI, fresh embryos were transferred and later, possible pregnancy was assessed. Patients participating in this study were divided into four groups on the basis of age and pregnancy outcome. RESULTS: SOD activity was not significantly different between the groups (P=0.218). GSH in the group whose participants were aged ≤35 years and were pregnant was higher than that in other groups. CAT activity in groups with younger participants was higher compared to the other groups. The mean TAC was higher in groups with pregnant participants compared to the non-pregnant women. Correlation analysis showed that: GSH level had a significant negative correlation with age (P<0.001, R -0.55) and a significant positive correlation with pregnancy (P=0.015, R=0.30). CAT level also had a significant negative correlation with age (P<0.001, R=-0.42) and the level of TAC had a significant positive correlation with pregnancy (P<0.001, R=0.59). CONCLUSION: According to our results, the levels of TAC, GSH and CAT in younger and pregnant women were higher compared with those undergoing ICSI cycles. Given the correlation of FF antioxidant activity with age and pregnancy, it is necessary to carry out more research on these compounds and the maintenance of pregnancy.

Assisted Reproductive Techniques and submucous myoma.

Submucous myomas have negative effects on fertility. To maintain fertility, conservative treatment should be suggested to women who wish to become pregnant, especially young patients. The patient was a 33-year-old woman, who had had secondary infertility for 3 years. Upon vaginal ultrasound, we noticed a submucous myoma measuring 26 mm x 31 mm with a compressive effect on the anterior surface of the endometrium. Ovarian reserve was low. The gold standard of myoma treatment is surgical intervention. But, for the following reasons: the adverse effects of surgery on the endometrium (intrauterine adhesion), the patient's refusal to undergo a myomectomy and her request for pregnancy, our strategy for treating was to reduce volume of submucous myoma and start the assisted reproductive techniques (ART) cycle, simultaneously. We administered three courses of Gonadotropin-releasing hormone analogues (GnRHa) and then induced controlled ovarian hyperstimulation. Ovum pick up was done. Finally, we transferred two embryos (4 and 6 cells). In subsequent patient visits, ßhCG was positive after 14 days. At the last patient visit, the heart of the embryo was formed. From this finding, it may be concluded that combined GnRHa and ART is the treatment of choice for infertile women with uterine submucous myoma, considering the reduced ovarian reserve and response.

A successful pregnancy following recurrent implantation failure with clinical laboratory strategy.

In treatment cycles of in vitro fertilization (IVF), 15% of the oocytes are immature and in the germinal vesicle (GV) phase. In rare occasions, more than 50% of the oocytes of a patient in a cycle are immature. Selecting fertility treatment for patients in this situation can be very challenging. The patient described in this report was a 35-year-old woman with primary infertility for 10 years. She had undergone fertility treatment six times, with no success. In her cycles, more than 50% of the oocytes were immature and in the GV phase. To address the situation, we developed a coordinated protocol involving medical and embryology procedures, analyzed the patient's medical records, and looked into the reasons of prior treatment failure. The development of a special and coordinated effort - instead of having medical and embryology teams working in isolation - proved efficient at providing better outcomes to the patient.

Expression of miR-302 in human embryo derived from in-vitro matured oocyte.

BACKGROUND: The expression of miR-302 over the period of early embryogenesis could possibly regulate the maternal transcript clearance. Zygotic transcription activation is mostly related to maternal messages degradation. OBJECTIVE: In this study, the effects of in-vitro maturation technique (IVM) on the expression of miR-302 in human embryo produced from immature and mature human oocytes (matured in vitro and in vivo, before sperm exposure) obtained from females under gonadotrophin therapy were evaluated for assisted reproduction. MATERIALS AND METHODS: Immature oocytes were cultured in vitro. The injection of oocytes-producing polar bodies was given using fresh sperm. Then, the embryo quality score was assessed in the IVM group compared with the control group. In both the groups, embryos with normal morphology were included in the molecular study. Only one blastomere was removed from three-day embryos and then the embryos were frozen. The expression of miR-302 in embryos was measured through quantitative real-time polymerase chain reaction. RESULTS: Our data showed a significant reduction of miR-302 expression in the IVM group vs. the control group (p = 0.02). The embryo quality score showed a significant difference between the two groups (p = 0.01). CONCLUSION: The present study demonstrated that the IVM process had a negative effect on the expression level of miR-302 in human pre-implantation embryos. Considering the major role of expression miR-302, a reduced potential in miR-302 expression could be related to a decrease in the early embryonic development.

The Effects of In Vitro Maturation Technique on The Expression of Genes Involved in Embryonic Genome Activation of Human Embryos.

OBJECTIVES: In vitro maturation technique (IVM) is shown to have an effect on full maturation of immature oocytes and the subsequent embryo development. Embryonic genome activation (EGA) is considered as a crucial and the first process after fertilization. EGA failure leads to embryo arrest and possible implantation failure. This study aimed to determine the role of IVM in EGA-related genes expression in human embryo originated from immature oocytes and recovered from women receiving gonadotrophin treatment for assisted reproduction. MATERIALS AND METHODS: In this experimental study, germinal vesicle (GV) oocytes were cultured in vitro. After intracytoplasmic sperm injection of the oocytes, fertilization, cleavage and embryo quality score were assessed in vitro and in vivo. After 3-4 days, a single blastomere was biopsied from the embryos and then frozen. Afterwards, the expression of EGA-related genes in embryos was assayed using quantitative reverse transcriptase-polymerase chain reaction (PCR). RESULTS: The in vitro study showed reduced quality of embryos. No significant difference was found between embryo quality scores for the two groups (P=0.754). The in vitro group exhibited a relatively reduced expression of the EGArelated genes, when compared to the in vivo group (all of them showed P=0.0001). CONCLUSIONS: Although displaying the normal morphology, the IVM process appeared to have a negative influence on developmental gene expression levels of human preimplanted embryos. Based on our results, the embryo normal morphology cannot be considered as an ideal scale for the successful growth of embryo at implantation and downstream processes.

Ovarian stimulation affects the population of mouse uterine NK cells at early pregnancy.

The aim of this study was to determine the influence of ovarian stimulation on endometrial mouse NK cell population. For superovulation, the female adult NMRI mice were injected i.p. with 10 IU of the pregnant mare serum gonadotropin followed 48 h later by an i.p. injection of 10 IU human chorionic gonadotropin hormone. Ovarian stimulated and nonstimulated mice were mated with fertile male. The presence of vaginal plug proved natural pregnancy, and this day was considered as day one of pregnancy. Tissue samples were prepared from the uterine horn and spleen of both groups of study on 7th day of pregnancy. Serum estradiol-17ß and progesterone were measured at the same time. The tissue cryosections were prepared and double stained for CD 161 and CD3 markers, and NK cells population was analyzed. Relative frequency of NK cells was significantly lower in stroma and myometrium in hyperstimulated mice compared with the control group. However, no difference was seen in percentage of NK cells in spleen. The ovarian stimulation influences the proportion of uterine NK cells and may affect the embryo implantation.