The p53 protein is known for performing essential functions in the maintenance of genomic stability in somatic cells and prevention of tumor formation. Studies of the p53 signaling pathway have suggested associations between some polymorphisms and infertility, post-in vitro fertilization implantation failure and recurrent abortions. The TP53 Pro72Arg polymorphism has been implicated as a risk factor for recurrent pregnancy loss (RPL); however, the association is controversial. In this study, our objective was to evaluate selected polymorphisms in genes of the p53 signalling pathway [TP53 c.215G>C (Pro72Arg), MDM2 c.14+309T>G (SNP309) and LIF c.1414T>G in the region 3' UTR] and determine their effect as risk factors for RPL. In a case-control study, we investigated 120 women with two or more pregnancy losses and 143 fertile control women reporting at least two live births and no history of pregnancy loss. When analyzed separately, the allele and genotype distributions of the polymorphisms in the two groups were not different. However, in a multivariate analysis adjusted for alcohol consumption, smoking, ethnicity, and number of pregnancies, the interaction between the genotypes TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) showed to be associated to RPL, increasing the risk for this condition (OR = 2.58, 95% CI: 1.31-5.07, p = 0.006). In conclusion, our study indicates that the combination of TP53 Arg/Arg (rs1042522) and MDM2 TT (rs2279744) genotypes may be a risk factor for RPL.

No abstract

Recurrent pregnancy loss (RPL) is defined as the occurrence of two or more consecutive pregnancy losses. It is an important reproductive condition with a complex etiology. In approximately 50% of RPL cases an explanation for the cause is not found and they are therefore classified as idiopathic RPL. One of the causes implicated in RPL is thrombophilia, which consists of hemostatic disorders that lead to an increase in thromboembolic processes. The aim of this study was to evaluate polymorphic variants in genes related to thrombophilia as a risk factor in women with RPL. We investigated 145 women with at least two consecutive pregnancy losses and 135 women with at least two children and no history of pregnancy loss. Genotypes for the polymorphisms MTHFR C677T, FVL, FII (prothrombin), eNOS T-786C, and eNOS Glu298Asp were determined using a real-time PCR. Information about the exposure to environmental risk factors was also collected. There was no significant association between the environmental risk factors assessed and the polymorphisms studied. We did not find statistically significant differences in genotypic or allelic frequencies for the polymorphisms studied, in either the women with RPL or in the control group. Such polymorphisms should therefore not be considered as risk factors for this condition in this population.

Recurrent pregnancy loss (RPL) affects ~3-5% of couples attempting to conceive and in around 50% of cases the aetiology remains unknown. Adequate vascularisation and placental circulation are indispensable for the development of a normal pregnancy. Prostaglandin-endoperoxide synthase 2 (PTGS2), vascular endothelial growth factor (VEGF) and the nitric oxide (NO) systems play important roles in reproductive physiology, participating in several steps including implantation and apoptosis of trophoblast cells. In this study we evaluated genetic polymorphisms in the inducible nitric oxide synthase (NOS2), PTGS2 and VEGFA genes as susceptibility factors for RPL. A case-control study was conducted in 149 women having two or more miscarriages and 208 controls. Allele and genotype distributions of the polymorphisms studied in the two groups were not statistically different. However, the dominant model showed that the presence of variant T (TT/GT) of rs2779249 (-1290G>T) of NOS2 was significantly associated with RPL (OR=1.58, CI 95%=1.03-2.44; P=0.037). The increased risk remained significant when adjusted for number of pregnancies, alcohol consumption and ethnicity (OR=1.92, CI95%=1.18-3.11; P=0.008). These results suggest that the variant genotypes of the functional polymorphism rs2779249 in the NOS2 promoter are a potential risk for RPL, possibly due to oxidative stress mechanisms.

IVM can be an advantageous technique when applied to PCOS (Polycystic Ovarian Syndrome) patients. The oocytes are retrieved from antral follicles of non-stimulated ovaries, specially preventing hyperstimulation syndrome. Apart from its role as a reproductive treatment, IVM has emerged as a promising tool for emergency fertility preservation, since it can be performed flexibly in either follicular or luteal phase. A 34-year-old patient with PCOS, high body mass index and tubal factor was submitted twice to IVM treatment. Her husband has low count spermatozoa. The first IVM cycle was in 2009, she transferred 3 fresh embryos and got pregnant giving birth to a healthy boy weighing 3.3 kg. In 2013, the patient returned for another IVM cycle and the embryos had to be vitrified because she failed to develop an adequate endometrium for transfer. In the next cycle, the endometrium was prepared using estrogen and progesterone and the two best embryos were warmed up and transferred. She became pregnant and after 36 weeks gave birth to a healthy girl weighing 2.7 kg. She still has four embryos left to transfer. IVM may be an alternative technique to be considered when dealing with PCOS patients. Although clinical outcomes are currently inferior when compared with conventional hormone driven ART (Artificial Reproductive Techniques), it does apply in some cases while preventing hyperstimulation risks. Thus, embryos obtained by IVM can also be vitrified with successful outcomes.

Background/Purposes A common observation in oocyte in vitro maturation (IVM) cycles is poor embryo quality. However, no study was dedicated to assess zygote and early cleavage embryo quality in IVM cycles. The objective of this study is to analyze fertilization outcome, embryo development and the resulting pregnancy and births in unstimulated IVM cycles. Methods IVM oocytes were collected 36 h post hCG and matured in vitro for 28-30 h. All oocytes were inseminated by ICSI. Resulting zygotes and embryos were assessed on day-1, 2 and 3, when transfers were made. Results The overall oocyte maturation and fertilization rates were 63% and 62%, respectively. Abnormal fertilization rate was 1.7%. Ninety five and 14.6% of the 2Pn zygotes reached the 2-cell and 8 -cell stage at day-2 and day-3, respectively. Embryo quality assessment on day-3 at transfer revealed that only 9% of the embryos were of very good quality. Most embryos showed developmental delay. An average of 3.29 embryos were transferred per patient resulting in implantation and clinical gestation rates of 16% and 32%, respectively. Overall 14 healthy babies were born and there is one ongoing pregnancy. ConclusionResults show a significant rate of abnormal fertilization and poor embryo quality after IVM, which is reflected in a higher than average number of embryos being transferred. However, pregnancy, implantation and birth rates are reasonably high and allow us to consider IVM a valuable approach for the treatment of infertility in PCO or PCOS patients.

ABSTRACT. Some cases of recurrent first trimester miscarriage have a thrombotic etiology. The aim of this study was to investigate the prevalence of the most common thrombophilic mutations -factor 2 R.O. Gonçalves et al. Genetics and Molecular Research 15 (3): gmr.15038156 V (FV) Leiden G1691A (FVL), prothrombin (FII) G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677T -in women with recurrent miscarriages. In this case-control study, we included 137 women with two or more consecutive first-trimester miscarriages (£12 weeks of gestation) and 100 healthy women with no history of pregnancy loss, and with at least one living child. DNA was extracted from the patient samples, and the relevant genes (FVL, FII, and MTHFR) were amplified by PCR, followed by restriction fragment length polymorphism, to assess the polymorphisms in these genes. The allelic frequencies of polymorphisms were not significantly different between the case and control groups. Polymorphisms in the MTHFR, FVL, and FII genes were not associated with recurrent miscarriage during the first trimester of pregnancy in Brazilian women (P = 0.479; P = 0.491 and P = 0.107, respectively). However, the etiologic identification of genetic factors is important for genetic counseling.

Non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) aiming to assess cell-free embryonic DNA in spent culture media is promising, especially because it might overcome the diminished rates of implantation caused by the inadequate performance of trophectoderm (TE) biopsy. Our center is part of the largest study to date assessing the concordance between conventional PGT-A and niPGT-A, and we report here the delivery of the first baby born in Brazil using niPGT-A. The parents of the baby were admitted to our center in 2018. They did not present history of infertility, and they were interested in using in vitro fertilization (IVF) and PGT-A in order to avoid congenital anomalies in the offspring. A total of 11 (3 day-5 and 8 day-6) expanded blastocysts were biopsied, and the spent culture media (culture from day-4 to day-6) from 8 day-6 blastocysts were collected for niPGT-A. Overall, 7 embryos yielded informative results for trophectoderm (TE) and media samples. Among the embryos with informative results, 5 presented concordant diagnosis between conventional PGT-A and niPGT-A, and 2 presented discordant diagnosis (1 false-positive and one false-negative). The Blastocyst 4, diagnosed as 46, XY by both niPGT-A and conventional PGT-A, was warmed up and transferred, resulting in the birth of a healthy 3.8 kg boy in February 2020. Based on our results and the recent literature, we believe that the safest current application of niPGT-A would be as a method of embryo selection for patients without an indication for conventional PGT-A. The approximate 80% of reliability of niPGT-A in the diagnosis of ploidy is superior to predictions provided by other non-invasive approaches like morphology and morphokinetics selection.