Although the causality between Zika virus, microcephaly, and other central nervous system disorders has been taken for granted by the scientific community, many uncertainties remain. The gap of knowledge at the moment is large enough to remove part of the confidence physicians have on the advice given to patients - and infertile women in particular - on their reproductive plans. Pretreatment serologic screening is a possible strategy to offer more confidence for individuals choosing to bear children regardless of the Zika virus, but the tests currently available do not seem to be sufficiently adequate. Until now, there is no formal recommendation to avoid pregnancy solely because of the Zika virus outbreak, and the choice of becoming pregnant has been regarded as a personal decision to be made by each woman and her family.
Objective: To assess the relationship between human blastocyst chromosomal ploidy established by niPGT-A and increasing age. Methods: This is a prospective multicenter study carried out by ten assisted reproduction centers after their embryologists acquired training and validated their results with the previous use of niPGT-A. A total of 94 couples with indication for niPGT-A due to increase maternal age, male factor, repeated implantation failures, recurrent abortion or because they requested niPGT-A were included in this study. The couples had no karyotype abnormalities. After ICSI, the embryos were cultured until blastocyst stage using one or two step culture systems, single or sequential media respectively, at 37ºC in an atmosphere of 6-7% CO2 and 5-20% O2 incubators. On day 3, we re-evaluated cleavage embryos to complete cumulus cells removal. The embryos were then cultured in individual well, with 20µl of medium under oil until they reached blastocyst stage. The blastocysts were vitrified and stored in liquid nitrogen. After that, the spent blastocyst culture medium (20µl) was transferred to a PCR tube and sent for analysis in the genetic laboratory, where it was stored at -80ºC until sequencing. A total of 243 samples of spent blastocyst culture medium were collected on the 5th/6th day. Cell-free DNA secreted on culture medium was amplified using NICS Sample Preparation Kit (Yikon Genomics), based on the MALBAC technology. After whole genome amplification, the DNA was measured using a Qubit 2.0 fluorometer and subjected to next generation sequencing (NGS) using Illumina MiSeq ® platform. The data were analyzed using the ChromGo ® software (Yikon Genomics). Results: The mean age of the patients was 38±4.08 years with an interval of 20-44 years. The euploid was diagnosed in 36.4% (80/220) of cases, aneuploidy in 31.3% (69/220), and mosaicism in 32.3% (71/220; with ≥60% aneuploidy) of blastocysts. Mosaic values ranged from 29.8% to 33.8% in different age groups. Individually, the most frequent chromosomal abnormality was XXY (Klinefelter Syndrome) occurring in 18 cases, followed by chromosome 21 (trisomy/monosomy) in 8 cases. The niPGT-A data showed a ≥60% incidence of aneuploid cells in all cases of chromosomal mosaicism (n=71). Conclusion: A high degree of mosaicism with aneuploidy cells was detected, and some hypotheses were suggested for this data (niPGT-A sensitivity in detecting the self-correction of chromosomal abnormalities phenomenon). However, it did not vary remarkably with age. On the other hand, euploidy levels had a negative correlation with age and aneuploidy levels had a positive relationship. This is the first report in the literature to relate chromosomal ploidy in blastocysts using niPGT-A and increasing patient age.
ObjectiveThis study aimed to analyze the results of hysterosonography performed prior to in vitro fertilization (IVF) and to correlate anomalous findings with hysteroscopy.MethodsFindings from 197 hysterosonograms of patients examined in an assisted reproduction clinic between January 2012 and August 2014 were included. Enrollment criteria: patients in preparation for IVF not recently submitted to uterine examination through hysterosalpingography or hysteroscopy referred to hysterosonography. Uterine cavity evaluation was considered anomalous when one or more of the following were found: polyps, submucous myomas, uterine synechiae, Müllerian duct anomalies. Individuals with cavity abnormalities that might interfere with IVF results were referred to hysteroscopy.ResultsNormal test results were seen in 170/197 of the cases (86.3%). Eighteen of the 197 cases (9.1%) were suspected for polyps, two (1%) for submucous myoma, six (3.5%) for synechiae, and one (0.5%) for Müllerian duct anomalies. Sixteen of the patients diagnosed with abnormalities underwent hysteroscopy to confirm or treat the suspected pathology. In only two cases there was no agreement between tests: one patient suspected for synechiae and another for polyps were not confirmed; another individual suspected for polyps was found to have focal endometrial thickening in hysteroscopy. The positive predictive value (PPV) in our study was 93.7%.ConclusionIn most cases, the diagnoses obtained by hysterosonography showed normal uterine cavities. The most common anomalous findings were polyps, followed by synechiae, submucous myoma, and Müllerian duct anomalies. Hysterosonography is a good option for evaluating the uterus and offers a high positive predictive value, while hysteroscopy stands as the gold standard.
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