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RESEARCH QUESTION: Is endometriosis detrimental to embryo implantation? DESIGN: A retrospective matched case-control study of women with a surgical or ultrasound diagnosis of endometriosis at Fondazione IRCCS Ca'Granda Ospedale Maggiore Policlinico di Milano between 2015 and 2021. Women with endometriosis who underwent a 'freeze-all' cycle during an IVF treatment were eligible to be included. They were matched to patients without the disease, who also underwent cryopreserved blastocyst transfer cycles, in a 1:1 ratio by age (±1 year), and number (=) and quality (±1 top versus low) of cryopreserved blastocysts. All women underwent single frozen embryo transfer, and assisted reproductive technology outcomes suggested by the Core Outcome Measure for Infertility Trials initiative were evaluated. The main outcome was the cumulative live birth rate per cycle. RESULTS: One hundred and one women with endometriosis and 101 matched unaffected women were included. Cumulative live birth rate per cycle did not vary between women with and without endometriosis (50% versus 58%, respectively; P = 0.32). On the basis of the Kaplan-Meier analysis, the predicted success rates over four embryos transferred were also similar (74% versus 82%, respectively; P = 0.67). CONCLUSION: In women with moderate or severe endometriosis, these retrospective results seem to indicate no or a limited effect of the disease on endometrial receptivity.

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OBJECTIVE: Embryo transfer on day-5 has been associated with higher success rates, therefore our IVF clinics started to extend embryo culture until blastocyst stage. This study aimed to compare the success rates of day-3 vs. day-5 embryo transfers. METHODS: We had 266 patients included, all having undergone ICSI, with 221 patients having undergone day-3 embryo transfers, and 45 patients having undergone day-5 embryo transfers. Patients with more than five good quality embryos on day-3 were chosen to prolong the culture of embryos into day-5. RESULTS: There were no significant differences in patient characteristics, including baseline LH, FSH, Prolactin and Estradiol hormone levels. In addition, there were also no significant differences in rFSH total dosage and duration of stimulation day. Final estradiol levels, number of follicles, retrieved oocytes, matured oocytes, fertilized oocytes and number of embryos were significantly higher in day-5 compared to day-3 embryo transfer groups. Number of embryos transferred on day-3, were significantly higher compared to day-5. Neither group showed any significant differences in clinical pregnancy, implantation, multiple pregnancy or living birth rates. There were no differences in birth weights and lengths, head circumstances and Apgar Scores between both groups either in singleton or twin group. CONCLUSIONS: Transferring embryos at day-3 may provide the same benefits as day-5 embryo transfers to patients. However, more embryos were required to be transferred to achieve these comparable results.

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OBJECTIVE: The recent improvement in sequential media has refocused its attention on the role of human blastocysts in ART, not only because of its advantages but also because of the possible cancellation of embryo transfer when relying on blastocyst transfer only. Hence, the idea of sequential transfer on day 3 and day 5 was proposed. Objective: To compare the pregnancy outcomes of sequential embryo transfer on day 3 and day 5, versus cleavage transfer on day 3 and blastocyst transfer on day 5 in cases of recurrent implantation failure. METHODS: This was a prospective and randomized trial, in which 210 qualified patients with recurrent implantation failures undergoing IVF/ICSI were randomized into three groups, each group included 70 patients. Embryo transfer was performed in day 3 in the first group, day 5 (blastocyst transfer) in the second group and sequential embryo transfer in days 3 and 5 in the third group. We assessed pregnancy outcomes from all the three groups. Results: Clinical pregnancy and live birth rates were significantly higher in the sequential group than either group day-3 or day-5 of embryo transfer in cases with recurrent implantation failures. CONCLUSIONS: Sequential embryo transfer in cases with recurrent implantation failures and adequate number of retrieved oocytes is associated with higher implantation and clinical pregnancy rates, and it is advocated for patients having an adequate number of good quality embryos.

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OBJECTIVE: To analyze the genetic and clinical outcomes of blastocysts derived from 0PN oocytes after IVF/ICSI. METHODS: This retrospective observational study included patients aged 40 years or younger submitted to IVF/ICSI with their own oocytes and with blastocysts derived from 0PN oocytes between January 2015 and April 2018. The clinical outcomes of 0PN blastocyst transfers were analyzed. Genetic tests were performed on biopsied 0PN blastocysts with Next Generation Sequencing. RESULTS: A total of 27 0PN blastocysts were transferred, yielding an implantation rate of 48.0% and an ongoing pregnancy rate of 50.0%. The transfers resulted in 13 live births (59.0% live birth rate). Genetic test results revealed that four of the 17 0PN blastocysts biopsied were 46XX; three were 46XY; and 10 were aneuploid embryos, awarding a diploid rate to 76.4% (13/17). CONCLUSION: Almost half of the 0PN blastocysts implanted (48.0%) and 13 healthy babies were born. More than three quarters (76.4%) of the 0PN blastocysts were diploid, thus ruling out the possibility of parthenogenetic activation. Our study indicated that the transfer of 0PN blastocysts is a safe, worthy option when the number of normal 2PN embryos is insufficient.

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OBJECTIVE: To study the value of oxygen consumption (OC) as a predictor of the developmental potential of D3 embryos in frozen embryo transfer (FET) cycles. METHODS: This observational study included 148 patients undergoing FET cycles with two embryos transferred per cycle. OC rates were examined by scanning electrochemical microscopy before embryo transfer. Implantation, clinical pregnancy, miscarriage, and live birth rates were calculated. RESULTS: A total of 296 embryos were transferred in 148 cycles, or two embryos per cycle. The embryos were divided into three groups based on OC: Group A included the cases in which the OC rate of each of the two transferred embryos was greater than 3.0 fmol/s; Group B included the cases in which the OC rate of one of the embryos was greater than 3.0 fmol/s and the OC rate of the other embryo was less than 3.0 fmol/s; and Group C included the cases in which the OC rates of the two embryos were less than 3.0 fmol/s. Higher live birth rates and lower miscarriage rates were observed in Group A (p<0.05). CONCLUSIONS: Our data suggest that OC is positively correlated with embryo developmental potential. Therefore, measuring the OC of human embryos may be useful in embryo assessment.

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OBJECTIVE: To describe a patient selection method for elective single embryo transfer (eSET), emphasizing inclusion criteria and results. METHODS: This retrospective study included all cases seen in a private clinic between June 2011 and December 2016, in La Paz, Bolivia (3600 meters above sea level). Elective single embryo transfer was the method of choice in 34 IVF/ICSI cycles, all in the blastocyst stage. Gardner's blastocyst classification criteria were used. Between the two stages of the study (July 2015), each embryo grade implantation rate was recalculated, which led to the expansion of the inclusion criteria. RESULTS: The clinical pregnancy rate of the 34 cases in the first transfer group was 55.9% (19/34). Twin or multiple pregnancies did not occur. The cumulative pregnancy rate to date is 64% [(19+3)/34]. The first stage comprised 2.56% (12/468) of the patients offered elective single embryo transfers; the implantation rate was 58.3% (7/12). In the second stage, 14.29% (22/154) of the patients were eligible, and the implantation rate was 54.55% (12/22). CONCLUSIONS: The implementation of an eSET program based on in-depth morphological embryo assessment combined with the calculation of the implantation potential of each embryo grade led to acceptable clinical outcomes and fewer multiple pregnancies in patients transferred two embryos. Each clinic should be aware of the implantation rates of each embryo grade in its own setting.

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OBJECTIVE: To define the appropriate number of embryos to be transferred at day 5. METHODS: Retrospective analysis of 784 consecutive fresh day-5 embryo transfers performed between 2007 and 2015, divided in three groups: Group A (N = 219): received the only 2 embryos that reached a transferable stage; Group B (N = 357): received 2 selected embryos among several that reached a transferable stage; Group C (N = 208): received the only 3 developing embryos. Clinical pregnancy, implantation, multiple pregnancy and delivery rates were registered. Kruskal-Wallis and Fisher Exact tests were applied as appropriate. RESULTS: Age and previous attempts were comparable in the 3 groups. Compared with Group A, Groups B and C had a higher oocyte recovery (10.7 ± 5.6 vs. 14.7 ± 8.0 vs. 13.8 ± 6.6), fertilization rate (75.97% vs. 81.60% vs. 83.29%) and percentage of embryos reaching a transferable stage on day 5 (39.98% vs. 63.99% vs. 60.97%), as well as a significantly higher clinical pregnancy (42.92% vs. 61.06% vs. 58.17%) and implantation rates (21.09% vs. 40.98% vs. 36.97%). The multiple pregnancy rate was higher in Groups B and C than in Group A (11.70% vs. 31.19% vs. 37.19%). The high order multiple pregnancy rate (> 2) was significantly increased in group C (1.06% vs. 0.92% vs. 14.05%). CONCLUSIONS: In patients with 3 or more day 5 developing embryos, delivery rates are similar if 2 or 3 embryos are transferred. The transfer of 3 embryos carries an unacceptable increase in the risk of high order multiple pregnancy, with its known consequences. According to our data, we should not exceed the number of 2 day-5 fresh embryos transferred.

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Leptin is a protein secreted by the adipocytes, which serves as a link between fat and brain. Its main action is to decrease appetite and increase energy expenditure, but it is also involved in the control of different neuroendocrine systems, including gonadal axis. Although the effects of leptin deficiency on reproduction are well recognized, the effect of excess leptin on male reproductive function is not clear. The aim of this study was to evaluate fertility and sperm parameters of male rats exposed to exogenous leptin. A group of adult male rats received exogenous leptin intraperitoneally (30 µg/kg/day) for 42 days, and a control group received only the vehicle during the same period. After the treatment, animals were evaluated for sperm count, sperm motility, and fertility after intrauterine artificial insemination. There was no statistically significant difference between the groups related to sperm production, sperm concentration, and sperm motility. However, fertility evaluation after artificial insemination showed a quantitative decrease in the uterus plus fetuses weight, number of implantation sites, and number of live fetuses. The fertility potential showed a reduction of about 40%, whereas the preimplantation loss rate increased more than 2-fold in leptin-treated animals. In conclusion, leptin administration to nonobese male rats impairs ability of treated animals to generate offspring, since the occurrence of implantation was diminished. So leptin can impair sperm quality, affecting the reproductive capacity.

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CAPSULE: Clinical outcomes using INVOcell device with ICSI. OBJECTIVE: Intravaginal culture of oocytes (INVO) procedure is an intravaginal culture system that utilizes the INVOcell device in which the fertilization and embryo culture occur. In this procedure, the vaginal cavity serves as an incubator for oocyte fertilization and early embryonic development. The objective of this study was to evaluate the clinical outcomes of this intravaginal culture system in intracytoplasmic sperm injection (ICSI). METHODS: A total of 24 cycles INVO-ICSI (study group) and 74 cycles of ICSI (control group) were included in the study. The cleaved oocytes at day 3/total injected oocytes, embryo quality, pregnancy rate (PR), implantation rate (IR), and miscarriage rate (MR) were compared between both groups. RESULTS: At day 3, there was no difference in the cleaved oocyte rate (78.7 and 76.1%) and embryo quality (77 and 86.8%) for the study and control groups, respectively. In the study group, more embryos were significantly transferred compared to the control group (2.63 ± 0.58 versus 1.93 ± 0.25; P < 0.05). PRs, IRs, and MRs were similar for the study group compared with the control group (PR: 54.2% versus 58.1%; IR: 31.7% versus 33.6%; MR: 7.7% versus 20.9%). CONCLUSIONS: Good PR and IR can be obtained using the INVOcell device, and the INVO-ICSI procedure can be considered as an alternative option to infertile patients.

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The Conference was an important achievement of the National Health Forum, a group set up by The National Council of Justice for monitoring and creating solutions concerning health claims. The Conference gathered BioLaw, public and private health care professionals, and manage a group of interdisciplinary discussion in the search for more uniform solutions to current issues. Seven statements of the 10 BioLaw proposals selected were approved.

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The aim of this study was to review the experience and outcomes of assisted reproduction cycles with embryos grown up to day 5 of development, comparing different parameters according to the ages of the patients. We retrospectively studied 1,874 assisted reproduction cycles where embryo culture was extended up to the fifth or sixth day of development. All IVF and ICSI cycles were included, comparing, according to patient age, the following rates: blastocyst formation, pregnancy, implantation and abortion. As control, we analyzed cycles with donated oocytes from young donors (OD). The number of embryos reaching the blastocyst stage is similar in all groups of patients. Only the OD group was different in terms of blastocyst formation, pregnancy and implantation rates. Patients over 39 years of age had an abortion rate of 59.1 percent, which is significantly higher than the other groups. Extended embryo culture up to the blastocyst stage can be implemented in programs of assisted reproduction in order to increase the pregnancy rate. The potential of blastocyst implantation is high, allowing us to transfer fewer embryos and reduce the probability of multiple pregnancies.

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