Evaluation of Oxidative Stress as a Contributor of Anesthetic Agents' Effects on Embryo Quality and ICSI Outcome in Iraqi Patients

Infertility is described as a couple's failure to conceive for at least a year, using unprotected sex. Even though all anesthetic chemicals have been revealed in the follicular fluids, general anesthesia is still used in many IVF centers for patients who want to get pregnant. This study included 60 infertile women randomly divided into two groups under general anesthesia. 30 patients were given a Ketamine dose of 0.5 mg/Kg and the other 30 patients were received Remifentanil dose of 0.5 µg/Kg for induction of anesthesia. We collected serum for Reactive Oxygen Species (ROS) levels assessment for all patients before and after starting general anesthesia. Routine ICSI procedures was performed on all participants, including clinical evaluation (history, examination, and investigation), controlled ovarian and ovulation stimulation, oocyte retrieval under general anesthesia, follicular fluid collection for postoperative anesthesia medication concentration (Remifentanil and Ketamine), oocyte stripping, oocyte maturity assessment, intra-oocyte sperm injection into mature cells (MII), fertilization and division evaluation and embryo categorization, embryo selection and transfer, luteal phase support, and beta-hCG determination. ROS concentrations were compared between the Remifentanil and Ketamine groups. There was no significant difference in embryo features between Remifentanil and Ketamine, indicating that neither one is superior to the other in this regard. When Remifentanil or Ketamine was taken, there was no significant difference in ROS levels in serum or follicular fluid.

Effect of GnRH Agonist Alone or Combined With Different Low-dose hCG on Cumulative Live Birth Rate for High Responders in GnRH Antagonist Cycles: a Retrospective Study

Background: There is insufficient evidence regarding the impact of dual trigger on oocyte maturity and reproductive outcomes in high responders. Thus, we aimed to explore the effect of gonadotropin-releasing hormone agonist (GnRHa) trigger alone or in combination with different low-dose human chorionic gonadotropin (hCG) regimens on rates of oocyte maturation and cumulative live birth in high responders who underwent a freeze-all strategy in GnRH antagonist cycles.Methods: A total of 1343 cycles were divided into three groups according to different trigger protocols: group A received GnRHa 0.2 mg (n=577), group B received GnRHa 0.2 mg and hCG 1000 IU (n=403), and group C received GnRHa 0.2 mg and hCG 2000 IU (n=363). Results: There were no significant differences in age, body mass index, and rates of oocyte maturation, fertilization, available embryo, and top-quality embryo between the groups. However, the incidence of moderate to severe ovarian hyperstimulation syndrome (OHSS) was significantly different between the three groups (0% in group A, 1.49% in group B, and 1.38% in group C). For the first frozen embryo transfer (FET) cycle, there were no significant differences in the number of transferred embryos and rates of implantation, clinical pregnancy, live birth, and early miscarriage between the three groups. Additionally, the cumulative ongoing pregnancy rate and cumulative live birth rate were not significantly different between the three groups. Similarly, there were no significant differences in gestational age, birth weight, birth height, and the proportion of low birth weight among subgroups stratified by singleton or twin.Conclusions: GnRHa trigger combined with low-dose hCG (1000 IU or 2000 IU) did not improve oocyte maturity and embryo quality and was still associated with an increased risk of moderate to severe OHSS. Therefore, for high responders treated with the freeze-all strategy, the GnRHa single trigger is recommended for final oocyte maturation, which can prevent the occurrence of moderate to severe OHSS and obtain satisfactory pregnancy and neonatal outcomes in subsequent FET cycles.

Fertilization and early embryonic development of in vitro matured metaphase I oocytes in patients with unexpected low oocyte maturity rate

Summary To determine the fertilization and embryonic potential of immature metaphase I (MI) oocytes in patients with low oocyte maturity rate in whom the percentage of mature oocytes obtained was less than 75% of the total retrieved ones. In vivo matured metaphase II (MII) oocytes (MII-ICSI, n = 244), and in vitro matured MI oocytes (MI-MII-ICSI, n = 202) underwent an intracytoplasmic sperm injection (ICSI) procedure. Maturation rate, fertilization rate and early embryonic development were compared in both groups. In total, 683 oocytes were collected from 117 ICSI cycles of 117 patients. Among them, 244 (35.7%) were mature MII and 259 (37.9%) were MI after the denudation process. Of those 259 MI oocytes, 202 (77.9%) progressed to MII oocytes after an incubation period of 18–24 h. The maturation rate was 77.9%. Fertilization rate was found to be significantly higher in the rescued in vitro matured MI oocyte group when compared with the in vivo matured MII oocyte group (41.6% vs 25.8%; P = 0.0006). However, no significant difference was observed in terms of cleavage rates on days 2 and 3 between the groups (P = 0.9126 and P = 0.5031, respectively). There may be unidentified in vivo factors on the oocyte maturation causing low developmental capacity in spite of high fertilization rates in the group of patients with low oocyte maturity rate. Furthermore, studies are needed to determine the appropriate culture characteristics as well as culture period and ICSI timing of these oocytes.

Maternal Underweight Does Not Adversely Affect The Outcomes of IVF/ICSI and Frozen Embryo Transfer Cycles or Early Embryo Development

Purpose To compare assisted reproductive technology (ART) outcomes and preimplantation embryo development between underweight and normal weight women. Methods This retrospective cohort study included 26 underweight women (body mass index [BMI] < 18.50 kg/m2) and 104 normal weight women (BMI > 20 and < 24.9 kg/m2) who underwent a total of 204 in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles and 358 fresh/frozen embryo transfers (ET) in our institution between January 2016 and December 2018. Statistical analyses compared selected ART outcomes (ovarian stimulation, fertilization, and pregnancy) between both weight groups. Morphokinetic and morphological parameters were also compared between 346 and 1467 embryos of underweight and normal weight women, respectively. Results The mean ± standard deviation age of the underweight and normal weight women was similar (31.6 ± 4.17 vs 32.4 ± 3.59 years; p = 0.323). There were no differences in the peak estradiol levels, the number of retrieved oocytes, the number of metaphase II oocytes, and the oocyte maturity rates between the two groups. The IVF/ICSI fertilization rates and the number of embryos suitable for transfer or cryopreservation were similar for both groups. All morphokinetic parameters that were evaluated by means of time-lapse imaging as well as the morphological characteristics were comparable between low and normal BMI categories. There were no significant differences in pregnancy achievement, clinical pregnancy, live births, and miscarriage rates between the suboptimal and optimal weight women. Conclusion Underweight status has no adverse impacts on the outcomes of IVF/ICSI with either fresh or frozen ET or on preimplantation embryo development and quality.

Compatibility of Distinct Label-Free Proteomic Workflows in Absolute Quantification of Proteins Linked to the Oocyte Quality in Human Follicular Fluid

We present two separate label-free quantitative workflows based on different high-resolution mass spectrometers and LC setups, which are termed after the utilized instrument: Quad-Orbitrap (nano-LC) and Triple Quad-TOF (micro-LC) and their directed adaptation toward the analysis of human follicular fluid proteome. We identified about 1000 proteins in each distinct workflow using various sample preparation methods. With assistance of the Total Protein Approach, we were able to obtain absolute protein concentrations for each workflow. In a pilot study of twenty samples linked to diverse oocyte quality status from four donors, 455 and 215 proteins were quantified by the Quad-Orbitrap and Triple Quad-TOF workflows, respectively. The concentration values obtained from both workflows correlated to a significant degree. We found reasonable agreement of both workflows in protein fold changes between tested groups, resulting in unified lists of 20 and 22 proteins linked to oocyte maturity and blastocyst development, respectively. The Quad-Orbitrap workflow was best suited for an in-depth analysis without the need of extensive fractionation, especially of low abundant proteome, whereas the Triple Quad-TOF workflow allowed a more robust approach with a greater potential to increase in effectiveness with the growing number of analyzed samples after the initial effort of building a comprehensive spectral library.

O-175 Impact of female chromosomal polymorphic variants on ovarian reserve markers and fertility treatments prognosis

Study question Could the presence of chromosomal polymorphisms in women undergoing fertility treatments influence ovarian reserve, oocyte preservation or IVF clinical outcomes? Summary answer Polymorphic variants of chromosomes seem to adversely influence the Anti-Müllerian Hormone (AMH) serum levels and the post-thawing oocyte survival rate. What is known already Infertile couples have been reported to have a higher incidence of karyotype abnormalities than the general population, which could lead to lower fertility rates. However, few studies have demonstrated the controversial relationship between these karyotype alterations in women and the specific involvement of these variants and their combinations in an IVF cycle. Therefore, there is a growing interest in categorizing chromosomal polymorphic variants and relating them to the subsequent evolution in ART cycles. Study design, size, duration Retrospective evaluation of a cohort of women undergoing IVF cycles in a private fertility center. The sample included 280 cycles performed between July 2017 and December 2020. The study explored the correlation between karyotype polymorphisms and IVF outcomes in terms of: Anti-Müllerian Hormone (AMH) level, Antral Follicle Count (AFC), Oocyte Maturity (MII), Oocyte Survival after Thawing (TS), Fertilization (FZ), Useful Embryos (UE), Biochemical (BP) and Clinical Pregnancy (CP), Miscarriage (M) and Live Birth (LB) rates. Participants/materials, setting, methods Women with karyotype performed before an IVF cycle. Chromosome analysis was carried out according to the International System for Human Cytogenetic Nomenclature guidelines (ISCN). Only own eggs cycles were included, and testicular sperm cycles were excluded. The normality of the distribution of the variables was assessed using the Shapiro-Wilk test. The association between IVF cycle parameters and the presence of polymorphisms was assessed by T-Student (parametric) or U-Mann-Whitney (non-parametric). Main results and the role of chance From a total of 280 IVF cycles, 198 met the inclusion criteria: Control Group (CG) with normal karyotype (94) and Study Group (SG) with presence of polymorphisms (104) were analyzed. Cycles with fresh (136) and warmed (62) oocytes were included. Mean female age was 37.66 ± 3.99 (CG) and 36.61 ± 3.72 (SG). The overall outcome rates were: 75.82% MII, 81.39% TS, 72.78% FZ, 49.07% UE on day 5, 21.82% BP, 78.18% CP, 14.53% M and 63.65% LB. Statistically significant differences were found between the presence of polymorphisms and mean AMH serum level between CG (2.36 ng/mL) and SG (1.74 ng/mL) (p = 0.04), especially when the type “qh+” was detected (1.01 ng/mL) (p = 0.02). Furthermore, statistically significant differences were found regarding oocyte thawing survival rate, which decreased in the SG (78.94%) compared to the CG (93.69%) (p = 0.02), mainly when the type “ps+” was detected (75.13%) (p &lt; 0.01). No statistically significant differences were found between the presence of polymorphism and the AFC (p = 0.25), MII (p = 0.10), FZ (p = 0.93) or UE (p = 0.52) rate on day 5. In the same way, from 106 embryo transfers (ET) performed, no significant differences were found for BP, CP, M and LB rates (p &gt; 0.05). Limitations, reasons for caution Larger prospective studies including homogeneous cohorts are needed in order to corroborate our initial results. Wider implications of the findings Our findings may represent a practical tool in order to advise these patients about their reproductive success, especially, in terms of post-thawing oocyte survival prognosis. Therefore, it could be provided more personalized prognostic information before embarking on IVF treatments. Trial registration number Not Applicable

Assessment of cGMP level in medium during in vitro growth period of murine preantral follicles with and without supplementation of C-type natriuretic peptide

Summary To enhance the developmental competency of murine ovarian follicles cultured in vitro, C-type natriuretic peptide (CNP) was supplemented in the culture system. Although the mechanism is not fully elucidated, it was reported that the effect of CNP supplementation was mediated by increased cyclic guanosine monophosphate (cGMP). In the present study, cGMP levels in media for murine preantral follicle culture were compared both between a control group without CNP supplementation and an experimental group with CNP supplementation and between days in each group. In addition, follicle growth patterns and oocyte maturity were assessed and compared between the two groups. Results demonstrated that along with in vitro culture, cGMP levels increased (P < 0.05) both in the control group and the experimental group, whereas cGMP levels were not significantly different between the two groups on the same day of in vitro culture (P > 0.05). The oocyte’s maturity was superior in the experimental group compared with the control group (P < 0.05). As ovarian follicles grew three-dimensionally in the experimental group but were flattened in the control group, CNP might improve oocyte maturity through maintaining the three-dimensional architecture of the ovarian follicle because of increased transzonal projections (TZP) and functional gap junctions between oocyte and surrounding granulosa cells.