Average Number Of Embryos Research Articles

Protective effects of engineered Lactobacillus crispatus strains expressing G-CSF on thin endometrium of mice.

Does recombinant Lactobacillus expressing granulocyte colony-stimulating factor (G-CSF) have a better protective effect than the current treatment of thin endometrium (TE)? This study suggested that the intrauterine injection of Lactobacillus crispastus (L. crispastus)-pPG612-G-CSF has a positive effect on preventing TE induced by 95% alcohol in mice. TE has a negative impact on the success rate of ART in patients, and is usually caused by intrauterine surgery, endometrial infection, or hormone drugs. Exogenous G-CSF can promote endometrial vascular remodelling and increase endometrial receptivity and the embryo implantation rate. Moreover, Lactobacillus plays a crucial role in maintaining and regulating the local microecological balance of the reproductive tract, and it could be a delivery carrier of the endometrial repair drug G-CSF. We constructed engineered L. crispastus strains expressing G-CSF. The mice were divided into five groups: (i) Control group (C, n = 28), uteri were treated with preheated saline solution via intrauterine injection on the third and sixth day of oestrus; (ii) Model group (M, n = 35), where uteri were treated with 95% alcohol on the third day of oestrus and preheated saline solution on the sixth day of oestrus via intrauterine injection; (iii) L. crispatus-pPG612-treatment group (L, n = 45), where uteri were treated with 95% alcohol on the third day of oestrus and 0.1 ml × 108 CFU/ml L. crispatus-pPG612 on the sixth day of oestrus via intrauterine injection; (iv) L. crispatus-pPG612-treatment group (LG, n = 45), where uteri were treated with 95% alcohol on the third day of oestrus and 0.1 ml × 108 CFU/ml L. crispatus-pPG612-G-CSF on the sixth day of oestrus via intrauterine injection; (v) G-CSF-treatment group (G, n = 52), where uteri were treated with 95% alcohol on the third day of oestrus and 30 µg/kg G-CSF on the sixth day of oestrus via intrauterine injection. Then, we compared the effects of L. crispastus, L. crispatus-pPG612-G-CSF and G-CSF on endometrial thickness, angiogenesis, fibrosis, and inflammation in the TE mouse. We collected uterine tissues for haematoxylin-eosin staining, immunohistochemical staining, Western blot and RT-PCR, as well as serum for ELISA and uterine flushing solution for high-throughput sequencing. Compared with those in the M group (the mice of the group were intrauterine injected 95% alcohol and treated with saline solution), the L. crispatus-pPG612-G-CSF strain increased the thickness of the endometrium (P < 0.001) and the number of blood vessels and glands (both P < 0.001), enhanced the expression of cytokeratin 19 (CK19) (P < 0.001), vimentin (Vim) (P < 0.001), vascular endothelial growth factor-A (P < 0.001), and CD34 (P < 0.001), and decreased fibrosis levels (P = 0.004). In addition, the high-throughput sequencing results indicated that the L. crispatus-pPG612-G-CSF strain could decrease the abundance of Pseudomonas (P = 0.044) and Actinomyces spp. (P = 0.094) in TE mice and increased the average number of embryos (P = 0.036). Finally, the L. crispatus-pPG612-G-CSF strain was preliminarily confirmed to activate the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signalling pathway and enhance the mRNA expression of hypoxia-inducible factor-1α (P < 0.001), vascular endometrial growth factor (P = 0.003), and endothelial cell nitric oxide synthase (P = 0.003) in mouse uterine tissue. N/A. Therapy with the L. crispatus-pPG612-G-CSF strain has tremendous potential to accelerate the reparative processes of TE. However, we have reported only the expression of genes and proteins related to the PI3K/AKT pathway, and numerous other mechanisms may also be involved in the restoration of the endometrium by L. crispatus-pPG612-G-CSF. The results from the study provide new ideas and suggest new methods for TE treatment. This work was financially supported by the Project of Science and Technology Development Plan of Jilin Province (grant number 20210101232JC), the Science and Technology Plan Item of Jilin Provincial Education Department (grant number JT53101022010), and the Doctoral Research Start-up Fund of Jilin Medical University (grant numbers JYBS2021014LK and 2022JYBS006KJ). The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as potential conflicts of interest.

Reproductive characteristics and success rate of intracytoplasmic sperm microinjection in spinal cord injury infertile men: Retrospective cohort study.

Thanks to modern methods of assisted reproduction (ART), parenthood has become an attainable goal for couples in which the male partner has experienced spinal cord injury (SCI). The aim of our study was to determine the success of the treatment of infertile patients with SCI with intracytoplasmic sperm injection (ICSI) of cryopreserved sperm obtained by the testicular sperm aspiration (TESA) procedure. In this retrospective study 156 infertile couples were included, in which the male partner is primarily infertile due to azoospermia. Infertile couples were divided into two groups. The first group (n= 82) includes men with SCI, and the second (n= 74) men with obstructive azoospermia (OA) as the cause of infertility. All infertile men were examined and processed in the diagnostic procedure, and based on the urological findings, surgical extraction of sperm from the testicles was indicated. Exclusion criteria were the age of women over 40 and men over 45. We found that the quality of sperm was worse in the group with SCI, compared to the group with OA, but without statistical significance. Zenica and Johnsen score (p= 0.001; p= 0.000) showed worse semen characteristics in the group with SCI. There were no significant differences in the average number of embryos (p= 0.698), pregnancy rates per cycle (p= 0.979) and pregnancy rates per embrio transfer (ET), clinical pregnancy rates per ET (p= 0.987) and delivery rates per ET (p= 0.804) in couples with SCI, compared to couples with OA. Based on the results of this research, the TESA and ICSI procedures can be recommended as a successful method in the treatment of male infertility caused by azoospermia due to SCI.

P-233 Embryo transfer timing and clinical pregnancy success: a retrospective study

Abstract Study question The aim of this study is to compare the clinical outcome of day5 versus day6 genetically normal blastocyst transfers. Summary answer 1000 oocytes were analyzed showing a highly significant difference in the number of normal embryos and clinical pregnancy rates in day5 versus day6 blastocysts What is known already studies have shown that day 5 embryos have a higher pregnancy rate compared to day 6 embryos, some studies had only euploid embryos transferred and others did not have any genetic testing done on embryos. No study had a sample size as large as 1760 and the number of embryos studied was a limiting factor in their results. Study design, size, duration The study is a retrospective cross-sectional chart review with multivariate analysis, data from January 2019 up to February 2023 were extracted. 1760 patients were recruited, 1000 in the day5 group and 760 in the day6 group. The data recorded for each patient included maternal age, body mass index (BMI), anti-Müllerian hormone (AMH) level, number, prenatal genetic diagnostic testing, frozen embryo transfer, sperm parameters, and pregnancy status. Participants/materials, setting, methods All embryos were genetically tested for aneuploidy before transfer, using next-generation sequencing technology to rule out 24-chromosome aneuploidies (1-22, X, Y). All patients who have single embryo transfer, frozen embryos and pre-implantation genetic diagnosis (PGD) were included in the study. Continuous variables were expressed as mean with standard deviation and categorical variables as numbers and frequencies (percentages). Statistical differences between groups were estimated using the chi-square or Fisher’s test. All analyses were performed using SAS studio. Main results and the role of chance 1,000 cycles were analyzed for Day 5 and 760 for Day 6 embryo. The number of blastocysts formed, and genetically diagnosed embryos significantly favor the Day 5 group. There’s a significant difference in the number of normal embryos, with the Day 5 group having more. There is a highly significant difference in clinical pregnancy rates. On the other hand, the differences in biochemical pregnancies are not statistically significant. The occurrence of ectopic pregnancies shows no significant difference. Additionally, there is no statistically significant difference in the rates of miscarriage. There is a highly significant difference in blastocyst formation rates, with Day 5 embryos. This indicates that Day 5 embryos have a significantly higher likelihood of forming blastocysts compared to Day 6 embryos. Implantation: Day 5 embryos have an implantation rate of 62.0% compared to 49.7% for Day 6 embryos. Day 5 embryos exhibit a significantly higher likelihood of successful implantation. Euploid: The difference in euploid rates is statistically significant, albeit with a lower p-value of 0.0236. Day 5 embryos have a rate of 59.6%, while Day 6 embryos have a rate of 56.0%. Day 5 embryos are associated with a lower likelihood of being aneuploid Limitations, reasons for caution Limitation of this study is that we do not have data available regarding complications during pregnancy in the day 5 versus day 6 groups and we had no data regarding the the live birth rate between the two groups. Wider implications of the findings This study has the potential to revolutionize worldwide discussions during IVF consultations. With the large sample size we can provide for the first time a statistically significant difference in the percentage of success when transferring a day5 (61%) versus day6 (49%) euploid embryo. Trial registration number not applicable

Polyembryony and morpho-genetic characterisation of zygotic seedlings through microsatellite markers in ten polyembryonic citrus rootstocks

Morpho-genetic characterisation of zygotic and nucellar seedlings of 10 citrus rootstocks was carried out to know the degree of polyembryony. 'Volkameriana' rootstock had maximum fruit and seed weight. Polyembryony, number of embryos per seed and clutch size was highest in 'Katazamir'. However, the seeds of 'Schaoub' took minimum days to germinate while 'Cleopatra' took maximum days. Significant positive correlation was observed between percent polyembryony and average number of embryos per seed, seed weight and number of seeds per fruit. Twenty SSR primers were utilised for detection of zygotic seedling from all rootstocks. None of the individual primers used in this study were able to consistently identify zygotic seedlings in all rootstocks tested. The maximum (26.7%) zygotic seedlings were identified in 'Katazamir' and minimum (5.0%) in 'X-639' rootstocks. There was not a significant correlation between seedling height and zygotic and nucellar seedlings, identified with SSR markers. Therefore, seedling height cannot be taken as a criteria to identify the nucellar and zygotic seedlings.

Neu5Gc regulates decidual macrophages leading to abnormal embryo implantation.

Repeated implantation failure (RIF) is one of the most prominent problems in the field of assisted reproduction. Neu5Gc on the surface of decidual macrophages (dMΦ) leads to different activation patterns of dMΦ, which affects embryo implantation and development. Cmah-/- (Neu5Gc-deficient) mice induced to produce anti-Neu5Gc antibodies in vivo were given a special diet rich in Neu5Gc and their fertility was monitored. The long-term diet rich in Neu5Gc induced the decrease of endometrial receptivity of female mice. The pregnancy rate of female mice fed the normal diet was 63.6% (n = 11) and the average number of embryos was 9.571 ± 1.272, while the pregnancy rate of female mice fed the diet rich in Neu5Gc was 36.4% (n = 11) and the average number of embryos in pregnant mice was 5.750 ± 3.304. The intake of Neu5Gc and the production of anti-Neu5Gc antibody led to M1 polarization of endometrial dMΦ and abnormal embryo implantation.

CHANGES IN INDICATORS OF EMBRYOTOXICITY OF CADMIUM CHLORIDE IN COMBINED ADMINISTRATION WITH ZINC AND COPPER SUCCINATE IN A CHRONIC EXPERIMENT ON RATS

The search for bioantagonists of cadmium toxicity, its negative impact on the body and the developing embryo remains a relevant and, at the same time, is understudied issue in modern science. In recent years, the effectiveness of the use of succinic acid salts for the prevention and treatment of diseases of various human organs and systems has been established.&#x0D; Objective: To determine changes in the level of embryotoxic effect of cadmium chloride in a dose of 2 mg/kg with isolated and combined with copper or zinc succinate intragastric administration in a chronic experiment on pregnant female rats.&#x0D; Objects and methods. This embryological study included female rats with a defined gestation period, which were categorized into four distinct experimental groups: the first group served as the control, the second group received an isolated administration of cadmium chloride solution, the third group underwent combined administration of cadmium chloride solution and zinc succinate solution, and the fourth group experienced combined administration of a solution containing cadmium chloride and copper succinate. The collection of embryonic material occurred on the 13th and 20th days of embryogenesis. Statistical analysis of the collected data was performed using the STATISTICA program. Statistically significant differences were determined at a p-value of less than 0.05 (a significance level of 5%). The description, assessment, and comparison of the mean indicators among the groups were conducted employing variance analysis and the Kruskal-Wallis test.&#x0D; Results and discussions. All female rats in the experiment survived. The obtained results demonstrate a significant increase in all indicators of embryonic mortality in the group exposed to cadmium chloride in comparison with the control group on both the 13th and 20th days of the experiment. At the same time, comparing the groups exposed to cadmium in combination with zinc and copper succinates with the group of isolated exposure, can visualize statistically significant differences in the main indicators of embryonic development.&#x0D; Conclusion. Isolated chronic administration of cadmium chloride to pregnant female rats in doses of 2.0 mg/kg leads to a decrease in the average number of embryos and an increase in embryonic mortality in the group, whereas the combined administration of cadmium chloride in the same dose with zinc and copper succinates, all indicators of embryotoxicity are reduced, which enables to consider copper succinate and especially zinc succinates as bioantagonists of cadmium.

#325 : A Comparison of Three Different IVF FSH Dosing Strategies for PCOS Patients; A Retrospective Analysis of 855 IVF Cycles

Background and Aims: IVF responses to gonadotrophins may be difficult to predict in patients with PCOS. Nowadays there are various options including using the strategy of GnRH agonist trigger and freeze all embryos. The follitropin delta dosing algorithm has been used to predict an ovarian response that reduces the effects of ovarian hyperstimulation syndrome. The aim of this study was to compare the outcomes of three different FSH dosing strategies in PCOS patients. Methods: This retrospective multi-centre trial enrolled 626 PCOS women. They underwent 855 stimulated IVF cycles using a GnRH antagonist protocol and 634 frozen embryo transfers with embryos from the original stimulated cycles. Patients were classified into 3 groups: Group 1 (underdosing), Group 2 (algorithm dosing) and group 3 (overdosing). The algorithmic dose was based on the follitropin delta algorithm using AMH and body weight. The main outcome measure was the live birth rate per cycle started. Results: The live birth rate per cycle was 40.8% for group 1, 42.1% for group 2 and 45.5% for group 3. Group 3 had the greatest average number of embryos frozen (1.55 embryos in group 1, 2.45 group 2 and 2.96 group 3). The average number of eggs collected was greatest in group 3 (8.7 eggs in group 1, 11.9 group 2 and 14.2 group 3). The most frozen of all cycles occurred in group 3 (45.6%). Conclusions: Higher FSH dosing resulted in more eggs and more live births per cycle started. It also resulted in less fresh embryo transfers. These results could be used when choosing an FSH dosing strategy for PCOS patients.

OR12-02 NRF2 Is Essential For Expanding Pancreatic Beta-Cell Mass During Pregnancy

Abstract Disclosure: S. Baumel-Alterzon: None. A. Garcia-Ocana: None. D.K. Scott: None. The late stages of the mammalian pregnancy are accompanied with increased insulin resistance due to the increased glucose demand of the growing fetus. Therefore, as a compensatory response, in order to maintain the maternal normal blood glucose levels, maternal beta-cell mass expands leading to increased insulin release. Beta-cell proliferation, beta-cell neogenesis, and decreased beta-cell apoptosis, are major contributors for the beta-cell adaptive response during pregnancy. Defects in these cellular processes can lead to gestational diabetes mellitus (GDM). GDM, which occurs in ∼7-8% of the total pregnancy cases within the U.S, results in adverse outcomes for both the mother and newborn. This highlights the urgent need for exploring the yet undefined intracellular mechanisms that regulate functional beta-cell mass expansion during pregnancy. GWAS analysis revealed several mutations in the Nrf2 antioxidant pathway that are associated with diabetes. Additionally, we previously found that Nrf2 is required for adaptive beta-cell expansion after high fat feeding via stimulation of beta-cell proliferation, increased beta-cell survival, and maintenance of beta-cell identity. Interestingly, Nrf2 expression levels are increased by 4.3-fold (+/- 6.60, p&amp;lt;0.05, n=3-4) in maternal beta-cells during pregnancy, peaking at gestational day (GD)19, while returning to normal levels postpartum. Based on these observations, we hypothesize that Nrf2 might play an important role in expanding beta-cell mass during pregnancy and that disruption of Nrf2 expression or function might lead to GDM. In order to test our hypothesis, we generated tamoxifen (Tam)-induced beta cell-specific deletion of Nrf2 in female mice by crossing MIP-CreERTTAM with Nrf2lox/lox mice (βNrf2KO). In order to exclude any effect of Tam on pregnancy outcomes, females mice were injected with Tam (or vehicle control) a month before gestation. Pancreata were harvested at GD15 or GD19 and immunolabeled with Ki67 and insulin (proliferation assay), TUNEL and insulin (apoptotic assay) or insulin alone (beta-cell mass analysis). Our results show that βNrf2KO mice exhibit 70% reduction (+/- 0.25, p&amp;lt;0.01, n=4-6) in beta-cell proliferation and a 0.13% increase in beta-cell death (+/- 0.03, p&amp;lt;0.05, n=4-6) at GD15 compared to littermate controls. Importantly, βNrf2KO pregnant mice display a remarkable decrease in beta-cell mass at GD19 (50%) (+/- 0.68, p&amp;lt;0.05, n=4) compared with control mice, accompanied by a trending decrease in plasma insulin and increase in blood glucose. No changes were observed in the average number of embryos between βNrf2KO and control pregnant mice. We conclude that Nrf2 is required for beta-cell mass expansion during pregnancy by controlling beta-cell proliferation and survival and alterations in Nrf2 action could potentially lead to development of GDM. Presentation: Friday, June 16, 2023

Comparison of somatic embryogenesis efficiency from in vitro explant sources of Ngoc Linh ginseng (Panax vietnamensis Ha et Grushv.)

This study on the efficiency of embryogenesis from various explant sources of Ngoc Linh ginseng (Panax vietnamensis Ha et Grushv.) was conducted to select the appropriate materials and optimise embryogenesis in vitro. In this study, in vitroexplants from leaf fragments, petioles, roots, and embryos (globular, heart-shaped, and cotyledon) were cultured on MS medium supplemented individually or in combination with 2,4-D (2,4-dichlorophenoxyacetic acid: 0.2, 0.5, and 0.7 mg/l) and TDZ (Thidiazuron: 0.1, 0.3, and 0.5 mg/l). The results showed that all explants had embryogenesis after 6 weeks of culture (100%). The average number of embryos per leaf fragment (51.00 embryos) on the optimal medium was significantly higher than that of petioles (29.67 embryos) and root samples (18.00 embryos). Interestingly, the globular embryo explant was noted as the most suitable material source for embryogenesis (68.33 embryos/sample). The culture medium with the combination of 2,4-D and TDZ increased the ability to create secondary embryos in globular embryos, the highest result was achieved at the concentration of 0.7 mg/l 2,4-D combined with 0.1 mg/l TDZ in the medium containing 0.5 mg/l NAA - 1-Naphthaleneacetic acid (73.33 embryos/sample).

P-231 The effect of warming blastocysts in Vitrolife media previously vitrified in Cook media on reproductive potential and clinical outcome

Abstract Study question Does the use of a different manufacturer’s media for vitrification and warming of blastocysts affect pregnancy, miscarriage and live birth rates. Summary answer Warming blastocysts in Vitrolife media, previously vitrified in Cook media, does not significantly affect the pregnancy or live birth rate. What is known already Current good practice is to vitrify and warm blastocysts in media manufactured by the same company. This is due to the optimisation of the protocols and media composition intending to maintain high survival rates and associated pregnancy rates. However, circumstances can arise, such as supply and production issues, whereby it is not possible to source the same media for warming vitrified blastocysts as the media they were vitrified in. Study design, size, duration This retrospective cohort study was conducted at TFP-Oxford Fertility. Frozen embryo transfer (FET) cycles included in the study were performed between 2018 and 2022. Embryos that had undergone genetic testing were excluded from the study. The mean women's age was 35 in the control group (Cook warming media) and 36 in the study group (Vitrolife warming media). The average number of embryos transferred was 1.06 vs 1.17 in control vs study group. Participants/materials, setting, methods For the present study 817 patients underwent FET between 2016 and 2018 whereby blastocysts were vitrified in cook media (Sydney IVF Blastocyst Vitrification Kit, K-SIBV-5000) and warmed in cook media (Sydney IVF Blastocyst Warming Kit, K-SIBW-5000). In comparison 704 patients underwent a FET between 2018 and 2022 whereby blastocysts were vitrified in cook media and warmed in vitrolife warming media (Rapidwarm™ Blast, 10120). The pregnancy, clinical pregnancy and live birth rates were compared. Main results and the role of chance Fisher’s exact test was used to calculate the p value and a level of 0.05 was used for significance. The results showed that there was no significant difference in the pregnancy rate, clinical pregnancy rate or live birth rate for blastocysts warmed in Vitrolife media vs Cook media (59.0% vs 57.0% p = 0.56, 47.2% vs 43.9% p = 0.44, 39.3% vs 37.0% p = 0.72) Limitations, reasons for caution This is a retrospective study at a single centre. The number of previous treatment cycles per couple was not included in the study. The study only looked at a comparison of Cook and Vitrolife warming media, no other commercially available media was tested. Wider implications of the findings Although good practice to vitrify and warm blastocysts in the same manufacturer media, our results show that the reproductive potential and clinical outcome may not be adversely affected by using a different manufacturer’s media. However, composition of the media and protocol should be considered and further studies should be conducted. Trial registration number not applicable

Comparison of Pregnancy Outcomes in Thyroid Antibody-Positive Women with Different Fertilization Methods

<i>Objective</i> To compare the pregnancy outcomes of thyroid antibody (ATA) positive women treated by different fertilization methods during in vitro fertilization-embryo transfer. Methods A total of 204 female patients with normal thyroid function and positive ATA in the reproductive Medicine Center of the First Hospital of Shanxi Medical University from January 2020 to January 2022 were selected. According to different fertilization methods, they were divided into IVF group and ICSI group, including 115 IVF cycles and 89 ICSI cycles. The general data, clinical indicators after ovulation induction, fertilization, embryo status, and pregnancy outcome of the two groups were compared. <i>Results</i> There were no significant differences in age, BMI, bE₂, FSH, FT₃, FT₄, AFC, Gn dosage, Gn stimulation time, E₂ on HCG day, P on HCG day, intrauterine thickness on HCG day, number of retrieved oocytes and average number of embryos transferred between the two groups (P>0.05). The fertilization rate of the ICSI group was higher than that of IVF group within the group, while the high-quality embryo rate was significantly lower than that of the IVF group, with statistical significance (P<0.05). There were significant differences in sperm motility and sperm density between the two groups (P<0.05), and there was no significant difference between sperm volume and normal morphology rate (P>0.05). There were no significant differences in embryo implantation rate, biochemical pregnancy rate, clinical pregnancy rate, early abortion rate and live birth rate between the two groups (P>0.05). Logistic regression analysis showed that the method of assisted pregnancy and semen quality were not independent factors affecting high-quality embryo (P>0.05). In IVF group, TGAb was negatively correlated with fertilization rate (r=-0.202, P<0.05); and in ICSI group, TPOAb was negatively correlated with high quality embryo rate (r=-0.349, P<0.05). <i>Conclusion</i> Thyroid autoimmunity (TAI) in infertile women undergoing assisted reproductive technology (ART), the semen quality should be considered as the priority criteria for ATA. ICSI should not be recommended as routine fertilization.

Effects of Group Size on Behavior, Reproduction, and mRNA Expression in Brains of Brandt’s Voles

For social animals, a moderate group size is greatly important to maintain their reproductive success. However, the underlying neurobiological mechanism of group size on behavior and reproduction has rarely been investigated. In this study, we examined the effects of group size (1, 2, 4 pairs of adult male and female voles raised per cage) on behavior and reproduction. Meanwhile, the mRNA expression of stress and reproduction response-related genes in male brains was detected. We found that Brandt's voles (Lasiopodomys brandtii) in the large-sized group fight more severely than those in the small-sized group. Meanwhile, male voles were more anxious than females. The average number of embryos and litters per female in the medium-sized group was significantly higher than that of large-sized group. In male voles, stress- or reproduction-response mRNA expressions were more related to final group size or final density due to death caused by fighting. Our results indicated that a moderate group size was beneficial to the reproductive output of Brandt's voles. Our study highlights the combined effects of stress- or reproduction-related gene expression or behavior in regulating the fitness of voles with different group sizes.

Efficiency of Multiple Ovulation and Embryo Transfer (MOET) in a Cattle Farm over a Period of Ten Years in Gujarat

The present study was conducted at Sabarmati Ashram Gaushala, Bidaj, Gujarat. A total of 427 donor animals were investigated over a ten-year period to assess the efficiency of Standard protocol of Multiple Ovulation and Embryo Transfer (MOET). Simultaneously a season wise and breed wise study was also conducted. Breeds studied were Gir, HF, HFCB, Jersey, JYCB, Red Sindhi and Sahiwal. Overall average number of embryos, average number of viable embryos, number of pregnancies and conception rate (%) were found to be 7.8, 4.8, 281 and 39.4, respectively. The year-wise number of pregnancies and conception rates varied from 09 to 42, and 24.6% in 2009-10 to the highest 61.3% in 20014-15, respectively. Further, breed-wise results showed average viable embryo in different breeds as 2.8 in Gir, 4.1 in HF, 5.1 in HFCB, 6.3 in Jersey, 5.8 in JYCB, 3.7 in Red Sindhi and 6.6 in Sahiwal, with corresponding conception rates of 67.7. 34.6, 28.8, 53.1, 39.4, 47.1 and 36.4 %, respectively. A trend was observed that the efficiency of MOET programme was less during monsoon (31.2%) as compared to summer and winter (41.5 and 41.4%).

Competition, insurance mandates, and clinical practices and outcomes at in vitro fertilization clinics.

The high cost of in vitro fertilization (IVF) procedures coupled with public availability of success rates may influence IVF practice patterns and success rates but may be mitigated by mandated insurance coverage for IVF procedures. This study examined associations of competition with IVF practice patterns and success rates by insurance mandate status. We used publicly available 2018 National Assisted Reproductive Technology Surveillance System data from the CDC. We defined competition as distance to nearest clinic and number of clinics within certain radii. We used linear regression to explore associations of competition, insurance mandate status, and interactions between competition and mandate status with clinical practice patterns (e.g., percentage of cycles among patients < 35years, percentage of cycles using ICSI, average number of embryos transferred) and IVF success rates (e.g., live birth rates). We also assessed the percentage of variation in outcomes explained by our models, R2. For practice patterns, the largest R2 value was 0.3518, meaning only 35.18% of the variability in the practice pattern variable was explained by competition and insurance mandate status. In most cases, the R2 values were less than 0.20, indicating little to no association. Less than 10% of the variability in success rates was explained by competition and insurance mandate status. The multiple regression analyses all yielded low R2 values, indicating weak associations. These encouraging results coincide with previous studies, suggesting that competition, even by insurance mandate status, does not have a strong association with IVF practice patterns or success rates.

P-525 PGT-A with embryo pooling: does it result in a higher yield of euploid embryos and better clinical pregnancy outcomes?

Abstract Study question Does PGT-A with embryo pooling result in a higher yield of euploid embryos and better clinical pregnancy outcomes compared to PGT-A done after one cycle? Summary answer There is no evidence to support that embryo pooling for PGT-A purposes is favorable in preference to PGT-A after one cycle. What is known already PGT-A has been the method of choice that is thought to optimize embryo selection and enhance clinical outcomes. However, the number of embryos available for biopsy can be a source of limitation, as the yield of euploid embryos suitable for transfer can be affected when lower number of embryos are available for screening. A protocol that is suggested to overcome this limitation is to undergo multiple IVF/ICSI cycles with blastocyst biopsy and embryo pooling, then proceed with PGT-A screening once a sufficient number of biopsied embryos is attained. Study design, size, duration This is a retrospective study that included ICSI cycles from June 2019 to December 2021, involving 157 patients (average maternal age 36.4) that underwent PGT-A after one ICSI cycle (group A), and 79 patients (average maternal age 35.6 ) that underwent PGT-A after embryo pooling from more than one ICSI cycle (Group B). Patients in group B underwent pooling due to cost implications on the number of embryos to screen. Participants/materials, setting, methods A total of 843 embryos were biopsied and screened for group A, and 536 were biopsied and screened for group B. Following PGT-A screening, a number of frozen embryo transfers (FET) were conducted (120, 67 respectively). Investigations included: number of embryos available for biopsy, number of euploid embryos and clinical pregnancy outcomes after FET. Statistical analysis was calculated (P &amp;lt; 0.05). Main results and the role of chance Patients in group A proceeded with PGT-A after only one ICSI cycle, while in group B patients underwent 2.3 cycles on average. The average number of embryos suitable for biopsy was 5.4, and 6.9 per patient respectively. The rate of euploid embryos was 42.1% and 43.8%, and the average number of euploid embryos per patient was 5.4 and 6.5 respectively. A total of 144 embryos were transferred in group A with an average of 1.2 embryos per patient. Clinical pregnancy rate (CPR) was 60%, implantation rate was 54.9% and live birth rate (LBR) was 35.8%. In Group B a total of 67 embryos were transferred with an average of 1.1 embryos per patient. Clinical pregnancy rate (CPR) was 67.7%, implantation rate was 68.7% and live birth rate (LBR) was 40%. No significant difference was observed between both groups for clinical outcomes (P &amp;gt; 0.05). Limitations, reasons for caution This study is retrospective and small in size. Data on live birth rate requires further follow up as a number of patients are still in ongoing pregnancies. A larger data set study is needed to support conclusion. Wider implications of the findings Results from this study does not support the adoption of a pooling strategy for patients undergoing PGT-A. Although the number of embryos available for biopsy may increase slightly, the yield of euploid embryos is not significantly increased and the clinical pregnancy rate is not enhanced. Trial registration number not applicable

P-557 Fresh Transfer with Supernumerary PGT-A Biopsy: The Best of Both Worlds

Abstract Study question What is difference in time to pregnancy for patients between 35-40 years who undergo fresh versus frozen-thawed embryo transfers with or without PGT-A? Summary answer Patients undergoing fresh embryo transfer (ET) had a shorter time to pregnancy within the same cycle compared to patients electing for freeze-all cycles. What is known already When comparing clinical pregnancy rates following fresh versus frozen-thawed embryo transfers, multiple studies have shown a higher clinical pregnancy rate among frozen-thawed embryo transfers compared to fresh embryo transfers. Particularly for patients over 35 years, there has been a growing national trend towards elective freeze-all cycles, for reasons such as to pursue preimplantation genetic testing for aneuploidy (PGT-A). While PGT-A testing can be effective in reducing miscarriage in patients age 38-40, there is limited studies evaluating time to pregnancy within this population, especially in the context of each passing month’s effect on fertility. Study design, size, duration Retrospective review was performed for 697 IVF cycles and 881 transfers from January 2016 – December 2021 at a single academic fertility center in Boston, Massachusetts. Two-tailed t-tests and analysis of variance (ANOVA) were used to compare differences, with p-value less than 0.05 set for statistical significance. All PGT-A testing as performed using a modified FAST-SeqS next generation sequencing method (Invitae, San Francisco, CA). Participants/materials, setting, methods Cycle characteristic of patients between 35-40 were categorized to four groups: Fresh ET with PGT-A of supernumerary embryos (82 transfers), Fresh ET without PGT-A (526 transfers), Freeze-all cycle with PGT-A (223 transfers), and Freeze-all cycle without PGT-A (50 transfers). All averages were calculated within one IVF cycle with a single cohort of embryos. Time to pregnancy was calculated in days from day of retrieval to positive serum beta-human chorionic gonadotropin (bHCG). Main results and the role of chance Among the 881 transfers, 478 had fresh ETs and 403 had frozen-thawed ETs with or without PGT-A respectively. When comparing fresh and freeze-all cycles, there was no difference in average numbers of transfers performed to achieve a clinical pregnancy (1.27 fresh vs 1.25 freeze-all, p = 0.56). For patients who didn’t elect for PGT-A, there was no difference in number of embryos transferred (1.86 for fresh versus 2.06 for freeze-all, p = 0.54). For patients who elected for PGT-A, the average number of embryos transferred was higher for patients that opted for fresh transfer over freeze-all (1.67 for fresh ET with PGT-A of supernumerary embryos versus 1.24 for freeze-all PGT-A, p = 0.0025). Average days from oocyte retrieval to pregnancy was significantly lower for all fresh ETs (28.8 days for fresh ET without biopsy, 26.6 days for fresh ET with biopsy of supernumerary embryos, p &amp;lt; 0.0001) compared to all freeze-all transfers (110.9 days for freeze-all ET without biopsy, 119.5 days for freeze-all ET with biopsy, p &amp;lt; 0.0001). This delay in time to pregnancy is partially attributed to pending genetic testing results, however, these results usually return in 2-3 weeks, which does not account for the near 3-month delay in time to pregnancy among freeze-all cycles. Limitations, reasons for caution These retrospective findings were of all women who achieved pregnancy from one retrieval. These results are not reflective of the patients who needed to undergo multiple cycles to achieve pregnancy, or patients who never achieved pregnancy. Wider implications of the findings These findings suggest that PGT-A may be offered to patients &amp;gt; 35 to minimize the number of embryos for transfer. Patients opting for a fresh transfer of an untested embryo and biopsy of supernumerary embryos had a nearly 3-month shorter time to pregnancy than those electing for a freeze-all cycle Trial registration number not applicable

Fertility preservation decisions and outcomes of young women with breast cancer.

e12520 Background: As breast cancer (BC) survival improves, fertility preservation (FP) counselling prior to initiating gonadotoxic chemotherapy for women of child-bearing age becomes more important. This study explores the factors that influence the decisions to pursue FP before and after breast cancer treatment and the FP outcomes. Methods: An IRB approved web-based survey examining decisions about FP, factors influencing decisions, and post-decision regret was administered to English and Spanish speaking women (age 18-45) diagnosed with stage I-III BC. All women received education and counselling by a MSK Fertility Nurse Specialist prior to initiating cancer treatment at MSKCC in 2009-2017. Additional data was extracted from the medical record. Results: 211 women with BC (82% ER+, 19% HER-2+, 12% triple negative) with a mean age of 34 (age range 20-45) participated in this study. At the time of diagnosis 128 (61%) women were married/living with partner and 46 (22%) women had children. 151 (72%) wanted (additional) children, 48 (23%) were unsure if they wanted (additional) children and 12 (6%) did not want (additional) children. Of the 151 women who desired (additional) children, 94 underwent FP pre-treatment and 3 post-treatment. Of 48 unsure women, 14 underwent FP. A total of 115 women underwent FP before treatment: 73 froze eggs and 49 froze embryos. After treatment 28 (13%) women met with a reproductive endocrinologist and 13 underwent FP: 9 froze eggs and 4 froze embryo. The average number of eggs frozen was 14 pre-treatment and 18 post-treatment. The average number of embryos frozen was 7 both pre- and post-treatment. After completing primary breast cancer treatment, 61 women opted to try to build their family and 42 had children: 35 gave birth and 7 used a surrogate. In a multivariate analysis lower socioeconomic status and non-white race were predictive of discordance between desire for FP and completion of FP before treatment (p &lt; 0.05). 65% of patients reported cost of freezing eggs/embryos was important in their decision. Insurance did not cover any FP treatment for 56/115 (49%) of women and 34% (39/115) of women paid $10,000 or more for their FP. Participants reported the ability to feel hopeful about their future (82%) and fear of regret (80%) were the most important factors when making their FP decision. 83% (176/211) of women believe they made the right FP decision and 80% would make the same decision again. Women had less regret if they underwent FP. Conclusions: Our study demonstrates the importance of discussing FP with women with BC of child-bearing age, regardless of race or economic status. Fertility preservation is often successful in women with BC. Regardless of pregnancy outcomes, women felt that having FP offered them a sense of control and hope for the future during a vulnerable time in their lives with low risk of regret.

Assisted Reproductive Technology Surveillance - United States, 2018.

Assisted Reproductive Technology Surveillance - United States, 2018.

CUMULATIVE CLINICAL PREGNANCY RATES PER OOCYTE RETRIEVAL AFTER IN VITRO FERTILIZATION

To investigate cumulative clinical pregnancy rates (CCPR) and to improve patient counseling with the prognostic pregnancy outcome per oocyte retrieval cycle. DESIGN: Retrospective study. All classical IVF or ICSI Cycles involving fresh embryo transfer carried out between January 2015 and December 2018 at our clinic were investigated, including fresh ET only and fresh ET followed by 1-6 time frozen-thawed embryo transfers (FET). Both fresh ET and FET were included to calculate the CCPR. Clinical pregnancy was confirmed by the observation of a gestational sac with a fetal heartbeat on ultrasound evaluation. Results were analyzed by using chi-square test (P=0.05). This study included 535 fresh ET cycles and 330 FET cycles. The average age of the patients was 35.9 (26-45). The ratio of fresh cycles with frozen embryos after transfer was 75% (402/535). The average number of embryos cryopreserved per cycle was 3.1. The average number of embryos transferred was 1.3. The CCPR per oocyte retrieval increased by each cycle of frozen-thawed embryo transfer (Table 1). The significant differences in CCPR were found among Fresh, FET1 and FET2 cycles. No significant differences in CCPR were observed among FET2, FET3, FET4, FET5 and FET6 cycles.Tabled 1Table 1. The cumulative clinical pregnancy rates per oocyte retrievalCycleClinical pregnancy rateCumulative clinical pregnancy rateFresh42.4% (227/535)42.4% (227/535) aFET146.8% (88/188)58.9% (315/535) bFET243.1% (34/80)65.2% (349/535) cFET342.9% (15/36)68.0% (364/535) cFET46.3% (1/16)68.2% (365/535) cFET537.5% (3/8)68.8% (368/535) cFET650.0% (1//2)69.0% (369/535) ca,b,c Values with different superscript letters within the same column are significant different (p<0.05) Open table in a new tab a,b,c Values with different superscript letters within the same column are significant different (p<0.05) Our data analysis indicated that the CCPR per oocyte retrieval increased until the second FET cycle with no benefit for additional FET cycles to have one pregnancy. The results provided here may be useful in counseling patients on the high pregnancy outcome (69.0%) per oocyte retrieval cycle.

Bazı Biber (Capsicum annuum L.) Genotiplerinin Anter Kültürü Performanslarının Belirlenmesi

Bu çalışmada, farklı tiplerde 23 biber genotipinde anter kültürü performansı belirlenmiştir. Çiçek tomurcuklarının dezenfeksiyonunda, tomurcuklar öncelikle % 70’lik etil alkolde 30 saniye bekletilmiş ve sonrasında 10 dakika % 15’lik sodyum hipoklorit çözeltisinde tutulmuşlardır. Anterler ilk aşamada; 30 g/L sakkaroz, 2.5 g/L aktif kömür, 15 mg/L gümüş nitrat (AgNO3), 4 mg/L 1-Naphthaleneacetic acid (NAA), 0.5 mg/L 6-Benzylaminopurine (BAP) ve 6.5 g/L agar içeren Murashige and Skoog (MS) besin ortamında kültüre alınmışlardır. Sonrasında anterler, 30 g/L sakkaroz ve 6.5 g/L agar içeren ve büyümeyi düzenleyici içermeyen MS besin ortamına aktarılmışlardır. Elde edilen embriyolarda, aynı besin ortamına alınmışlardır (ikinci ortama). Anterler, ön uygulama olarak, +35ºC’de ve karanlık koşullarda 2 gün süresince bekletilmişlerdir. Çalışma sonuçları değerlendirildiğinde, genotiplere göre 100 anter için ortalama embriyo sayıları 0.83 ile 44.44 arasında değişmiştir. En yüksek oran, FT-509 genotipinde gözlemlenmiş ve bu genotipi FT-508 (23.61 embriyo/100 anter), FT-1181 (23.37 embriyo/100 anter) ve FT-905 (22.89 embriyo/100 anter) genotipleri takip etmiştir. En düşük performansı, 0.83 adet embriyo/100 anter ile FT-1178 genotipi göstermiştir. Oluşan embriyoların tamamı, bitkiye dönüşebilmiştir. Bu nedenle, 100 anter için ortalama bitki oluşturan embriyo sayısı değerleri, 100 anter için ortalama embriyo sayıları ile aynı olmuştur. En yüksek ortalama bitki sayısı 20 adet ile FT-508 no’lu genotipte gözlemlenmiş ve FT-509 (17.5 bitki), FT-1181 (16 bitki), FT-905 (12 bitki), FT-263 (10.5 bitki) ile FT-507 (8.25 bitki) genotipleri tarafından takip edilmiştir. En düşük bitki sayısı ise 0.5 adet ile FT-1178 no’lu genotipte kaydedilmiştir.