Day Of Embryo Transfer Research Articles

P-357 Downregulation (DR) with combined gonadotropin-releasing hormone agonist (GnRHa) and aromatase inhibitor (AI) optimizes the frozen-thawed embryo transfer (FET) results in women with adenomyosis

Abstract Study question Does combined GnRHa & AI therapy for downregulation in women with adenomyosis optimize the FET success rates / outcomes on par with women without adenomyosis? Summary answer GnRHa combined with AI used for downregulation in women with adenomyosis optimized the outcomes of frozen–thawed embryo transfer cycle on par with women without adenomyosis. What is known already Women with adenomyosis have lower rates of successful implantation via altered molecular expressions in the endometrium due to local hyperestrogenism & also an increased risk of early pregnancy loss (Munro et al 2019). DR improves the clinical pregnancy rate by reducing the endometrial inflammatory reaction and/or myometrial contractility and their impact on uterine receptivity in women with adenomyosis undergoing FET (Sania Latif et al 2021). Combined treatment for uterine adenomyosis with Anastrazole plus GnRHa showed better results than dienogest treatment with a higher reduction of symptoms & higher pregnancy rates (M Sbracia & F Scarpellini, 2018). Study design, size, duration A Retrospective cohort study conducted at a tertiary care fertility unit. Data for 326 women with/without adenomyosis undergoing frozen–thawed embryo transfer after IVF was was retrieved from the hospital's database and analyzed for a period between September 2021 to November 2022. Participants/materials, setting, methods Women with adenomyosis (n = 107) received Anastrozole 1mg/day for 2months plus 3doses of Inj. Goserelin 3.6mg subcutaneously at 28days interval between 2successive doses. Hormone replacement therapy (HRT) was started 2weeks after the 3rddose of Inj. Goserelin & FET was performed after an optimum endometrial thickness (EMT) was achieved. For women without adenomyosis (n = 219) HRT was started on cycle Day-2 & FET was performed after achieving an optimum EMT. Serum beta-hCG was performed on Day15 after FET. Main results and the role of chance Statistical analysis was performed using SPSS20version. Normally distributed continuous variables were compared using a student t-test, and categorical variables were compared by χ2 and Fisher's exact test, where appropriate. To reduce selection bias, propensity score matching was used, and propensity matching yielded 99 pairs. Baseline characters like age (p-value=0.36), BMI (p-value=0.12), duration (p-value=0.28), type (p-value=1) & cause (p-value=0.3) of infertility, endometrial thickness (p-value=0.37), day of embryo transfer (p-value=0.57) were comparable for the two groups. FET results in terms of positive pregnancy test (Serum beta-hCG > 50mIU/ml) were found to be 68.21% for the DR- FET group and 67.71% for the HRT group which were comparable(p-value=0.17). This suggests that downregulation in women with adenomyosis helps achieve success rates similar to women without the disease. Pregnancy outcomes like miscarriage (15 vs 13.13 %, p-value=1.00), biochemical pregnancy (3.03 vs 2.02 %, p-value=1.00) and ectopic pregnancy (1.01 vs 0 %, p-value=0.49) rates analyzed between the DR FET and HRT groups showed no statistically significant difference. Clinical pregnancy rates were almost similar for the 2 groups (55.71 Vs 57.46%, p-value=0.65). Thus, we conclude that downregulation with combined GnRHa and AI optimizes the frozen-thawed embryo transfer results in women with adenomyosis on par with women without adenomyosis. Limitations, reasons for caution This is a retrospective study and hence randomized comparison was not possible. Women were followed up for 12 weeks of pregnancy, hence live birth rates were not analyzed in our study. Wider implications of the findings Combining the two treatment modalities (GnRHa + AI) which work at different sites optimizes IVF success rates & pregnancy outcomes. Hence we suggest that well-designed prospective randomized studies are needed to further analyze the synergistic role of this drug combination for downregulation in women with varied severity of adenomyosis. Trial registration number Not applicable

P-521 A gene expression risk signature of endometrial failure for prognosis in In Vitro Fertilization (IVF) patients

Abstract Study question It is possible to identify a characteristic pattern of endometrial gene expression indicative of implantation failure, which is independent of implantation window displacements? Summary answer An endometrial transcriptomic signature was able to identify patients with a > 3-fold increased risk of implantation/pregnancy failure with 95% accuracy. What is known already Implantation failure of endometrial origin is a complex and multifactorial symptom with diverse causes, diagnosed in IVF patients after repeated failures with good quality embryos. A generation of gene expression tools have assumed that Window of implantation (WOI) displacement is the principal cause of this condition, but strategies seeking to counteract this problem by adjusting the day of embryo transfer have not yielded convincing improvements in outcomes. However, it is conceivable that other forms of endometrial disruptions, relevant to embryo implantation, could exist. New endometrial diagnostic strategies are needed to understand, diagnose and potentially treat patients affected with such problems. Study design, size, duration A prospective multicenter study between January 2018 and December 2021 recruited 281 Caucasian IVF patients (mean age of 39.4±4.8 years and BMI of 22.9±3.5 kg/m2) undergoing hormone replacement therapy and encompassing 114.5±7.2 h of progesterone administration at time of endometrial biopsy. Following experimental quality controls and clinical follow up, 186 patients who had a good quality embryo transferred in the cycle after endometrial biopsy collection were included for gene discovery and prediction performance. Participants/materials, setting, methods The expression of 404 genes selected for their potential to distinguish endometrial timing and/or endometrial disruption was measured. Transcriptomic variation related to progression of the menstrual cycle was removed using transcriptomic endometrial dating (TED) and linear models. Study groups were established according to clinical and gene expression parameters through a semi-supervised artificial intelligence procedure. Gene signature discovery and a cross-validation processes were undertaken to define predictive expression patterns. Reproductive outcomes were compared between prognosis profiles. Main results and the role of chance We developed a procedure called Clinically Acute Transcriptomic Stratification (CATranS), combining clinical parameters and deep transcriptomic molecular characterisation to stratify patients according to endometrial prognosis: ‘poor’ (n = 137) or ‘good’ (n = 49). These two transcriptomic profiles were associated with differing reproductive outcomes in the single embryo transfer following biopsy: pregnancy rate (45.1% vs 79.6%, poor vs good prognosis, respectively, p = 3.8E-5); live birth rate (56.4% vs 97.5%, p = 3.0E-06); clinical miscarriage rate (27.9% vs 2.6%, p = 0.0020); biochemical pregnancy rate (20.4% vs 0%, p = 0.0023). Patients with a poor prognosis profile had 3.3-times higher relative risk of a transferred embryo failing to implant or a pregnancy not being sustained to term, compared with good prognosis patients. Initially, a reference dataset was used to build a prototype for diagnosing endometrial failure, revealing that a gene expression signature consisting of 135 genes was the most predictive. Prediction performance was estimated using a 5-fold 100-times cross-validation process, resulting in a median accuracy of 0.92, median sensitivity of 0.96, and median specificity of 0.84. From these 135 predictive genes, 122 were differentially expressed (FDR<0.05) in endometrial poor prognosis, 59 up- and 63 down-regulated, most involved in functional processes such as regulation (17%), metabolism (8.4%), immune response and inflammation (7.8%). Limitations, reasons for caution We describe a potential new strategy for evaluating endometrial competence. However, to confirm predictive value, validation using additional samples from patients independent of signature discovery set is required. Further research to identify potential treatments for patients classified as poor prognosis is needed, providing a tailored clinical pathway for these patients. Wider implications of the findings The prototype described is a novel concept, potentially leading to development of a new generation of tools for diagnosis of fertility problems related to endometrial factors. The ‘poor’ prognosis profile is not caused by asynchronies in menstrual cycle progression, opening the possibility of finding new treatment pathways for these patients. Trial registration number NA

P-785 Promising Reproductive Outcomes In Patients With Refractory Thin Endometrium After Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) – A Case Series

Abstract Study question Does Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) improve endometrial thickness and receptivity and restoration of fertility in refractory cases of thin endometrium? Summary answer Selective Autologous Bone Marrow Regenerative Cell Therapy from Seragen (ABM-RCT) improves endometrial thickness, pregnancy, and live birth rate in refractory cases of thin endometrium. What is known already The impaired endometrial function can limit implantation due to insufficient tissue regeneration. Clinical investigation has proved that stem cells existing in the endometrium might originate from bone marrow and provides evidence that bone marrow-derived cells have a non-hematopoietic physiologic contribution to decidual stroma and play a vital role in implantation and pregnancy maintenance and non-hematopoietic BMSCs are able to impact decidual molecular milieu and overcome implantation defects. ABM-RCT is the most commonly used cell therapy and its safety and efficacy are well-established and documented. This forms the basis for exploring and standardizing ABM-RCT in the management of refractory thin endometrium. Study design, size, duration Eleven patients (from age 29 to 43 with refractory thin endometrium resistant to conventional treatment modalities were recruited and obtained informed consent with detailed explanation that the therapy is still experimental and the risk of failure was given to the couple as part of gynec-oncology treatment. Five patients had asherman’s syndrome, two patients had genital tuberculosis, two patients had thin endometrium hypo-responsive/unresponsive to estrogens, with RIF and two patients underwent chemo and radiotherapy Participants/materials, setting, methods BM aspiration was performed under local anesthesia, from the iliac crest using a disposable BM aspiration needle (Jamshidi, 11 G) and collected in heparinized syringes. Progenitor cell enrichment was done by Seragen’s Selective Enrichment Protocol. Peripheral blood was collected to enrich growth factor concentrate. A 2.9mm Hysteroscope was used with an operating channel and egg pickup needle attached to it and cells were implanted in subendometrial zone in all four walls of the uterine cavity. Main results and the role of chance All patients presented to us with refractory thin endometrium in HRT cycle despite giving all possible medications. Hence the decision was taken for Autologous Bone Marrow Cell Concentrate Therapy.Post ABM-RCT, endometrial thickness showed increase 100% (11/11) of more than 8 mm in all cases on day of embryo transfer with average endometrial thickness improvement was 1.8 mm than previous cycles, with uniform triple layer pattern.10 out of 11 (91%) patients conceived after autologous stem cell injection. 9 Out of 10 (90%) patients conceived had delivered healthy babies and 1 patient (9%) had a miscarriage at 12th week of pregnancy. 1 patient did not conceive will be assessed for immunoprofiling and one more rejuvenation cycle. Limitations, reasons for caution Our study outcomes are consistent with several previous studies and the feasibility of treatment at an IVF setup and encouraging results. Well-planned study with more patients is warranted to evaluate safety, effectiveness, and cost of this modality before it becomes integrated in treatment of this frustrating condition during IVF procedures Wider implications of the findings Our results demonstrates selectively enriched ABM-RCT enhances endometrial thickness, tissue remodeling, uneventful gestation, improved pregnancy and live birth rate. Need for Surrogacy in Non-Responsive Thinendometrium May Be Re-Evaluated. Role of hematopoetic stem cell research and knowledge generated is representing reliable strategy in IVF that may be routinely used in clinicalpractice. Trial registration number Not applicable

P-330 Is 17 alpha hydroxyprogesterone caproate an adequate alternative to improve reproductive outcomes in artificially prepared frozen embryo transfer cycles (FET)?

Abstract Study question Is 17 alpha-hydroxyprogesterone caproate an adequate option to improve reproductive outcomes in patients with low serum progesterone (P4) level the day of frozen embryo transfer? Summary answer Patients with serum P4<10ng/ml may benefit with the addition of intramuscular 17alpha-hydroxyprogesterone caproate injections, and have similar ongoing pregnancy rates than patients with serum P4≥10ng/ml. What is known already Indications of embryo cryopreservation have expanded in the last years. Recently published papers highlight that low serum progesterone levels before frozen embryo transfers decrease pregnancy and live birth rates and increase miscarriage rates, even in cases of euploid embryos transfers. Other studies have published a beneficial effect with the addition of daily subcutaneous progesterone injections in patients with low serum progesterone levels around the time of embryo transfers. In Argentina subcutaneous progesterone has been available since October 2022, not before. 17alpha-hydroxyprogesterone caproate is a synthetic progestin used for luteal phase support and approved for the prevention of preterm birth. Study design, size, duration This prospective cohort study was performed between May 2021 and August 2022 in 114 patients undergoing 138 FET cycles after an artificial endometrial preparation with estradiol valerate 4mg daily and micronized vaginal progesterone 200mg, three times daily. Participants/materials, setting, methods Patients <50 years old with triple-layer endometrium ≥7mm underwent frozen embryo transfer. Progesterone was measured immediately before the embryo transfer. Group 1 (N = 47): P4>10ng/ml; Group 2 (N = 46): P4<10ng/ml with addition of 17 alpha-hydroxyprogesterone caproate after embryo transfer; Group 3 (N = 46): P4<10ng/ml without addition of 17 alpha-hydroxyprogesterone. Primary endpoint was to compare ongoing pregnancy rate beyond week 12 between the three groups. Secondary endpoints were pregnancy rate and miscarriage rate. Main results and the role of chance Parameters were comparable between the groups in terms of age, body mass index, estradiol, P4 levels at the beginning of endometrial preparation and number of embryos transferred. Ongoing pregnancy rate was: Group 1 = 38.3% (18/47); Group 2 = 39.1% (18/46); Group 3 = 28.3% (13/46) (p = 0,337). Pregnancy rate was: Group 1 = 48,9% (23/47); Group 2 = 50% (23/46); Group 3 = 50% (23/46) (p>0,05). Miscarriage rate was: Group 1 = 21.73% (5/23), Group 2 = 26.1% (6/23) and Group 3 = 43.47% (10/23) (p = 0,208). Although there were no significant differences between the groups, both, Group 1 and Group 2, had higher ongoing pregnancy rates and lower miscarriage rates than group 3. This is probably due to the limited sample size. Limitations, reasons for caution The main limitation of our study is the sample size and the fact that most of the embryos were not genetically evaluated before being transferred, which would eliminate a variable that could interfere with the results obtained. Only micronized progesterone and 17alpha-hydroxyprogesterone caproate were studied. Wider implications of the findings The addition of exogenous 17alpha-hydroxyprogesterone caproate on the embryo transfer day may improve ongoing pregnancy rates in cases of low serum progesterone levels, being a simple and safe strategy to individualize luteal phase support when no other form of progesterone administration is available. Trial registration number NOT APPLICABLE

O-260 The influence of the vaginal microbiome on clinical outcomes in patients undergoing a frozen embryo transfer: a prospective pilot study

Abstract Study question Are Lactobacillus-dominant vaginal microbial compositions advantageous for clinical outcomes and is the specific Lactobacillus species relevant? Summary answer Favourable clinical outcomes are increased in patients with a Lactobacillus-dominant vaginal microbiome, particularly in those patients with microbial communities dominated by L.crispatus and L.jensenii. What is known already The clinical relevance of the urogenital microbiome is emerging as a topic in female reproductive health. Recent evidence suggests that the composition of the vaginal microbiome may have an impact on clinical outcomes in patients undergoing assisted reproductive treatment. A Lactobacillus-dominant microbiome (LDM), typically composed of L.crispatus, L.gasseri, L.iners or L.jensenii, is considered to be associated with favourable outcomes. In contrast, a non-Lactobacillus-dominated microbiome (NLDM), consisting predominantly of anaerobe species, may be linked to poor outcomes. The evidence surrounding this relationship is controversial and the importance of relative Lactobacillus abundance in the vagina in relation to clinical outcomes remains unclear. Study design, size, duration A single-centre prospective pilot study included 81 patients (mean age: 38.2) undergoing frozen embryo transfer using their own (n = 52) or donated (n = 29) oocytes. A vaginal swab was taken on the embryo transfer day and the microbiome analysed. Implantation, ongoing pregnancy and early miscarriage rates were compared between different microbiome compositions. To minimise the confounding effect of aneuploidy on clinical outcomes, a patient sub-group (n = 54) receiving donor oocytes or a euploid embryo transfer was also considered. Participants/materials, setting, methods Quantitative PCR was utilised to determine the relative abundance of the four predominant Lactobacillus species as well as 15 species associated with vaginal dysbiosis. Microbiome compositions were grouped according to the relative Lactobacillus abundance and patients divided into an LDM (>80%) or NLDM (<80%) group. Patients were further sub-divided into community state types (CSTs) according to the dominant species present in their sample (CST-I: L.crispatus, CST-II: L.gasseri, CST-III: L.iners, CST-V: L.jensenii and CST-IV: dysbiotic species). Main results and the role of chance Relative abundance of Lactobacillus was significantly higher in samples from patients achieving an ongoing pregnancy (80.7% vs. 61.7%; p = 0.05). Implantation rates were comparable between LDM and NLDM patients (66.0% and 64.3%), but the ongoing pregnancy rate showed an apparent increase in LDM patients (58.5% vs. 39.3%; p = 0.11), concomitant with a significant decrease in the miscarriage rate (11.4% vs. 38.9%; p = 0.03). Limiting the analysis to patients at low risk of aneuploidy (young donor and PGT-A cycles), Lactobacillus abundance appeared higher in patients with ongoing pregnancies (74.8% vs. 58.0%; p = 0.34). Likewise, LDM patients showed higher ongoing pregnancy (61.3% vs. 43.5%; p = 0.27) and lower miscarriage (5.0% vs. 33.3%; p = 0.06) rates, although significance was not achieved in this restricted sample. Comparison of CST groups indicated better outcomes in CST-I and CST-V compared to other groups. The ongoing pregnancy rate was significantly higher (65.7% vs. 41.3%; p = 0.04), alongside a lower miscarriage rate (8.0% vs. 32.1%; p = 0.04). When only considering patients with low aneuploidy risk, similar results were obtained (implantation rate: 78.9% vs. 57.1%, p = 0.14; ongoing pregnancy rate: 73.7% vs. 42.9%, p = 0.04; miscarriage rate: 6.7% vs. 25.0%, p = 0.21). Of note, there were no differences in baseline characteristics (age, ethnicity & BMI) or embryo morphologies between groups. Limitations, reasons for caution All patients were from a single fertility clinic and the study population was predominantly composed of white women. Consequently, results may not be applicable to women of different ethnicities. Patients with a low risk of aneuploid embryo transfer represented a small subgroup where meaningful statistical analysis was not always possible. Wider implications of the findings This study identifies a correlation between Lactobacillus species colonisation in the vagina and successful clinical outcomes, suggesting that the vaginal microbiome modulates the chances of IVF success. The results are promising, providing motivation for further, larger studies, involving more diverse populations in order to draw definitive conclusions. Trial registration number not applicable

P-500 Relationship of midluteal serum progesterone levels and pregnancy outcomes after day-3 embryo-transfer(ET) in hormonal replacement therapy (HRT) cycles with intramuscular progesterone: a prospective cohort study

Abstract Study question Does midluteal(day-9 of progesterone) serum progesterone level in HRT cycles with intramuscular progesterone affect the clinical pregnancy rate (CPR)/early miscarriage rate(MR) and live birth rate(LBR)? Summary answer Midluteal serum progesterone between 32.2-43.7ng/ml results in significantly greater CPR versus that with lower/higher levels and significantly greater LBR compared to that with lower levels. What is known already Evidence points to low serum progesterone levels at the time of FET being associated with lower ongoing pregnancy rates. Recently, the relation of midluteal serum progesterone levels in HRT cycles with vaginal progesterone as luteal phase support (LPS) has been elaborated indicating better pregnancy outcomes with an optimum range of serum progesterone/higher levels. There is no data establishing the association of pregnancy outcomes with midluteal serum progesterone levels when intramuscular progesterone is used as LPS in HRT cycles. Study design, size, duration A prospective cohort study conducted between December 2019-March 2022 at a tertiary centre with 196 patients(204 cycles), both-autologus(AO)-56.3% and donor oocytes(DO)- 43.6%: fresh ET-16.6% and frozen ET(FET)-83.3% with similar ICSI, vitrification and ET protocols(2/3 Grade A day-3 embryos). CPR, early MR (≤13 weeks miscarriage/clinical pregnancy) and LBR (≥28 weeks, LBs/ETs) were compared. Serum progesterone was measured on the morning of day-5 post-ET(day 9th from the start of progesterone). Most patients delivered at the same institute. Participants/materials, setting, methods Patients were non-smokers with normal hysteroscopy. Oral estradiol 6-8mg was initiated from Day 1-2 (for 10-17 days). When ET ≥ 7mm and serum progesterone <1ng/ml, intramuscular progesterone 100mg/day was started (daily till day-4 post-ET, then every 3rd day till week 8). Vaginal progesterone gel was added from day 5 post-ET till week 10. Serum progesterone levels were categorized in tertiles. Chi-square, t-tests and multiple regression analysis was used for statistical analysis. Two-tailed P<.05 indicated statistical significance. Main results and the role of chance Mean age was 34.76±5.36 years with a mean BMI 26.4±4.38kg/m2 Overall, CPR was 106/204(51.9%), LBR 78/204(38.2%) and early MR 28/106(26.4%) Midluteal serum progesterone levels ranged from 11.1-261ng/ml and were divided into tertiles(T) T1<32.0ng/ml(n = 68),T2 32.2-43.7ng/ml(n = 68),T3 >43.7ng/ml (n = 68) Median progesterone levels of T1,T2,T3 were 27.7ng/ml(Interquartile range [IQR] 7.0), 38.4ng/ml (IQR 5.0) and 51.2ng/ml (IQR 12.6) respectively. Mean age and mean BMI was not significantly different across the tertiles P=.27, P=.20 respectively CPR was significantly different across T1 32/68(47%),T2 45/68(66.1%)and T3 29/68(42.6%) P=.014. CPR was significantly more in T2 versus T1 and T3, (OR:2.2, 95%CI:1.1-4.4 P =.02) and (OR:2.6,95%CI:1.3-5.3, P=.006), respectively. T3 versus T1, P =.60 LBR was also significantly different across the tertiles [T1 20/68(29.4%), T2 34/68(50%),T3 24/68(35.3%) P=.03] LBR in T2 was significantly more than in T1(OR: 2.4,95% CI:1.2-4.9, P=.02). There was no significant difference betweenT3 versus T1 and T2 versus T3, P=.46, P=.08, respectively. Early MR wasn’t significantly different across T1 12/32(37.5%), T2 11/45(24.4%) and T3 5/29(17.2%), P=.18 Serum progesterone levels were not significantly affected by age r = 0.0257, P=.71 or BMI r = 0.0097, P=.89 Multivariate logistic regression showed that age affected CPR after adjusting for BMI, midluteal serum progesterone levels, origin of oocytes, fresh ET/FET(AOR 0.92: 95% CI:0.86-0.98, P=.01). Limitations, reasons for caution This is a single centre study with limited sample size. Including the measurement of serum progesterone at the time of ET would have given a better perspective of the patient specific behavior of progesterone as LPS. Larger studies with different LPS protocols are needed to validate the results. Wider implications of the findings In HRT cycles with intramuscular progesterone, an optimum midluteal serum progesterone level is related to better pregnancy outcomes. Factors causing the wide variation of progesterone levels need to be evaluated so as to individualize the dose. Identifying the appropriate time frame for optimising the midluteal progesterone levels should be explored. Trial registration number not applicable

P-475 Effect of transdermal estradiol on endometrial preparation in frozen-thawed embryo transfer: A randomized control study.

Abstract Study question Is patient usage of transdermal estradiol more effective than oral estradiol on endometrial preparation in frozen-thawed embryo transfer cycles? Summary answer This study showed no significant differences in clinical pregnancy rates between the 2 groups. Transdermal gel have advantage of better patient comfort, lesser side effects. What is known already Estrogen and progesterone are crucial factors for endometrial preparation in frozen embryo transfer (FET) cycles. Studies assessing different forms of estradiol in FET have already been published. However, literature and studies in Vietnam are still limited to evaluating, research evaluating transdermal estrogen in ART. This study aimed to evaluate the use of transdermal estradiol gel and compare it with oral estradiol for the endometrium preparation in frozen-thawed embryo transfer (FET) cycles. Study design, size, duration This randomized clinical trial (RCT) included 380 frozen embryo transfer (FET) cycles from February 2020 to August 2021 at the IVF Department, Hung Vuong Hospital. Participants/materials, setting, methods They were randomized into one of the two groups. Group A (n = 190) received oral estradiol and group B (n = 190) received estradiol gel. In both groups, medication was started on day 2 of the menstrual, and endometrial thickness was monitored by ultrasound. Main outcomes were the following: serum estradiol level , endometrial thickness,.... Secondary outcomes were CPR, MR, and side effects in the 2 groups. Statistical testing was performed with Stata 14. Main results and the role of chance There was a significant difference in estradiol level on the day of progesterone administration and embryo transfer between the two groups (270.5 pg/ml versus 186.5 pg/ml, p, p. <0.001). According to the comparison, the two groups were not significantly different in the endometrial thickness on the day of progesterone administration (p = 0.85). as the data suggest, the total dose of estradiol used and the total number of days of dosing were similar in the two groups. The biochemical and clinical pregnancy rates were similar between the two groups and did not a significantly differ (56.2% vs. 52.2%, p = 0.474). The rate of miscarriage was higher than in the study group, but it was not statistically significant. Almost 20.3% of patients (n = 37) in group A had mild adverse effects when compared with only 10.1% (n = 18) in group B, and this was clinically significant (P = 0.007). Limitations, reasons for caution The limitation of our study was not asses ongoing pregnancy rate and live-birth rate. Wider implications of the findings Patients who estradiol valerate is contraindicated such as, high risk for deep vein thrombosis, hyperlipidemia, and clotting disturbance we can use oestrogel to be safe. It is suggested that estradiol transdermal patches be used instead of oral estradiol in FET cycles Trial registration number ISRCTN15301227

In vitro fertilization outcomes In infertile women with adenomyosis

Objectives: This study aimed to investigate the efficacy of in vitro fertilization (IVF) in infertile patients with adenomyosis and to identify relevant factors. Subjects and methods: Retrospective descriptive study of infertile cases with adenomyosis who received IVF therapy and embryo transfer from November 2013 to October 2022 at the Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy Hospital, excluding cases of oocyte donation or surrogacy. The β-hCG test was examined two weeks following embryo transfer. Then, women with hCG positive test were followed the pregnancy at 6 weeks, 8 weeks, and 12 weeks, and examined some factors that influence the clinical outcome of pregnancy. Results: Among 61 cycles of IVF treatment for infertile patients with adenomyosis, the average number of retrieved oocytes was 10.9±6.6 oocytes. The percentage of mature oocytes was 82.7%, the fertilization rate was 79.5%, the implantation rate was 16.7%, the clinical pregnancy rate was 19.7%, the miscarriage rate was 6.6%, and the ongoing pregnancy rate was 13.1%. In the group with GnRH agonist administration before embryo transfer, the pregnancy rate was greater than in the group without therapy (29.2% vs. 13.5%), and th e pregnancy rate in the group with > 10 oocytes was higher than in the group with ≤ 10 oocytes (28.0% vs 13.5%). However, these differences were not statistically significant (with p > 0.05). The cut-off point of endometrial thickness on the day of hCG injection, 10.25 mm, had a sensitivity of 58.3%, a specificity of 81.6%, an area under the curve (AUC) of 75.6%, and a p-value of 0.006; the cut-off point of endometrial thickness on the day of embryo transfer, 9.75 mm, had an accuracy of 9.75 mm. sensitivity 75%, specificity 71.4%, AUC 78.50%, p = 0.002 for clinical pregnancy prognosis. Conclusion: The IVF treatment of infertile patients with adenomyosis remains challenging, and additional research is required to explain the influence of this disorder on IVF outcomes. Key words: in vitro fertilization (IVF), adenomyosis, β-hCG.

Association between serum progesterone levels on the day of frozen-thawed embryo transfer and pregnancy outcomes after artificial endometrial preparation

BackgroundPrevious studies have examined that a range of optimal serum P level during the implantation period was associated with optimal live birth rates. However, those results obtained with vaginal or intramuscular route of progesterone administration for luteal phase support (LPS) alone. Is there a relationship between the serum progesterone (P) on the day of frozen-thawed embryo transfer (FET) with the likelihood of a live birth (LB) in artificial cycles (AC) when using a combination of oral dydrogesterone and vaginal progesterone for LPS?MethodsThis was a retrospective study of 3659 FET cycles with artificial endometrial preparation in a Chinese tertiary-care academic medical centre from January 2015 to February 2017. Endometrial preparation was performed using estradiol (E2) valerate (Fematon-red tablets) 8 mg/d beginning on day 3 of the cycle, followed by administration of P both orally (8 mg/d Fematon-yellow tablets) and vaginally (400 mg/d; Utrogestan). The primary endpoint was live birth rate (LBR). The association between the serum P level on the embryo transfer day and pregnancy outcomes was evaluated by univariable and multivariable logistic regression analysis.ResultsThe LBRs according to the serum P quartiles were as follows: Q1: 35.7%; Q2: 37.4%; Q3: 39.1% and Q4: 38.9%. Logistic regression analysis showed that the odds of a LB were not significantly different between the low (P < 7.9 ng/mL) and high (P ≥ 7.9 ng/mL) progesterone groups before or after adjustment (crude OR = 0.89, 95% CI: 0.76–1.04; adjusted OR = 0.89, 95% CI: 0.75–1.04).ConclusionThe present study suggests that the serum P levels on the day of embryo transfer (ET) do not correlate with the likelihood of a LB in artificial cycles when using a combination of oral dydrogesterone and vaginal progesterone for luteal phase support.

Are systemic progesterone levels in true natural cycle euploid frozen embryo transfers with luteal phase support predictive for ongoing pregnancy rates?

Are serum progesterone (P4) levels on the embryo transfer (ET) day predictive of ongoing pregnancy (OP) following a single euploid blastocyst transfer in a natural cycle (NC) when luteal phase support is routinely given? In single euploid frozen ETs in NC, P4 levels on ET day are not predictive for OP, when luteal phase support (LPS) is routinely added after the ET. In an NC frozen embryo transfer (FET), P4 produced by the corpus luteum initiates secretory transformation of the endometrium and maintains pregnancy after implantation. There are ongoing controversies on the existence of a P4 cutoff level on the ET day, being predictive for the chance of OP as well as of the possible role of additional LPS after ET. Previous studies in NC FET cycles, evaluating and identifying P4 cutoff levels did not exclude embryo aneuploidy as a possible reason for failure. This retrospective study analyzed single, euploid FET in NC, conducted in a tertiary referral IVF centre between September 2019 and June 2022, for which measurement of P4 on the day of ET and the treatment outcomes were available. Patients were only included once into the analysis. Outcome was defined as OP (ongoing clinical pregnancy with heartbeat, >12 weeks) or no-OP (not pregnant, biochemical pregnancy, early miscarriage). Patients with an ovulatory cycle and a single euploid blastocyst in an NC FET cycle were included. Cycles were monitored by ultrasound and repeated measurement of serum LH, estradiol, and P4. LH surge was identified when a rise of 180% above the previous level occurred and P4 levels of ≥1.0 ng/ml were regarded as confirmation of ovulation. The ET was scheduled on the fifth day after P4 rise and vaginal micronized P4 was started on the day of ET after P4 measurement. Of 266 patients included, 159 (59.8%) patients had an OP. There was no significant difference between the OP- and no-OP-groups for age, BMI, and day of embryo biopsy/cryopreservation (Day 5 versus Day 6). Furthermore, P4 levels were not different between the groups of patients with OP (P4: 14.8 ng/ml (IQR: 12.0-18.5 ng/ml)) versus no-OP (P4: 16.0 ng/ml (IQR: 11.6-18.9 ng/ml)) (P = 0.483), and no differences between both groups, when P4 levels were stratified into categories of P4 levels of >5 to ≤10, >10 to ≤15, >15 to ≤20, and >20 ng/ml (P = 0.341). However, both groups were significantly different for the embryo quality (EQ), defined by inner cell mass/trophectoderm, as well as when stratified into three EQ groups (good, fair, and poor) (P = 0.001 and 0.002, respectively). Stratified EQ groups remained the only significant parameter influencing OP in the uni- and multivariate analyses (P = 0.002 and P = 0.004, respectively), including age, BMI, and P4 levels (each in categories) and embryo cryopreservation day. Receiver operator characteristic curve for the prediction of an OP revealed an AUC of 0.648 when age, BMI and EQ groups were included into the model. The inclusion of P4 measurement on ET day into the model did not add any benefit for OP prediction (AUC = 0.665). The retrospective design is a limitation. Monitoring serum P4 levels can be abandoned in NC FET cycles with routine LPS as they do not seem to be predictive of live birth. No external funding was used for this study. The authors state that they do not have any conflicts of interest. N/A.

Single Dose of Granulocyte-Colony Stimulating Factor Use to Improve Intracytoplasmic Sperm Injection Outcomes

Receptive endometrium is essential for successful embryo implantation and is created by the synergistic activities of estrogen and progesterone, as well as support from other endocrine, paracrine, and autocrine pathways. Granulocyte Colony Stimulating Factor (G-CSF) may play a crucial role in modulating the immune response to enhance implantation. To estimate the role of granulocyte-colony stimulating factor on the clinical intracytoplasmic sperm injection (ICSI) outcome parameters. A prospective comparative study with random sample selection was carried out from 2021 to 2023 at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. It included 80 women with infertility, with or without previous failure trials of intracytoplasmic sperm injection, who completed IVF and ICSI protocols and reached the day of embryo transfer. On the day of embryo transfer, patients were divided into two groups: 49 patients received no additional treatment (group 1), and 31 patients received granulocyte-colony stimulating factor subcutaneous injection within one hour of embryo transfer (group 2). The implantation rate was 13.3% in group 2 compared to 7.6% in group 1. The chemical and clinical pregnancy rates were 29.0% and 25.8% in group 2, while group 1 had 18.4% for both chemical and clinical pregnancy rates. The pregnancy and implantation rates were slightly higher in group 2, but the difference was not significant. More research is needed to support the use of G-CSF as a standard treatment in ICSI patients.

İn Vitro Fertilizasyon/İntrasitoplazmik Sperm Enjeksiyonu Uygulanan Çiftlerde Paternal Yaşın Düşük Oranlarına Etkisi

Aim: The aim of this study was to determine the effect of paternal age on miscarriage rates in couples undergoing in vitro fertilization/intracytoplasmic sperm injection treatment cycles. Material and Methods: Patients were classified into two groups. The study group consisted of the patients whose pregnancy resulted in a miscarriage (n=73) and a control group in which the patients had a live singleton birth (n=256). Demographic characteristics, treatment indications, duration of infertility, menstruation day 3 follicle stimulating hormone, estradiol, luteinizing hormone, total antral follicle count, anti-Mullerian hormone levels and controlled ovarian stimulation parameters, day of trigger, estradiol and progesterone levels on the day of trigger, the total number of oocytes retrieved, the number of mature oocytes, the number and quality of the embryo, endometrial thickness on the day of trigger, oocyte pick up and embryo transfer, the distance of embryo-fundus and the day of embryo transfer were recorded. Results: Totally 329 women were included in the present study. The number of patients with the diagnosis of unexplained infertility was statistically significantly higher in the study group than in the control group (p=0.020). Maternal age was found statistically significantly higher in the study group than in the control group (p=0.025). When maternal age increased by 1 unit, the risk of miscarriage increased by 8.7% and those with unexplained infertility had a 75.6% higher risk of miscarriage than those without unexplained infertility. Conclusion: Paternal age was not associated with miscarriage whereas maternal age and unexplained infertility had a positive correlation with miscarriage rate.

Role of intramuscular progesterone supplementation on the day of embryo transfer in artificial frozen cycles

BackgroundSufficient endometrial preparation with or without progesterone supplementation is crucial in artificial cycles with frozen embryo transfer (FET). We aimed to study the effect of intramuscular progesterone supplementation on the day of embryo transfer (ET) in artificial frozen cycles.MethodsA clinical cohort study involved women undergoing FET with artificially prepared endometrium. Serum progesterone levels were assessed on the day of ET. Accordingly, we recruited 177 women with progesterone levels more than 9.2 ng/ml in group 1, and 177 other women with progesterone levels less than 9.2 ng/ml in group 2. Women in group 1 received only 400 mg vaginal progesterone twice-daily after ET, while women in group 2 received additional intramuscular progesterone supplementation. The chemical, clinical, and ongoing pregnancy rates, as well as the pregnancy loss rate, were assessed in both groups.ResultsExpectantly, both groups showed a significant difference regarding the serum progesterone level on the day of ET (13.43 ± 4.65 vs 4.62 ± 2.77, P = 574). However, with additional intramuscular progesterone supplementation in group 2, both groups showed no significant difference regarding the chemical pregnancy rate (68.93% in group 1 vs 63.84% in group 2, P = 0.311), the clinical pregnancy rate (61.02% in group 1 vs 58.76% in group 2, P = 0.664), ongoing pregnancy rate (56.50% in group 1 vs 53.11% in group 2, P = 0.522), and pregnancy loss rate (7.41% in group 1 vs 9.62% in group 2, P = 0.564).ConclusionsIntramuscular progesterone supplementation in women with decreased serum progesterone levels could improve pregnancy outcomes in artificial frozen cycles.Trial registrationIt was first registered at ClinicalTrials.gov on 8/4/2021 with registration number NCT04837768.

Histological Characteristics of the Endometrium and its Affecting Factors in Dairy Cattle (Bos taurus) on Embryo Transfer Day

Abstract In cows embryo transfer occurs on the 7th day of the oestrous cycle. Embryo survival and successful pregnancy depend on many factors, including early interaction between embryo and endometrium. Therefore, the aim of our research was to describe histo-morphological characteristics of endometrium in donor and recipient cows on obtaining and transfer day, respectively (7th day of the oestrus cycle). In addition, progesterone and oestradiol levels, the ipsilateral and contralateral corpus luteum, animal age, lactation number and days postpartum were determined. The results of the study revealed a significant difference in endometrial gland cell size, vacuolisation of glandular epithelium cytoplasm, and stromal oedema in the donor and recipient cows. However, these parameters were related only to the level of progesterone in the blood, and no other investigated factors in this study.

A pilot study using unique targeted testing of the urogenital microbiome has potential as a predictive test during IVF for implantation outcome

PurposeThis pilot study aimed to develop a methodology characterising the urogenital microbiome as a predictive test in the IVF workup.MethodsUsing unique custom qPCRs, we tested for the presence of specific microbial species from vaginal samples and First Catch Urines from the male. The test panel included a range of potential urogenital pathogens, STIs, ‘favourable bacteria’ (Lactobacillus spp.) and ‘unfavourable bacteria’ (anaerobes) reported to influence implantation rates. We tested couples attending Fertility Associates, Christchurch, New Zealand for their first round of IVF.ResultsWe found that some microbial species affected implantation. The qPCR result was interpreted qualitatively using the Z proportionality test. Samples from women at the time of Embryo Transfer who did not achieve implantation had significantly higher percent of samples that were positive for Prevotella bivia and Staphylococcus aureus compared to women who did achieve implantation.DiscussionThe results provide evidence that most other microbial species chosen for testing had little functional effect on implantation rates. The addition of further microbial targets (yet to be determined) could be combined in this predictive test for vaginal preparedness on the day of embryo transfer. This methodology has a substantial advantage of being affordable and easily performed in any routine molecular laboratory. This methodology is most suitable as a foundation on which to develop a timely test of microbiome profiling. Using the indicators detected to have a significant influence, these results can be extrapolated.ConclusionUsing a rapid antigen test, a woman can self-sample prior to embryo transfer and obtain an indication of microbial species present which could influence implantation outcome.

Successful induction of pseudopregnancy using sonic vibration in mice

Embryo transfer (ET) is an essential reproductive technology for the production of new animal strains and maintenance of genetic resources. We developed a method, named Easy-ET, to induce pseudopregnancy in female rats by artificial stimulation using sonic vibration instead of mating with vasectomized males. This study examined the application of this method for the induction of pseudopregnancy in mice. Offspring were obtained from two-cell embryos transferred into females with pseudopregnancy induced using sonic vibration in proestrus on the day before embryo transfer. Furthermore, high developmental rates of offspring were observed when pronuclear and two-cell embryos were transferred to females in estrus that were stimulated on the day of embryo transfer. Genome-edited mice were also obtained using frozen-warmed pronuclear embryos with clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated system (Cas) nucleases introduced using the technique for animal knockout system by electroporation (TAKE) method, which were transferred to females with pseudopregnancy induced on the day of embryo transfer. This study demonstrated that induction of pseudopregnancy by sonic vibration was also possible in mice.

Effect of administration of atosiban at day of embryo transfer on in-vitro fertilization out come

Background: The most crucial step of assisted reproduction is embryo implantation, which is impacted by a variety of factors, including the patient's age, embryo quality, and endometrial receptivity. The embryo transfer (ET) procedure, which induces uterine contractions, may hinder embryo implantation. Objectives: On the day of embryo transfer, examine the effects of Atosiban administration on implantation, clinical pregnancy, miscarriage, and live birth rates. Material and Methods: The study included 60 individuals with a diagnosis of primary or secondary infertility due to male or female factor, tubal factors, moderate endometriosis, or other unknown reasons. Two groups of patients were selected at random. The first group received 6.75 milligrammes of atosiban intravenously 30 minutes before ET, whereas the second group received no medication. Pregnancy outcomes in both groups were compared. Results: Significantly higher clinical pregnancy and implantation rates were observed (60% versus 30%) and (37.8% versus 17.8%) respectively, with non-significantly higher live birth rates (94.4% versus 66.7%), while the miscarriage rate was lower in the Atosiban group but not significantly (5.6% versus 33.3%). Conclusion: These findings imply that atosiban therapy prior to embryo transfer improves embryo implantation.

Evaluation of the ideal vaginal Progesterone effectiveness doses for luteal support in embryo thawing cycles after endometrial preparation without using the GnRh analogue.

Frozen-thawed embryo transfer (FET) cycles require the use of luteal phase support (LPS) for supporting implantation, endometrial and embryo maturity. Individualized LPS should be chosen according to the used endometrial preparation protocol. The aim of the study was to analyze the effectiveness of two different vaginal Progesterone doses for women who underwent FET cycle and the same endometrial preparation without using the GnRh analogue. 607 women who underwent FET cycle were included in the study. 305 patients received luteal support with 600 mg/day vaginal Progesterone and 302 patients were treated with 800 mg/day of vaginal Progesterone. In the 800 mg/day group, the mean serum Progesterone concentration on the day of embryo transfer was higher than in the 600 mg group (14.00±6.18 ng/mL and 12.22±5.39, respectively, p < 0.001). Moreover, human chorionic gonadotrophin (hCG) positive and ongoing pregnancy rates were higher in the group of patients who received LPS with 800 mg/day of Progesterone than in the group of patients treated with 600 mg/day of Progesterone. CONCLUSIONS: In patients undergoing FET cycles following endometrial preparation made without previously using the GnRh analogue, 800 mg doses of vaginal Progesterone as LPS improve reproductive outcomes.

Circulating maternal prorenin and oocyte and preimplantation embryo development: the Rotterdam Periconception Cohort.

Could circulating maternal prorenin serve as a proxy for oocyte and preimplantation embryo development, assessed by time-lapse parameters and clinical treatment outcomes? High circulating maternal prorenin concentrations after ovarian stimulation associate with a larger oocyte area, faster cleavage divisions from the five-cell stage onwards and increased chance of successful implantation. After ovarian stimulation, circulating prorenin (renin's precursor), is largely ovary-derived. Prorenin may contribute to ovarian angiotensin synthesis, which is relevant in reproduction given its role in follicular development and oocyte maturation. Prospective observational cohort study including couples requiring fertility treatment from May 2017 as a subcohort of the ongoing Rotterdam Periconception Cohort conducted in a tertiary referral hospital. Between May 2017 and July 2020, 309 couples with an indication for IVF treatment or ICSI were included. Resulting embryos (n = 1024) were submitted to time-lapse embryo culture. Time of fertilization (t0), pronuclear appearance (tPNa), and fading (tPNf) as well as the exact timing of reaching the two- to eight-cell stage (t2-t8), the start of blastulation (tSB), reaching the full (tB), and expanded blastocyst (tEB) were retrospectively recorded. Oocyte area was measured at t0, tPNa, and tPNf. Prorenin was determined at the day of embryo transfer. After adjustment for patient- and treatment-related factors, linear mixed modeling showed that higher prorenin concentrations associate with a larger oocyte area at tPNa (β 64.45 µm2, 95% CI 3.26; 125.64, P = 0.04), and faster progression from five-cell stage onwards (e.g. β8-cell -1.37 h, 95% CI -2.48; -0.26, P = 0.02). Prorenin associated positively with pre-transfer outcomes (e.g. βfertilized oocytes 2.09, 95% CI 1.43; 2.75, P < 0.001) and implantation (odds ratio+β-hCG-test: 1.79, 95% CI 1.06; 3.08, P = 0.03), but not with live birth. This prospective observational study provides associations and therefore residual confounding cannot be excluded and causality has to be shown in intervention studies. Theca cell-derived factors, such as prorenin, may help to clarify the underlying endocrine mechanism of oocyte maturation and embryo development, with a special focus on the (patho)physiological reproductive role of prorenin and the identification of factors influencing its secretion and activity, which is of great added value for improving embryo selection and predicting implantation and pregnancy outcomes. This will bring us to investigate which determinants of oocyte quality and embryo development should take center stage in developing preconception care strategies. This research was funded by the Department of Obstetrics and Gynecology of the Erasmus MC, University Medical Center, Rotterdam, the Netherlands, and the Erasmus MC Medical Research Advisor Committee's 'Health Care Efficiency Research' program (OZBS72.16080). The authors have no competing interests to disclose. N/A.

Effect of inducing accessory corpus luteum formation with gonadotropin-releasing hormone or human chorionic gonadotropin on the day of embryo transfer on fertility of recipient dairy heifers and lactating cows

The objective was to determine the effect of inducing an accessory corpus luteum (CL) using GnRH or human chorionic gonadotropin (hCG) on the day of in vitro-produced (IVP) embryo transfer (ET) on pregnancy per ET (P/ET) and calving/ET in dairy heifers and lactating cows. Dairy heifers (11–15 mo of age; n = 1,547) and lactating cows (n = 1,480) detected in estrus by tail chalk (d 0) were used as recipients. Before ET, the presence of a CL was evaluated by transrectal palpation from d 6 to 9 of the estrous cycle. Animals with a CL were randomly assigned to receive 1 of 3 treatments immediately before ET: control (no treatment; n = 1,009), GnRH (86 µg of GnRH; n = 1,085) and hCG (2,500 IU; n = 1,069). Embryos were implanted in the uterine horn ipsilateral to the ovary with a CL (fresh IVP embryos, n = 2,544; vitrified IVP embryos n = 545; slow-freezing IVP embryos, n = 74). Pregnancy diagnosis was performed on d 37 ± 3 of gestation by transrectal palpation. Pregnancy loss data and calving records were collected from the dairy farm management software. Treatment did not affect P/ET, calving/ET, or pregnancy loss either overall or within parity. When treatments inducing CL formation were combined (GnRH + hCG), heifers tended to have greater P/ET than controls (67.7 vs. 63.5%, respectively). Yet, calving/ET were similar. Response variables were also analyzed within embryo type and parity. For heifers receiving stage 6 (blastocyst) fresh IVP embryos, hCG had greater P/ET than controls (74.5 vs. 51.1%, respectively). In addition, GnRH tended to have greater P/ET than controls (67.8 vs. 51.2%, respectively). However, calving/ET in heifers receiving blastocyst fresh IVP embryos was similar among treatments. When only stage 7 (expanded blastocyst) fresh IVP embryos were considered, primiparous GnRH cows had greater P/ET (59.3 vs. 47.1%) and calving/ET (48.6 vs. 38.1%) than hCG. Moreover, hCG showed decreased calving/ET compared with controls in primiparous cows transferred with expanded blastocyst fresh IVP embryos. In summary, the effects of hCG or GnRH at ET on P/ET and calving/ET were inconsistent according to different embryo characteristics (e.g., embryo stage) and parity of recipients. Furthermore, treatment did not improve the overall fertility outcomes for recipient animals receiving IVP embryos.