Progesterone Exposure Research Articles

Comparison of live birth rates following the transfer of day-6 blastocysts on the 6th versus 7th day of progesterone exposure in hormone replacement treatment–frozen embryo transfer cycles

Objective: The duration of progesterone exposure is critical for establishing and maintaining a pregnancy in hormone replacement treatment (HRT)–frozen embryo transfer (FET) cycles. This study compared the live birth rates (LBRs) of patients undergoing day-6 warmed blastocyst transfer on either the 6th or 7th day of progesterone administration in HRT-FET cycles.Methods: A retrospective cohort study was conducted using data from the central registry of Indira IVF Hospital Private Limited. In total, 2,633 patients who underwent day-6 blastocyst transfer cycles with HRT-FET were identified. The cycles were categorized based on the timing of the day-6 blastocysts transfer: P+6 vs. P+7 (initiation date: P+1). Propensity scores were calculated. The primary outcome measure was the LBR.Results: Following propensity score matching, a total of 1,401 patients were included in the final analysis (P+6: n=1,212; P+7: n=189). The number of previous attempts was significantly higher in the P+6 group, whereas the rate of preimplantation genetic testing for aneuploidy was significantly higher in the P+7 group. The LBRs were 59.2% and 54.5% in the P+6 and P+7 groups, respectively (p=0.21). Multivariate regression analysis revealed that the number of previous in vitro fertilization attempts (odds ratio [OR], 3.246; 95% confidence interval [CI], 2.429 to 4.337; p<0.001), the number of blastocysts transferred (OR, 2.011; 95% CI, 1.461 to 2.768; p<0.01), and endometrial thickness on the day of progesterone initiation (OR, 1.143; 95% CI, 1.022 to 1.28; p=0.02) were independent predictors of live birth.Conclusions: The LBRs were comparable in patients who underwent day-6 warmed blastocyst transfer on day 6 or 7 post-progesterone initiation in HRT-FET cycles.

Microbial Coadaptation Drives the Dynamic Stability of Microecology in Mainstream and Sidestream Anammox Systems Under Exposure of Progesterone

Microbial cooperation determines the efficacy of wastewater biological treatment, and the adaptability of microorganisms to environmental stresses varies. Recently, extensive use of hormones results in their inevitable discharge into aquatic environment. Therefore, mainstream and sidestream anammox reactors were constructed in this study to evaluate their removal performance of progesterone and nitrogen simultaneously, the adaptability of anammox consortia to progesterone stress and the corresponding regulation mechanism. Both anammox processes had the resilience to progesterone stress, with the average nitrogen removal efficiency exceeding 90%. At the same time, progesterone removal efficiency also exceeded 70%. In contrast, microbial community in the mainstream reactors was more susceptible to progesterone interference. The adaptation of anammox consortia mainly depended on microbial cooperation and molecular regulation. Initially, bacteria secreted more extracellular polymeric substances to detain progesterone. Biodegradation also contributed to mitigating the side effect of progesterone, which was demonstrated by the proliferation of potential degrading bacteria such as Bacillus salacetis, Bacillus wiedmannii and Rhodococcus erythropolis. In addition, the enhancement of microbial interaction intensity drove their cooperation to enhance adaptability and maintain stable performance. Combined with metagenomic and metatranscriptomic analyses, such microbial adaptability was enhanced through molecular regulations, including the energy redistribution for amino acid synthesis and alteration of key metabolic pathways. Related functional gene expressions and microbial interactions were, in turn, regulated by quorum sensing. This work verifies the feasibility of anammox process in hormone-containing wastewater treatment and provides a holistic understanding of molecular mechanism of microbial interaction and coadaptation to stress.

Detrimental effect of prenatal progesterone exposure on anxiety and depressive-like responses in adult male and female rat offspring: role of plasma, hippocampal corticosterone and hippocampal progesterone receptors

In clinical practice, the use of exogenous progesterone (Pro) is often required in assisted reproduction programs due to reduced levels of the hormone and the risk of miscarriage. Exposure to the hormone reduces anxiety in rodents, but the long-term effects of prenatal exposure in adult offspring are unknown. Therefore, the present study was designed to investigate the effect of prenatal Pro treatment on anxiety- and depression-like behavior and the effect on plasma, hippocampal corticosterone (CORT) and hippocampal progesterone receptor (PR) in young adult male and female rat offspring. The behavioral responses of offspring of both sexes were tested in the open field, and the elevated plus maze tests, and for depressive-like behavior in the sucrose preference test, the forced swimming test and the splash test. CORT levels and PR expression were measured by ELISA. The results indicate that prenatal Pro exposure at low and high doses (10 and 50 mg/kg, s.c. during G0-G10) induces anxiogenic and depressive-like effects compared to vehicle-treated controls, which are associated with high plasma and hippocampal CORT levels and upregulated hippocampal PR in male and female adult offspring. Our results demonstrate that prenatal Pro exposure has detrimental effects on the emotional status of male and female adult offspring, which may be associated with long-term changes in hormonal homeostasis.

O-208 The Programmed Ovulatory Frozen-Thawed Embryo Transfer Cycle (PO-FET): a suggested FET protocol integrating aspects of embryo transfer scheduling, efficacy optimization and maternal/fetal safety

Abstract Study question What is the effect of 30mg oral dydrogesterone (DYD), given to induce endometrial receptivity in natural-proliferative-phase-FET-cycles (NPP-FET), on ovulation, progesterone and DYD/DHD-levels, respectively, and outcomes? Summary answer Oral DYD 30mg allows scheduling endometrial receptivity in natural cycles while not interfering with ovulation in the vast majority of patients. What is known already Globally, approximately 60% of all ETs are performed after cryopreservation. Hormone Replacement Treatment (HRT)-FET cycle-regimens have come under scrutiny, because of i) risk of insufficient progesterone exposure with conventional dosing-schemes and ii) maternal/fetal risks stemming from the iatrogenic lack of a corpus luteum. Oral DYD 10mg (tid) supposedly does not interfere with ovulation/corpus luteum formation, and does not cross-react with progesterone in ELISA. It could thus be used for inducing endometrial receptivity (scheduling, in a natural cycle, the time point of ET) and luteal phase support, while allowing ovulation dissociated from the window-of-implantation and monitoring of endogenous progesterone serum levels. Study design, size, duration Within a multi-centric, prospective, clinical cohort study (NCT03507673), 559 normally cycling women from the routine care population who underwent FET after IVF/ICSI in a natural cycle (2/2021 – 8/2023) had serum/plasma samples prospectively collected/stored on day of FET for measurement of E2, progesterone and DYD/DHD in Roche Elecsys®-Immunoassay and liquid chromatography/tandem mass spectroscopy (LC-MS/MS), respectively. FET was performed on day 2/3 (cleavage-stage) or day 4/5 (morula/blastocyst) in 17.4% and 82.6% of FETs, respectively. Participants/materials, setting, methods From cycle day 10, women underwent endocrine (E2, LH and P) and sonographic monitoring until E2 >180pg/ml, leading follicle >16mm and endometrial thickness was >6mm. Physicians could then initiate oral DYD10mg (tid) or–in absence of LH rise–postpone initiation, under 2-daily monitoring, to meet the organizational preferences of the patient and/or center (for example, avoiding weekends). FET was performed for day 2 (cleavage), 3, 4 or 5 (blastocyst) on day 3-6 of DYD-intake, respectively. Main results and the role of chance Previously reported PK of 10mg DYD (tid) are confirmed/extended. While log-levels of DYD and DHD were highly correlated (0.95), P-levels were not (0.02, 0.02). In 94.4% of patients, ovulation could be confirmed by serum-P levels on the day of FET or in late luteal phase. Medians (IQR) of DYD and P on day 3 and days 5, respectively, were 0.93ng/ml (0.93ng/ml) and 2.96ng/ml (3.19ng/ml), and 1.13ng/ml (1.09ng/ml) and 8.50ng/ml (4.40ng/ml). Serum-P levels increased with the FET day (medians 2.0, 3.0, 6.6, and 8.5 ng/ml on days 2, 3, 4, and 5) and only very weakly correlated with LH, E2 levels, and follicular diameter at last monitoring. 42.5% and 34.8% of patients had positive hCG and ongoing clinical pregnancy, respectively. Ongoing pregnancy was independent from plasma-DYD levels (< 25th percentile [<0.66 ng/ml] 37.9% vs. ≥25thperc. [≥0.66 ng/ml] 34.3%, RD 1.5%, 95%CI: -16.5% to 17.4%, p = 0.334), yet dependent from serum-P levels (< 25thperc [<4.85 ng/ml] 25.9% vs. ≥25thperc [≥4.85 ng/ml] 35.3%, RD 9.4%, 95% CI 0.4% to 17.6%, p = 0.0161).However, neither of the two serum hormone values is predictive of a dichotomous outcome in isolation, nor do they demonstrate predictive interaction when considered together in logistic regression modelling. Limitations, reasons for caution DYD, like hCG or any other progestin given for LPS, may negatively interfere with LH surge, ovulation and luteal phase characteristics. Controlled studies on different progestin types and doses are necessary to better understand how to optimize luteal phase and thereby optimally support embryo nidation, pregnancy maintenance and maternal/fetal health. Wider implications of the findings The PO-FET protocol induces the implantation window, thus offers (some) flexibility in FET timing, shows little interference with ovulation/corpus luteum formation, provides double gestagenic support with no need for monitoring P or DYD levels, however allows corpus luteum activity monitoring in luteal phase and early pregnancy, is injection-free, and low-cost. Trial registration number NCT03507673

P-410 A modified artificial cycle-frozen embryo transfer(mAC-FET) protocol with evidence of corpus luteum(CL) formation as determined by serum relaxin-2 in early pregnancy :a prospective proof-of-concept study

Abstract Study question Can a mAC-FET protocol for endometrial preparation in ovulatory patients lead to formation of a CL as evidenced by serum relaxin-2 in the pregnant patients? Summary answer CL forms in the mAC-FET protocol as serum relaxin-2 in early pregnancy was comparable to that in spontaneous conception.Late luteal rise of relaxin-2 wasn’t demonstrated. What is known already Clinical evidence points to increased incidence of preeclampsia in pregnancies by AC-FET cycles compared to ovulatory FET cycles-natural cycle(NC) and mNC. The absence of CL and it’s secretory products like relaxin-2 is a contributing factor. FET protocols for endometrial preparation in ovulatory patients need to be optimised. NC/mNC-FET protocols need frequent monitoring. Also there is a temporal variability between LH/hCG and progesterone levels. We studied a mAC-FET protocol in ovulatory patients with a delayed start(day3-5 of the cycle),low dose transdermal 17β estradiol gel to see the formation of CL as elucidated by the presence of serum relaxin-2 in early pregnancy. Study design, size, duration A proof-of- concept prospective study was conducted at a tertiary centre (July2023-Dec2023). Serum relaxin-2(R&D/Quantikine/ELISA) was determined in 4 groups of patients- Group-A. mAC-FET(n = 34) 1.On the day of start of progesterone 2.Post-ET day5-9 (along with serum progesterone) 3.In pregnancy(6-9 weeks of gestation) Group-B. AC-FET(n = 8) 1.On the day of start of progesterone 2.In pregnancy(6-9 weeks) Group-C. Spontaneous conception (n = 9) 1.In pregnancy(6-9 weeks) Group-D. Infertility patients ( n = 24) 1.Preovulatory. All patients were ≤40 years with a BMI ≤30kg/m2. Participants/materials, setting, methods In the AC-FET cohort-oral estradiol 6mg/day was started from day1/2 of the cycle. When endometrial thickness was>7mm,progesterone<1ng/ ml then intramuscular progesterone was started- 50mg/day-3 days,100mg on day4-the day of ET(Day-3 embryos) and continued every alternate day.Progesterone gel was added post-ET. In the mAC-FET cohort-17β transdermal estradiol gel(0.06%) 3mg/day was started from day3-5,rest of the protocol was similar. Statistical analysis was performed using appropriate non-parametric tests.Pearson coefficient(r) was used to assess linear-relationship.Two-tailed P<.05 indicated statistical significance. Main results and the role of chance Mean age and BMI of the patients with mAC-FET was 33.3±4.7 years and 24.4±3.7 kg/ m2 which was comparable across all groups (P=.14,P=.52 respectively). Clinical pregnancy rate in the mAC-FET group was 52.94%(18/34). 3 patients had an early abortion. Baseline relaxin-2 levels, median[Interquartile range(IQR)] were comparable- AC-FET cohort-9.1pg/mL(8.6-16.8), mAC-FET cohort-11.5pg/mL(9.6-13.7) and the infertility cohort-10.9pg/mL(8.4-14.5) (P=0.74) The primary outcome was the comparable levels of serum relaxin-2, in early pregnancy in the mAC-FET group and spontaneous conception group - median (IQR) 449.8pg/mL(73.1-492.3) and 487.9pg/mL(419.1-497.8) respectively (P=.12) The relaxin-2 levels, in early pregnancy in the mAC-FET group were significantly higher than the AC-FET group median(IQR) 449.8pg/mL(73.1-492.3) and 7.9pg/mL(7.8-9.5) respectively (P=.0005). As secondary outcomes, we studied the late luteal rise of relaxin-2 in the mAC-FET group and the relationship of late luteal progesterone with relaxin-2 in early pregnancies in the mAC-FET cohort. A late luteal rise of relaxin-2 was not demonstrated in the mAC-FET cohort. The serum relaxin-2, on the day of start of progesterone and on day5-9 post ET- median(IQR) 11.5pg/mL(9.6-13.7) and 11.4pg/mL(9.0-18.5), respectively was comparable (P=.18). The relationship between serum relaxin-2 in early pregnancy and the progesterone levels on day5-9 post ET in the mAC-FET cohort was not significant r = -.059 (P=.07) Limitations, reasons for caution This was a small proof- of-concept single centre study. Due to the limited numbers and short duration of the study, we could not compare efficacy, cancellation rates and pregnancy outcomes of the mAC-FET protocol with other FET protocols. Wider implications of the findings The mAC-FET protocol studied for endometrial preparation was similar to the AC -FET protocol regarding monitoring,flexibility and consistent progesterone exposure with the added advantage of the presence of the CL.This may decrease the risk of maternal/neonatal complications. Larger studies are needed to confirm it’s efficacy and impact on pregnancy outcomes. Trial registration number Not Applicable

Hormonal regulation of miRNA during mammary gland development.

The mammary gland is a unique organ as most of its development occurs after birth through stages of proliferation, differentiation and apoptosis that are tightly regulated by circulating hormones and growth factors. Throughout development, hormonal cues induce the regulation of different pathways, ultimately leading to differential transcription and expression of genes involved in this process, but also in the activation or inhibition of post-transcriptional mechanisms of regulation. However, the role of microRNAs (miRNAs) in the different phases of mammary gland remodeling is still poorly understood. The objectives of this study were to analyze the expression of miRNA in key stages of mammary gland development in mice and to determine whether it could be associated with hormonal variation between stages. To do so, miRNAs were isolated from mouse mammary glands at stages of adulthood, pregnancy, lactation and involution, and sequenced. Results showed that 490, 473, 419, and 460 miRNAs are detected in adult, pregnant, lactating and involuting mice, respectively, most of them being common to all four groups, and 58 unique to one stage. Most genes could be divided into six clusters of expression, including two encompassing the highest number of miRNA (clusters 1 and 3) and showing opposite profiles of expression, reaching a peak at adulthood and valley at lactation, or showing the lowest expression at adulthood and peaking at lactation. GO and KEGG analyses suggest that the miRNAs differentially expressed between stages influence the expression of targets associated with mammary gland homeostasis and hormone regulation. To further understand the links between miRNA expression and hormones involved in mammary gland development, miRNAs were then sequenced in breast cells exposed to estradiol, progesterone, prolactin and oxytocin. Four, 38, 24 and 66 miRNAs were associated with progesterone, estradiol, prolactin, and oxytocin exposure, respectively. Finally, when looking at miRNAs modulated by the hormones, differentially expressed during mammary gland development, and having a pattern of expression that could be correlated with the relative levels of hormones known to be found in vivo, 16 miRNAs were identified as likely regulated by circulating hormones. Overall, our study brings a better understanding of the regulation of miRNAs throughout mammary gland development and suggests that there is a relationship between their expression and the main hormones involved in mammary gland development. Future studies will examine this role more in detail.

242 Comparison of GnRH administration day in the 7&7 Synch versus 8&6 Synch Protocol for fixed-timed artificial insemination in beef heifers

Abstract The research aimed to investigate the impact of reducing the time interval for progesterone exposure on the ovulatory follicle just before performing fixed-timed artificial insemination (FTAI). This was achieved by adjusting the timing of administering a gonadotropin-releasing hormone agonist (GnRH; specifically, 100 µg gonadorelin, known as Cystorelin). The study assessed the effectiveness of two estrous synchronization protocols, the “7 & 7 Synch” and a modified protocol, by examining estrus expression (EE) and the resulting pregnancy rates (PR) following FTAI. The research utilized frozen conventional Angus semen from Select Sires, Inc. (Plain City, OH), and involved 66 Angus-cross heifers from the beef cattle research unit near Fayetteville at the University of Arkansas. To ensure consistency, the heifers were divided into two treatment groups based on their cyclicity status (CYC) and age. CYC status was determined by assessing a reproductive tract score (RTS) 1 wk before the synchronization process. RTS scores ranged from 1 to 5, with RTS 1-3 indicating pre- or peripubertal status and scores of 4 and 5 indicating cycling. Heifers with an RTS of 1 and 2 were excluded from the study before synchronization commenced. Both treatment groups involved the insertion of a controlled internal drug-releasing device (CIDR; specifically, Eazi-breed, containing 1.38 g of progesterone) into the vagina on d -14. Simultaneously, a prostaglandin F2α analogue (PGF; 500 µg cloprostenol sodium, known as Synchsure) was administered. In the first treatment group (7&7), GnRH was administered on d -7, while in the second group (8&6), GnRH was given on d -6. In both cases, PGF was administered on d 0 when the CIDR was removed, precisely 72 ± 1 h before FTAI, along with another dose of GnRH. Estrus expression (EE) was assessed during FTAI by examining Estrotect patches (Rockway Inc., Spring Valley, WI). EE was considered present when a patch had 50% or more of its color rubbed off, and it was deemed absent when less than 50% of the color had worn off. Pregnancy rates (PR) were determined through ultrasound examination on d 55. EE and PR were analyzed by treatment using the GLIMMIX procedure on SAS 9.4 (SAS Institute Inc., Cary, NC). Statistical differences were declared at α ≤ 0.05 and tendencies declared between 0.05 < P ≤ 0.1. Heifers on the 8&6-d protocol had greater rectal temperatures at breeding compared with 7&7-d heifers (P < 0.0001). AI pregnancy rates were greater in heifers on the 8&6-d protocol compared with the heifers on 7&7-d protocol (P = 0.625) with average pregnancy rates of 39.4% and 30.3%, respectively. Overall, pregnancy rate was greater as well for the 8&6-d heifers compared with the 7&7-d heifers (P = 0.044), with mean overall pregnancy rates of 78.8% and 54.5%, respectively.

Natural Killer Cell Dysfunction in Premenopausal BRCA1 Mutation Carriers: A Potential Mechanism for Ovarian Carcinogenesis.

Tissue-specificity for fimbrial fallopian tube ovarian carcinogenesis remains largely unknown in BRCA1 mutation carriers. We aimed to assess the cell autonomous and cell-nonautonomous implications of a germline BRCA1 mutation in the context of cancer immunosurveillance of CD3- CD56+ natural killer (NK) cells. Premenopausal BRCA1 mutation carriers versus age-matched non-carriers were compared. Daily urinary 5β-pregnanediol levels were used to determine progesterone metabolomics across an ovarian cycle. Using peripherally acquired NK cells the cell-mediated cytotoxicity of tumor targets (OVCAR-3, K-562) was determined using live cellular impedance (xCELLigence®) and multicolor flow cytometry. Hypoxia-inducible factor 1-alpha (HIF-1α) immunohistochemistry of cancer-free fallopian tube specimens allowed a comparison of proximal versus distal portions. Utilizing these findings the role of environmental factors relevant to the fimbrial fallopian tube (progesterone, hypoxia) on NK cell functional activity were studied in an ovarian phase-specific manner. BRCA1 mutation carriers demonstrate a differential progesterone metabolome with a phase-specific reduction of peripheral NK cell functional activity. Progesterone exposure further impairs NK cell-mediated cytotoxicity in a dose-dependent manner, which is reversed with the addition of mifepristone (1.25 µM). The fimbrial fallopian tube demonstrated significantly higher HIF-1α staining, particularly in BRCA1 mutation carriers, reflecting a site-specific 'hypoxic niche'. Exposure to hypoxic conditions (1% O2) can further impair tumor cytotoxicity in high-risk carriers. Phase-specific differential NK cell activity in BRCA1 mutation carriers, either systemically or locally, may favor site-specific pre-invasive carcinogenesis. These cumulative effects across a reproductive lifecycle in high-risk carriers can have a detrimental effect further supporting epidemiological evidence for ovulation inhibition.

Differential effect of estrogen and progesterone on <i>in vitro</i> growth of uterine leiomyoma cells with chromosome 7 deletions

BACKGROUND: The studies on how sex steroid hormones affect growth of uterine leiomyoma cells with chromosomal abnormalities is highly relevant for development of personalized tumor therapy.
 AIM: To study in vitro the isolated and combined effects of estrogen and progesterone on uterine leiomyoma cells with chromosomal aberrations — deletions in 7q.
 MATERIALS AND METHODS: The study was performed on 15 uterine leiomyomas, excised from 15 women of 26–44 years of age who were not treated with hormones. Uterine leiomyoma cells were cultured in hormone-free medium, in the medium supplemented with estrogen, progesterone or both hormones. The chromosome preparations were made and stained with QFH/AcD to perform conventional karyotyping and fluorescence in situ hybridization (FISH) to accurately describe chromosomal rearrangements. The frequency of uterine leiomyoma cells with chromosomal aberrations was assessed by interphase FISH.
 RESULTS: Deletions in 7q were identified in 6 out of 15 karyotyped uterine leiomyomas; four of them had one clone with deletion in 7q whereas two others comprised two clones with 7q deletions of different length. The frequency of cells carrying deletions in 7q greatly varied in uterine leiomyoma samples cultured in hormone-free medium: from 3.5% to 93.6%. Exposure of cell cultures to estrogen and progesterone resulted in a fold change frequency increase in some of the uterine leiomyomas and decrease in the others. The most significant changes in the frequency of cells with deletions in 7q were registered in response to the isolated estrogen and, to a lesser extent, to progesterone exposure; less significant changes were observed after combined hormonal effect.
 CONCLUSIONS: In uterine leiomyomas with deletions in 7q, the frequency of abnormal cells may either increase or decrease in response to estrogen and progesterone in vitro supplementation. The isolated effect of estrogen or progesterone on the frequency of uterine leiomyoma cells with deletion in 7q is more pronounced compared to the combined one.

#198 : When to Transfer Day 6 Vitrified Embryos? On Day 5 vs Day 6 of Progesterone, a Retrospective Analysis

Background and Aims: The age of cryopreserved embryos impacts the ART outcome of frozen embryo transfer cycles. The development potential of embryos is not uniform. Though the days of progesterone exposure before transfer of D5 blastocyst is so far standardized, there is lack of standardization on transfer of Slow growing embryo i.e. D6 blastocysts. So, this study aims to compare the clinical outcomes of D6 blastocyst transferred on the 5th day or 6th day of progesterone supplementation. Method: The retrospective data were collected from the central registry of Indira IVF Hospital Private Limited to analyze 62227 frozen cycles from April 2019 to July 2021, divided into two groups: D6-Frozen embryos transferred on P+5 (Group A) vs P+6 (Group B). Results: There were 1212 cycles in Group A, and 189 cycles in the Group B arm in the final analysis). The propensity scores were calculated using binary logistic regression and the nearest neighborhood technique based on the following variables at baseline: Wife Age, Husband Age, Body max Index (BMI), Anti mullerian hormone (AMH), and Endometrial thickness. The clinical pregnancy rate was significantly higher in Group A than in Group B (CPR 70.5% vs. 63.0%, p=0.037). No significant differences were observed in the biochemical pregnancy rate (72.6% vs. 66.1%; p=0.066), biochemical loss rate (3.0% vs. 4.8%; p=0.271), clinical miscarriage rate (15.9% vs. 13.4%; p=0.485), live birth rate (59.2% vs. 54.5%; p=0.218) and multiple live birth rate (16.5% vs. 14.8%; p=0.559). Conclusion: In this study Frozen Embryo Transfer of D-6 blastocysts on the day-5 of progesterone results in better pregnancy outcomes than the day-6 of progesterone.

Exogenous Progestogen Hypersensitivity and its Increasing Association with Assisted Reproductive Techniques (ART)/in vitro Fertilization (IVF).

Progestogen hypersensitivity (PH) also known as autoimmune progesterone dermatitis is a rare clinical entity that may be triggered by endogenous progesterone (menstrual cycles and pregnancy) or exogenous progestin exposure (examples: contraceptive medicines, in vitro fertilization treatments). It is a poorly recognized syndrome due to its heterogeneous clinical presentation. The pathomechanism of PH is believed to be primarily IgE mediated but less commonly other immune responses may be involved. Management is usually focused on symptomatic control with medications. Recently, with the increasing use of exogenous progestins for in vitro fertilization more cases of hypersensitivity to exogenous progestins have been reported. Progesterone is an essential drug in the luteal phase support improving chances of implantation and pregnancy rates, and hence, PH is an important and difficult challenge to manage in these patients. Because patients require IVF and there is no alternative to progesterone, desensitization is suggested as an approach to endure fertility treatments and provides symptom control in refractory cases. Here, we will review the different aspects of PH.

Induction of lactation in a patient with complete androgen insensitivity syndrome.

With rising rates of adoption and surrogacy, induced lactation is likely to become increasingly relevant, allowing women who did not undergo pregnancy to breastfeed. We describe the case of a woman with complete androgen insensitivity syndrome (CAIS) on conventional oestrogen therapy who was expecting a child via surrogacy and who wished to breastfeed. The woman was commenced on supplementary oestrogen therapy, domperidone and breast stimulation by mechanical breast pump 8 weeks prior to the delivery of her child. Following delivery, the patient produced a small, unquantified amount of milk, allowing her to suckle the infant for a short period of time. Induced lactation is possible in chromosomally XY individuals. It has been most successful in cis-women and transwomen, both of whom have had progesterone/progestogen exposure to the breast. We suggest that the addition of a progestogen to our patient's treatment regimen, either as part of her original hormone therapy or part of the lactation induction program, would have improved her changes of establishing successful lactation. Induced lactation is possible in chromosomally XY individuals with the use of pharmacological and non-pharmacological therapies. There are no standardised guidelines regarding the optimal regimen for induced lactation. Progesterone exposure to the breast is essential for ductal branching and alveolar maturation. In the published literature, induced lactation is more successful in transwomen and other XY individuals who have had prior progesterone exposure. The addition of progestogen to our patient's treatment regimen would have improved her chances of establishing successful lactation.

P-468 Personalized embryo transfer guided by transcriptomic endometrial receptivity analysis (ERA) in infertile patients: a systematic review and meta-analysis

Abstract Study question What is the effectiveness of a personalized embryo transfer (pET) guided by an endometrial receptivity test in assisted reproduction? Summary answer We have not found evidence supporting the routine use of a personalized embryo transfer guided by an endometrial receptivity test in any case yet. What is known already Currently, the outcomes of good quality embryo transfers are far from ideal. Clinical focus is shifting towards other actors that could play a key role in the good-quality embryos implantation failure, such as the endometrial preparation and the achievement of an embryo-endometrium synchrony, so that the embryo has a better chance to implant during the window of implantation (WOI). Today, there is a wide range of different endometrial receptivity tests that use different sets of genes to identify patients with displacements of their WOI in order to adjust the individual length of progesterone exposure in a personalized embryo transfer (pET). Study design, size, duration This is systematic review and meta-analysis were conducted following the Cochrane methods and the GRADE approach to rate the certainty of evidence (CoE). The protocol has been sent for registration to PROSPERO (CRD42022299827). We included studies that compared women that were tested with transcriptomic endometrial receptivity tests versus those that were not tested. We only included comparative studies, including randomized controlled trials (RCTs), and retrospective and prospective cohorts. Participants/materials, setting, methods Participants were women undergoing an infertility treatment (both with own or donor gametes), with or without repeated implantation failure (RIF), with or without preimplantation genetic testing for aneuploidies (PGT-A). Each of the above-mentioned populations was analyzed separately as different subgroups. Electronic searches in CENTRAL via the Cochrane Register of Studies Online (CRSO), PubMed, and Embase were performed from inception to October 2022. Screening, study inclusion and data extraction was performed by pairs of independent reviewers. Main results and the role of chance We included 35 studies. Two of them were RCTs and 25 were cohort studies that compared pET guided by an endometrial receptivity tests guided versus sET in women with non-tested endometrium. Besides, we included 16 studies that compared women with sET with a receptive endometrium versus pET in women with non-receptive endometrium. And we included three studies that compared pET guided by an endometrial receptivity test in a specific population of women (RIF or adenomyosis) versus a control population. The two RCTs included women with no history of RIF. In non-RIF women, no important differences (moderate-CoE) were found in live birth rate (LBR) and clinical pregnancy rate (CPR). In that population, cumulative CPR may be higher with pET (RR 1.15, 95%CI 1.00-1.34; low-CoE) but no differences were found in cumulative LBR. Seven cohort studies that adjusted by confounding were meta-analyzed. In RIF, pET may improve the CPR (OR 2.50, 95% CI 1.42-4.40; low-CoE) but, in agreement with the RCTs, no benefits were found in non-RIF. In summary, routine use of pET guided by endometrial receptivity tests is not supported by current published evidence in non-RIF women. However, low-CoE suggests that it may be useful in RIF. Limitations, reasons for caution Most data come from non-RCTs which increases the risk of bias. Besides, the large heterogeneity found in the population, interventions and comparisons led to large imprecision due to the few studies per analyzed strata. It is important to avoid the extrapolation of the results to any different setting. Wider implications of the findings Routine use of pET guided by endometrial receptivity tests is not supported by current published evidence in non-RIF women. However, low-CoE suggests that it may be useful in RIF. Better designed studies are needed to find out if any of these tests work in any specific population. Trial registration number not applicable

P-618 Morning or evening start of exogenous progesterone five days before blastocyst transfer does not have an impact on ongoing pregnancy rates in artificial cycles.

Abstract Study question Does morning or evening start of exogenous progesterone five days before a blastocyst transfer have an impact on pregnancy rates in artificial cycles? Summary answer Ongoing pregnancy rate is comparable when exogenous progesterone is started either in the morning or evening on the first day of progesterone exposure before ET. What is known already Luteal phase support (LPS) with exogenous progesterone (P) is mandatory in artificial cycles due to the absence of corpus luteum. The time of P exposure needed to optimize the outcome remain uncertain. Some endometrial receptivity tests claim for individualization of duration of LPS before ET to optimize the implantation potential of the embryo. As routine practice, 5 days of P exposure are needed for a blastocyst transfer in general population, but it is not clear yet if exogenous P has to be started in the morning or evening of the first day of P administration. Study design, size, duration Single-centre retrospective cohort study of 6493 artificial cycles for blastocyst transfer between December 2018 and July 2022. Endometrial preparation was performed with oestrogens and LPS with micronized vaginal progesterone (400 mg/12h MVP). LPS was given from five days before blastocyst transfer. Ongoing pregnancy rate (>12w) was analysed according to the moment of starting progesterone (morning or evening) and hours of P exposure. Cases in which LPS duration was different to 5 days were not included. Participants/materials, setting, methods Infertile patients undergoing ET in artificial cycles. Until March 2021, exogenous P was started in the evening of day 0 of P exposure (“evening start”); since April 2021, P was started in the morning of day 0 (“morning start”), before blastocyst ET. ET are performed between 12 and 18pm. Serum P was measured at the time of ET (+2h). Routinely, patients with suboptimal P levels were added subcutaneous P (25mg/day) from the day of ET. Main results and the role of chance LPS was started in the evening in 3993 patients and in the morning in 2500 patients, all started five days before ET. Of note, there were no differences in these baseline/cycle characteristics: age, BMI, serum P in the proliferative phase, mid-luteal serum P, oocyte origin, EMT, number of embryos transferred. Patients with morning start had a significantly higher estradiol level (320.4 pg/mL vs 282.2pg/mL, p = 0.001). Mean number of hours of P exposure until ET was 113.8 + 2.1h after evening start of MVP and 125.5 + 2.1h after morning start, p = 0.000. The minimum was 108.2 hours and maximum of 132.9 hours of P exposure. Quartiles of P exposure (hours) before ET were: Q1:113.2h, Q2:115.8h, Q3:124.7h. No differences in OPR were observed according to these quartiles (<Q1:47.4%; Q1-Q3:45.7%; >Q3:47.1%, p = 0.46). Likewise, evening or morning start of LPS did not exert any impact either in OPR (46.2% vs 46.8%, p = 0.65) or clinical miscarriage rate (17.2% vs 18.1%, p = 0.53). Binary logistic regression showed no differences in OPR (aOR (95%CI): 0.96 (0.85-1.09; p = 0.52) after adjusting for potential confounding variables. Suboptimal mid-luteal P levels were observed in 23% (evening) vs 19.4% (morning start) of patients (p = 0.01). Having low P didńt impact on OPR (45.7%vs46.7%, p = 0.55) as rescue treatment was performed. Limitations, reasons for caution The limitation is the retrospective study design, although sample size is large enough to draw conclusions and control for potential confounding factors. Moreover, study period was different in both groups, although protocols and general results were comparable. If these results are similar in other ways of P administration, remains unknown. Wider implications of the findings These findings suggest that a difference of 12 hours in the initiation of exogenous P does not impact on the cycle outcome in artificial cycles as far as it is started 5 days before ET. Thus, LPS can be started either in the morning or evening without hampering the results. Trial registration number not applicable

P-125 Day 5 endometrium, Is the pregnancy rate following day 5 or day 6 vitrified/thawed blastocysts transfer affected?

Abstract Study question Are ongoing pregnancy rates of embryos vitrified on day 5 and day 6 when transferred into a day 5 endometrium comparable? Summary answer Day 5 blastocysts implant better in day 5.5 endometrium than day 6, whether they were vitrified on day 6 or 5 and cultured 24 hours. What is known already Preparation of the endometrium for transfer of cryopreserved embryos is usually done so that transfer takes place on endometrial day 5, although embryos may be at day 5 or 6 of development, so there may be asynchrony between the endometrium and the embryo if a day 6 embryo is transferred. Fresh transfer of day 5 blastocysts has been reported to have better results than day 6 blastocysts, predictably due to this endometrial asynchrony. However, data from frozen blastocyst transfers are less clear, possibly due to heterogeneity of the patient population and/or embryo quality. Study design, size, duration In this study we aim to analyse whether ongoing pregnancy after vitrified/thawed embryo transfers is affected if day 5 or day 6 embryos are transferred into day 5 endometrium. For this purpose, we examined 2266 cryotransfers performed in three Next Fertility clinics in Spain between 2016 and 2022. Day 5 (n = 1583) and 6 (n = 519) blastocyst transfers and day 5 embryos that devitrified one day before transfer (n = 164) were retrospectively compared. Participants/materials, setting, methods In this work blastocyst transfers were performed from donor or autologous oocytes, with or without PGT-A. Embryos were vitrified on day 5 or 6. Endometria were prepared with hormone replacement therapy or in natural cycle. All FET were performed after 5.5 days of progesterone exposure, regardless of embryo stage (day 5 or 6). The results were analysed considering all the above groups. Main results and the role of chance Ongoing pregnancy are superior with day 5 blastocysts versus day 6 and day 5 cultured for 24 hours prior to transfer. However, day 6 blastocysts were similar to cultured ones. Own oocytes: Day 5 44.98% (n = 538) vs Day 6 35.75% (n = 207) (p < 0.03); Day 5 44.98% vs Day 5 cultured 32.58% (n = 89) (p < 0.03); Day 6 35.75% vs Day 5 cultured 32.58% (p > 0.05). Donated Oocytes: Day 5 45.86% (n = 726) vs Day 6 28.51% (n = 221) (p < 0.00001); Day 5 44.98% vs Day 5 cultured 39.34% (n = 61) (p > 0.05); Day 6 28.51% vs Day 5 cultured 39.34% (p > 0.05). PGT-A Own oocytes: Day 5 51.72% (n = 203) vs Day 6 25.39% (n = 63) (p < 0.003); Day 5 51.72% vs Day 5 cultured 40.00% (n = 10) (p > 0.05); Day 6 25.39% vs Day 5 cultured 40.00% (p > 0.05). PGT-A Donated Oocytes: Day 5 42.24% (n = 116) vs Day 6 35.71% (n = 28) (p > 0,05); Day 5 42.24% vs Day 5 cultured 40.00% (n = 4) (p > 0.05); Day 6 35.71% vs Day 5 cultured 50.00% (p > 0.05). Miscarriage rates were similar across all groups. Limitations, reasons for caution Not all cohorts in the study were equally large, so some results have less statistical power, especially in abortion rates. Wider implications of the findings Day 5 blastocysts should always be transferred into a 5.5-day endometrium on the day they are devitrified, as this is when they are most synchronous with the endometrium. Further studies are needed to assess whether day 6 embryos implant better in a 6.5-day endometrium than in a 5.5-day endometrium. Trial registration number not applicable

Personalized embryo transfer guided by endometrial receptivity analysis: a systematic review with meta-analysis.

Does a personalized embryo transfer (pET) guided by tests for endometrial receptivity (TER) increase the effectiveness of ART procedures? The use of TER-guided pET is not supported by current published evidence in women without repeated implantation failure (RIF), while in women with RIF more research is needed to assess a potential benefit. Implantation rates are still far from ideal, especially in some patients that have RIF with good-quality embryos. As a potential solution, a wide range of diverse TER use different sets of genes to identify displacements of the window of implantation to adjust the individual length of progesterone exposure in a pET. A systematic review with meta-analysis was performed. Search terms included endometrial receptivity analysis, ERA, personalized embryo transfer. CENTRAL, PubMed, Embase, reference lists, clinical trials registers, and conference proceedings (search date October 2022) were searched, with no language restrictions. Randomized controlled trials (RCTs) and cohort studies comparing a pET guided by TER vs standard embryo transfer (sET) in different subgroups that undergo ART were identified. We also investigated pET in non-receptive-TER vs sET in receptive-TER, and pET in a specific population vs sET in a general population. Risk of bias (RoB) was assessed with the Cochrane tool and ROBINS-I. Only those with low/moderate RoB underwent meta-analysis. The GRADE approach was used to evaluate the certainty of evidence (CoE). We screened 2136 studies and included 35 (85% used ERA and 15% used other TER). Two studies were RCTs comparing endometrial receptivity analysis (ERA)-guided pET vs sET in women with no history of RIF. In women without RIF, no important differences (moderate-CoE) were found in live birth rates and clinical pregnancy rates (CPR). We also performed a meta-analysis of four cohort studies that were adjusted for confounding. In agreement with the RCTs, no benefits were found in women without RIF. However, in women with RIF, low CoE suggests that pET might improve the CPR (OR 2.50, 95% CI 1.42-4.40). We found few studies with low RoB. Only two RCTs in women without RIF were published, and none in women with RIF. Furthermore, the heterogeneity observed in populations, interventions, co-interventions, outcomes, comparisons, and procedures limited the pooling of many of the included studies. In the population of women without RIF, in agreement with previously published reviews, pET did not prove to be more effective than sET and, therefore, it precludes the routine use of this strategy in this population until more evidence is available. However, more research is advisable in women with RIF as low-certainty evidence from observational studies adjusted for confounders suggests that the CPR might be higher with pET guided by TER in this population. Although this review presents the best available evidence, it is still insufficient to change current policies. No specific funding was obtained for this study. There are no conflicts of interest to declare. PROSPERO CRD42022299827.

Agmatine prevents the manifestation of impulsive burying and depression-like behaviour in progesterone withdrawn female rats

Premenstrual dysphoric disorder (PMDD) is characterized by various physical and affective symptoms, including anxiety, irritability, anhedonia, social withdrawal, and depression. The present study investigated the role of the agmatinergic system in animal model of progesterone withdrawal in female rats. Chronic progesterone exposure of female rats for 21 days and its abrupt withdrawal showed enhanced marble burying, increased immobility time, and reduced no. of entries in open arm as compared to control animals. The progesterone withdrawal-induced enhanced marble burying anxiety and immobility time was significantly attenuated by agmatine (5–20 mg/kg, i.p.), and its endogenous modulators like L-arginine (100 mg/kg, i.p.), amino-guanidine (25 mg/kg, i.p.) and arcaine (50 mg/kg, i.p.) by their once-daily administration from day 14-day 21 of the protocol. We have also analysed the levels of agmatine, progesterone, and inflammatory cytokines in the hippocampal region of progesterone withdrawn rats. There was a significant decline in agmatine and progesterone levels and an elevation in cytokine levels in the hippocampal region of progesterone withdrawn rats compared to the control animals. In conclusion, the present studies suggest the importance of the endogenous agmatinergic system in progesterone withdrawal-induced anxiety-like and depression-like behaviour. The data also projects agmatine as a potential therapeutic target for the premenstrual dysphoric disorder.

B-cell lymphoma 6 expression significantly differs by the uterine preparation method used for frozen embryo transfer

To determine whether B-cell lymphoma 6 (BCL6), an endometriosis-associated marker postulated to predict poor pregnancy outcomes, is differentially expressed in the window of implantation with various uterine preparation regimens commonly used for frozen embryo transfers. A retrospective cohort study. An academic center. Patients with infertility who underwent endometrial biopsy for BCL6 evaluation INTERVENTION(S): Exogenous estradiol and/or progesterone. Endometrial BCL6 histological score (HSCORE) and overexpression (HSCORE >1.4) RESULT(S): Two hundred and forty-four patients were included in the analysis: 76 patients were sampled in a natural menstrual cycle without exogenous hormone exposure (NC), 25 under a modified natural cycle embryo transfer protocol with choriogonadotropin alfa injection followed by luteal phase vaginal progesterone supplementation (mNC), and 143 under a programmed cycle embryo transfer protocol, with estradiol administration followed by addition of intramuscular progesterone-in-oil injections (PC). Median HSCORE (interquartile range) was the highest in NC (3.0 [1.8-3.6]). BCL6 expression was significantly lower in mNC (1.1 [0.4-2.1]) and PC groups (0.8 [0.3-1.3]) compared with NC. In addition, BCL6 overexpression (HSCORE >1.4) was observed in 80.3% of NC, 40.0 % of mNC, and 23.1 % of PC. After adjusting for covariates, the endometrium exposed to exogenous progesterone had significantly lower odds of BCL6 overexpression than that of a natural menstrual cycle (adjusted odds ratio, 0.12 [95% CI 0.04-0.35] for mNC; and odds ratio, 0.08 [95% CI 0.04-0.17] for PC). BCL6 expression differs by the type of uterine preparation method, with lower levels observed with exogenous progesterone exposure. The validity and utility of BCL6 testing under medicated endometrial state warrants further investigation.

Daily ovarian hormone exposure and loss of control eating in adolescent girls: A registered report.

The daily biobehavioral factors that precipitate loss of control eating (LOCE) in adolescent girls are not well known. Ovarian hormone levels are key biological factors associated with the etiology of eating disorders in adolescent girls. Yet, models on how daily ovarian hormone exposure predicts LOCE in adolescent girls are underdeveloped. The goal of this study is to examine the daily patterns and mechanisms of ovarian hormone levels on LOCE across the menstrual cycle in adolescent girls and the mediating roles of food-related reward anticipation and response inhibition. Ecological momentary assessment (EMA) paired with daily hormonal sampling will be used to examine (1) daily associations between within-person hormones and LOCE, and (2) the mediating role of within-person food-related reward anticipation and response inhibition. Normally cycling adolescent girls who have reached menarche will provide daily saliva samples for hormone analysis and complete EMA for 35 days. During EMA, girls will report LOCE and will complete task-based and self-report measures of food-related response inhibition and reward anticipation. This work has implications for the development of new real-world biobehavioral models of LOCE in adolescent girls, which will guide theory improvements and treatment for LOCE. Results will provide preliminary evidence for treatment targets for novel interventions for adolescent girls-for example, a response inhibition intervention. Adolescent eating disorders are severe mental health conditions, often marked by loss of control eating. Estrogen and progesterone play a role in the development and persistence of loss of control eating. The current study will examine how daily exposure to estrogen and progesterone predicts loss of control eating in adolescent girls and identify possible daily mechanisms linking estrogen and progesterone exposure and loss of control eating.

Dietary supplementation with nicotinamide riboside improves fetal growth under hypoglycemia

Nicotinamide riboside (NR) is considered a super-supplement that prevents obesity and diabetes. While NR has been investigated for various effects depending on nutritional conditions, metabolic research on women and pregnant women has rarely been discussed. In this study, we focused on the glycemic control of NR in females and found the protective role of NR in pregnant animals under hypoglycemic conditions. Metabolic-tolerance tests were performed in vivo under progesterone (P4) exposure after ovariectomy (OVX). NR enhanced resistance to energy deprivation, such as starvation, and showed a slight increase in gluconeogenesis in naïve control mice. However, NR reduced hyperglycemia and significantly induced gluconeogenesis in OVX mice. While NR reduced hyperglycemia in the P4-treated OVX mice, it reduced insulin response and substantially increased gluconeogenesis. Similar to animal experiments, NR increased gluconeogenesis and mitochondrial respiration in Hep3B cells. The gluconeogenic function of NR is mediated by tricarboxylic acid cycle (TCA) cycle enrichment, as residual pyruvate could induce gluconeogenesis. NR recovered fetal growth by increasing blood glucose levels when artificial hypoglycemia was induced during pregnancy. Our study revealed the glucose-metabolic function of NR in hypoglycemic pregnant animals, suggesting NR as a dietary supplement to improve fetal growth. Because diabetic women suffer from hypoglycemia due to insulin therapy, NR has therapeutic potential for use as a glycemic control pill.