High Progesterone Levels Research Articles

NOTCH1- and CD117-positive stem cells in human endometrium and their implications for successful implantation

To investigate the quantity of 2 stem cell types in the endometrial stroma of women undergoing invitro fertilization and their association with steroid hormone signaling and implantation success after embryo transfer. Prospective cohort study. Private hospital. A total of 109 patients undergoing invitro fertilization. Not applicable. Immunohistochemistry staining of endometrial biopsies taken during the midluteal phase using antibodies against NOTCH1 and CD117 was performed. The percentage of endometrial stromal cells positive for these markers was determined. The link of these stem cell percentages with the serum progesterone and estradiol levels and the endometrial expression of their respective receptors were assessed. After embryo transfer, the quantity of stained cells for each marker was also compared according to implantation outcome. The percentage of NOTCH1+ stromal cells ranged from 0.003%-2.112% (median, 0.062%) and was significantly higher than that of CD117+ cells, which ranged from 0.000%-0.210% (median, 0.020%) (Z = -7.035). The percentage of NOTCH1+ stem cells showed no difference between the studied serum hormone level groups and no relationship with the expression of their receptors in the endometrium. In contrast, the number of CD117+ cells significantly differed between patients with high and low levels of serum progesterone (cutoff, 14.9 ng/mL) and estradiol (cutoff, 135.6 pg/mL). Furthermore, the quantity of CD117+ stem cells was positively correlated with the progesterone receptor (R = 0.277) and estradiol receptor (R= 0.318) expression levels in the endometrium. Although the quantity of NOTCH1+ cells did not differ between the 2 implantation groups, the median percentage of CD117+ cells was significantly higher in patients with successful implantation than in those with unsuccessful implantation (0.03% vs. 0.01%, respectively). The cutoff value for the percentage of CD117+ cells predicting successful implantation was 0.018% (area under the curve, 0.66; 95% confidence interval, 0.56-0.77; sensitivity, 63.1%; specificity, 61.4%). This study indicates that the quantity of certain stem cell types (CD117+), but not others (NOTCH1+), in the functional endometrium is associated with implantation success and sex hormone signaling during the midluteal phase. These findings highlight the role of CD117+ cells in preparing the endometrium for embryo implantation, and their quantity may be an indirect indicator of endometrial receptivity.

Variation in Progesterone Levels and Urinary Tract Infections in Pregnant Women Attending Moi Teaching and Referral Hospital, Eldoret, Kenya

Background and Aims: Urinary tract infections (UTIs) are common among pregnant women and major predisposing factors for pyelonephritis linked to obstetrical complications including preterm labour and low infants’ birth weights. This study sought to determine the relationship(s) between pregnancy trimesters, UTIs and changes in progesterone levels among pregnant women. Materials and Methods: The study was conducted in 2016 at Moi Teaching and Referral Hospital (MTRH) antenatal clinic which is a referral facility that attends to patients from most Counties in western region of Kenya. A cross-sectional study design was used to collect blood and urine specimens from 78 participants. Blood was used to determine progesterone levels using ELISA technique and urine cultures with bacterial colony counts ≥ 105 were appropriately identified to species level. Trimester periods and participants’ demographic information were obtained using a structured questionnaire. Results: Culture results showed that the most abundant bacterial species isolated in urine from the pregnant women was Escherechia coli (63.7%). The more affected age-group was women between 30 - 39 years during trimester three, suggesting that bacterial colonization of genital track occurred more frequently in older compared to the younger women. There was an exponential increase in progesterone levels among the pregnant women during trimester three compared to other trimesters, although these increases occurred independent of age. However, high levels of progesterone among pregnant women in third trimester corresponded with increased number of E. coli causing UTI. Conclusion: The results showed that progesterone levels increase with trimester and the most prevalent bacteria associated with this was E. coli even though age and increase in progesterone levels had no significant impact on E. coli infection.

Hormonal indexes as predictors of porcine reproductive traits.

The aim of the present study was to examine the applicability of several hormonal indexes for early prediction of puberty and reproductive state in pigs. For this purpose, we have compared the level of hormones leptin, estradiol, progesterone, and IGF-I in the blood of gilts at 150 days of age and their indexes of puberty and ovarian state at the age of 200 days. The association between blood leptin, estradiol, progesterone, and IGF-I and indexes of future reproductive state has been demonstrated. High blood concentrations of leptin and IGF-I levels were associated with relatively low reproductive traits, while high levels of estradiol and progesterone were associated with future high reproductive indexes. These observations are the first demonstration of the applicability of these endocrine indexes for prediction of porcine reproductive traits.

Effect of High Progesterone Level on Day of Human Chrionic Gonadotropin Triggering on Pregnancy Rate in Frozen Embryo Transfer

Effect of High Progesterone Level on Day of Human Chrionic Gonadotropin Triggering on Pregnancy Rate in Frozen Embryo Transfer

Ratlarda Gebeliğin Farklı Periyotlarında İnce Bağırsaklarda Mast Hücrelerinin Dağılımı

The gastrointestinal tract has different characteristics. The gastrointestinal tract is affected by the enlarging uterus as pregnancy progresses. The enlargement of the uterus changes the digestive organs, especially the metabolism of the stomach and intestines. Anatomical changes produce acute abdominal changes. In addition to the mechanical effects caused by the growing uterus, high progesterone levels cause delayed gastric emptying and increased gastrointestinal transit time. Stomach and intestinal motility are also affected, resulting in lower transit times. Mast cells are cells that are activated and show degranulation under all kinds of stress conditions. These cells are considered to be cells that play an important role in the physiological and pathological responses of the immune system by showing both proinflammatory and immunosuppressive functions. Maternal malnutrition adversely induces fetal bowel development and function through the maternal intestinal barrier, intestinal content, structure, and peripheral circulation. In this study, samples were taken from the small intestine in 3 different trimesters of pregnancy. Tissue samples taken from duodenum, jejunum and ileum were fixed in Carnoy solution for 12 hours. Paraffin blocks were prepared after tissue follow-up without washing. Paraffin blocks were formed from the samples and 5-6 µ thick sections were taken and stained with 0.5% Toluidine Blue to determine the distribution of mast cell number. After staining, the distribution of mast cells in the lamina propria + submucosa and tunica serosa layers was examined. It was determined that mast cells changed numerically in the duodenum, jejunum and ileum in the small intestine according to the layers and according to the gestational periods. It is thought that this study will contribute to the literature on this subject.

Delayed blastocyst development is influenced by the level of progesterone on the day of trigger.

To evaluate the association between progesterone (P) level on the day of trigger and time to blastulation in IVF cycles. This was a retrospective cohort study with autologous IVF cycles performed at our Institution from January 2019 to December 2021. A total of 1109 IVF cycles were included. The primary outcome was to compare time to blastulation in terms of percentage of expanded (grade 3) blastocysts on day 5 according to progesterone level at trigger. A total of 3517 blastocysts were analyzed. After dividing progesterone level in quartiles (Q1, P < 0.50ng/ml; Q2 0.50ng/ml ≤ P ≤ 0.78ng/ml; Q3, 0.79ng/ml ≤ P ≤ 1.15ng/ml; Q4, P > 1.15ng/ml), we observed a delay in blastocyst development according to the increasing level of progesterone at trigger (analysis by rank, P-value = 0.01). After adjusting for confounding factors at the multivariate analysis, the percentage of day 5 blastocysts was reduced for Q3 (- 13.8%, 95% CI from - 20.5 to - 7.0%, p < 0.001) and Q4 (- 7.7%, 95% CI from - 15.5 to 0.0%, p = 0.05) compared to Q1 (reference). Progesterone levels on day of trigger correlate to the percentage of expanded (grade 3) blastocysts on day 5 and a delayed blastocyst development day 5 is expected for high progesterone levels.

The estrous cycle modulates early-life adversity effects on mouse avoidance behavior through progesterone signaling

Early-life adversity (ELA) increases the likelihood of neuropsychiatric diagnoses, which are more prevalent in women than men. Since changes in reproductive hormone levels can also increase the probability of anxiety disorders in women, we examined the effects of ELA on adult female mice across the estrous cycle. We found that during diestrus, when progesterone levels are relatively high, ELA mice exhibit increased avoidance behavior and increased theta oscillation power in the ventral hippocampus (vHIP). We also found that diestrus ELA mice had higher levels of progesterone and lower levels of allopregnanolone, a neurosteroid metabolite of progesterone, in the vHIP compared with control-reared mice. Progesterone receptor antagonism normalized avoidance behavior in ELA mice, while treatment with a negative allosteric modulator of allopregnanolone promoted avoidance behavior in control mice. These results suggest that altered vHIP progesterone and allopregnanolone signaling during diestrus increases avoidance behavior in ELA mice.

Toxic effect of heavy metals on ovarian deformities, apoptotic changes, oxidative stress, and steroid hormones in rainbow trout

BackgroundAs is well known, the pollution in the aquatic environment in which fish grow has a direct impact on aquaculture practices. Pollution in aquatic systems because of multiple adverse effects on fish metabolic processes, especially the reproductive systems. AimThe goal of this study was to assess the severity of pollution impact in two different hatcheries, Verinag hatchery, Site 1 (S1) and Panzath hatchery, Site 2 (S2) in Anantnag region, using histopathological, ultrastructural, oxidative stress, genotoxic, and hormonal analysis in rainbow trout gonad (ovary).M&M: Fish were collected between May 2018 and April 2019 from two locations, Verinag hatchery (S1) and Panzath hatchery (S2), which were affected by heavy metals. ResultsThe histological and ultrastructural examination of rainbow trout ovaries from the Verinag hatchery (S1) revealed normal structure in growing oocytes in rainbow trout at various stages based on morphological features while the fish ovaries in the Panzath hatchery (S2) showed various deformities and irregularly shaped oocytes. The surfaces of some of these oocytes were wrinkled, rough, or distorted. Apoptotic studies revealed that the frequency of apoptotic cells collected from S2 water was significantly increased in ovarian cells (P < 0.05). The activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were found to be increased in fish collected from S1 but decreased in fish collected from S2. In S2 caught fish, malondialdehyde (MDA) levels were found to increase gradually, and the degree of heavy metal stress was positively correlated (p < 0.05). The comet assay was used to determine the induction of DNA damage in ovarian cells. The induction of DNA damage was found to be significantly higher (p < 0.05) in S2 fish specimens compared to fish from S1. On comparing the DNA damage of the rainbow trout from the two sampling sites, it was revealed that the fish is much more sensitive to aquatic contaminants. Regarding steroid hormones, higher levels of progesterone and estrogen were reported in the fish samples collected from S1 as compared to S2 captured fish. ConclusionIn conclusion, the comparative study of fish from two different sites viz. Verinag hatchery (S1) and Panzath hatchery (S2) revealed that S2 sampled fish suffered more heavy metal damage, including cellular deformities, apoptosis, oxidative damage, and altered steroid hormones.

Kombinasi Aromaterapy Jasmine Sambac dan Loving Yoga terhadap Disfungsi Seksual pada Akseptor KB Suntik DMPA

One of the side effects of the 3-Month Injectable KB is impaired sexual function caused by the high level of the hormone progesterone, resulting in vaginal drying, which causes pain during sexual intercourse, which can lead to a decrease in sexual desire or dysfunction in women. Jasmine Sambac is an effective and gentle way to promote a healthy body and for relaxation. Loving yoga is a type of yoga developed by researchers, with a focus on Kegel exercises and pelvic muscles that function to increase blood flow, so that the body feels relaxed which can increase libido in sexual intercourse. The purpose of this study was to identify the effect of the combination of Aromatherapy Jasmine Sambac and Loving Yoga on sexual dysfunction in DMPA injection family planning acceptors. The research method used is a quasi-experimental approach with a one group pre-post test design approach. Measurement of pre-test and post test was carried out using the FSFI (Female Sexual Function Index) instrument. The results showed that jasmine sambac aromatherapy and loving yoga had a significant effect on increasing the total libido, including its 6 aspect domains, on DMPA injection acceptors.

Does the Ketogenic Diet Improve the Quality of Ovarian Function in Obese Women?

Background: Polycystic ovarian syndrome (PCOS) is the most common endocrine disorder in women of reproductive age, the prevalence of which ranges from 8 to 13%. It is characterized by metabolic, reproductive, and psychological alterations. PCOS prevalence is related to body mass index (BMI). Women with BMI < 25 kg/m2 have a prevalence of 4.3%, whereas women with BMI > 30 kg/m2 have a prevalence of 14%. Moreover, women with PCOS have a risk of type 2 diabetes mellitus (T2DM) two-fold higher than controls, independently of BMI. Both PCOS and T2DM are also consequences of lower serum sex-hormone-binding globulin (SHBG) levels, which is currently considered a biomarker of metabolic disorders, in particular T2DM. Aim: To evaluate the effect of the very-low-calorie ketogenic diet (VLCKD) on markers suggested to be predictive of metabolic and ovulatory dysfunction. These comprehend SHBG, anti-Mullerian hormone (AMH), and progesterone levels on day 21 of the menstrual cycle in a cohort of obese non-diabetic women with PCOS and regular menses. Methods: Twenty-five patients (mean age 25.4 ± 3.44 years) with obesity and PCOS underwent VLCKD for 12 weeks. Each of them underwent measurements of anthropometric parameters (body weight, height, and waist circumference) and blood testing to evaluate serum levels of SHBG, AMH, and progesterone before and after 12 weeks of VLCKD. Results: At enrollment, all patients had high BMI, WC, and AMH, whereas SHBG and progesterone levels were low. After VLCKD, the patients showed a significant reduction in BMI, WC, and HOMA index. In particular, 76% of patients (19/25) switched from obesity to overweight, and the HOMA index normalized, reaching values lower than 2.5 in 96% (24/25) of patients. In addition, serum AMH levels significantly decreased, and progesterone and SHBG significantly increased after VLCKD. Conclusions: This is the first study documenting the effects of VLCKD on ovarian reserve and luteal function in women with PCOS. VLCKD could be used to improve metabolic and ovulatory dysfunction in women with PCOS. Further studies are needed to understand the reasons for the AMH reduction.

Lnc2300 is a cis-acting long noncoding RNA of CYP11A1 in ovarian granulosa cells.

The high level of progesterone and 17β-estradiol ratio (P4/E2) in follicular fluid has been considered as a biomarker of follicular atresia. CYP11A1, the crucial gene encoding the rate-limiting enzyme for steroid hormone synthesis, has been reported differently expressed in the ovary during follicular atresia. However, the regulation mechanism of CYP11A1 expression during follicular atresia still remains unclear. Here, we have demonstrated that lnc2300, a novel pig ovary-specific highly expressed cis-acting long noncoding RNA (lncRNA) transcribed from chromosome 7, has the ability to induce the expression of CYP11A1 and inhibit the apoptosis of porcine granulosa cells (GCs). Mechanistically, lnc2300, mainly located in the cytoplasm of porcine GCs, sponges and suppresses the expression of miR-365-3p through acting as a competing endogenous RNA (ceRNA), which further relieves the inhibitory effects of miR-365-3p on the expression of CYP11A1. Besides, CYP11A1 is validated as a direct functional target of miR-365-3p in porcine GCs. Functionally, lnc2300 is an antiapoptotic lncRNA that reduces porcine GC apoptosis by inhibiting the proapoptotic function of miR-365-3p. In summary, our findings reveal a cis-acting regulation mechanism of CYP11A1 through lncRNA, and define a novel signaling pathway, lnc2300/miR-365-3p/CYP11A1 axis, which is involved in the regulation of GC apoptosis and follicular atresia.

Fetal programming: in utero exposure to acrylamide leads to intergenerational disrupted ovarian function and accelerated ovarian aging.

In this study we investigated the effects of multigenerational exposures to acrylamide (ACR) on ovarian function. Fifty-day-old Wistar albino female rats were divided into the control and ACR-treated groups (2.5, 10, and 20 mg/kg/day) from day 6 of pregnancy until delivery. The obtained females of the first (AF1) and second generation (AF2) were euthanized at 4 weeks of age, and plasma and ovary samples were collected. We found that in utero multigenerational exposure to ACR reduced fertility and ovarian function in AF1 through inducing histopathological changes as evidenced by the appearance of cysts and degenerating follicles, oocyte vacuolization, and pyknosis in granulosa cells. TMR red positive cells confirmed by TUNEL assay were mostly detected in the stroma of the treated groups. Estradiol and IGF-1 concentrations significantly decreased as a result of decreased CYP19 gene and its protein expression. However, ACR exposure in AF2 led to early ovarian aging as evidenced by high estradiol and progesterone levels among all treated groups compared to control group, corresponding to the upregulation of the CYP19 gene and protein expression. The apoptotic cells of the stroma were greatly detected compared to that in the control group, whereas no significant difference was reported in ESR1 and ESR2 gene expression. This study confirms the developmental adverse effects of ACR on ovarian function and fertility in at least two consecutive generations. It emphasizes the need for more effective strategies during pregnancy, such as eating healthy foods and avoiding consumption of ACR-rich products, including fried foods and coffee.

Assessment of bacterial diversity associated with assisted reproductive technologies through next-generation sequencing

BackgroundTo assess bacterial diversity in infertile couples with their biochemical pregnancy outcomes. Using a retrospective case-control study design, participants were recruited for collection of vaginal swab, follicular fluid, endometrial fluid, and semen samples. The microbial composition was analyzed by 16S ribosomal RNA (rRNA) gene amplification with (MinION) Oxford Nanopore Ltd.ResultsOur findings revealed that age and endometrial thickness had a significant impact on the pregnancy success rate of pregnant (P) and non-pregnant (NP) patients receiving IVF, with high levels of luteinizing hormone, estrogen, and progesterone in the P group. In addition, the partial least squares discriminant analysis (PLS-DA) revealed a difference in microbial composition between the P and NP groups, as well as a higher microbial abundance in non-pregnant patients compared to pregnant patients. After comparison between pregnant patients and non-pregnant patients, pregnant patients had a higher abundance of Firmicutes and Proteobacteria and a lower abundance of Actinobacteria, Fusobacterium, and Bacteroidetes at the phylum level. Non-pregnant patients had a lower abundance of the probiotics lactobacillus and a higher abundance of infections Prevotella and Gardnerella at the genus level. As a result, a disordered microbiota in non-pregnant patients, characterized by a decrease in probiotics and an increase in pathogens, could be used as a possible marker for a higher IVF failure rate.ConclusionAlteration of the microbiota of the reproductive tract or the presence of certain microbes, regardless of the degree of pathogenicity that can affect fertilization, as well as implantation and subsequent embryonic development. This could result in failed fertility treatments and a lower live birth (LBR) rate.

Study of progesterone level during first trimester of pregnancy in cows.

Measurement of progesterone is an indirect method for pregnancy diagnosis in many livestock species including cattle, buffaloes, sheep, and goats. Conception extends the life of the corpus luteum (CL) by preventing the luteolytic mechanism from being triggered, thus prolonging and maintaining its functional characteristics, ensuring continued high progesterone levels (Spencer, et.al., 1995). (Shemesh et al., 1973) proposed that the difference in peripheral plasma progesterone levels between pregnant and non pregnant cows, 19 days after insemination, can form the basis for a very early pregnancy test. (Laing and Heap 1972) first documented this in milk to diagnose cows in early pregnancy Progesterone maintains the uterine endometrium in a state which supports embryonic development, implantation, and foetoplacental development. Progesterone concentrations vary with the stage of the estrous cycle which makes it one of the most commonly studied reproductive hormones in bovine ruminants for pregnancy detection and ovarian activity (Kaneko,et.al.,1994). Studies in the bovine estrous cycle indicate that the milk or serum progesterone concentrations reach a maximum value 13-14 days after estrus, and if the animal is pregnant, these continue to remain elevated up to day 21 after fertilization (Parkinson, et al.,1994) . These high levels of progesterone in serum or milk between days 18 and 24 after insemination form the basis of establishment of pregnancy in cattle (Sasser, and Ruder,1987; Shemesh,1973). Interferon-???? exerts its antiluteolytic effect by inhibiting the endometrial expression of oxytocin receptors, through which oxytocin stimulates pulsatile PGF2???? release (Wolf,2003). Although low progesterone concentrations at 18 to 24 days after breeding can accurately predict non pregnancy, high progesterone concentrations during this period are not the specific indicators of pregnancy due to variations among cows in duration of the estrous cycle as well as the incidence of early or late embryonic mortality. The advantages of progesterone assay for pregnancy diagnosis include noninvasive collection of milk sample and the feasibility to conduct the test on the farm using commercial cow-side milk progesterone test kits [Pennington,1985 ; Gowan,1982 ; Wimpy,et.al.,1986), though the sensitivity gets compromised to some extent with these assay kits.&#x0D; Key Word: progesterone, pregnancy, cows

New insights on hyperglycemia in 17-hydroxylase/17,20-lyase deficiency.

ObjectiveThe adrenal glands of patients with 17-hydroxylase/17,20-lyase deficiency (17OHD) synthesize excessive 11-deoxycorticosterone(DOC) and progesterone, and produce less amount of sex steroid production. Mineralocorticoids and sex hormones play an important role in regulating glucose homeostasis. This study aimed to describe the glucose metabolism in 17OHD patients diagnosed at Peking Union Medical College Hospital (PUMCH).Design/methodsA total of 69 patients diagnosed with 17OHD after adolescence in PUMCH from 1995 to June in 2021. Among them 23 patients underwent a 3-hours oral glucose tolerance test (3hOGTT) after being diagnosed with 17OHD. Insulin response in patients with normal glucose tolerance (NGT) were further compared between the study two groups with different kalemia status. Another 19 patients were followed up to 30 years and older. All clinical data were obtained from the hospital information system of PUMCH.Results Baseline: (1) The average body mass index(BMI) of all patients at baseline was 20.3 ± 3.7kg/m2. Twenty-three patients underwent 3hOGTT, of whom three were diagnosed with diabetes mellitus, and one with impaired glucose tolerance (IGT). Positive correlation between the ratio of progesterone to upper limit of normal range (P times) and hyperglycaemia was exist(r=0.707, P=0.005). (2) In 19 NGT patients, the insulin concentrations at 0 minute, results of the homeostasis model assessment for β-cell function and insulin resistance were lower in the hypokalaemia group than in the normal kalemia group(7.0(5.8-13.2) vs 12.4(8.9-14.9) μIU/ml, P=0.017; 115.5(88.2-240.9) vs 253.1(177.2-305.8), P=0.048; 1.54(1.17-2.61) vs 2.47(1.91-2.98), P=0.022, respectively). Follow-up: Four patients had IGT, while seven patients had diabetes mellitus. Of the 19 patients,11 had hyperglycaemia. P times was significantly higher(7.6(5.0-11.0) vs 3.75(2.2-5.3), P=0.008) in hyperglycemia group than in the normal glucose group.ConclusionsAbnormal glucose metabolism was common in 17OHD patients, which was possibly associated with hypokalaemia and high progesterone levels. Routine monitoring on glucose metabolism in 17OHD patient should be conducted.

Influence of refeeding on production, blood biochemistry parameters, and reproduction in underfed Kacang goat does.

This research aimed to investigate the effect of refeeding on production, blood biochemical parameters, and reproduction in underfed Kacang goat does. Twelve 2-3-year-old Kacang goats scored (body condition score [BCS]: 1-1.5) with poor body condition but appeared healthy, gave normal births 3-4 months before but no longer nursing their youngs, and anestrus were used in this study. The experiment consisted of 110 days with 14 days of adaptation period and 96 days of feeding treatment and data collection. After the adaptation period, the animals were divided into two groups (A and B), with six animals each. During the first 40 days (underfeeding period), they were fed with kangkong (Ipomoea reptans) straw at a level of 2% of body weight (BW). For the following 56 days (refeeding period), each animal in group A was fed at the level of 4% BW per day while group B was fed at the level of 5% BW per day consisting of 50% kangkong straw and 50% concentrate. Feed intake, average daily gain, body condition, and estrus were assessed in the period of underfeeding and refeeding. Blood samples were collected at the end of the underfeeding and refeeding period for blood biochemical parameter analyses. During the underfeeding period, the animals were anestrous and had poor body condition (BCS: 1.17-1.33) and high serum levels of cortisol, progesterone, and estradiol. Refeeding resulted in a significant increase in BCS (2.75-3.0), restoring estrous cycle, and reduced serum cortisol and progesterone level (p < 0.05). The Kacang goats had good compensatory growth. It could be concluded that refeeding at the level of 4-5% BW would restore undernourished goats' clinical condition, productivity, and reproductive performance after 8 weeks of refeeding.

P-683 High Serum Progesterone Levels on Day of Embryo Transfer for Pregnancy Outcomes in Patients Undergoing Artificial Frozen-thawed Blastocyst Transfer; Is There a Ceiling Effect?

Abstract Study question Is there a ceiling effect of high serum progesterone levels on day of embryo transfer for pregnancy outcomes in patients undergoing artificial frozen-thawed blastocyst transfer? Summary answer There is no ceiling effect of high serum progesterone levels on day of embryo transfer(&amp;gt;40 ng/mL)for pregnancy outcomes in patients undergoing artificial frozen-thawed blastocyst transfer. What is known already In recent studies, 10 ng/mL was usually used as the lower threshold value for serum progesterone (P4) level on the day of embryo transfer in patients undergoing artificial frozen-thawed blastocyst transfers and pregnancy outcomes were compared between patients with P4 levels above and below this threshold. The main reason for choosing this threshold in the studies is that it reflects the healthy corpus luteum function. Although many studies have investigated the lower threshold value, there is paucity of data on the high threshold value that can indicate the possible ceiling effect of P4 in the literature. Study design, size, duration This was a retrospective cohort study including 595 patients undergoing artificial frozen-thawed blastocyst transfer between 2017 to 2021. In order to eliminate the age-related bias, only patients performed euploid embryo transfer according to preimplantation genetic testing for aneuploidy from patients above aged 35 years were included. Participants/materials, setting, methods When we evaluated the percentiles of according to the progesterone levels,40.6 ng/ml corresponded to 90th percentile and 23.9 ng/ml corresponded to 50th percentile. Based on this finding, rounding up the numbers, we determined the progesterone levels cut-off as &amp;lt; 20ng/mL,n=220(37.0%);20-40ng/mL,n=312(52.4%) and ≥40ng/mL,n=63(10.6%). The main outcome measure was pregnancy outcomes as clinical pregnancy and live birth rate. We compared pregnancy outcomes according to age, BMI, blastocyst expansion, trophectoderm and inner cell mass grade and progesterone cut-off levels . Main results and the role of chance The median age of the patients was 31 (Range 20-46) and the mean body mass index (BMI) of the patients was 24.5±4.0 kg/m2. The overall clinical pregnancy rate and live birth rate were 61.8% (368/595) and 52.9% (315/595), respectively. Patients were subdivided into two different groups according to clinical pregnancy and live birth rate outcomes as follows; positive and negative. Age and BMI of patients were comparable between groups. Blastocyst morphology grade as expansion, trophectoderm and inner cell mass grade are statistically significantly associated with clinical pregnancy (p&amp;lt;.001 for all) and progesterone level between 20-40ng/mL is associated with higher clinical pregnancy rate (p:0.043). In multivariate analysis; only blastocyst expansion and inner cell mass grade were the independently and significantly factors that are associated with clinical pregnancy rate (p:0.011, OR: 1.6, CI95%: 1.13-2.39 and p:0.007, OR: 1.65, CI95%: 1.14-2.39, respectively). Progesterone level and trophectoderm grade were not fount statistically significant (p: 0.310 and p:0.489, respectively). In evaluation of the association between factors and live birth rate, only blastocyst expansion grade equal or over 4 and trophectoderm grade A or B were associated with statistically significant live birth rates. Limitations, reasons for caution The main limitations of current study were the retrospective design and the limited sample size of the ≥40 ng/mL cohort of serum progesterone levels. Wider implications of the findings According to this data, we speculated that if the serum P4 level is above 40 ng/mL on the day of embryo transfer in patients undergoing artificial frozen-thawed blastocyst transfer, there is no need to reduce the dose of progesterone used. Trial registration number N/A

P-275 Delayed blastocyst development is associated with a higher level of progesterone: the missing link in the chain

Abstract Study question Is there an association between progesterone (P) level on the day of trigger and blastulation rate on day 5 versus day 6? Summary answer Higher level of progesterone on the day of trigger is associated with lower blastulation rate on day 5 versus day 6. What is known already Premature progesterone elevation is known to determine a premature advanced endometrium by worsening endometrial-embryo asynchrony and affecting live birth rate during fresh cycles. To overcome the detrimental effect on the implantation window shift, freeze-all-strategy was introduced. Questions have been raised about the possible impact of progesterone also on oocyte’s quality and embryo’s euploidy rate. Studies on preimplantation genetic testing (PGT) cycles have demonstrated no differences in terms of fertilization, blastulation, euploidy and pregnancy rates. However, no studies have evaluated whether progesterone may impact blastocyst development in terms of blastulation rate. Study design, size, duration This was a retrospective cohort study with autologous IVF cycles performed at our institution from January 2019 to December 2021. 2819 IVF cycles were screened for inclusion. A total of 888 IVF cycles were included. The primary outcome was to compare blastulation rate on day5 versus day6 in terms of percentage of formed blastocysts on day5 versus day6 according to progesterone level at trigger. Participants/materials, setting, methods Patients between 18 and 42 years of age who underwent blastocyst culture of the whole cohort of embryos formed were included. Blastocyst evaluation was performed by a team of expert embryologists according to the Istanbul Consensus. We performed the Kruskal-Wallis test to assess differences between groups (blastulation rate according to progesterone level at trigger) and a multiple logistic regression analysis to evaluate association between the presence/absence of full blastocyst on day5 and progesterone levels. Main results and the role of chance A total of 3249 blastocysts were analyzed. The blastulation rate per total number of cycles was 39.4% for day-5 blastocyst, 54.0% for day-6 blastocyst, and 6.6% for day-7 blastocyst. After dividing progesterone level in quartiles (Q1, P &amp;lt; 0.51 ng/ml; Q2, 0.51 ng/ml ≤ P &amp;lt; 0.82 ng/ml; Q3, 0.82 ng/ml ≤ P &amp;lt; 1.21 ng/ml; Q4, P ≥ 1.21 ng/ml), we observed a progressive delay in blastocyst development according to the increasing level of progesterone at trigger (with a median blastulation rate on day5 of 50.00% for Q1 versus 40.00% for Q2, 30.00% for Q3 and 30.00% for Q4, p &amp;lt; 0.001). After adjusting for confounding factors, the the probability to observe at least one full blastocysts on day5 was significantly reduced for Q3 (OR = 0.44, CI 0.28-0.70, p &amp;lt; 0.001) and Q4 (OR = 0.31, CI 0.19-0.48, p &amp;lt; 0.001) compared to Q1 (reference). Limitations, reasons for caution The retrospective nature of the study and potential variability between our study center’s laboratory protocol(s) and blastocyst culture medium used compared to those of other reproductive centers may limit the external validity of our findings. A well-designed multicenter study is needed to further extend and validate our results. Wider implications of the findings This study suggests that progesterone level on the day of induction may affect blastocyst development. Thus, embryologists should not be worried by a slow growing embryo and should expect different blastulation rate on day 5/6 according to progesterone level. Trial registration number Not applicable

O-241 Elevated serum progesterone levels before frozen embryo transfer do not negatively impact reproductive outcomes: a large retrospective cohort study

Abstract Study question Do patients with high serum progesterone levels before frozen embryo transfer (FET) under hormonal replacement therapy (HRT) present worse reproductive outcomes? Summary answer Elevated serum progesterone levels before FET in artificially prepared cycles with vaginal or vaginal plus subcutaneous progesterone do not impair reproductive outcomes. What is known already Low serum progesterone levels before FET do negatively affect reproductive outcomes in terms of live birth rate. However, there is not robust data regarding the impact of high serum progesterone levels in the luteal phase of patients who undergo HRT for FET. Study design, size, duration Retrospective cohort study of 3183 blastocyst FET cycles under HRT performed in a university-affiliated fertility centre between March 2009 and December 2020. All the cycles presented adequate serum progesterone levels before FET (≥10.6 ng/ml). A total of 1360 cycles corresponded to frozen homologous embryo transfer (ET) (hom-FET), 1024 were euploid ET (eu-FET) after preimplantational genetic testing for aneuploidies (PGT-A), and 799 cycles were frozen heterologous ET (het-FET). The primary objective was live birth rate (LBR). Participants/materials, setting, methods Standard HRT was used. Luteal phase was covered with vaginal progesterone 200 mg/8h, or vaginal plus a daily subcutaneous injection of progesterone (25 mg). Serum progesterone levels were measured the day before FET. Elevated progesterone levels were considered in the 90th and 95th centiles. A generalized additive model (GAM) was performed to study the functional relationships between progesterone and LBR. A multivariable logistic regression was used to evaluate the effect of high progesterone over LBR. Main results and the role of chance Mean serum progesterone level before FET was 16.77±8.43 ng/ml. Progesterone levels were significantly higher in the group under vaginal plus subcutaneous progesterone (21.87±14.17 vs. 15.56±5.72, p &amp;lt; 0.001). No differences in clinical pregnancy, miscarriage and LBR were found according to the use of vaginal or vaginal plus subcutaneous progesterone for each of the groups (hom-FET, eu-FET and het-FET). Live birth rates were comparable among patients in the highest centile of serum progesterone levels (≥p90, ≥22.33 ng/ml) and the rest of patients (p &amp;lt; 90) (43.9 vs 41.3%; p = 0.381). Patients with progesterone levels ≥p90 presented lower BMI compared to those in the lower centiles (&amp;lt;p90) (22.62±3.82 vs. 23.32±4.06; P = 0.009). After dividing patients in deciles according to serum progesterone levels before, no differences in LBR were observed among groups (P = 0.938). No association was observed with GAM model between progesterone levels and LBR. A multivariable logistic regression adjusted by oocyte age, type of treatment and number of embryos transferred was applied for centile 90 and centile 95 of progesterone, and showed that serum progesterone in their highest levels did not negatively impact LBR. Limitations, reasons for caution The main limitation of this study is its retrospective design. The results only apply for patients under HRT with vaginal micronized progesterone alone or plus subcutaneous progesterone. Progesterone determination was measured before blastocyst FET. Extrapolation to other HRT protocols or timings of progesterone measurement needs to be validated. Wider implications of the findings The results of this study suggest that once a threshold of serum progesterone before FET is achieved, progesterone levels are not predictive of the clinical outcome. Actually, LBR are not negatively affected when progesterone levels are found in their highest centiles after luteal phase rescue with vaginal plus subcutaneous progesterone. Trial registration number Not applicable

O-217 Associations between endometrial stem cells quantities, serum levels of progesterone and estradiol and expression of their respective receptors in endometrial tissue during the mid-luteal phase

Abstract Study question Is the percentage of endometrial stem cells linked to serum progesterone, serum estradiol, and their receptors expression in human endometrium during the mid-luteal phase? Summary answer We found an association between CD117+ endometrial stem cells and serum progesterone, estradiol, as well as with the expression of their receptors in the endometrium. What is known already Various types of stem cells including NOTCH1+ and CD117+ stromal cells have been found in the endometrium. Their quantities have been hypothesised to fluctuate with the menstrual cycle and to have a link with endometrial receptivity. It has been well established that progesterone and estradiol play a crucial role in the preparation of the endometrium for implantation and the expression of their receptors also varies with the menstrual cycle. Nonetheless, the relationship between stem cells and other established markers of endometrial receptivity, such as the sex hormones signaling system, constitutes a novel field of research. Study design, size, duration The present observational study included 87 women aged 26-56 (mean 39.5 years old) undergoing in-vitro fertilisation and for whom a peripheral blood sample and an endometrial biopsy were obtained during the mid-luteal phase (LH + 7) of a natural cycle. Immunohistochemical (IHC) markers for NOTCH1 (E-AB-12815, Elabscience), CD117 (CD117/c-Kit/SCF-Receptor RB-9038-RQ, Epredia), progesterone (PR) and estrogen receptors (ER) (1-PR026-07, 1-ES006-07, Quartett) were used to stain endometrial tissue. The investigation was carried out between March 2020 and January 2022. Participants/materials, setting, methods Serum progesterone and estradiol were measured via electrochemiluminescence using Cobas e411 analyser (Roche Diagnostics, Germany) on the day of obtaining the endometrial biopsy. Stem cells (count and percentage) and the expression of hormone receptors (6-point scale) were determined using ImageJ. Stem cells percentage was compared between groups with high and low progesterone (analogously for estradiol) using Wilcoxon signed-rank test. To assess the link between stem cell percentages and receptor expression, Spearman correlation was used. Main results and the role of chance The mean percentage of positively stained NOTCH1 endometrial stromal stem cells was 0.13 ± 0.29% and CD117+ cells was 0.034 ± 0.039%. The hormone serum levels ranged from 0.2 to 60 ng/mL (median of 14.9 ng/mL) for progesterone and from 12 to 1985 pg/mL (median of 135.6 pg/mL) for estradiol. While the quantities of the two stem cell types demonstrated a positive correlation with each other (R = 0.379, p &amp;lt; 0.001), the percentage of NOTCH1+ stem cells showed no difference between the studied serum hormone levels (p &amp;gt; 0.05), nor revealed a relationship with the expression of their receptors (p &amp;gt; 0.05). In contrast, the amount of CD117+ cells differed significantly between patients with high and low serum progesterone (cutoff 14.9 ng/mL, p = 0.001) and estradiol levels (cutoff 135.6 pg/mL, p = 0.019). Furthermore, the quantity of stem cells positive for CD117 correlated positively with the two hormone receptor levels in the endometrium – PR (R = 0.277, p = 0.019) and ER (R = 0.318, p = 0.007). Limitations, reasons for caution A limitation of the study was the sample size. These findings should be confirmed using a larger population. In addition, confounding variables such as the immunological profile of the tissue should also be taken into account. Wider implications of the findings This study shows that certain endometrial stem cell types (CD117+) but not others (NOTCH1+) are associated with sex hormone signaling during the mid-luteal phase. This relationship highlights the role these cells play in preparing the endometrium for embryo implantation and their quantity might be an indirect indicator of endometrial receptivity. Trial registration number not applicable