Number Of Preovulatory Follicles Research Articles

P-631 Extended letrozole regimen versus routine letrozole regimen in women with polycystic ovary syndrome undergoing their first ovulation induction cycle

Abstract Study question Whether an extended letrozole regimen could result in a higher ovulatory rate than a routine regimen in PCOS women undergoing their first ovulation induction cycle. Summary answer PCOS women using the extended letrozole regimen failed to obtain a statistically higher rate of ovulation in comparison with the routine letrozole regimen. What is known already Letrozole has become the first-line agent for ovulation induction, however, nonresponsive cycles occurred in 10-58.8% of PCOS women during their ovulation induction therapy with letrozole alone. The extended letrozole regimen has been demonstrated to be a feasible method for inducing ovulation in those nonresponders. However, whether it could be applied to all the PCOS women as a first choice in the induction of ovulation remains to be explored. Study design, size, duration This is a prospective randomized controlled trial including 148 women with PCOS undergoing their first ovulation induction cycle with letrozole from January 2021 to October 2022. Participants/materials, setting, methods Participants were randomly assigned to receive an extended letrozole regimen (5 mg letrozole daily for 7 days) or a routine regimen(5 mg letrozole daily for 5 days) for one treatment cycle. Ovulation rate was the primary outcome. Secondary outcomes included the clinical pregnancy rate, the number of preovulatory follicles, the rate of multiple pregnancies. Main results and the role of chance The ovulation rate among women receiving a 7-day regimen was slightly higher than the rate with a 5-day regimen, but it did not reach a statistical significance in both the intention-to-treat analysis (90.54%[67/74]vs. 82.43%[59/74], P = 0.065) and per-protocol analysis (90.54%[67/74]vs. 84.29%[59/70], P = 0.257). The number of preovulatory follicles was nearly identical in the two groups (1.39±0.62 vs.1.37±0.59, P = 0.956), and no cases of ovarian hyperstimulation syndrome were observed. In the per-protocol analysis, the rates of clinical pregnancy (20.27%[15/74]vs.16.95%[10/70], P = 0.343) and live birth (13.51%[10/74]vs.11.43%[8/70], P = 0.976) did not differ significantly between treatment groups. Moreover, all the conceptions were singletons without neonatal defects. Limitations, reasons for caution The major concern of the current research is its single-centre and open label nature. Additionally, the limited number of lean PCOS women with a mean body mass index of 23-25 kg/m2 enrolled in our trial also restrict the generalizability of our findings. Wider implications of the findings Letrozole 5-day regimen remains to be the first choice for PCOS women undergoing ovulation induction therapy. Additional prospective trials with a larger sample size and different subgroups of PCOS are needed to assess the ovulatory effects of different letrozole treatment duration. Trial registration number ChiCTR2100042082

P-619 AMH/AFC ratio as a predictive factor for ovarian responsiveness to gonadotropins during ovarian stimulation

Abstract Study question Does AMH/AFC ratio correlate to the number of follicles on the day of trigger and to the number of oocytes retrieved after ovarian stimulation? Summary answer The AMH/AFC ratio correlates significantly to the number of pre-ovulatory follicles and the number of oocytes collected after ovarian stimulation (OS). What is known already Although AFC is a robust marker of oocyte yield after OS, its combination with Anti-Mullerian hormone (AMH) seems to improve the predictability. Besides, Follicular Output Rate [FORT: (number of pre-ovulatory follicles on day of trigger/AFC at baseline) x 100] and Follicle-to-Oocyte Index [FOI: (number of retrieved oocytes/AFC) × 100] evaluate the ovarian responsiveness to gonadotropins during OS, representing its sensitivity to exogenous FSH. Both parameters take antral follicle count (AFC) only into consideration. AMH and AFC as a ratio might optimize the accuracy of FORT and FOI metrics, helping to predict which patients are at higher risk of suboptimal results. Study design, size, duration This retrospective study analyzed 2,163 ovarian stimulation cycles for IVF/ICSI, performed between May 2015 and October 2023 in a tertiary referral IVF centre. For all the cycles, measurement of AMH using Elecsys (Roche®) no longer than 6 months before starting OS, AFC at baseline, number of pre-ovulatory follicles on the day of trigger, and the treatment outcomes were available. Participants/materials, setting, methods Couples with primary/secondary infertility who underwent OS with antagonist protocol, starting on day 2/3 for IVF/ICSI were included, independent of their age or ovarian reserve parameters. Exclusion criteria were patients with body mass index (BMI) >40 kg/m2, non-vaginal ultrasound measurement for AFC and/or follicles during OS, cycles with previous estradiol priming or oral contraceptives. Ethical approval was obtained from the Research Ethics Committee. Main results and the role of chance 2,163 ovarian stimulation cycles were included in the analysis. The mean age, BMI, AMH, AFC, FOI and FORT of the included women were: 35.6±6.1 years, 27.5±4.7 kg/m2, 2.4±2.1 ng/mL, 12±7 follicles, 1.02±0.66 and 1.38±0.79. The correlation of AMH/AFC ratio (coefficient: 0.62, 95% CI: 0.59 to 0.65, p<0.001) to FOI was stronger than the correlation of AMH (coefficient: 0.31, 95% CI: 0.27 to 0.34, p<0.001) or AFC alone (coefficient: -0.09, 95% CI: -0.14 to -0.05, p<0.001). The correlation of AMH/AFC ratio (coefficient: 0.62, 95% CI: 0.59 to 0.64, p<0.001) to FORT was also stronger than the correlation of AMH (coefficient: 0.21, 95% CI: 0.16 to 0.25, p<0.001) or AFC alone (coefficient: -0.23, 95% CI: -0.27 to -0.19, p<0.001). AMH/AFC ratio was better at predicting stimulation efficiency parameters, FOI and FORT, compared to joint use of AMH and AFC (p>0.001 for both). However, regression analysis showed stand-alone use of the AMH/AFC ratio did not provide a better prediction for collected oocyte count compared to joint use of AMH and AFC. In fact, prediction errors were significantly larger for the AMH/AFC ratio (median: -0.4, range: -11 to 11) compared to the model using AMH and AFC separately (median: -0.2, range: -6 to 7). Limitations, reasons for caution Although clinical assessment was performed in the same centre following the same methodology, inter-observer variability for AFC is a limitation. Additionally, AMH levels used in the study were not obtained in the same cycles, which may adversely affect the performance of the ratio. Wider implications of the findings While AMH/AFC ratio was better at predicting stimulation efficiency compared to FOI and FORT parameters, it was not significantly better at predicting absolute oocyte yield. This may imply AMH/AFC ratio can represent the ovarian sensitivity to exogenous FSH. Trial registration number Not applicable

The average gonadotrophin dosage per follicle is predictive of ovarian response and cumulative live birth chances after in vitro fertilization: a retrospective cohort study

BackgroundWith the development of assisted reproduction technology (ART), many indicators have been proposed to evaluate ovarian response, and then predict pregnancy outcomes. In general, the predictive values remain limited.ObjectiveTo further explore the indicators to evaluate ovarian sensitivity to gonadotrophin (Gn) stimulation more accurately.MethodsThis retrospective cohort study included 330 women who underwent an entire ART cycle. We aimed to assess whether a new index, termed as average Gn dosage per follicle, could be used as a marker for ovarian response and pregnancy outcomes. It was calculated as the ratio of total Gn dose during ovarian stimulation and the number of pre-ovulatory follicles (PFC) on the trigger day. Patients were divided into three subgroups according to the average Gn dosage per follicle: below the 33rd percentile (Group A), between 33rd and 67th percentiles (Group B), and above the 67th percentile (Group C). Then stimulation data, laboratory and clinical outcomes were compared among the groups.ResultsThe results showed patients in Group A had the best ovarian response, the number of retrieved oocytes was significantly higher than in Group B and C. A multivariate regression analysis showed that average Gn dosage per follicle was an independent predictor of cumulative live birth rates (CLBRs) [adjusted odds ratio (OR): 0.96, 95% confidence interval (CI): 0.95–0.98, P < 0.01].ConclusionsThe present study showed that average Gn dosage per follicle appears to be a highly reliable index of ovarian response to exogenous Gn and can be useful to estimate CLBR.

Evaluation of correlation between antral follicle diameters with Follicular Output Rate (FORT) in women under controlled ovarian hyperstimulation for assisted reproductive techniques

Abstract Background This study aimed to determine the association of diametrical antral follicles with the ovarian response by Follicular Output Rate (FORT) ratio in 100 females undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). This study selected 100 women undergoing controlled ovarian hyperstimulation (COH). The number of antral follicles (diameter of 3–10 mm) was calculated with two-dimensional vaginal sonography on days 1–3 of the cycle. Then, on a triggering day with human chorionic gonadotropin (HCG), the number of follicles (with a diameter of 16–22 mm) and the ratio of FORT were determined. The correlation among FORT with age, antral follicle count (AFC), AFC ≤ 5, AFC &gt; 5, number of preovulatory follicles (16–20 mm), number of metaphase II (MII) oocytes, body mass index (BMI), infertility period, and anti-Mullerian hormone (AMH) was assessed. Results There was a significant correlation between FORT and total AFC, AFC &gt; 5, number of preovulatory follicles (16–20 mm), and number of MII oocytes retrieved. There is no significant relationship between FORT and examined two variables (AMH and AFC ≤ 5). Multiple linear regression analysis showed no significant relationship between FORT and examined two variables (AMH and AFC &gt; 5). There was a significant correlation between MII oocytes retrieved and age, total AFC, AFC ≤ 5, AFC &gt; 5, number of preovulatory follicles (16–20 mm), and AMH. A significant positive relationship existed between MII oocytes retrieved and examined two variables (AFC ≤ 5 and AFC &gt; 5). Conclusion There was not much difference in the correlation between the AFC ≤ 5 and AFC &gt; 5, and both positively correlated with the number of MII oocytes retrieved.

Superovulatory response, anti-Müllerian hormone concentration and antral follicle count in Holstein cattle with short or long anogenital distance

To understand the mechanisms underlying the inverse relationship between anogenital distance (AGD) and fertility in dairy cattle, we determined if embryo yield and quality differed between cattle of short- and long-AGD (Study 1), and whether anti-Müllerian hormone (AMH), antral follicle count (AFC) and superovulation responses differed by AGD group (Study 2). In Study 1, records of embryo yield and quality in Holstein heifers (n = 43) and cows (n = 14) from two commercial dairy herds were analyzed, retrospectively. Short- and long-AGD groups were based on the median AGD cut-points of 113 and 134 mm, for heifers and cows, respectively. The mean numbers of total (7.4 vs. 9.0; 9.4 vs.12.9), fertilized ova (5.6 vs. 6.2; 6.6 vs. 6.9), and viable embryos (4.4 vs. 4.8; 5.9 vs. 5.3) per heifer and cow did not differ between cattle of short vs. long AGD. Short-AGD cows, however, had greater proportions of fertilized ova (69.7 vs. 53.3%; P = 0.07) and viable embryos (62.1 vs. 41.1%; P = 0.03) than long-AGD cows. The odds of short-AGD cows yielding fertilized ova and viable embryos were 2.0 and 4.0 times greater, respectively, than the odds of long-AGD cows. In Study 2, lactating cows (n = 24) of a research herd had their AGD categorized as in Study 1, and AFC, AMH, and superovulation responses (i.e., no. of preovulatory follicles [≥ 10 mm] and CL) were determined. Mean AFC (27 ± 4.5 vs. 21 ± 4.0) and CL (9 ± 1.6 vs. 7 ± 1.4) per cow did not differ between short- and long-AGD groups. Serum AMH concentration (pg/mL) was lesser in short-AGD cows compared with long-AGD cows (114 ± 30.4 vs. 200 ± 26.8; P = 0.05), but the no. of preovulatory follicles was greater (15 ± 1.9 vs. 8 ± 1.7; P = 0.01) in short-AGD than in long-AGD cows. The overall proportion of follicles ≥10 mm was also greater in short-AGD cows than in long-AGD cows (56 vs 44%; P = 0.03). Preovulatory follicle number was affected by a parity x AGD group interaction (P = 0.04), with multiparous short-AGD cows accounting for the largest number of follicles. No associations were found among AGD, AFC and AMH. The associations between AFC and superovulation responses (follicles: r = 0.67, and CL: 0.58; P < 0.01) were moderate but AMH was not associated with superovulation responses. In summary, whereas AGD-associated differences in the yield and quality of embryos were not evident in heifers, the proportions of fertilized ova and viable embryos were greater in short-AGD than in long-AGD cows. In addition, the proportion of preovulatory follicles, an indicator of superovulatory response, was greater in short-AGD cows than in long-AGD cows.

Immuno-Neutralization of Follistatin Bioactivity Enhances the Developmental Potential of Ovarian Pre-hierarchical Follicles in Yangzhou Geese.

Simple SummaryFollistatin involves in the regulation of ovarian follicular development in mammals; however, the role of follistatin in goose ovarian follicular development has not been investigated. In this study, following immuno-neutralization of follistatin bioactivity in geese, the number of ovarian pre-ovulatory follicles significantly increased, and mRNA levels of genes involved in ovarian steroidogenesis and yolk deposition were upregulated in the granulosa layer of pre-hierarchical follicles. These results suggest that follistatin plays a limiting role in the development of ovarian pre-hierarchical follicles into pre-ovulatory follicles. These results also expand our understanding of the mechanism of follistatin on ovarian follicular development in geese.In order to explore the role of follistatin (FST) in ovarian follicular development and egg production in Yangzhou geese, sixty-four egg laying geese of the same genetic origin were selected and divided into two groups with equal numbers. One group was immunized against the recombinant goose FST protein by intramuscular injection, whereas the control group received bovine serum albumin (BSA) injection. Immunization against FST significantly increased the number of pre-ovulatory follicles. Furthermore, immunization against FST upregulated Lhr, Star, Vldlr, Smad3, and Smad4 mRNA levels in the granulosa layer of pre-hierarchical follicles. The results suggest that FST plays a limiting role in the development of ovarian pre-hierarchical follicles into pre-ovulatory follicles by decreasing follicular sensitivity to activin in geese. The mechanism may be achieved by regulating the SMAD3 signaling pathway, which affects progesterone synthesis and yolk deposition in pre-hierarchical follicles.

O-299 Analyzing the impact of simple wash versus density gradient sperm preparations on intrauterine insemination outcomes. Is the cost-effective and time-efficient option just as good?

Abstract Study question How do different sperm preparations, such as simple-wash (SW) and density-gradient (DG), impact intrauterine insemination (IUI) outcomes among women seeking fertility treatments? Summary answer Using a population from a large, academic medical center, this study discerned no differences in pregnancy outcomes between SW and DG sperm preparations. What is known already IUI with or without ovulation induction (OI) is often a first-line treatment among couples seeking fertility services. SW and DG are two common methods used to prepare sperm for IUI. In comparison to its latter counterpart, the SW technique is lesser-used, yet is more time-efficient, and cost-effective due to its utilization of only a single centrifugation step. Since the impact of sperm preparation techniques on the post-processing sperm yield and its parameters varies by the method used, the cycle outcomes might differ as well. However, limited data exists on various sperm preparations’ impact on IUI clinical outcomes. Study design, size, duration Data from 3378 IUI+OI cycles (from 1503 women of all diagnoses seeking IUI with fresh-ejaculated sperm) that took place at a large academic fertility center between 9/2014 and 3/2021 were retrospectively reviewed. Cycles were either unstimulated (natural) or stimulated with either oral OI agents (clomiphene-citrate &amp; letrozole) or gonadotropins. Cycles were divided in two groups based on sperm preparation technique: SW (n = 1691) and DG (n = 1687) and outcomes were compared between them. Participants/materials, setting, methods Sperm preparation: SW semen were mixed in 10ml MHM (FujiFilm) and centrifuged for 10min. DG semen were layered over 45:90 gradient of Isolate (FujiFilm), centrifuged for 20min, and washed twice (10min) in 10ml MHM. Outcome measures: hCG-positivity (posHCGR), clinical pregnancy (CPR), spontaneous abortion (SABR), and livebirth rates/cycle (LBR). Statistics: Logistic regression with Odds Ratios (OR) adjusted for both partners’ ages, day-3 FSH, stimulation, and sperm score (poor, fair, good, excellent). Sub-analysis limited cohort to first-cycles only. Main results and the role of chance Groups were comparable in patient [age (maternal, paternal), BMI, day-3 FSH, infertility diagnosis], and cycle characteristics [follicular response (measured as number of preovulatory follicles), and endometrial thickness]. Preprocessing sperm parameters differed slightly with higher mean sperm concentrations and lower total motility among SW cycles (75.3 + 57.0 vs. 71.0 + 51.3 million, p = 0.02; 48.6 + 19.6 vs. 52.5 + 20.2, p &amp;lt; 0.001, for SW and DG, respectively). posHCGR, CPR, SABR, and LBR per cycle did not differ between groups (15.8% vs. 15.4%, p = 0.76;13.7% vs. 13.2%, p = 0.62;18.1% vs. 18.5%, p = 0.93; 9.5 vs. 8.9%, p = 0.56; for SW and DG, respectively). Odds for posHCG, CP, SAB, or LB did not differ between groups [adjOR(95%CI): 1.05(0.87-1.26), p = 0.65; 1.10(0.67-1.83), p = 0.71; 0.98(0.60-1.60), p = 0.94; 1.08(0.85-1.37), p = 0.66, respectively]. When cycles were stratified by type of ovarian stimulation, rather than adjusted for it, no difference was seen in any of the clinical outcomes within individual strata{adjOR(95%CI): [Oral OI: 1.00(0.74-1.37), p = 0.98; 1.78(0.68-4.61), p = 0.25; 0.97(0.40-2.38), p = 0.95; 1.05(0.72-1.53), p = 0.81], [Gonadotropins: 0.99(0.78-1.28), p = 0.96; 0.93(0.49-1.77), p = 0.83; 0.97(0.52-1.80), p = 0.96; 1.03(0.75-1.41), p = 0.87], [Natural: 2.36(0.97-5.76), p = 0.06; 0.08(0.001-6.84), p = 0.26; 0.20(0.003-11.02), p = 0.43; 2.52(0.63-10.00), p = 0.19], for posHCG, CP, SAB, and LB, respectively}. Similarly, no difference was seen in any of the clinical outcomes when cycles were stratified by sperm score or when analysis was limited to first-cycles only. Limitations, reasons for caution LBR were calculated excluding pregnancies with no information after discharge to obstetrics (approximately 16%). Although not significant, there might be minor variations in individual provider’s practices between time frames in which these techniques were implemented. Wider implications of the findings SW is a much simpler, time-efficient, and cost-effective sperm processing technique for IUI compared to DG, however remains infrequently utilized. Adoption of SW, over DG, could yield comparable clinical efficacy, yet optimize teamwork flow and lower healthcare costs, due to its non-labor-intensive and inexpensive nature. Trial registration number not applicable

O-069 Does dual stimulation with continuous FSH administration increase the total number of metaphase II oocytes in poor responders?

Abstract Study question Is there an increase in the total number of metaphase II (MII) oocytes between a conventional ovarian stimulation and a dual uninterrupted stimulation? Summary answer Dual stimulation with continuous follicle stimulating hormone (FSH) administration is not superior to conventional stimulation in terms of number of MII oocytes. What is known already In the last decade, the concept of multiple follicular waves during one menstrual cycle has gained a lot of interest. Translated into clinical practice, combining two stimulations in the same ovarian cycle has appeared beneficial with regard to the number of retrieved oocytes, without affecting the embryo quality or ploidy status. Usually, this so-called dual stimulation approach is characterized by a discontinuation of FSH administration for approximately 5 days between two consecutive stimulations. The role of dual uninterrupted ovarian stimulation has not yet been studied. Study design, size, duration This was an open-label randomized controlled trial (RCT) with superiority design, performed in a single tertiary center. Power analysis indicated a required sample of 46 patients to detect superiority (defined as two surplus MII oocytes) of the dual stimulation group. Between October 2020 and September 2021, 70 patients were screened, 48 were randomized and 46 completed the study. Participants/materials, setting, methods Women aged 25-40 with a serum anti-Müllerian hormone (AMH) level of ≤ 1.5 ng/mL, antral follicle count (AFC) of ≤ 6, or ≤ 5 oocytes after a previous stimulation, were eligible for inclusion. Randomization occurred only in case of ≤ 9 follicles of ≥ 11mm on the trigger day. In the control group, patients underwent one round of ovarian stimulation and oocyte retrieval only, while the study group had two uninterrupted rounds of ovarian stimulation and two retrievals. Main results and the role of chance Baseline characteristics were similar between both groups. The cumulative number of COC and MII oocytes after completion of the second oocyte retrieval was similar in the control and study group [5.3 ± 2.7 versus 5.3 ± 3.1 [difference 95%CI (-1.7 to 1.7), p = 0.92] and 4.1 ± 2.5 versus 4.3 ± 2.7 [difference 95%CI (-1.7 to 1.3), p = 0.82]. Likewise, a comparable number of good quality embryos at day 3 was available (3.0 ± 2.0 versus 2.7 ± 2., p = 0.63). In the study group, the cancellation rate due to insufficient response to the second round of stimulation was 39.1%. When focusing on the first stimulation in both groups, there were no significant differences regarding basal FSH, FSH consumption and the number of preovulatory follicles. After the first oocyte retrieval, the mean number of COC and MII oocytes was significantly higher in the control group (who had human chorionic gonadotropin (hCG) triggering), compared to the study group (who had Gonadotropin Releasing Hormone (GnRH) agonist triggering) (5.3 ± 2.7 versus 3.3 ± 2.2, p = 0.004 and 4.2 ± 2.4 versus 2.9 ± 2.2, p = 0.05). Likewise, the number of good quality embryos on day 3 was significantly higher (3.0 ± 2.0 versus 1.9 ± 1.7, p = 0.04). Limitations, reasons for caution This study was powered to demonstrate superiority for number of MII oocytes. Investigating the impact of dual stimulation on pregnancy rates would have required a larger sample size. Wider implications of the findings The observed suboptimal oocyte yield after agonist triggering in poor responders is a reason for concern and further scrutiny, given that previous RCTs have shown similar outcomes in normal and high responders after hCG and GnRH agonist trigger. Trial registration number NCT03846544

Association of AMH and AMHR2 gene polymorphisms with ovarian response and pregnancy outcomes in Indian women.

To evaluate the association of single-nucleotide polymorphisms (SNPs) in the anti-Müllerian hormone (AMH) and AMH type II receptor (AMHR2) genes with ovarian response and clinical pregnancy outcomes in women undergoing controlled ovarian hyperstimulation. In this prospective study, we genotyped AMH polymorphisms (c. -649T > C, c. 146T > G, c. 252 G > A, and c. 303 G > A) in 365 women and AMHR2 polymorphisms (c. -482 A > G, c. 622-6 C > T, c. 4952 G > A, c. 10 A > G) in 80 women undergoing controlled ovarian hyperstimulation for IVF. Higher doses of exogenous FSH and lower numbers of preovulatory follicles were noted in women having AMH c. -649T > C and AMH c. -146T > G polymorphisms, respectively. Overall, we found that the presence of a polymorphic genotype (homozygous or heterozygous) at positions c. -649T > C, c. 146T > G, c. 252 G > A, and c. 303 G > A in the AMH gene was associated with higher doses of FSH for ovulation induction (p < 0.001). Interestingly, a higher live birth rate was noted in women with a homozygous polymorphic genotype for all four AMH SNPs investigated while none of the women showing a homozygous polymorphic genotype at all AMHR2 SNPs investigated in this study had a live birth. Our results show that presence of AMHR2 SNPs (c. 482 A > G, c. 622-6 C > T, c. 4952 G > A, and c. 10 A > G) negatively correlate with live birth rate. However, these findings need to be validated by using larger sample size.

Serum Scavenging Capacity and Folliculogenesis Impact following Flaxseed Consumption in the First-Generation Mice Pups.

Flaxseed is a source of antioxidants utilized for female infertility treatment in traditional medicine. This study investigated the effects of flax hydroalcoholic extract and flaxseeds during prenatal and postnatal (PND) periods on folliculogenesis and serum total antioxidant capacity (TAC). Pregnant NMRI mice received 500 and 1000 mg/kg of flax extract (LE) and the same doses of flaxseed (LS). Female pups received the same regimen for 56 days. The body, ovarian morphometry, follicle development, and TAC levels were evaluated. The ovarian weight significantly increased in the LE1000 group compared to the LS500 group. The LE500 group had a considerably lower number of primary and antral follicles compared to the CTL and LS1000 groups. The number of antral follicles significantly increased in the LE1000 group compared to the LS500 and LE500 groups. The number of preovulatory follicles was higher in the LE1000 group. A significant increase in the TAC levels was detected in the LS500, LS1000, and LE1000 groups. LE showed a dose-dependent protective effect on the folliculogenesis in F1, which is more evident with the dosage of 1000 mg/kg. This could be related to the strongest antioxidant property of LE1000, as shown by the highest levels of TAC.

Bisphenol S Alters the Steroidome in the Preovulatory Follicle, Oviduct Fluid and Plasma in Ewes With Contrasted Metabolic Status.

Bisphenol A (BPA), a plasticizer and endocrine disruptor, has been substituted by bisphenol S (BPS), a structural analogue that had already shown adverse effects on granulosa cell steroidogenesis. The objective of this study was to assess the effect of chronic exposure to BPS, a possible endocrine disruptor, on steroid hormones in the ovary, oviduct and plasma using the ewe as a model. Given the interaction between steroidogenesis and the metabolic status, the BPS effect was tested according to two diet groups. Eighty adult ewes were allotted to restricted (R) and well-fed (WF) groups, that were further subdivided into two subgroups. Ewes were exposed to 50 µg BPS/kg/day in their diet (R50 and WF50 groups) or were unexposed controls (R0 and WF0 groups). After at least 3 months of BPS exposure, preovulatory follicular fluid, oviduct fluid and plasma were collected and steroid hormones were analyzed by gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). A deleterious effect of restricted diet on the volume of oviduct fluid and numbers of pre-ovulatory follicles was observed. Exposure to BPS impaired estradiol concentrations in both follicular and oviduct fluids of well-fed ewes and progesterone, estradiol and estrone concentrations in plasma of restricted ewes. In addition, a significant interaction between metabolic status and BPS exposure was observed for seven steroids, including estradiol. In conclusion, BPS acts in ewes as an endocrine disruptor with differential actions according to metabolic status.

Are There Ovarian Responsive Indexes That Predict Cumulative Live Birth Rates in Women over 39 Years?

Objective: Ovarian response indexes have been proposed in assisted reproductive technology (ART) in order to optimize live birth rates (LBR), adjusting ovarian stimulation (OS), and minimizing risks. Gonadotropin doses are commonly adjusted according to ovarian reserve parameters, including antral follicle count (AFC), anti-Mullerian hormone (AMH), and basal follicle stimulating hormone (FSH) levels. The retrospective assessment of ovarian responses allows one to identify three primary indexes: (i) follicular output rate (FORT), the ratio of the number of pre-ovulatory follicles obtained at OS completion over AFC; (ii) follicle oocyte index (FOI), the ratio of oocytes retrieved over AFC; (iii) ovarian sensitivity index (OSI), the ratio of oocytes retrieved over the total gonadotropin dose administered. In recent publications, these indexes were reported to predict ART outcome. In the present study, we assessed the ability of these indexes to predict cumulative ART outcome in women ≥39 years. Materials and Methods: Retrospective cohort study. All patients ≥39 years who performed their first ART cycle with an antagonist protocol in our center between 01/2018 and 04/2020 were included. Patients with basal FSH > 20 IU/l, AMH < 0.1 ng/mL and severe male factors (azoospermia with testicular biopsy) were excluded. All patients received both recombinant FSH and human menopausal gonadotropin (hMG). Cumulative live birth rate (cLBR) was the primary outcome. Secondary outcomes included: the number of MII oocytes, cumulative implantation (cIR), and usable blastulation rates. Logistic regressions were performed to assess the predictive values of FORT, FOI, and OSI in cLBR and embryo culture success. For each parameter, the ability of the logistic regression models to predict embryo culture success was quantified by the area under the ROC curve (AUC). Only the significant findings related to FORT, FOI, and OSI were included in the multiple logistic regression model. Linear regression models were performed between cIR, cLB, FORT, FOI, and OSI. Each statistic model was adjusted for age. Concerning OR for OSI, values were multiplied *100 due to the very low value. Results: 429 patients met the inclusion criteria. There were 298 obtained usable blastocysts after ART treatment. Age-adjusted OSI was significantly associated with cLBR [OR = 17.58 95% CI (5.48–56.40), AUC = 0.707 95% CI (0.651–0.758)) and cIR (beta = 30.22 (SE: 7.88), p < 0.001, R2= 0.060). Both FOI (OR = 6.33 95% CI (3.27–12.25), AUC = 0.725 95% CI (0.675–0.771), R2 = 0.090, p < 0.001) and OSI (OSI*100; OR = 1808.93 95% CI (159.24–19,335.13), AUC = 0.790 95% CI (0.747–0.833), R2 = 0.156, p < 0.001) were independently, when age adjusted, associated with embryo culture success. OSI showed a main performance to explain successful embryo culture than FOI (R2 = 0.156 vs. R2 = 0.090, p < 0.001). In the age-adjusted linear regression model, FOI (R2 = 0.159, p < 0.001), OSI (R2 = 0.606, p < 0.001), and FORT (r2 = 0.030, p < 0.001) were predictive of the number of MII oocytes collected. Furthermore, for OSI (r2 = 0.759, p < 0.001) and FOI (r2 = 0.297, p < 0.001), the correlation with the number of metaphase II oocytes collected was significantly higher in the non-linear regression model. Conclusions: Our findings suggest that the best index, among those analyzed, to predict cIR and cLBR, is OSI. Both OSI and FOI predict embryo culture with success, but OSI is more accurate. OSI, FOI, and FORT are significantly related to the number of MII oocytes obtained.

Differences in egg yolk precursor formation of Guangxi Ma chickens with dissimilar laying rate at the same or various ages

Egg yolk formation and deposition are quite vital for oocyte maturation and egg production in hens. There are great differences in egg production and number of preovulatory follicles among individuals in Guangxi Ma chickens at the same or various ages, however, the causes of this phenomenon remain unclear. This study aimed to elucidate the differences in yolk precursors formation among Guangxi Ma hens at the same or various ages with dissimilar egg laying rate via comparing the synthesis and transportation of yolk precursors, serum estradiol (E2) levels, as well as the antioxidant capacity of the liver and serum from hens of 32-week-old (32W), 50-week-old with higher laying rate (50WH), 50-week-old with lower laying rate (50WL), 72-week-old with higher laying rate (72WH), and 72-week-old with lower laying rate (72WL). The results showed that the contents of triglyceride (TG), total cholesterol (T-CHO), high density lipoprotein cholesterol (HDL-C) and very low density lipoprotein cholesterol (VLDL-C) in the liver, serum and ovarian stroma, serum E2 levels, expression levels of proteins and genes related to yolk precursors formation, and the antioxidant capacity of liver and serum decreased significantly during aging process. TG, HDL-C and VLDL-C contents in serum and ovarian stroma, as well as HDL-C and VLDL-C levels in liver tissues of hens in 50WL and 72WL were significantly lower compared to hens in 50WH and 72WH, respectively. Meanwhile, expression levels of estrogen receptor ɑ and transcription levels of genes related to lipid transportation in liver tissues, and the antioxidant capacity of livers were remarkably lower in hens in 50WL and 72WL than those in 50WH and 72WH, respectively. However, no significant difference was detected in liver TG levels, serum E2 levels, genes related to fatty acids synthesis and serum antioxidant capacity between hens in 50WL and 50WH. In conclusion, the age-related decline of egg production in Guangxi Ma chicken maybe related to the age-related decreases in the synthesis and transportation of liver yolk precursor as well as the antioxidant capacity of liver and hens. And the non-age related decline of egg production may mainly attribute to the decreases in yolk precursors transportation ability and hepatic antioxidant capacity.

Women in Advanced Reproductive Age: Are the Follicular Output Rate, the Follicle-Oocyte Index and the Ovarian Sensitivity Index Predictors of Live Birth in an IVF Cycle?

(1) Background: Several researchers have investigated alternative markers related to ovarian responsiveness in order to better predict IVF outcomes, particularly in advanced reproductive-aged women. The follicular output rate (FORT), the follicle-oocyte index (FOI) and the ovarian sensitivity index (OSI) are among the most promising. However, these three metrics have not been investigated as independent predictors of live birth in women of advanced reproductive age; neither have they been compared to the two ‘component’ characteristics that are used to calculate them. (2) Methods: A logistic regression model containing all relevant predictors of ovarian reserve or response was used to evaluate the potential of FORT, FOI and OSI as predictors of live birth. After, the non-linear associations between FORT, FOI and OSI and the probability of live birth were evaluated. Finally, we fitted multiple logistic regression models to compare whether FORT, FOI and OSI were more informative predictors than their components. (3) Results: 590 couples received a total of 740 IVF cycles, after which, 127 (17.5%) obtained a live birth. None of FORT, FOI and OSI showed a strength of association or a p-value even close to female age (odds ratio for live birth (95% confidence interval) 1.00 (0.99–1.01), 1.00 (0.99–1.01), 0.98 (0.88–1.11) and 0.58 (0.48–0.72), respectively). The three models comparing FORT, FOI and OSI with the number of oocytes retrieved, the AFC, the number of preovulatory follicles and the FSH total dose were not more informative. (4) Conclusions: In a population of women of advanced age with unexplained infertility, none of FORT, FOI and OSI were predictive of live birth or more predictive than the two ‘component’ characteristics that were used to calculate them. We suggest clinicians and researchers still use female age as the most reliable predictor of an IVF treatment.

ARE THERE OVARIAN RESPONSIVE INDEXES THAT PREDICT CUMULATIVE LIVE BIRTH RATES IN WOMEN OVER 39 YEARS?

Ovarian response indexes have been proposed in Assisted Reproductive Technology (ART) in order to optimize live birth rates (LBR), adjusting ovarian stimulation (OS) and minimizing risks. Gonadotropin doses are commonly adjusted according to ovarian reserve parameters, including antral follicle count (AFC), antimülerian hormone (AMH) and basal Follicle Stimulating Hormon (FSH) levels. The retrospective assessment of ovarian responses allows to identify three primary indexes: (i) Follicular output rate (FORT), the ratio of the number of pre-ovulatory follicles obtained at OS completion over AFC; (ii) Follicle oocyte index (FOI), the ratio of oocytes retrieved over AFC, and (iii) Ovarian sensitivity index (OSI), the ratio of oocytes retrieved over the total gonadotropin dose administered.

Predictive value of sperm motility before and after preparation for the pregnancy outcomes of intrauterine insemination.

ObjectiveThis study aimed to investigate sperm motility and its changes after preparation as predictors of pregnancy in intrauterine insemination (IUI) cycles.MethodsIn total, 297 IUI cycles from January 2012 to December 2017 at a single tertiary hospital were retrospectively analyzed. Patient and cycle characteristics, and sperm motility characteristics before and after processing were compared according to clinical pregnancy or live birth as outcomes.ResultsThe overall clinical pregnancy rate per cycle was 14.5% (43/297) and the live birth rate was 10.4% (30/289). Patient and cycle characteristics were similar between pregnant and non-pregnant groups. Sperm motility after preparation and the total motile sperm count before and after processing were comparable in terms of pregnancy outcomes. Pre-preparation sperm motility was significantly higher in groups with clinical pregnancy and live birth than in cycles not resulting in pregnancy (71.4%±10.9% vs. 67.2%±11.7%, p=0.020 and 71.6%±12.6% vs. 67.3%±11.7%, p=0.030, respectively). The change in sperm motility after processing was significantly fewer in the non-pregnant cycles, both when the comparison was conducted by subtraction (post-pre) and division (post/pre). These relationships remained significant after adjusting for the female partner’s age, anti-Müllerian hormone level, and number of pre-ovulatory follicles. According to a receiver operating characteristic curve analysis, an initial sperm motility of ≥72.5% was the optimal threshold value for predicting live birth after IUI.ConclusionInitial sperm motility, rather than the motility of processed sperm or the degree of change after preparation, predicted live birth after IUI procedures.

P–714 Insulin-like growth factor-I as a mediator of the effect of transdermal testosterone in poor responder patients

Abstract Study question Is insulin-like growth factor-I (IGF-I) a mediator of the effect of transdermal testosterone (TT) in poor responder (PR) patients? Summary answer IGF-I might be a mediator of the effect of TT in PR patients who undergo an IVF cycle What is known already Many strategies have been tried to improve the results in PR patients. Androgen supplementation with TT is the only that has significantly increased live birth rate in these patients. The mechanism by which TT might influence on the better results remains unclear but it is likely mediated or facilitated by IGF-I. Testosterone increases the number of primordial follicles, increase IGF-I by threefold and increase IGF-I receptor mRNA by fivehold in primordial follicles in primates. Some studies have suggested that IGF-I could be a parameter that reflects the endocrinological environment of mature follicles, which is correlated with oocyte and embryonic quality Study design, size, duration This prospective cohort study of 93 women PR according Bologna criteria treated with TT and IVF/ICSI was conducted between May 2015 and December 2016 Participants/materials, setting, methods Exogenous andogenization with TT for 5 days prior to ovarian stimulation was carried out. Hormonal parameters were evaluated: basal FSH, LH and Estradiol, AMH, IGF–1 pre and post TT. Ultrasound parameterswere also analysed: antral follicle count (AFC) and number of pre-ovulatory follicles the day of HCGr. We compared these parameters according to the ovarian response: adequate (&amp;gt; 4 oocytes) or insufficient (&amp;lt;3 oocytes), as well as the pregnancy was achieved or not. Main results and the role of chance Baseline characteristics of the patients were: 36.9 years, FSH 11.8, AMH 0.86 and RFA 5.3. In 83% of the patients the oocyte retrieval was carried out, obtaining an average of 3.8 MII oocytes and 2.9 embryos of 2pn with a clinical pregnancy rate per transfer of 33.3%. The FORT Test (AFC/pre-ovulatory follicles x100) was 70%, higher than that observed in other studies with patients with PR without TT (55%). In cases in which an insufficient response was obtained (&amp;lt;3 oocytes) or the cycle was canceled, a higher age and FSH and lower AMH were observed (p &amp;lt; 0.05). There were no differences in the rest of the parameters. Evaluating the hormonal and ultrasound parameters depending on whether or not pregnancy was achieved, a significant increase in IGF1 pre and post-TT was observed in the cases of pregnancy (31.5%) compared to those cases where there was no pregnancy (10.9%) (p = 0’01). There were no differences in the rest of the parameters. A significant correlation was found between AMH, AFC and increase in IGF-I levels (p &amp;lt; 0’05). Limitations, reasons for caution This a prospective cohort study with limited number of patients included. Wider implications of the findings: The significant increase in serum levels of IGF–1 in pregnant patients would indicate the existence of a more favorable clinical setting for the administration of testosterone, probably related to a more favorable ovarian reserve as demonstrated by its correlation with serum levels of AMH and with the AF. Trial registration number Not applicable

Pregnancy outcomes following different protocols of controlled ovarian hyperstimulation in couples undergoing intrauterine insemination.

Clomiphene citrate (CC), letrozole and cetrorelix acetate are frequently used agents in controlled ovarian hyperstimulation (COH). However, these three agents have not yet been compared to one another regarding their pregnancy outcomes. The present study was designed to retrospectively compare pregnancy outcomes among the three aforementioned agents. This study involved infertile couples with an infertility duration of at least 2 years, ages 18 to 42years and who were referred to have their first intrauterine insemination (IUI) treatment cycle. All patients underwent COH with recombinant follicle-stimulating hormone (rFSH) plus CC (n=118), letrozole (n=81), or cetrorelix acetate (n=62), followed by IUI. Using the one-way multivariate analysis of covariance to control female patients' ages, patients stimulated with cetrorelix acetate/rFSH or CC/rFSH had higher numbers of preovulatory follicles than women stimulated with letrozole/rFSH (P<.02), whereas women stimulated with cetrorelix acetate/rFSH had a thicker endometrium than women stimulated with CC/rFSH (P<.0005). Biochemical pregnancy rates were similar among the three protocols of COH. However, women stimulated with letrozole/rFSH showed clinical pregnancy rates higher than those stimulated with CC/rFSH (P=.003) or cetrorelix acetate/rFSH (P=.03) and subclinical abortion rates lower than those stimulated with CC/rFSH or cetrorelix acetate/rFSH (P=.009). Of the different protocols of COH, the odds of having a clinical pregnancy was 3.1 times greater for women stimulated with letrozole/rFSH than women stimulated with CC/rFSH (P=.004) and 2.8 times greater for women stimulated with letrozole/rFSH than women stimulated with cetrorelix acetate/rFSH (P=.03). Our observations show that increased numbers of preovulatory follicles or endometrium thickness do not necessarily improve pregnancy outcomes, because pregnancy outcomes are also subjected to the type of COH used agent. In this regard, letrozole produced fewer preovulatory follicles and did not significantly increase endometrium thickness, but significantly improved pregnancy outcomes in comparison to CC and cetrorelix acetate.

Impact of chronic variable stress on neuroendocrine hypothalamus and pituitary in male and female C57BL/6J mice.

Chronic stress exerts multiple negative effects on the physiology and health of an individual. In the present study, we examined hypothalamic, pituitary and endocrine responses to 14days of chronic variable stress (CVS) in male and female C57BL/6J mice. In both sexes, CVS induced a significant decrease in body weight and enhanced the acute corticosterone stress response, which was accompanied by a reduction in thymus weight only in females. However, single-point blood measurements of basal prolactin, thyroid-stimulating hormone, luteinising hormone, growth hormone and corticosterone levels taken at the end of the CVS were not different from those of controls. Similarly, pituitary mRNA expression of Fshb, Lhb, Prl and Gh was unchanged by CVS, although Pomc and Tsh were significantly elevated. Within the adrenal medulla, mRNA for Th, Vip and Gal were elevated following CVS. Avp transcript levels within the paraventricular nucleus of the hypothalamus were increased by CVS; however, levels of Gnrh1, Crh, Oxt, Sst, Trh, Ghrh, Th and Kiss1 remained unchanged. Oestrous cycles were lengthened slightly by CVS and ovarian histology revealed a reduction in the number of preovulatory follicles and corpora lutea. Taken together, these observations indicate that 14days of CVS induces an up-regulation of the neuroendocrine stress axis and creates a mild disruption of female reproductive function. However, the lack of changes in other neuroendocrine axes controlling anterior and posterior pituitary secretion suggest that most neuroendocrine axes are relatively resilient to CVS.

Proof of concept use of progesterone/estradiol ratio to investigate late follicular progesterone in women with low number of preovulatory follicles.

The aim of this study was to investigate late follicular progesterone (P) serum levels in women with a low number of preovulatory follicles in the assisted reproductive technologies (ART) setting. Fifty-five consecutive women having four or fewer preovulatory follicles of >14 mm on the day of human chorionic gonadotropin (hCG) administration were prospectively evaluated. Spearman correlation tests were performed between serum estradiol (E<inf>2</inf>) level, serum P level, P/E<inf>2</inf> ratio, number of preovulatory follicles, oocytes and embryos. Women enrolled were further divided into two groups in accordance with the P/E<inf>2</inf> ratio on the day of hCG administration and compared. Serum E<inf>2</inf> level correlated positively with P serum level (r<inf>s</inf>=0.36, P<0.01), number of mature follicles (r<inf>s</inf>=0.50, P<0.01) and number of oocytes retrieved (r<inf>s</inf>=0.36, P<0.05), whereas negatively with P/E<inf>2</inf> ratio (r<inf>s</inf>=- 0.68, P<0.01). Likewise, number of preovulatory follicles correlated positively with E<inf>2</inf> level (r<inf>s</inf>=0.50, P<0.01), P level (r<inf>s</inf>=0.27, P<0.05) and number of oocytes retrieved (r<inf>s</inf>=0.33, P<0.05), while it correlated negatively with P/E<inf>2</inf> ratio (r<inf>s</inf>=-0.33, P<0.05). Furthermore, women with P/E<inf>2</inf> ratio >1 on the day of hCG administration received considerably higher total follicular stimulating hormone (FSH) dosage and achieved significantly lower number of oocytes and embryos as compared to controls. The reverse relationship between number of preovulatory follicles and P/E<inf>2</inf> ratio implies that P rise is not only the result of increased steroidogenic activity, but other oocyte-follicle disrupted mechanisms seem to be involved. An exaggerated FSH stimulation appears to disrupt further these mechanisms.