Follicular fluid content and oocyte quality: from single biochemical markers to metabolomics

The assessment of oocyte quality in human in vitro fertilization (IVF) is getting increasing attention from embryologists. Oocyte selection and the identification of the best oocytes, in fact, would help to limit embryo overproduction and to improve the results of oocyte cryostorage programs. Follicular fluid (FF) is easily available during oocyte pick-up and theorically represents an optimal source on non-invasive biochemical predictors of oocyte quality. Unfortunately, however, the studies aiming to find a good molecular predictor of oocyte quality in FF were not able to identify substances that could be used as reliable markers of oocyte competence to fertilization, embryo development and pregnancy. In the last years, a well definite trend toward passing from the research of single molecular markers to more complex techniques that study all metabolites of FF has been observed. The metabolomic approach is a powerful tool to study biochemical predictors of oocyte quality in FF, but its application in this area is still at the beginning. This review provides an overview of the current knowledge about the biochemical predictors of oocyte quality in FF, describing both the results coming from studies on single biochemical markers and those deriving from the most recent studies of metabolomic

Evolution of the use of corticosteroids for the treatment of hospitalised COVID-19 patients in Spain between March and November 2020: SEMI-COVID national registry

Objectives: Since the results of the RECOVERY trial, WHO recommendations about the use of corticosteroids (CTs) in COVID-19 have changed. The aim of the study is to analyse the evolutive use of CTs in Spain during the pandemic to assess the potential influence of new recommendations. Material and methods: A retrospective, descriptive, and observational study was conducted on adults hospitalised due to COVID-19 in Spain who were included in the SEMI-COVID- 19 Registry from March to November 2020. Results: CTs were used in 6053 (36.21%) of the included patients. The patients were older (mean (SD)) (69.6 (14.6) vs. 66.0 (16.8) years; p < 0.001), with hypertension (57.0% vs. 47.7%; p < 0.001), obesity (26.4% vs. 19.3%; p < 0.0001), and multimorbidity prevalence (20.6% vs. 16.1%; p < 0.001). These patients had higher values (mean (95% CI)) of C-reactive protein (CRP) (86 (32.7-160) vs. 49.3 (16-109) mg/dL; p < 0.001), ferritin (791 (393-1534) vs. 470 (236- 996) µg/dL; p < 0.001), D dimer (750 (430-1400) vs. 617 (345-1180) µg/dL; p < 0.001), and lower Sp02/Fi02 (266 (91.1) vs. 301 (101); p < 0.001). Since June 2020, there was an increment in the use of CTs (March vs. September; p < 0.001). Overall, 20% did not receive steroids, and 40% received less than 200 mg accumulated prednisone equivalent dose (APED). Severe patients are treated with higher doses. The mortality benefit was observed in patients with oxygen saturation </=90%. Conclusions: Patients with greater comorbidity, severity, and inflammatory markers were those treated with CTs. In severe patients, there is a trend towards the use of higher doses. The mortality benefit was observed in patients with oxygen saturation </=90%

Pregnancy after drug-free in vitro activation of follicles and fresh tissue autotransplantation in primary ovarian insufficiency patient: a case report and literature review

Abstract Background The aim of this report was to describe a case of pregnancy after drug-free in vitro activation (IVA) of follicles and fresh tissue autotransplantation in primary ovarian insufficiency (POI) patient and to review the pertinent literature. Methods We present a case in wich a 32 - years old patient with POI became pregnant after IVA without tissue culture and with ovarian tissue transplantation. We also reviewed the literature using Pubmed database. Case presentation Pretreatment with estradiol/progesterone stopped the day before surgery. The removal of the ovarian cortex and autotransplantation were performed by laparoscopy in the same surgical act. Ovarian fragments were transplanted in contralateral ovary and peritoneal pocket near to the ovary. Immediately after surgery GnRH agonist together HMG injections started, leading the growth of 3 preovulatory follicles and the retrieval of two mature eggs. After IVF two embryos were transferred and singleton pregnancy was established and currently she is 25 weeks pregnant. Results A total of 51 patients with POI in whom an in vitro activation of ovarian tissue was performed, were collected from the revieew of the literature. In 29.4% of them, follicular development was obtained and in 4 of them a pregnancy. In all of them, a combined technique (fragmentation and activation) was performed in two laparoscopies. No case has been reported successfully after drug-free in vitro activation. Conclusions This is the first report about a case with pregnancy after drug-free in vitro activation of follicles and fresh tissue autotransplantation in POI patient

P-605 Biomechanical characteristics of the ovarian cortex in POI patients and functional outcomes after drug-free IVA

Abstract Study question Is there a relationship between the biomechanical characteristics of ovarian cortex of POI patients and the resumption ovarian function after Drug-Free IVA? Summary answer Physical properties of ovarian cortex in POI are different. A significant ovarian stiffness increase was observed when resumption of ovarian function occurred after Drug-Free IVA. What is known already There is increasing evidence that the ovarian extracellular matrix (ECM) plays a critical role in follicle development. Primordial follicles are localized at the collagen-rich ovarian cortex, which offers a rigid physical environment that supports follicular architecture and increases survival. Therefore, premature loss of ovarian function would be associated with a physically less rigid ovarian cortex. Study design, size, duration Prospective observational cohort study at a tertiary-care university hospital including POI patients according ESHRE criteria who underwent Drug-Free IVA by laparoscopy between January 2018 and December 2019 and were follow-up for a year after intervention. Participants/materials, setting, methods Nineteen patients were included. Resumption of ovarian function was defined as resumption of menstrual cycles (at least three consecutive episodes of menstrual bleeding) and/or the presence of follicles &amp;gt; 10 mm on ultrasound. A sample of ovarian cortex taken during the intervention was analyzed by atomic force microscopy (AFM) in order to quantitatively measure mechanical properties at the nanometer scale (Young’s elastic modulus, E). Main results and the role of chance Median data at intervention were: age = 35 years (28–39); length of amenorrhea = 2 years (1–10); FSH level = 104.4 mIU/ml (38.9–176); and AMH = 0.02 ng/ml (0.01–0.1). Resumption of ovarian function was observed in 10 patients (52.6 %), achieving 2 pregnancies (one spontaneous and one after an IVF cycle). Median stiffness (E) measured by AFM was 2583 Pa (999–11296). There were no differences in clinical and hormonal parameters as a function of resumption of ovarian activity. Remarkably, ovarian cortex stiffness was significantly increased in patients with ovarian activity after Dug-Free VIA: 5519 (2260–11296] vs 1501 [999–3474], p value &amp;lt;0.001. Limitations, reasons for caution The main limitations of the study are the unavailability of ovarian tissue samples from a control group of patients without POI and the inability to perform histological studies on the same samples in which the biomechanical study was carried out. Wider implications of the findings These findings show the high variability of ovarian cortex stiffness in POI and that the increase of this stiffness entails a more favorable status after Drug-Free IVA. This may be related to an ovary with more residual follicles, which would explain a greater chances of ovarian follicular reactivations after treatment. Trial registration number not applicable </jats:sec

P-614 Predicting if or when ovarian activation occurs after Drug-Free <i>in vitro</i> Activation (IVA) in primary ovarian insufficiency (POI) patients

Abstract Study question Can we predict if or when ovarian activation resumption occurs after Drug-Free in vitro Activation (IVA) in primary ovarian insufficiency (POI) patients? Summary answer Duration of amenorrhea alone could predict if ovarian activity resumption occurs. Regarding to when: as early as 21 days until 330 days after surgery. What is known already Primary ovarian insufficiency occurs before age 40, leading patients to infertility. Although there are a few remaining follicles, their only chance to achieve pregnancy is through egg donation. Recently, studies have been focusing on the determinants and intracellular signaling pathways involved in the activation of primordial follicles, as the maintenance of a correct ovarian reserve. A new surgical technique known as Drug-Free IVA would activate ovarian function by mechanical fragmentation. For selected patients whose ovaries still contain residual secondary follicles, the fragmentation step alone, which implies a Hippo signaling pathway disruption, would be sufficient to promote follicle growth. Study design, size, duration Our study is a prospective observational cohort of patients recruited in a tertiary-care university hospital approved by the Ethics Research Committee from Hospital Clinic of Barcelona (HCB/2017/0856). The diagnostic criteria of POI followed the European Society of Human Reproduction and Embryology (ESHRE) Guideline from 2016. All patients provided written informed consent.Thirty-two women included in our study underwent Drug-Free IVA by laparoscopy between January 2018 and December 2019. After surgery, a year-long follow-up was carried out. Participants/materials, setting, methods All patients were younger than 40 years of age; presented oligomenorrhoea or amenorrhea for at least 4 months; and FSH level greater than 25 IU/mL . We also restrict inclusion criteria to low estradiol serum levels (&amp;lt;20 pg/ml) absence of antral follicles in transvaginal ultrasound; and anti-Müllerian hormone (AMH) levels less than 0.10 ng/mL. Regardless of the time from POI diagnosis, all patients had duration of amenorrhea greater than 1 year. Main results and the role of chance As for prediction of ovarian activation, duration of amenorrhea was the only factor that could significantly differentiate patients' response to Drug-Free IVA. Since a shorter time of amenorrhea (less than 2 years) was related with ovarian activation during a year follow-up. No differences were found in age at Drug-free IVA, age of menarche, baseline FSH or AMH levels; nor presence of follicles in ovarian tissue biopsy. Regarding to time to response, in 12 patients ovarian activation occurred in the first 6 months after surgery with a total of 5 pregnancies achieved (22.7%)- resulting in 4 heathy live births and, unfortunately, one miscarriage. The other 10 patients showed ovarian activation resumption once the 180 days passed, resulting in 3 pregnancies (pregnancy rate of 30%): 2 healthy live births and one neonatal death due to extreme prematurity. An important concept here is that activation before 6 months after surgery could mean an effect on primary and/or secondary follicles, while beyond 6 months the effect might occurred on primordial follicles. Sugesting that maybe the disruption of the Hippo pathway would not only be able to act in the activation of the secondary follicles but also act in the earlier phases of the folliculogenesis. Limitations, reasons for caution The lack of a control group is the main limitation of our study because it would be interesting to seek for the placebo effect or intensive medical attention/care consequences in those patients, whose reported pregnancy rate is as low as 4% in observational studies. Wider implications of the findings Our findings define the clinical profile of patients with POI in which Drug-Free IVA would be more effective and report late ovarian follicular resumption. Meaning that disruption of the Hippo signaling pathway by ovarian fragmentation could be capable of acting in the initial phases of folliculogenesis prior to secondary follicles. Trial registration number not applicable </jats:sec

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 </jats:sec