Assisted Reproduction Protocols Research Articles

Sperm Toolbox-A selection of small molecules to study human spermatozoa.

Male contraceptive options and infertility treatments are limited, and almost all innovation has been limited to updates to medically assisted reproduction protocols and methods. To accelerate the development of drugs that can either improve or inhibit fertility, we established a small molecule library as a toolbox for assay development and screening campaigns using human spermatozoa. We have profiled all compounds in the Sperm Toolbox in several automated high-throughput assays that measure stimulation or inhibition of sperm motility or the acrosome reaction. We have assayed motility under non-capacitating and capacitating conditions to distinguish between pathways operating under these different physiological states. We also assayed cell viability to ensure any effects on sperm function are specific. A key advantage of our studies is that all compounds are assayed together in the same experimental conditions, which allows quantitative comparisons of their effects in complementary functional assays. We have combined the resulting datasets to generate fingerprints of the Sperm Toolbox compounds on sperm function. The data are included in an on-line R-based app for convenient querying.

Deformities and their potential control in larviculture of the European eel Anguilla anguilla

With the current assisted reproduction protocols, yolk-sac larvae of the European eel can be regularly produced. However, high mortality and deformity rates may exist during the first week after hatching with few larvae reaching the exogenous feeding stage. Recently, we suggested that larval survival could be impaired by pathogen pressure. Therefore, in this study, we tested the effects of antibiotics (rifampicin and ampicillin 50 mg L−1) and egg surface disinfection treatment (povidone iodine 25 ppm) on hatching success, larval survival and the occurrence of deformities in wild and feminized eels. Differences in egg quality aspects existed between offspring of wild and feminized eels. Eggs produced by the wild females were more buoyant than the ones released by the feminized eels. Larvae of the wild females hatched at higher rates and survived longer than the ones produced by the feminized eels. Disinfection treatment negatively affected hatching success and larval survival but the application of antibiotics improved larval survival. While larvae survived until 5 days post-hatch (dph) in the untreated controls, larvae continued to survive up to 21 dph when treated with antibiotics. Despite the beneficial effects of antibiotics on larval survival, a wide diversity of deformities could be observed during early development of eels that have not been reported before. The deformity determination key for young eel larvae that is presented in this paper fills a gap in the knowledge about larval deformities during early ontogeny. The larvae showed pericardial oedemas, microcephaly, an enlarged-yolk sac, necrosis and notochord deformities. The treatment with antibiotics decreased the deformity rates although they remained high (75–85%). The causes behind each deformity need to be clarified in order to prevent them in future trials.

P-639 Are our advanced maternal age population being affected by the shortage of HP-hMG?

Abstract Study question Could we stimulate patients of advanced maternal age in the face of a shortage of HP-hMG without impacting outcomes? Summary answer Given the current low availability of HP-hMG, it is safe to stimulate only with recombinant FSH in women of advanced maternal age without affecting results. What is known already . Studies on the role of luteinizing hormone (LH) supplementation in patients undergoing assisted reproductive treatments use different sources of LH bioactivity. It is well known that the addition of LH supplementation was associated with a tendency towards improved IVF outcome in specific profiles of patients undergoing Assisted Reproductive treatment. However, during the last quarter of 2022, we are facing a temporary fall in the global supply of HP-hMG, which has hCG-driven LH activity, and may impact ovarian stimulation outcomes in women of advanced maternal age undergoing a preimplantation genetic testing treatment (PGT). Study design, size, duration Retrospective and observational study during November-December 2022 in any of the 11 clinics belonging to the IVIRMA group in Spain. Study population included women ≥38 years undergoing PGT treatment who received monotherapy with recombinant FSH (n = 73), a combination of FSH + HMG containing an FSH: LH activity in a ratio 1:1 (n = 103), or a fertility drug with recombinant FSH + recombinant LH in a 2:1 ratio (n = 112). The study was approved by an IRB (2003-MAD-020-AR) Participants/materials, setting, methods All women underwent short GnRH antagonist protocol. Initial doses were adjusted based on weight and body mass index according to the clinician’s experience. Daily doses of 0.25 mg GnRH antagonist were started on day 6 of stimulation. Finally, a single dose of 0.2 mg GnRH agonist was administered to trigger final oocyte maturation. Embryo biopsy was performed on day 5 with a subsequent freeze-all. A frozen single embryo transfer was accomplished in a subsequent cycle. Main results and the role of chance Assuming the limitations of the study design, we observed significant differences in gonadotropin doses but not in retrieved oocytes, metaphase II oocytes or euploidy rate: the results were as follows for recombinant FSH, FHS + hMG and FSH + LH respectively: gonadotropin doses [2254 (692) vs. 2033 (680) vs. 2000 (660), p = 0.0035]; retrieved oocytes [10.1 (5.4) vs. 11.1 (8.0) vs. 9.2 (6.0), p = 0.131], metaphase II oocytes [8.7 (4.7) vs. 9.8 (7.1) vs. 8.3 (5.1), p = 0.147]; and euploidy rate (38.7% vs. 36.3% vs. 31.5%, p = 0.182). Moreover, recombinant FSH in monotherapy in this specific population could guarantee the efficiency of the cycle since there are no significant differences in the usable blastocyst rate. Finally, clinical results were analyzed in those cases in which a frozen single embryo had been performed in a subsequent artificial cycle, in which the effect of LH on the endometrium is obviated; once again, no significant differences were observed between the study groups neither for the implantation rate (50% vs. 56% vs. 45%, p = 0.780) or the clinical pregnancy rate (50% vs. 56% vs. 45%, p = 0.772). Limitations, reasons for caution This is a retrospective and observational study and thus possible confounders cannot be completely excluded. More data are needed to draw firm conclusions and it will be critical to increase the sample size to check if the results observed in this work remains in the general population. Wider implications of the findings In view of tailoring assisted reproductive protocols to meet patient needs and improve the likelihood of positive outcomes, these data suggest that it is optimal to perform an ovarian stimulation protocol only with recombinant FSH in a frozen embryo program in advanced maternal age context without clinical outcomes being affected. Trial registration number Not applicable

Dominant follicle growth patterns and associated endocrine dynamics in anovulatory and ovulatory waves in women.

Growth patterns and associated endocrine profiles were compared between dominant anovulatory (ADF) and ovulatory follicles (OvF) developing from different waves within and between menstrual cycles in women. Follicular mapping profiles of 49 healthy women of reproductive age and blood samples were obtained every 1-3 days. Sixty-three dominant follicles were classified into wave 1 (W1ADF; n = 8) and wave 2 (W2ADF; n = 6) anovulatory follicles and wave 2 (W2OvF; n = 33) and wave 3 (W3OvF; n = 16) ovulatory follicles. Comparisons were made between W1ADF and W2ADF, W2ADF and W2OvF, and W2OvF and W3OvF. The waves were numbered 1, 2, or 3 based on when the waves emerged relative to the preceding ovulation. W1ADF emerged closer to the preceding ovulation, and W2ADF emerged in the late luteal or early follicular phase. The interval from emergence to maximum diameter was shorter for W2ADF than W1ADF and for W3OvF than W2OvF. Selection of W3OvF occurred at a smaller diameter compared to W2OvF. W1ADF regressed at a faster rate than W2ADF. Also, W1ADF were associated with lower mean FSH and higher mean estradiol than W2ADF. In contrast, W3OvF were associated with higher FSH and LH compared to W2OvF. However, W2OvF were associated with higher progesterone than W3OvF. This study contributes to the understanding of the physiologic mechanisms underlying selection of the dominant follicle, ovulation, and pathophysiology of anovulation in women, as well as optimization of ovarian stimulation protocols for assisted reproduction.

Steroid implants for the induction of vitellogenesis in feminized European silver eels (Anguilla anguilla L.).

Assisted propagation of the European eel will lead to a closed production cycle supplying the aquaculture industry with juvenile glass eels. Females require long-term weekly treatment with pituitary extract (PE), which is stressful and causes abnormalities in oogenesis. We tested the effects of 17α-methyltestosterone (17 MT), as potent androgen activating the androgen receptor, and 17β-estradiol (E2), as an inducer of vitellogenesis, to shorten the duration of PE treatment.Four groups of feminized eels were subjected to a simulated migration and subsequent injection with implants containing 17 MT (17 MT-group), E2 (E2-group) or 17 MT plus E2 (17 MT + E2-group) to test for synergistic effects, or without any steroids as controls (C-group). The effects of a 2-months treatment were investigated by determining the eye index (EI), hepatosomatic and gonadosomatic index (HSI and GSI, respectively), plasma steroid concentrations by liquid chromatography mass spectrometry (LCMS), gonadal histology, expression of androgen receptors a and b (ara, arb); estrogen receptor 1 (esr1); FSH receptor (fshr); vitellogenin receptor (vtgr) and aromatase (cyp19), and the required number of weekly PE injections to fully mature. For many parameters, both the 17 MT and E2 groups showed an increase vs. controls, with the 17 MT + E2 group showing a synergistic effect, as seen for EI, GSI (3.4 for 17 MT and for E2, 6.6 for 17 MT + E2), oocyte diameter and ara, arb and esr1 expression. Concentrations of almost all focal steroids decreased with simulated migration and steroid treatment. Only eels of the 17 MT-group showed increased expression of cyp19 and of fshr, while fshr expression increased 44-fold in the 17 MT + E2 group, highlighting that co-implantation is most effective in raising fshr mRNA levels. Specific for eels of the E2 groups were vitellogenesis-associated changes such as an increase of HSI, plasma E2, and presence of yolk in the oocytes. Steroid treatments reduced the duration of PE treatment, again synergistically for co-implantation. In conclusion, E2 is necessary to start vitellogenesis, but 17 MT has specific effects on cyp19 and fshr expression. The combination is necessary for synergistic effects and as such, steroid implants could be applied in assisted reproduction protocols for European eel to improve oocyte quality leading to the production of more vital larvae.

O-009 Controlled ovarian stimulation (COS) protocols for assisted reproduction: a Cochrane systematic review and network meta-analysis

Abstract Study question What is the relative effectiveness and safety of existing COS protocols for women undergoing assisted reproductive technology (ART) treatment? Summary answer There was no difference in live birth between all protocols, but short antagonist protocols may reduce ovarian hyperstimulation syndrome in women with predicted normal/high response. What is known already Controlled ovarian stimulation is an essential step in most ART cycles. It involves the administration of exogenous gonadotrophins to induce multifollicular growth, usually in addition to drugs that prevent untimely ovulation by suppressing the pituitary gland. Different treatment combinations may be used in COS. These vary according to the type of drugs administered for pituitary suppression (e.g., gonadotrophin-releasing hormone [GnRH] agonists, antagonists) and ovarian stimulation (e.g., urinary or recombinant gonadotrophins). Drug dosages, timing and routes of administration also vary between different regimens. However, there is no consensus on how the existing COS protocols rank according to their effectiveness and safety. Study design, size, duration We searched the following databases to November 2021: MEDLINE, EMBASE, CINAHL, CENTRAL and ClinicalTrials.gov. We included randomised controlled trials (RCTs) comparing at least two COS protocols using GnRH agonists or antagonists for pituitary suppression; and human menopausal gonadotrophin (hMG), urinary or recombinant follicle-stimulating hormone (u/rFSH), with or without luteinising hormone (LH) for ovarian stimulation. The primary outcomes were the rates of live birth (LBR) and ovarian hyperstimulation syndrome (OHSS) per participant after one stimulation cycle. Participants/materials, setting, methods Two reviewers independently selected studies and extracted data. We conducted pairwise and network meta-analyses (NMA) according to participants’ predicted response to COS (normal, high and low). Using the Cochrane-RoB-1 tool, we restricted our primary analyses to RCTs at low risk of selection and other biases. We presented effect estimates as risk ratio (RR) with 95% confidence interval (CI) and considered I2>50% as representing substantial heterogeneity. For each outcome, we generated ranking plots comparing different interventions. Main results and the role of chance In total, our searches identified 9464 studies. The primary analysis included 68 RCTs assessing 17861 women and 34 different COS protocols. The evidence showed that in women with predicted normal or high response, the use of short GnRH antagonist protocols may result in little to no difference in LBR (RR 0.98, 95% CI 0.85 to 1.13; 6 studies; 2063 women; I2 = 0%; low-certainty evidence) and a reduction in OHSS (RR 0.88, 95% CI 0.78 to 0.99; 7 studies; 2246 women; I2 = 0%; low-certainty evidence) compared with long GnRH agonist protocols. The rankogram comparing different COS protocols showed a probability of 98% that short GnRH antagonist regimens are the best treatment to prevent OHSS. Sensitivity analyses including all studies showed that in women with predicted normal response undergoing long GnRH agonist cycles for pituitary suppression, the use of rFSH for ovarian stimulation may result in decreased fresh-cycle LBR compared to hMG (RR 0.80, 95% CI 0.68 to 0.95; 7 studies; 1575 women; I2 = 1%; low-certainty evidence). For the remaining interventions (e.g., agonist flare or progestogens for pituitary suppression, in combination with various gonadotrophin regimens) the evidence was uncertain of an effect or insufficient for quantitative synthesis. Limitations, reasons for caution The high number of interventions resulted in disconnected networks, limiting our ability to perform NMA for some comparisons. The certainty of the evidence was limited by serious risk of bias. Finally, the lack of data on cumulative LBR and differences in oocyte yield made comparisons between FSH preparations potentially unbalanced. Wider implications of the findings Our findings suggest that the use of short GnRH antagonist protocols may result in reduced OHSS rates in women with predicted normal or high ovarian response without compromising live birth rates. There is a paucity of high-quality RCTs comparing different gonadotrophin preparations (e.g., hMG versus rFSH) for COS. Trial registration number N/A

Comparative Study of Semen Parameters and Hormone Profile in Small-Spotted Catshark (Scyliorhinus canicula): Aquarium-Housed vs. Wild-Captured.

Simple SummaryComprehensive knowledge of chondrichthyan reproductive biology is crucial for the development of reproductive technologies. For that reason, a male reproductive evaluation was performed on the basis of a comparison of samples collected from wild-captured and aquarium-housed small-spotted catshark (Scyliorhinus canicula). Semen quality, sperm morphometry, and reproductive hormones were assessed. The results demonstrate good in vitro semen quality in aquarium-housed sharks, although there was lower plasma testosterone. Several chondrichthyan species are threatened, and we must increase our knowledge of their reproductive biology in order to establish assisted reproductive protocols for ex situ or in situ endangered species. The small-spotted catshark (Scyliorhinus canicula) is one of the most abundant shark species of the Mediterranean coast and is easy to maintain in aquaria; therefore, it is considered an ideal reproductive model. This study aimed to compare S. canicula male reproductive function in aquarium-housed (n = 7) and wild-captured animals, recently dead (n = 17). Aquarium-housed animals had lower semen volume (p = 0.005) and total sperm number (p = 0.006) than wild-captured animals, but similar sperm concentrations. In terms of sperm parameters, aquarium-housed sharks showed higher total sperm motility (p = 0.004), but no differences were observed regarding sperm viability, mitochondrial membrane potential, or membrane integrity. A morphometric study pointed to a significantly longer head (p = 0.005) and acrosome (p = 0.001) in wild-captured animals. The results of the spermatozoa morphological study of S. canicula were consistent with previous results obtained in other chondrichthyan species. With regard to sex hormones, testosterone levels were significantly lower in aquarium-housed animals (p ≤ 0.001), while similar levels of 17β-estradiol and progesterone were found. In short, the present study provides evidence of good in vitro semen quality in S. canicula housed in an aquarium, underlining their excellent potential for application in reproductive technologies for this and other chondrichthyan species.

Determining the Effects of Serial Injections of Pregnant Mare Serum Gonadotropin on Plasma Testosterone Concentrations, Testicular Dynamics, and Semen Production in Leopard Geckos (Eublepharis macularius).

Simple SummaryCurrent estimates have determined that one in five reptile species are already threatened with extinction. The aim of this study was to determine if pregnant mare serum gonadotropin could be used in leopard geckos (Eublepharis macularius) to stimulate the production of sperm and increase their testosterone concentrations. This information may aid in the development of breeding programs for endangered gecko species in the future that will be useful for conservation efforts. Our results demonstrated that this hormone did stimulate sperm production and increased testicular size in leopard geckos. However, it did not increase testosterone concentrations under the current conditions between October–December in the Northern hemisphere. Ultimately, future studies are needed to further characterize the annual reproductive cycle of leopard geckos.Reptiles are highly susceptible to anthropogenic activities as a result of their narrow geographical ranges and habitat specialization, making them a conservation concern. Geckos represent one of the mega-diverse reptile lineages under pressure; however, limited assisted reproductive technologies currently exist for these animals. Exogenous pregnant mare serum gonadotropin (PMSG) has been found to exhibit follicle stimulating hormone-like action and has been routinely used to alter reproductive hormones of vertebrates in assisted reproductive protocols. The purpose of this study was to determine the effects of serial injections of 20 IU and 50 IU PMSG on circulating testosterone concentrations, testicular dynamics, and semen production in a model species of gecko. Twenty-four captive-bred, adult, male leopard geckos (Eublepharis macularius) were divided into three treatment groups and administered a once-weekly injection of either PMSG or saline for a total of nine weeks. Ultrasonographic testicular measurements, electrostimulation for semen collection, and venipuncture were performed on days 0, 21, 42, and 63. Right unilateral orchidectomies and epididymectomies were performed in all animals on day 63; tissues were submitted for histopathology. PMSG treated geckos had significantly higher testicular volumes and weights, spermatozoa motility, and spermatozoa concentrations compared with controls. However, there were no significant differences in testosterone concentrations by treatment or time. Under the conditions outlined, PMSG is effective at stimulating spermatogenesis and increasing testicular size, but not effective at increasing testosterone concentrations in the leopard gecko between October–December in the Northern hemisphere.

Тактика ведения пациенток с синдромом доброкачественных узловых образований молочной железы на фоне беременности (обзор литературы с собственными клиническими наблюдениями)

The paper presents data on the nodular formations during pregnancy and diagnosis principles of the mammary glands nodular formations during pregnancy. The data of 53 women survey with mammary glands benign tumors that appeared at different pregnancy stages after the assisted reproductive technology protocols use are presented. The tactics of these patients management are proposed. Unresolved issues that require further discussion are identified.

Obtaining Ejaculates through Operant Conditioning of the Golden Eagle (Aquila chrysaetos)

Background: The golden eagle, Aquila chrysaetos is one of the 65 species of birds of prey found in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora. It has been estimated that the world population is 250 thousand specimens. The operant conditioning to obtain ejaculates of A. chrysaetos is described and to contribute to assisted reproduction protocols in captivity.Methods: In the reproductive season, the procedure was performed twice a week on 5 adult birds, in order to obtain ejaculates.Result: The study lasted 17 weeks, in which 160 contacts were made in total. It was possible to obtain 20 ejaculates, which were obtained from 3 birds. 4% success was achieved in obtaining ejaculates. Additionally, behaviors of clinical utility for diagnosis, care and well-being were conditioned in birds. The above should be considered in assisted reproduction programs, since, as demonstrated by the results of this study, not all the birds provide sufficient ejaculates, for which reason, the evaluation of the individuals that present the greatest advance in operant conditioning to include them in assisted reproduction programs.

A human-based assisted reproduction protocol for the menstruating spiny mouse, Acomys cahirinus.

The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like menstruation and spontaneous decidualisation. We consider from these, and its other, human-like characteristics that this species will be a more useful and appropriate small animal model for human reproductive studies. Based on this, there is a need to develop specific laboratory-based assisted reproduction protocols including superovulation, in-vitro fertilisation, embryo cryopreservation and transfer to expand and make this model more relevant. Because standard rodent superovulation has not been successful in the spiny mouse, we have selected to test a human protocol. Female spiny mice will receive a subcutaneous GnRH agonist implant and be allowed to recover. Menstrual cycle lengths will then be allowed to stabilize prior to ovarian stimulation. After recovery, females will be injected IP once a day for 4 days with a FSH analogue, to induce follicular growth, and on day 5 will be injected IP with a hCG analogue to trigger ovulation. Females will either be culled 36hrs after trigger to collect oocytes or immediately paired with a stud male and two cell embryos collected 48hrs later. Mature oocytes will be inseminated using fresh spiny mouse spermatozoa and all in-vitro grown and in-vivo collected two cell embryos will be cryopreserved using methods developed in a close spiny mouse relative, the Mongolian gerbil. For embryo transfer, vitrified embryos will be rapidly warmed and non-surgically transferred to surrogate mice. Surrogates will be monitored until pregnancy is apparent (roughly 30 days) and then left undisturbed until birth, 38–40 days after transfer. By successfully developing robust assisted reproduction protocols in A. cahirinus we will be able to use this rodent as a more effective model for human reproduction.

Progesterone: History, facts, and artifacts.

Progesterone and its related molecules are a crucial tool in modern clinical practice, particularly in the fields of reproductive medicine. Its history is old, but still under development. Presently, the pharmacokinetic and pharmacodynamic profiles of progesterone is well-known and knowledge on natural progesterone (P4) and other molecules with progestational activity, namely progestogens or gestagens, are improved and their interest is still alive. Topics of great and current interest are progesterone and its role in assisted reproductive protocols, threatened and recurrent pregnancy loss, threatened preterm birth with favorable results on pregnancy, and perinatal outcomes. Moreover, progesterone provides several other positive effects on women's health. This paper describes the main chronological steps that characterized the history of progesterone, where scientific research and clinical practice are arrived and WHICH are the future perspectives on this hormone with a "never-ending history."

La libéralisation de la recherche sur l’embryon humain et sur les cellules souches issues du corps humain dans le projet de loi relatif à la bioéthique

Since the beginning of last autumn, the bioethics law reform bill has been debated in Parliament. One of the major challenges of this text concerns research carried out on the embryo and on stem cells from the human body. The rules currently in force should be profoundly changed, whether it is research carried out for scientific purposes or that carried out within the framework of an assisted reproduction protocol.

Improvement of Sperm Quality in Hyperviscous Semen following DNase I Treatment.

Semen hyperviscosity impairs sperm motility and can lead to male infertility. This prospective study aimed at assessing the ability of exogenous DNase in improving sperm quality, taking into consideration that DNase has been found in the seminal plasma of several species and that neutrophils release chromatin in order to trap bacteria. A total of seventy-seven semen samples with high seminal viscosity (HSV) as the study group and sixty-two semen samples with normal seminal viscosity (NSV) as the control group were compared in this analysis. These semen samples were divided into three groups of receiving treatment (a) with DNase I at 37°C for 15 min, (b) by density gradient centrifugation, and (c) with a combination of the above two methods. Following a fifteen-minute treatment of hyperviscous semen, the motility of spermatozoa in 83% of semen samples increased to a statistically significant degree. On the contrary, DNase treatment of semen with normal viscosity had no such effects. The above treatment was also accompanied by a significant increase in the percentage of normal spermatozoa, resulting in a major decrease of the teratozoospermia index. Comparison between semen samples that underwent density gradient centrifugation following DNase I treatment, to those collected after density gradient treatment alone, showed that in the first case the results were more spectacular. The evaluation of each preparation in terms of yield (% total progressively motile sperm count after treatment in relation to the initial total sperm count) revealed that the combined approach resulted in 29.8% vs. 18.5% with density treatment alone (p=0.0121). DNase I treatment results in an improvement of sperm motility and morphology and could be beneficial to men with hyperviscous semen in assisted reproduction protocols.

Unique Associations between Insulin-Like Growth Factor Binding Protein-1, Insulin-Like Growth Factor-1 and T Cell Immunoglobulin Mucin 3 in Successful Twin Pregnancies Conceived with Donor Oocytes.

Background and Objectives: To investigate if pregnancies conceived using an oocyte donor necessitate an alteration in immune regulation, we compared concentrations of insulin-like growth factor binding protein (IGFBP)-1, insulin-like growth factor (IGF)-1 and T cell immunoglobulin mucin-3 (Tim-3) in women with ongoing successful twin pregnancies conceived spontaneously, using assisted reproductive technologies that utilized homologous oocytes or with donor oocytes. Differences in levels of these immune modulatory proteins may be magnified and easier to detect in twin as compared to singleton pregnancies. Methods: In this prospective study IGFBP-1 and IGF-1 were measured in sera and Tim-3 in lysates of peripheral blood mononuclear cells (PBMCs) by ELISA. Results: Median IGFBP-1 levels were lower in women with donor oocytes (41.4 ng/ml) as compared to those with a spontaneous conception (51.2 ng/mL) or who conceived with various assisted reproduction protocols using homologous oocytes (52.4 ng/mL) (p < 0.001). IGF-1 and Tim-3 levels were comparable in each group. The IGFBP-1 level was inversely correlated to the IGF-1 concentration only in women with donor oocytes (p = 0.032). IGFBP-1 and Tim-3 levels were similarly negatively correlated in the donor oocyte group (p = 0. 012). Women in the assisted reproduction group who conceived following intracytoplasmic sperm injection were the only other group in which IGFBP-1 and Tim-3 were negatively correlated (p = 0.018). Conclusions: Down-regulation of IGFBP-1 production in pregnancies conceived with donor oocytes may reduce the extent of pro-inflammatory immunity and contribute to successful outcome in totally allogeneic pregnancies.

PO-01-004 Phytotherapy could improve sperm collection rate in psychogenic anejaculation affecting couples involved in assisted reproduction protocols

PO-01-004 Phytotherapy could improve sperm collection rate in psychogenic anejaculation affecting couples involved in assisted reproduction protocols

Small Molecule Follicle-Stimulating Hormone Receptor Agonists and Antagonists.

The follicle-stimulating hormone receptor (FSHR) has been targeted therapeutically for decades, due to its pivotal role in reproduction. To date, only purified and recombinant/biosimilar FSH have been used to target FSHR in assisted reproduction, with the exception of corifollitropin alfa; a modified gonadotropin in which the FSH beta subunit is joined to the C-terminal peptide of the human choriogonadotropin beta subunit, to extend serum half-life. Assisted reproduction protocols usually entail the trauma of multiple injections of FSH to initiate and promote folliculogenesis, which has prompted the development of a number of orally-available low molecular weight (LMW) chemical scaffolds targeting the FSHR. Furthermore, the recently documented roles of the FSHR in diverse extragonadal tissues, including cancer, fat metabolism, and bone density regulation, has highlighted the potential utility of LMW modulators of FSHR activity. Despite these chemical scaffolds encompassing a spectrum of in vitro and in vivo activities and pharmacological profiles, none have yet reached the clinic. In this review we discuss the major chemical classes of LMW molecules targeting the FSHR, and document their activity profiles and current status of development, in addition to discussing potential clinical applications.

Endometrial receptivity revisited: endometrial transcriptome adjusted for tissue cellular heterogeneity.

Does cellular composition of the endometrial biopsy affect the gene expression profile of endometrial whole-tissue samples? The differences in epithelial and stromal cell proportions in endometrial biopsies modify the whole-tissue gene expression profiles and affect the results of differential expression analyses. Each cell type has its unique gene expression profile. The proportions of epithelial and stromal cells vary in endometrial tissue during the menstrual cycle, along with individual and technical variation due to the method and tools used to obtain the tissue biopsy. Using cell-population specific transcriptome data and computational deconvolution approach, we estimated the epithelial and stromal cell proportions in whole-tissue biopsies taken during early secretory and mid-secretory phases. The estimated cellular proportions were used as covariates in whole-tissue differential gene expression analysis. Endometrial transcriptomes before and after deconvolution were compared and analysed in biological context. Paired early- and mid-secretory endometrial biopsies were obtained from 35 healthy, regularly cycling, fertile volunteers, aged 23-36 years, and analysed by RNA sequencing. Differential gene expression analysis was performed using two approaches. In one of them, computational deconvolution was applied as an intermediate step to adjust for the proportions of epithelial and stromal cells in the endometrial biopsy. The results were then compared to conventional differential expression analysis. Ten paired endometrial samples were analysed with qPCR to validate the results. The estimated average proportions of stromal and epithelial cells in early secretory phase were 65% and 35%, and during mid-secretory phase, 46% and 54%, respectively, correlating well with the results of histological evaluation (r = 0.88, P = 1.1 × 10-6). Endometrial tissue transcriptomic analysis showed that approximately 26% of transcripts (n = 946) differentially expressed in receptive endometrium in cell-type unadjusted analysis also remain differentially expressed after adjustment for biopsy cellular composition. However, the other 74% (n = 2645) become statistically non-significant after adjustment for biopsy cellular composition, underlining the impact of tissue heterogeneity on differential expression analysis. The results suggest new mechanisms involved in endometrial maturation, involving genes like LINC01320, SLC8A1 and GGTA1P, described for the first time in context of endometrial receptivity. The RNA-seq data presented in this study is deposited in the Gene Expression Omnibus database with accession number GSE98386. Only dominant endometrial cell types were considered in gene expression profile deconvolution; however, other less frequent endometrial cell types also contribute to the whole-tissue gene expression profile. The better understanding of molecular processes during transition from pre-receptive to receptive endometrium serves to improve the effectiveness and personalization of assisted reproduction protocols. Biopsy cellular composition should be taken into account in future endometrial 'omics' studies, where tissue heterogeneity could potentially influence the results. This study was funded by: Estonian Ministry of Education and Research (grant IUT34-16); Enterprise Estonia (EU48695); the EU-FP7 Eurostars program (NOTED, EU41564); the EU-FP7 Marie Curie Industry-Academia Partnerships and Pathways (SARM, EU324509); Horizon 2020 innovation program (WIDENLIFE, EU692065); MSCA-RISE-2015 project MOMENDO (No 691058) and the Miguel Servet Program Type I of Instituto de Salud Carlos III (CP13/00038); Spanish Ministry of Economy, Industry and Competitiveness (MINECO) and European Regional Development Fund (FEDER): grants RYC-2016-21199 and ENDORE SAF2017-87526. Authors confirm no competing interests.

Highly Purified Human Menopausal Gonadotropin (Menopur®): A Profile of Its Use in Infertility

Menopur® is a highly purified, urine-derived, human menopausal gonadotropin containing both follicle stimulating hormone (FSH) and luteinizing hormone (LH) activity. It is an effective option for controlled ovarian stimulation (COS) in assisted reproductive technology protocols and for ovulation induction (OI) in anovulatory infertility, and is associated with a different endocrine profile from that of recombinant (r) FSH in these settings (in terms of serum levels of FSH, androgens and/or estradiol). When used for COS in women undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI), Menopur® was as good as rFSH in terms of pregnancy rates (despite being associated with a lower oocyte yield) and was found to improve some aspects of embryo quality in the IVF (but not ICSI) setting; using Menopur® in combination with highly purified urinary FSH resulted in similar reproductive outcomes as Menopur® alone. Data for Menopur® in OI are limited, but suggest ovulation rates may be as good as those with rFSH + rLH (in type 1 anovulation) and rFSH (in type 2 anovulation). Moreover, compared with rFSH, Menopur® appeared to be associated with a less pronounced follicular response and a lower risk of ovarian overstimulation.

Effect of Mechanical Microvibration on the Quality of Human Embryos during In Vitro Culturing and Outcomes of Assisted Reproduction Technologies.

We summarized the data on the role and efficiency of mechanical microvibration during embryo culturing in assisted reproduction protocols. The efficiency of culturing systems with microvibration in assisted reproduction programs and possible mechanisms of microvibration effect on the preimplantation development of human embryos are discussed.