Recombinant Luteinizing Hormone Research Articles

Effects of Vaccination against Recombinant FSH or LH Receptor Subunits on Gonadal Development and Functioning Male Rats.

Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) play key roles in regulating testosterone secretion and spermatogenesis in male mammals, respectively, and they maintain the fertility of male animals by binding to their corresponding receptors. We designed and prepared a recombinant LH receptor (LHR) subunit vaccine and a recombinant FSH receptor (FSHR) subunit vaccine and used male Sprague Dawley (SD) rats as a model to examine their effects on testicular development, spermatogenesis, and testosterone secretion in prepubertal and pubertal mammals. Both vaccines (LHR-DTT and FSHR-DTT) significantly decreased the serum testosterone level in prepubertal rats (p < 0.05) but had no effect on the testosterone secretion in pubertal rats; both vaccines decreased the number of cell layers in the seminiferous tubules and reduced spermatogenesis in prepubertal and pubertal rats. Subunit vaccine FSHR-DTT decreased the sperm density in the epididymis in both prepubertal and pubertal rats (p < 0.01) and lowered testicular index and sperm motility in pubertal rats (p < 0.05), whereas LHR-DTT only reduced the sperm density in the epididymis in pubertal rats (p < 0.05). These results indicate that the FSHR subunit vaccine may be a promising approach for immunocastration, but it still needs improvements in effectiveness.

Maternal MicroRNA Profile Changes When LH Is Added to the Ovarian Stimulation Protocol: A Pilot Study.

While the use of follicle-stimulating hormone (FSH) in ovarian stimulation for in vitro fertilization (IVF) is an established practice, the use of luteinizing hormone (LH) remains debatable. MicroRNAs (miRNAs) are short, endogenous, non-coding transcripts that control a variety of cellular functions, such as gonadotrophin production and follicular development. The goal of this pilot study was to investigate whether the employment of recombinant LH (rLH) in ovarian stimulation protocols results in changes in the miRNA profiles in human oocytes. Patients were divided into two groups: seven received recombinant FSH (rFSH, 225 IU), and six received rFSH (150 IU) plus rLH (75 IU). MiRNA predesigned panels and real-time PCR technology were used to analyze the oocytes retrieved from the follicular ovarian retrieval. Among the miRNAs evaluated, a series of them evidenced upregulation or downregulation in their expression in the FSH plus LH group compared to the FSH group. Considering the results obtained from the functional and network analysis, the different maternal miRNA profiles in the two groups revealed a differential modulation of pathways involved in numerous biological functions. Overall, based on the pathways associated with most of these maternal miRNAs, the presence of LH may result in a different modulation of pathways regulating survival under the control of a Tp53-related mechanism. Interestingly, among the miRNAs differentially expressed in oocytes of the two groups, we have found miRNAs already investigated at ovarian, follicular, oocyte, and embryonic levels: hsa-miR-484, hsa-miR-222, hsa-miR-520d-5p, hsa-miRNA-17, hsa-miR-548, and hsa-miR-140. Thus, investigation into the role of these miRNAs in oocyte molecular pathways may help determine how LH affects oocyte competence and eventually leads to the clinical improvement of IVF.

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

Recombinant Gonadotropins to Induce Oocyte Development In Vitro and In Vivo in the European Eel Anguilla anguilla

Commonly, female European eels are injected weekly with pituitary extract (PE) from carp (CPE) or salmon (SPE) to induce sexual maturation. However, a PE is a mixture of gonadotropins and other hormones that are not specific for eel and rapidly cleared from circulation. The aim of this study was therefore to test the effects of highly stable eel-specific recombinant gonadotropins (rGTHs) on oocyte development in vitro and in vivo in European eels. For the in vitro trial, the dose–effect responses of maturing eel oocytes on CPE and recombinant luteinizing hormone (rLH) were studied before and after 12 and 18 h of incubation. For the in vivo experiment, sexual maturation was stimulated by treatment with (i) CPE, (ii) recombinant follicle-stimulating hormone (rFSH) followed by CPE and (iii) rFSH followed by rLH. For the in vitro experiment, the expression of the nuclear progestin receptor 2 (pgr2) was induced by rLH, implying that rLH was preparing the oocyte for ovulation. For the in vivo experiment, the females treated with rGTHs had high gonadosomatic index (GSI) values (rFSH-CPE: 75, 77; rFSH-rLH: 80) in comparison with the females injected with CPE (50–60), suggesting that rFSH strongly induced vitellogenic growth. Larvae were produced for all treatment groups and for the first time by rGTH treatment alone but dose and timing still need optimization.

Recombinant luteinizing hormone as a tool towards understanding reproductive dysfunction in captive Macquarie perch Macquaria australasica

The Macquarie perch Macquaria australasica is an endangered freshwater fish endemic to the Murray-Darling Basin in Australia. Captive breeding is considered an essential measure to save the species from extinction, yet its reproductive biology is not fully understood. We produced a recombinant single-chain Macquarie perch luteinizing hormone (rmpLh) in the methylotrophic yeast Pichia pastoris, confirmed by Western blot analysis and mass spectrometry. We developed and validated a heterologous competitive enzyme-linked immunosorbent assay (ELISA) for M. australasica Lh using antibodies generated against the giant grouper Epinephelus lanceolatus Lhb, which is 91% identical to the M. australasica sequence at the amino acid level. Measurement of Lh in plasma samples collected at 2 timepoints (June vs. August 2020) from captive F1 broodstock did not show a significant difference. Plasma Lh in samples from wild fish collected in September 2020, a month prior to the spawning season, were almost 10-fold higher than those of the F1 broodstock. The tools generated here will help us to understand the reproductive biology of M. australasica and develop reliable assisted reproduction techniques for the species.

Comparative Effects of Human Chorionic Gonadotropin and Recombinant Luteinizing Hormone Supplementation during the Late Follicular Phase

In selected patients, addition of LH activity can improve the outcome of ovarian stimulation. Two common LH activity supplementation strategies are recombinant human LH (rLH) and human chorionic gonadotropin (hCG). The respective effectiveness of hCG versus rLH using various daily dosages and modes of administration is still discussed. In this study, we compared the outcomes of hCG versus rLH supplementation among patients with slow response to recombinant follicle stimulating hormone (rFSH).

A retrospective, matched case-control study of recombinant LH versus hMG supplementation on FSH during controlled ovarian hyperstimulation in the GnRH-antagonist protocol.

BackgroundThe role of luteinizing hormone (LH) in controlled ovarian hyperstimulation (COH) requires more evidence for its efficacy. Several studies compared recombinant human LH (r-hLH) or human menopausal gonadotropin (hMG) in combination with recombinant human follicle-stimulating hormone (r-hFSH) but lack the results with GnRH-antagonist protocol and in Asians.MethodsThis is a retrospective, single-center study inspecting women receiving GnRH antagonist protocol and r-hFSH+hMG or r-hFSH+r-hLH regimen for over five days for COH in the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycle in Taiwan from 2013 to 2018. The outcomes of IVF/ICSI cycles were analyzed after propensity score matching between the two groups. A subgroup analysis was conducted in cycles in which women underwent their first embryo transfer (ET), including fresh ET and frozen ET (FET).ResultsWith a total of 503 cycles, the results revealed that the r-hFSH+r-hLH group performed better in terms of numbers of oocytes retrieved (r-hFSH+hMG vs. r-hFSH+r-hLH, 11.7 vs. 13.7, p=0.014), mature oocytes (8.7 vs. 10.9, p=0.001), and fertilized oocytes (8.3 vs. 9.8, p=0.022), while other outcomes were comparable. The analysis of first ET cycles also showed similar trends. Although the implantation rate (39% vs. 43%, p=0.37), pregnancy rate (52% vs. 53%, p=0.90), and live birth rate (39% vs. 45%, p=0.19) were not significantly different, the miscarriage rate was higher in the r-hFSH+hMG group than the r-hFSH+r-hLH group (26% vs. 15%, p<0.05) in first ET cycles. The cumulative pregnancy rate was significantly higher in the r-hFSH+r-hLH group (53% vs. 64%, p=0.02). No significant difference in rates of ovarian hyperstimulation syndrome (OHSS) was observed.ConclusionThe results support the hypothesis that the treatment of r-hLH+r-hFSH improves COH clinical outcomes in the IVF/ICSI cycle.

Effect of recombinant LH supplementation timing on clinical pregnancy outcome in long-acting GnRHa downregulated cycles

BackgroundTimely and moderate luteinizing hormone (LH) supplementation plays positive roles in in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) cycles with long-acting gonadotropin-releasing hormone agonist (GnRHa) pituitary downregulation. However, the appropriate timing of LH supplementation remains unclear.MethodsWe carried out a retrospective cohort study of 2226 cycles at our reproductive medicine centre from 2018 to 2020. We mainly conducted smooth curve fitting to analyse the relationship between the dominant follicle diameter when recombinant LH (rLH) was added and the clinical pregnancy outcomes (clinical pregnancy rate or early miscarriage rate). In addition, total cycles were divided into groups according to different LH levels after GnRHa and dominant follicle diameters for further analysis.ResultsSmooth curve fitting showed that with the increase in the dominant follicle diameter when rLH was added, the clinical pregnancy rate gradually increased, and the early miscarriage rate gradually decreased.ConclusionsIn long-acting GnRHa downregulated IVF/ICSI-ET cycles, the appropriate timing of rLH supplementation has a beneficial impact on the clinical pregnancy outcome. Delaying rLH addition is conducive to the clinical pregnancy rate and reduces the risk of early miscarriage.

The Association of Serum Progesterone Levels on Day of Oocyte Retrieval with Pregnancy Outcome

Aims and Objectives: To establish the level of serum progesterone (P4) on the day of oocyte retrieval beyond which it can affect the outcome of in vitro fertilisation (IVF), and to further establish the incidence of serum P4 rise in an agonist and antagonist cycle. Methods: This prospective observational cohort study was conducted from November 2020 to November 2021 at the Sarvodaya Fertility and IVF Centre, Delhi, India. For this study, the author recruited 352 couples with infertility who were treated with IVF/intracytoplasmic sperm injection-embryo transfer, of which 279 patients completed an IVF/intracytoplasmic sperm injection-embryo transfer cycle during the study period and were included in the final analysis. The standard gonadotropin-releasing hormone antagonist (fixed or variable) and long gonadotropin-releasing hormone agonist controlled ovarian stimulation protocols were used in all patients. Participants were recruited if they were undergoing controlled ovarian stimulation with all gonadotropins, recombinant follicle-stimulating hormone/urinary human menopausal gonadotropin, or recombinant luteinising hormone. The study population was sub-grouped into two groups according to their P4 level on day of oocyte retrieval (calculated according to receiver operating characteristics curve): Group A (p≤11.6 ng/dL; n=247 out of 27; 88.5%) and Group B (p&gt;11.6 ng/dL; n=32 out of 279; 11.5%). Statistical analysis was performed with the Statistical Package for Social Sciences (SPSS) 17.0 version (IBM, New York City, New York, USA). Results: The percentage of patients with a rise in P4 on the day of oocyte retrieval were found significantly more in the antagonist protocol (13.3% [24 out of 181]) than in the agonist protocol (8.2% [8 out of 98]; p=0.04). Pregnancy rate was significantly higher in Group A (39.3% [97 out of 247]) compared with Group B (12.5% [4 out of 32]). The clinical pregnancy rate was also significantly higher in Group A (34.4% [85 out of 247]) compared with Group B (6.3% [2 out of 32]). Conclusion: Patients with higher levels of P4 (&gt;11.6 ng/mL) were associated with lower pregnancy and clinical pregnancy rates.

Expression and function of the luteinizing hormone choriogonadotropin receptor in human endometrial stromal cells

The human luteinising hormone choriogonadotropin receptor (LHCGR) is a G-protein coupled receptor activated by both human chorionic gonadotropin (hCG) and luteinizing hormone (LH), two structurally related gonadotropins with essential roles in ovulation and maintenance of the corpus luteum. LHCGR expression predominates in ovarian tissues where it elicits functional responses through cyclic adenosine mononucleotide (cAMP), Ca2+ and extracellular signal-regulated kinase (ERK) signalling. LHCGR expression has also been localized to the human endometrium, with purported roles in decidualization and implantation. However, these observations are contentious. In this investigation, transcripts encoding LHCGR were undetectable in bulk RNA sequencing datasets from whole cycling endometrial tissue and cultured human endometrial stromal cells (EnSC). However, analysis of single-cell RNA sequencing data revealed cell-to-cell transcriptional heterogeneity, and we identified a small subpopulation of stromal cells with detectable LHCGR transcripts. In HEK-293 cells expressing recombinant LHCGR, both hCG and LH elicited robust cAMP, Ca2+ and ERK signals that were absent in wild-type HEK-293 cells. However, none of these responses were recapitulated in primary EnSC cultures. In addition, proliferation, viability and decidual transformation of EnSC were refractory to both hCG and LH, irrespective of treatment to induce differentiation. Although we challenge the assertion that LHCGR is expressed at a functionally active level in the human endometrium, the discovery of a discrete subpopulation of EnSC that express LHCGR transcripts may plausibly account for the conflicting evidence in the literature.

Androgen Flare after LHRH Initiation Is the Side Effect That Makes Most of the Beneficial Effect When It Coincides with Radiation Therapy for Prostate Cancer.

Simple SummaryProstate cancer tumor growth is stimulated by androgens. Surgical castration or medical castration using long-acting luteinizing hormone-releasing hormone (LHRH) agonists or antagonists is the backbone of the treatments of metastatic disease. Treatment of locally advanced prostate cancer was accomplished with radiation therapy alone until multiple studies showed that combining radiation therapy with LHRH agonists results in significant survival benefit. While the goal of the use of LHRH agonists was to suppress testosterone levels during radiation, we show, through review of previous studies, that survival benefit was achieved only when LHRH was initiated during the course of radiation, and thus androgen flare during the first 1–3 weeks after the initiation of LHRH is most likely the reason for higher survival. Androgens drive tumor cells into mitosis, and mitotic death is the dominant mechanism of tumor cell kill by radiation.Treatment of metastatic prostate cancer was historically performed via bilateral orchiectomy to achieve castration. An alternative to surgical castration is the administration of subcutaneous recombinant luteinizing hormone-releasing hormone (LHRH). LHRH causes the pituitary gland to produce luteinizing hormone (LH), which results in synthesis and secretion of testosterone from the testicles. When LHRH levels are continuously high, the pituitary gland stops producing LH, which results in reduced testosterone production by the testicles. Long-acting formulations of LHRH were developed, and its use replaced surgical orchiectomy in the vast majority of patients. Combining LHRH and radiation therapy was shown to increase survival of prostate cancer patients with locally advanced disease. Here, we present a hypothesis, and preliminary evidence based on previous randomized controlled trials, that androgen surge during radiation, rather than its suppression, could be responsible for the enhanced prostate cancer cell kill during radiation. Starting LHRH agonist on the first day of radiation therapy, as in the EORTC 22863 study, should be the standard of care when treating locally advanced prostate cancer. We are developing formulations of short-acting LHRH agonists that induce androgen flare, without subsequent androgen deprivation, which could open the door for an era in which locally advanced prostate cancer could be cured while patients maintain potency.

Recombinant follicular stimulating hormone plus recombinant luteinizing hormone versus human menopausal gonadotropins- does the source of LH bioactivity affect ovarian stimulation outcome?

ObjectiveLuteinizing hormone (LH) and human chorionic gonadotropin (hCG) activate distinct intracellular signaling cascades. However, due to their similar structure and common receptor, they are used interchangeably during ovarian stimulation (OS). This study aims to assess if the source of LH used during OS affects IVF outcome.Patients and methodsThis was a cross sectional study of patients who underwent two consecutive IVF cycles, one included recombinant follicular stimulating hormone (FSH) plus recombinant LH [rFSH+rLH, (Pergoveris)] and the other included urinary hCG [highly purified hMG (HP-hMG), (Menopur)]. The OS protocol, except of the LH preparation, was identical in the two IVF cycles.ResultsThe rate of mature oocytes was not different between the treatment cycles (0.9 in the rFSH+rLH vs 0.8 in the HP-hMG, p = 0.07). Nonetheless, the mean number of mature oocytes retrieved in the rFSH+rLH treatment cycles was higher compared to the HP-hMG treatment cycles (10 ± 5.8 vs 8.3 ± 4.6, respectively, P = 0.01). Likewise, the mean number of fertilized oocytes was higher in the rFSH+rLH cycles compared with the HP-hMG cycles (8.5 ± 5.9 vs 6.4 ± 3.6, respectively, p = 0.05). There was no difference between the treatment cycles regarding the number of top-quality embryos, the ratio of top-quality embryos per number of oocytes retrieved or fertilized oocytes or the pregnancy rate.ConclusionThe differences in treatment outcome, derived by different LH preparations reflect the distinct physiological role of these molecules. Our findings may assist in tailoring a specific gonadotropin regimen when assembling an OS protocol.

Luteal-phase support in assisted reproductive technology: An ongoing challenge.

It has been shown that in controlled ovarian hyper stimulation cycles, defective luteal phase is common. There are many protocols for improving pregnancy outcomes in women undergoing fresh and frozen in vitro fertilization cycles. These approaches include progesterone supplements, human chorionic gonadotropin, estradiol, gonadotropin-releasing hormone agonist, and recombinant luteinizing hormone. The main challenge is luteal-phase support (LPS) in cycles with gonadotropin-releasing hormone agonist triggering. There is still controversy about the optimal component and time for starting LPS in assisted reproductive technology cycles. This review aims to summarize the various protocols suggested for LPS in in vitro fertilization cycles.

Improving Implantation Rate in 2nd ICSI Cycle through Ovarian Stimulation with FSH and LH in GNRH Antagonist Regimen.

To investigate whether patients with a previous recombinant follicle stimulating hormone (rFSH)-stimulated cycle would have improved outcomes with rFSH + recombinant luteinizing hormone (rLH) stimulation in the following cycle. For the present retrospective case-control study, 228 cycles performed in 114 patients undergoing intracytoplasmic sperm injection (ICSI) between 2015 and 2018 in an in vitro fertilization (IVF) center were evaluated. Controlled ovarian stimulation (COS) was achieved with rFSH (Gonal-f, Serono, Geneva, Switzerland) in the first ICSI cycle (rFSH group), and with rFSH and rLH (Pergoveris, Merck Serono S.p.A, Bari, Italy) in the second cycle (rFSH + rLH group). The ICSI outcomes were compared among the groups. Higher estradiol levels, oocyte yield, day-3 high-quality embryos rate and implantation rate, and a lower miscarriage rate were observed in the rFSH + rLH group compared with the rFSH group. In patients < 35 years old, the implantation rate was higher in the rFSH + rLH group compared with the rFSH group. In patients ≥ 35 years old, higher estradiol levels, oocyte yield, day-3 high-quality embryos rate, and implantation rate were observed in the rFSH + rLH group. In patients with ≤ 4 retrieved oocytes, oocyte yield, mature oocytes rate, normal cleavage speed, implantation rate, and miscarriage rate were improved in the rFSH + rLH group. In patients with ≥ 5 retrieved oocytes, higher estradiol levels, oocyte yield, and implantation rate were observed in the rFSH + rLH group. Ovarian stimulation with luteinizing hormone (LH) supplementation results in higher implantation rates, independent of maternal age and response to COS when compared with previous cycles stimulated with rFSH only. Improvements were also observed for ICSI outcomes and miscarriage after stratification by age and retrieved oocytes.

P–659 Artificial intelligence (AI) as an assisting tool in generating patient-friendly corifollitropin alfa ovarian stimulation protocol during in vitro fertilization

Abstract Study question How machine learning assisted in generating patient-friendly corifollitropin alfa protocol in normal responders? Summary answer In retrospective experiments, our machine learning model integrated physiological measurements of patients and clinical experience to generate a patient-friendly corifollitropin alfa protocol. What is known already Long-acting corifollitropin alfa can simplify the regimen, minimizing injections during the whole cycle. The previous study has described the patient-friendly protocol using corifollitropin alfa without routine pituitary suppression in normal responder can result in non-compromised clinical outcomes. Some studies showed machine learning can help with making clinical decisions and have the ability to learn from physiological measurements. Those methods effectuate certain points throughout short-acting menotropin protocols, however, there are still no robust AI tools for long-acting corifollitropin alfa protocols. Study design, size, duration 1,309 cycles were collected at Stork Fertility Center from November 2016 to October 2019, and 1,221 cycles were available after data cleaning and applying exclusion criteria, which Anti-Mullerian Hormone (AMH) is lower than 2. The data from electronic medical records (EMRs) consisted of age, AMH, body weight, luteinizing hormone (LH), and estradiol (E2) concentrations measured on revisit. Evaluation is performed by one physician who has more than 20 years of experience in IVF. Participants/materials, setting, methods: The protocol generator consisted of 5 parts: doses of Elonva, trigger type, doses of recombinant follicle-stimulating hormone (rFSH), doses of recombinant luteinizing hormone (rLH), and day of oocyte retrieval. The protocol was predicted by age, AMH, and weight firstly, then fine-tuned by LH and E2 after the first revisit. We used the gradient boosting decision tree algorithm to learn the protocol. The dataset was randomly split into 80% for training and 20% for testing. Main results and the role of chance In classification, the model predicted the dose of Elonva achieved an accuracy of 0.913 and an AUC of 0.946, and trigger type got an accuracy of 0.901 and AUC of 0.852 only using features on stimulation day (SD) 1 and gained 0.012 and 0.056 in accuracy and AUC correspondingly after adding features on the first revisit day. In regression, the mean absolute error (MAE) of rFSH dose, rLH dose, and oocytes retrieved day was 156.30 IU, 232.75 IU, and 0.80 days respectively, and after refining, the MAE dropped to 92.37 IU, 100.07 IU, and 0.46 days. The error of predictions in rFSH and rLH was almost equal to half increments of rFSH (150 IU) and one increment rLH (75 IU). This indicated that our model could provide a better prediction of these clinical decisions with one revisit only. Limitations, reasons for caution The present study was a single-center retrospective, and only analyzed the data from normal responders, whose AMH was equal or greater than 2. Though, the recommendations of our system act as references, the physician will make the final decision. Wider implications of the findings: Our result showed the potential of machine learning in generating protocols is promising. Recommendations generated by our model can provide the junior clinical teams to optimize the clinical plans and learn from the experience of experts. We look forward to applying our machine learning model to different protocols. Trial registration number Not applicable

P–594 Ovarian stimulation with luteinizing hormone supplementation: the impact of timing on ovarian response and ICSI outcomes

Abstract Study question Is there an impact of recombinant luteinizing-hormone (rLH) administration timing during controlled ovarian stimulation (COS) on ovarian response and intracytoplasmic sperm injection (ICSI) cycles outcomes? Summary answer rLH supplementation in patients with poor ovarian response (POR) improves laboratorial and clinical outcomes when started in the mid-follicular phase, in GnRH antagonist ICSI cycles. What is known already Meta-analyses demonstrated that the use of rLH combined with rFSH for COS may lead to more ongoing pregnancies than rFSH alone. However, there is limited evidence that the timing of rLH addition to rFSH may impact the ovarian response or the outcomes of ICSI, based on a limited casuistic, which demonstrated improved ovarian response, embryo quality and pregnancy rate with LH supplementation from GnRH antagonist administration day, in estimated POR patients. The objective of the present study was to further investigate this hypothesis in a larger population, and in subpopulations of patients stratified by age and response to COS. Study design, size, duration This historical cohort study included data obtained via chart review of 1278 ICSI cycles performed in 1278 patients between 2015 and 2018, in a private university-affiliated in vitro fertilization center. Post hoc power analysis was calculated, given α of 5%, sample size of 1278, and effect size for implantation rate. The achieved power was superior to 99%. Participants/materials, setting, methods Two groups were formed according to timing of LH administration: Group LH-start (n = 323), in which LH was started on day–1; and Group LH-mid (n = 955), in which LH was started with GnRH antagonist. Then, data were stratified according to female age (&amp;lt;35 years-old, n = 283, and ≥35 years-old, n = 995) and response to COS (poor response (POR): ≤4 retrieved oocytes, n = 423, and normal response: &amp;gt;5 retrieved oocytes, n = 855). Ovarian response and ICSI outcomes were compared among the groups. Main results and the role of chance In POR patients, significantly higher fertilization rate (68.3% ± 2.5 vs. 78.6% ± 3.7, p = 0.023), blastocyst development rate (22.5% ± 7.2 vs. 44.7% ± 6.2, p = 0.022) and implantation rate (17.6% ± 59.1 vs. 20.2% ± 43.2, p &amp;lt; 0.001) were observed in Group LH-mid, even though the amount of LH used in these patients was not significant different from that used in Group LH-mid from patients with normal response to COS (1062.35 IU ± 54.33 vs. 925.81 IU ± 414.41, p: 0.431, respectively). For the general group and in patients aged ≥ 35 years, higher blastocyst development rates were observed in Group LH-mid compared to Group LH-start (33.0% ± 31.9 vs. 40.8% ± 32.6, p = 0.012, and 28.8% ± 30.4 vs 38.5% ± 32.3, p = 0.006, respectively). In patients aged &amp;lt; 35 years and in those with normal response to COS, similar outcomes were obtained irrespective of timing of LH administration. Limitations, reasons for caution The limitations included the retrospective design and limited sample size in subpopulations. In addition, the reduced clinical outcomes related to POR patients may hamper the true estimation of the differences between the stimulation groups in terms of pregnancy and miscarriage rates. Wider implications of the findings: In POR patients, mid-follicular phase LH supplementation starting with 150 IU daily doses, may rescue the ongoing cycle by compensating an initial slow response, and balancing the deprivation of endogenous LH in GnRH antagonist cycles, with no need of expending more gonadotropin compared to patients with normal response to COS. Trial registration number Not applicable

Effect of luteinizing hormone concentration on transcriptome and subcellular organelle phenotype of ovarian granulosa cells.

Granulosa cells (GCs) surrounding oocytes are crucial for follicular growth, oocyte development, ovulation, and luteinization under the dynamic co-stimulation of follicle stimulating hormone (FSH) and luteinizing hormone (LH). This study aimed to investigate the effect of LH levels on GCs in preovulatory follicles under gonadotropin releasing hormone antagonist-based ovarian stimulation. In vitro experiments were also conducted to study the direct effect of LH on GCs. Twelve infertile women were divided into low (L), medium (M), and high (H) LH groups according to their serum LH levels during ovarian stimulation. RNA-sequencing (RNA-seq) was conducted to examine the transcriptome profiles of GCs obtained from the above patients during the oocyte retrieval. The activity of mitochondrial dehydrogenase was measured under the stimulation of recombinant LH (rLH) concentration gradient combined with recombinant FSH. The ultrastructures of subcellular organelles were observed. Bioinformatic analyses showed that compared with the M group, molecule and pathway changes in the L group and in the H group were similar. In cultured GCs, both insufficient and excessive rLH impaired the activity of mitochondrial dehydrogenase. With the medium rLH concentration, numerous cell connections and abundant mitochondria and liposomes were observed. Compared with the medium concentration, GCs showed smaller and rounder mitochondria, more autophagosomes, and massive organelles damages with excessive rLH, and swollen, circular, or forked mitochondria were observed with inadequate rLH. RNA-seq provided a novel spectrum of transcriptome characteristics of GCs potentially affected by serum LH levels during ovarian stimulation. In vitro, rLH could directly affect GCs at the subcellular level.

Development of a giant grouper Luteinizing Hormone (LH) Enzyme-Linked Immunosorbent Assay (ELISA) and its use towards understanding sexual development in grouper

A recombinant giant grouper Luteinizing Hormone (LH) consisting of tethered beta and alpha subunits was produced in a yeast expression system. The giant grouper LH β-subunit was also produced and administered to rabbits for antibody development. The recombinant LH and its antibody were used to develop an Enzyme Linked Immunosorbent Assay (ELISA). This ELISA enabled detection of plasma LH levels in groupers at a sensitivity between 391 pg/ml and 200 ng/ml. Different species of grouper were assayed with this ELISA in conjunction with gonadal histology and body condition data to identify links between circulating LH levels and sexual development. We found that circulating levels of LH decreased when oocytes began to degenerate, and sex-transition gonadal characteristics were apparent when LH levels decreased further. When circulating LH levels were related to body condition (body weight/ body length), transitioning-stage fish had relatively high body condition but low plasma LH levels. This observation was similar across multiple grouper species and indicates that plasma LH levels combined with body condition may be a marker for early male identification in the protogynous hermaphrodite groupers.

Induced multiple follicular growth and ongoing twin pregnancy by IVF in a woman with premature menopause

Menopause, natural or premature, was long been considered to be incompatible with conception with the own eggs of the women affected. Here we describe a case of a 32-year-old woman with premature menopause, characterized by amenorrea, the absence of antral follicles in the ovaries, high basal follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and extremely low antimullerian hormone (AMH) levels. After preliminary treatment with contraceptive pill for two months, and an injection of recombinant human chorionic gonadotropin (HCG) to boost intrinsic androgen production, ovarian stimulation, using a customized protocol with a patient-tailored combination of recombinant FSH and LH, in addition to daily injections of growth hormone, resulted in the recovery of 3 metaphase II eggs after double triggering with recombinant HCG. After transfer of two embros, obtained by intracytoplasmic sperm injection, an ongoing twin clinical pregnancy was achieved. These data show that oocyte recovery, fertilization, embryo development and pregnancy can be achieved, with the use of a customized ovarian stimulation protocol, even in young women with clear signs of advanced premature menopause.

The Effect of Recombinant LH Addition to Recombinant FSH on Assisted Reproductive Technologies Outcomes in Overweight and Obese Patients without Polycystic Ovary Syndrome

Background: Obesity and overweight negatively affect fertility. Protocols need to be developed in ART treatments for obese and overweight women. Objectives: To investigate the effect of recombinant luteinizing hormone (rLH) addition to recombinant follicle stimulating hormone (rFSH) on treatment outcomes in assisted reproductive technologies (ART) cycles in obese and overweight women without polycystic ovary syndrome. Methods: This retrospective cohort study was carried out in Kocaeli University Faculty of Medicine, Assisted Reproductive Techniques Clinic between January 2016 and March 2019. To analyze the impact of rLH addition to rFSH in GnRH antagonist cycles in overweight and obese, the patients were divided into four groups according to body mass index (BMI) and gonadotropin type; Group 1: patients with BMI ≥ 25 stimulated with rFSH alone (n: 37), Group 2: patients with BMI ≥ 25 stimulated with rFSH + rLH (n: 37), Group 3: patients with BMI between 18.5 - 24.99 stimulated with rFSH alone (n: 33), Group 4: patients with BMI between 18.5 and 24.99 stimulated with rFSH + rLH (n: 30). Patients with polycystic ovary syndrome were excluded. Results: Basal LH levels were found to be significantly lower in obese and overweight patients compared to normoweight patients (p = 0.01). Grade 1 embryo ratio in obese and overweight patients was higher in rLH added obese group than in group LH not included (64.9%, p = 0.005). Ongoing pregnancy rates (OPR) in obese and overweight patients were significantly higher in rLH added group compared to rFSH only group (43.2% vs 18.9% respectively, p = 0.044). However, OPR did not differ significantly in rLH added and rFSH only groups in normoweight patients (p = 0.588). Conclusion: This study has shown that obese and overweight non-PCOS patients have lower endogenous LH levels. It has also shown that rLH supplementation in GnRH antagonist cycles in obese and overweight women improves embryo quality and ongoing pregnancy rates. However rLH addition to rFSH doesn’t seem to have a value in normoweight patients.