Combination Of Gonadotropin-releasing Hormone Agonist Research Articles

Gamete Collection, Artificial Fertilization and Captive-Rearing of Eggs in a Terrestrial-Breeding Anuran with Parental Care: Alytes obstetricans.

The genus Alytes consists of six primitive terrestrial-breeding species (five of them endangered or present in restricted distributions) with unique male parental care. Alytes obstetricans was used as a model for the development of assisted reproductive technologies for gamete collection, artificial fertilization and captive-rearing of embryos. Sperm collection was achieved with human chorionic gonadotropin (hCG), but only 17 to 50% of males responded to the hormone. In addition, an effect of captivity on sperm quality was observed. A combination of gonadotropin-releasing hormone agonist (GnRHa) and hCG was tested for induction of ovulation, and 100% of females responded to the treatment. Recently collected sperm was used in artificial fertilization, and 10% of eggs showed complete embryo development. To design a method for captive-rearing of embryos, natural clutches (n = 13) were removed from males and divided into two sub-clutches. One was kept with all eggs together and in the other, the egg string was cut and eggs were split individually. All sub-clutches with eggs kept separated presented embryo survival (ranging from 27 to 61%) that hatched normally, while only one sub-clutch with embryos maintained together had some embryo survival (36%). These results may contribute to the management of this and related species.

A combination of GnRH-hCG in aged mares improves embryo recovery

Older embryo donor mares typically have proven offspring, representing the largest population in most embryo transfer programs. However, aged embryo donors typically have poor embryo recovery due to uterine, cervical, and ovarian problems. In addition, older mares are thought to have deficient oocyte nuclear and cytoplasm maturation, leading to poor fertility. It has been proposed that better fertility results from combining LH-like (e.g. hCG) and GnRH-agonists to induce ovulation for potentially different mechanisms of action, as the LH-like molecule would presumably promote better nuclear maturation matching with the cytoplasm. Alternatively, the authors have been using a protocol in clinical practice, which consists of administering low doses of hCG and low doses of GnRH agonists before induction of ovulation to promote oocyte maturation; thus this study was undertaken to assess the usefulness of this protocol by assessing embryo recovery of commercial, aged embryo donor mares with subpar fertility. The study was conducted with six sub-fertile embryo donor mares (20±1.7, ranging 18-23yrs-old). In experiment 1, estrous cycles were assigned as control-cycles (n=13) or deslorelin-hCG cycles (n=12). Control-cycles received saline (0.5mL, i.m., q12h), and deslorelin-hCG cycles received deslorelin (125µg, i.m., q12h) starting when a dominant follicle achieved 20mm, and then once the same follicle achieved 25mm, mares received hCG (500units, i.m., q24h) until the dominant follicle achieved a periovulatorystage (i.e., ≥35mm with uterine edema ≥1). None of the mares received more than four doses of hCG. Then, ovulation was induced with hCG (2,500units). In experiment 2, thirty-cycles of the same mares were randomly assigned to control-cycles (identical to experiment 1, n=19) and histrelin-hCG cycles (n=11), which followed a similar design to experiment 1, except that instead of administering deslorelin, histrelin was given (125µg, i.m.). Ovulation was induced and confirmed by daily monitoring. In both experiments, mares were bred with cooled semen from fertile stallions. Embryo flushing was carried out eight days post-ovulation, and embryos graded. Statistical analyses were carried out with Fisher's exact test. Significance was set as P<0.05. Overall, 25% of cycles did not grow follicles in response to the treatment, double ovulation happened on three-treated-cycles. One mare had double ovulations in the control- and treated-cycles. None of the mares failed to ovulate after hCG. In experiment 1, the embryo recovery was 58.8% (7/12) for deslorelin-hCG cycles vs. 7.7% (1/13) for control-cycles (P<0.05). In experiment 2, the embryo recovery for histrelin-hCG cycles was 72.8% (8/11) in comparison with control-cycles 21% (4/19 cycles; (P<0.05). In conclusion, the results of this field study suggest that a combination of GnRH-agonist and hCG early in the cycle improves embryo recovery in sub-fertile aged mares and that the majority of cycles (75%) will respond to treatment namely grow follicles and ovulate.

Ovulation triggering with hCG alone, GnRH agonist alone or in combination? A randomized controlled trial in advanced-age women undergoing IVF/ICSI cycles.

Is a dual ovulation trigger with a combination of GnRH agonist (GnRHa) and hCG superior to single hCG and/or single GnRHa trigger in improving treatment outcomes in advanced-age women (aged ≥ 35 years) undergoing IVF/ICSI treatment? Co-administration of GnRHa and hCG as a dual trigger increases the number of good-quality embryos but it is not associated with a higher number of oocytes retrieved, compared with single hCG or GnRHa trigger. Many studies have demonstrated that a dual trigger has positive impact on oocyte maturation, retrieval rate and pregnancy rate without increasing the risk of ovarian hyperstimulation syndrome (OHSS) in some groups of IVF patients, when compared with single hCG trigger. Few studies have however been conducted to compare a dual trigger with a single GnRHa trigger, and insufficient evidence exists to support which trigger can achieve the best outcomes in IVF patients aged ≥35 years. This was an open-label randomized controlled trial of 510 participants conducted at single reproductive medical center from January 2019 to December 2021. After a sample size calculation performed by retrospectively analyzing our previous clinical data, we planned to recruit 170 patients in each group and 510 patients in total for the study. Women aged ≥35 years undergoing IVF/ICSI treatment, receiving a non-pituitary down-regulation protocol, and with low risk of OHSS, were enrolled in this trial. On the trigger day, patients were randomized into three groups: hCG alone (who received 6000 IU of hCG), GnRHa alone (who received 0.2 mg of triptorelin) and dual trigger (who received 0.2 mg of triptorelin plus 2000 IU of hCG) groups. The primary outcome parameter was the number of retrieved oocytes. The secondary outcome parameters included, among others, the number and rates of mature oocytes, two pronuclei (2PN) embryos and good-quality embryos, as the rates of OHSS, clinical pregnancy, miscarriage and live birth. There were no significant differences in the baseline demographic characteristics among the three groups. The dual trigger was associated with a higher retrieval rate (87.9% vs 84.1% in the hCG group, P = 0.031; 87.9% vs 83.6% in the GnRHa group, P = 0.014). However, the number of retrieved oocytes in the dual trigger group was comparable with those in the hCG group (4.08 ± 2.79 vs 3.60 ± 2.71, P = 0.080) and the GnRHa group (4.08 ± 2.79 vs 3.81 ± 3.38, P = 0.101); comparable data between the groups were also found when analyzing the number of 2PN embryos and the 2PN rate. In the dual trigger group, the numbers of good-quality embryos and viable embryos were both significantly higher than in the hCG group (1.74 ± 1.90 vs 1.19 ± 1.45, P = 0.016 and 2.19 ± 2.11 vs 1.56 ± 1.66, P = 0.008, respectively) and the GnRHa group (1.74 ± 1.90 vs 1.20 ± 1.67, P = 0.003 and 2.19 ± 2.11 vs 1.45 ± 1.75, P = 0.001, respectively). Pregnancy outcomes after fresh embryo transfer (ET) were comparable between the groups. The live birth rate and ongoing pregnancy rate after frozen ET in the dual trigger group were significantly higher than those in the GnRHa group (32.6% vs 14.1%, P = 0.007 and 34.8% vs 17.6%, P = 0.013, respectively), but not superior to those in the hCG group (32.6% vs 27.9%, P = 0.537 and 34.8% vs 27.9%, P = 0.358, respectively). Women of advanced age are quite a heterogeneous population and overlap with poor ovarian responders or patients with diminished ovarian reserve. We therefore could not entirely exclude selection biases or confounding factors. This study was also not a double-blinded trial; the patients in the GnRHa and dual trigger groups could have been affected by the placebo effect. The results of this study suggest that in advanced-age women with low risk of OHSS, a dual trigger or even a single hCG trigger may be a better choice than a single GnRHa trigger. This study was supported by the Shanghai Municipal Health Commission of Science and Research Fund (20184Y0289). The authors declare no conflict of interest. This trial was registered in the Chinese Clinical Trial Registry (ChiCTR-1800016285). 24 May 2018. 2 January 2019.

Effect and Management of Excess Weight in the Context of Fertility-Sparing Treatments in Patients With Atypical Endometrial Hyperplasia and Endometrial Cancer: Eight-Year Experience of 227 Cases.

ObjectiveTo investigate the oncologic and reproductive outcomes of fertility-sparing treatments (FSTs) in atypical endometrial hyperplasia (AEH) and endometrial cancer (EC) patients with excess weight (EW).MethodsThis retrospective study comprised patients with AEH or EC who achieved a complete response (CR) after FST from 2010 to 2018. The clinical characteristics, oncological and reproductive outcomes were compared between the excess weight (EW) group (body mass index (BMI)≥25 kg/m2) and normal weight (NW) group (BMI<25 kg/m2). The risk factors associated with recurrence and unsuccessful pregnancy in patients with EW were analyzed.ResultsOverall, 227 patients were enrolled, including 139 (61.2%) in EW group and 88 (38.8%) in NW group. In patients with EW, the pregnancy rate, the live birth rate and the relapse rate were 29.8%, 23.4%, and 30.9%, respectively. In patients with NW, these rates were 61.1%, 47.2%, and 31.8%, respectively. No significant differences were observed in the time to remission (P=0.865) and disease-free survival (DFS) (P=0.750). Patients in NW group achieved a better pregnancy rate than patients in the EW group (P=0.034). The patients with EW using ovulation induction to increase fertility tended to have a shorter time to pregnancy (P=0.042). However, no significant risk factors associated with unsuccessful pregnancy were identified after the multivariate analysis. In terms of DFS, the combination of gonadotropin-releasing hormone agonist (GnRH-a) and LNG-IUD was better for patients with EW than GnRH-a or oral progestin therapy alone (P=0.044, adjusted hazard ratio (HR)=0.432, 95% confidence interval (CI): 0.152-1.229), especially for patients with EW diagnosed with EC (P=0.032).ConclusionFSTs for overweight and obese patients should be more individualized. GnRH-a and/or LNG-IUD may be options prior to FSTs in patients with EW. Further prospective studies are needed.

Dual trigger with the combination of gonadotropin-releasing hormone agonist and standard dose of human chorionic gonadotropin improves in vitro fertilisation outcomes in poor ovarian responders

The study aimed to evaluate the impact of the dual trigger with the combination of GnRH agonist and standard dose of recombinant hCG on IVF outcomes in poor ovarian responders with GnRH antagonist protocol. 1283 cycles of 1010 poor responder patients according to Bologna criteria were retrospectively analysed in terms of final oocyte maturation: dual trigger group (250 μg hCG + 0.2 mg triptorelin) or standard group (250 μg hCG). Primary outcome measures were the number of retrieved and mature oocytes. The secondary outcome measures were clinical pregnancy rates and live birth rates. The number of retrieved oocytes, mature oocytes, and the top-quality embryos transferred were significantly higher in the dual trigger group (p < .001). Fertilisation rates (73.6% vs 69.6%, p = .009), implantation rates (18.7% vs 14.6, p = .039), clinical pregnancy rate per embryo transfer (27.5% vs. 19.9%, p = .010) and live birth rate per embryo transfer (21.6% vs. 14.9%, p = .011) were also significantly higher in the dual trigger group as compared to the hCG trigger group. The usage of dual trigger with a GnRH agonist and a standard dosage of hCG could improve clinical pregnancy rates and live birth rates in poor ovarian responders undergoing GnRH antagonist IVF/ICSI cycles. IMPACT STATEMENT What is already known on this subject? Dual trigger with standard dose of hCG has been the subject of trials in normal responders to optimise IVF outcomes. The results of these studies showed significant improvements in implantation and pregnancy rates with an increase in the number of mature oocytes retrieved. As a result, dual trigger has become a popular ovulation trigger option in GnRH antagonist cycles. What do the results of this study add? There is limited data about the use of dual trigger in poor ovarian responders (PORs). According to our study, increasing the number of retrieved oocytes, mature oocytes, the number of fertilised oocytes, the number of transferred embryos and top quality embryos transferred by using dual trigger in patients with PORs have a positive impact on pregnancy outcomes. What are the implications of these findings for clinical practice and/or further research? These findings implies potential advantages of dual trigger usage for improving IVF outcomes in PORs. With large sample sized prospective randomised trials, dual trigger with combination of GnRHa and a standard dose of hCG might replace the traditional ovulation trigger with hCG in PORs.

Application of vaginoscopy in the diagnosis and treatment of occult vaginal high-grade squamous intraepithelial lesions

Objective: To investigate the application value of vaginoscopy in the diagnosis and treatment of vaginal squamous intraepithelial lesions post hysterectomy. Methods: From May 2017 to June 2020, patients with high-risk (HR)-HPV infection and (or) cytological abnormalities after hysterectomy in Obstetrics and Gynecology Hospital Affiliated to Fudan University were examined by colposcopy, and those who were not satisfied with exposure under colposcopy were further examined by vaginoscopy. The role of vaginoscopy in the detection and treatment of occult vaginal squamous intraepithelial lesions was analyzed. Results: A total of 153 patients with HR-HPV infection and (or) cytological abnormalities, and inadequate colposcopy were enrolled in this study. The average age was (49.8±8.1) years. All cases were successfully performed vaginoscopy with no vaginal perforation, no bladder and intestinal injury. During vaginoscopy, 11 (7.2%, 11/153) cases with unclear high-grade squamous intraepithelial lesion (HSIL) boundary under colposcopy were found with clear HSIL boundary, and new HSIL was found in 23 (15.0%, 23/153) cases. According to colposcopy, there were 89 cases of normal or inflammation, 45 cases of vaginal low-grade squamous intraepithelial lesion (LSIL) and 19 cases of vaginal HSIL. According to vaginoscopy, there were 56 cases of normal or chronic inflammation, 55 cases of vaginal LSIL, 40 cases of vaginal HSIL (including 2 cases of vaginal HSIL could not exclude cancer) and 2 cases of vaginal carcinoma. There were significant difference between colposcopy and vaginoscopy (P<0.01). The missed diagnosis rate of vaginal apex blind curettage under colposcopy was 54.8%. For the 40 cases with vaginal HSIL under vaginoscopy, 15 cases were completely curettaged with vaginal endoscopic claw forceps, and 22 cases were vaporized by Versapoint. Among the 37 cases of vaginal HSIL treated by vaginoscopy, 34 cases were followed up for 6 months with 31 cases of vaginal HSIL cured. The cure rate was 91.2% (31/34), the positive rate of HR-HPV decreased from 100.0% (34/34) to 79.4% (27/34). Conclusion: For patients with inadequate exposure of vaginal apex during colposcopy posthysterectomy, vaginoscopy is helpful to detect the occult lesions in the vaginal apex, and it could also be used for the treatment of vaginal squamous intraepithelial lesions.

Evaluation of Dual Trigger with Combination of Gonadotropin-Releasing Hormone Agonist and Human Chorionic Gonadotropin in İmproving Oocyte-Follicle Ratio in Normo-Responder Patients

Our aim was to compare the efficacy of two triggering method one with dual triggering with gonadotropin-realising hormon (GnRH) agonist plus standard dosage human chorionic gonadotropin (hCG) and the other with hCG only for final oocyte maturation on oocyte/follicle ratio and pregnancy rates in normoresponders in GnRH antagonist cycles in invitro fertilization-intrastoplasmic sperm injection (IVF-ICSI). In this retrospective study, all patients underwent GnRH antagonist protocol. When at least ≥3 follicles reached ≥17 mm diameter, 116 patients received dual trigger with GnRH agonist plus hCG (1mg Leuprolide acetate plus 10.000 IU uhCG) and 178 patients received uhCG (10.000 IU u hCG) for final oocyte maturation. All follicles ≥10 mm diameter were aspirated. Number of oocytes and metaphase II oocytes retrieved per aspirated follicles, implantation rate, and clinical pregnancy rate per cycle was recorded. There was no statistically significant difference in terms of metaphase II oocyte ratio per aspirated follicle, implantation rate and clinical pregnancy rate between the dual trigger group and hCG only group (45.7% vs. 51%; 35.4% vs.30.3% and 45%vs. 40% respectively). Oocyte/ follicle ratio was significantly higher in dual trigger group (68.2%vs 63.8% p=0,028). Dual triggering in normal responders with a GnRH-agonist and a standard dosage of hCG is superior to hCG only protocol in terms of oocyte/follicle ratio but does not improve metaphase II oocyte, implantation and clinical pregnancy rates in GnRH-antagonist cycles. Dual triggering method may be beneficial in patients with immature oocytes and emty follicle syndrome.

Long-term follow-up of 60 incarcerated male sexual offenders pharmacologically castrated with a combination of GnRH agonist and cyproterone acetate

ABSTRACTHerstedvester Prison is a penal institution and center for treating violent sexual offenders in Denmark. From 1989 to 2014, 60 dangerous sexual offenders were treated with the combination of a GnRH agonist and cyproterone acetate, providing pharmacological castration and androgen receptor blockade. Five years after initiation of treatment, 81.7% (95% CI 74.4, 92.1) of offenders were released. Twenty years after treatment initiation, 30.6% (95% CI = 19.6, 47.6) of offenders had a relapse to any crime, including sexual offense, and 13.6% (95% CI = 11.5, 33.3) had a relapse to sexual crime. Treatment was paused or discontinued for 44.1% (95% CI = 31.7, 61.4) of offenders. Cessation was not associated with higher recidivism to any crime (hazard ration [HR] = 2.2; 95% CI = 0.8, 6.3; p = 0.14), but the risk of a new sexual offense was 7.7 (95% CI = 1.4, 42.8; p = 0.02) times higher among those who stopped treatment, compared to those who continued it.

Pregnancy predictors in the fresh cycle using dual trigger protocol

Dual trigger protocol using a combination of GnRH agonist and hCG for final oocyte maturation has been shown to minimize ovarian hyperstimulation syndrome (OHSS) risk without compromising fresh embryo transfer outcomes. Therefore, we sought to determine if any cycle characteristics were associated with predictive of pregnancy outcomes in fresh cycles that utilized this protocol for in-vitro fertilization.

Dual Triggering With Gonadotropin-Releasing Hormone Agonist and Standard Dose Human Chorionic Gonadotropin in Patients With a High Immature Oocyte Rate.

In in vitro fertilization (IVF) cycles, some patients show a high rate of immature oocytes retrieved after controlled ovarian stimulation. In vitro oocyte maturation is an experimental technique, with poorer results than conventional IVF. For this reason, improving in vivo maturation could meliorate the reproductive outcome of these patients. We performed a retrospective, not interventional, study analyzing the difference in the number and percentage of mature oocytes retrieved in patients with more than 50% immature oocytes in a previous IVF cycle triggered with human chorionic gonadotropin (hCG) compared to the number and rate of mature oocytes retrieved in subsequent cycles, triggered with both gonadotropin-releasing hormone agonist (GnRH-a) and hCG. The number of mature oocytes retrieved with a dual trigger was significantly higher than that for hCG alone: 5.3 ± 3.6 (4.4-6.1) versus 2.4 ± 2.2 (2-2.9). The proportion of mature oocytes showed the same tendency (79.6% vs 43.6%). The implantation, clinical, and ongoing pregnancy rates were 17.3%, 26.9%, and 15.3%, respectively, for the hCG trigger and 30.8%, 43.6%, and 31.6%, respectively, for the dual trigger. In patients with a low percentage of retrieved mature oocytes, who were triggered with a combination of GnRH-a and hCG, the number and percentage of retrieved mature oocytes improved. The dual trigger also seemed to meliorate gestational outcomes after IVF.

Optimal usage of dual trigger to prevent OHSS in a long-protocol IVF cycle

When a patient undergoing an long protocol IVF cycle has high risk of severe OHSS, rescuing the cycle by withdrawing the agonist, replacing (adding) it with an antagonist and triggering with a GnRH agonist has been considered. However, we underwent lower the number of oocytes retrieval in GnRH agonist trigger group. Therefore we aimed to compare outcomes between trigger with GnRH agonist and dual trigger (GnRH agonist and low dose hCG). In addition, optimal hCG dosage is investigated in dual trigger with combination of gonadotropin-releasing hormone agonist and human chorionic gonadotropin. A prospective clinical study. We evaluated total 26 high responder patients with a long protocol during 2009∼2013. We monitored by measuring the follicle number and serum E2 level on day 7 of the cycle. On the first day of monitoring, we observed that serum E2 was 500∼1500pg/mL and 10∼20 follicles were >10mm in each ovary. For these patients, we applied that agonist was interrupted and the antagonist (0.25mg/day) administrated 1-3 times (depending on a patients) until the day of hCG trigger. When at least two follicles had reached 18mm diameter, final follicular maturation was induced with GnRH agonist (Group I, n=7), dose of 1000 IU hCG and GnRH agonist (Group II, n=8) or 2000 IU hCG and GnRH agonist (Group III, n=10). The number of retrieved oocytes in Group I was significantly lower than those in Group II and Group III (4.7±1.5 vs. 10.0±3.7 vs. 20.2±6.8, p<0.01). The oocyte retrieval rate was the highest the Group III (p<0.01). However, no significant difference was observed in fertilization rate (66.7% vs. 67.5% vs. 61.4%) and cleavage rate (86.4% vs. 90.7% vs. 93.5%). Clinical pregnancy rate was significantly higher in group II (75.0%) and III (70.0%) than that in group I (14.3%, P<0.05) However, group II and III didn’t find significant differences. And there was 1 case of mild OHSS in the group III but none in the group II. For higher responders of a long protocol IVF cycle, dual trigger with a combination of GnRH agonist and low dose hCG (1000 IU) is induced the prevention of OHSS, successful high oocyte retrieval rate and ultimately increased pregnancy outcome.

Dual trigger with gonadotropin-releasing hormone agonist and standard dose human chorionic gonadotropin to improve oocyte maturity rates

To evaluate the percentage (%) of mature oocytes retrieved in patients with a previous history of >25% immature oocytes retrieved who were triggered with gonadotropin-releasing hormone agonist (GnRH-a) and human chorionic gonadotropin (hCG) to induce oocyte maturation. Retrospective cohort study. A university-based tertiary fertility center. Patients with a history of >25% immature oocytes retrieved in a prior invitro fertilization cycle who were triggered with GnRH-a and hCG 5,000 IU or 10,000 IU in a subsequent cycle from January 2008 through February2012. Dual trigger of GnRH-a and hCG 5,000 or 10,000 IU. Percent of mature oocytes retrieved and fertilization rate. The proportion of mature oocytes retrieved was significantly higher with a dual trigger compared with the subject's previous cycle (75.0%, interquartile range 55.6%-80.0% vs. 38.5%, interquartile range 16.7%-55.6%). The odds of a mature oocyte retrieved for patients who received a dual trigger was 2.51 times higher after controlling for stimulation protocol, hCG dose, gonadotropin dose, and oocyte retrieval time interval (odds ratio 2.51; confidence interval 1.06-5.96). The implantation, clinical, and ongoing pregnancy rates for the dual trigger were 11.8%, 26.1%, and 17.4%, respectively. In patients with a low percentage of mature oocytes retrieved who are triggered with a combination of GnRH-a and hCG, the % of mature oocytes retrieved improved. invitro fertilization outcomes, however, remain poor, suggesting an underlying oocyte dysfunction.

Dual trigger with combination of gonadotropin-releasing hormone agonist and human chorionic gonadotropin significantly improves the live-birth rate for normal responders in GnRH-antagonist cycles

To investigate whether dual triggering of final oocyte maturation with a combination of gonadotropin-releasing hormone (GnRH) agonist and human chorionic gonadotropin (hCG) can improve the live-birth rate for normal responders in GnRH-antagonist in vitro fertilization/intracytoplasmic sperm injection (IVF-ICSI) cycles. Retrospective cohort study. Infertility unit of a university-affiliated medical center. Normal responders to controlled ovarian hyperstimulation who were undergoing IVF-ICSI with a GnRH antagonist protocol. Standard dosage of hCG trigger (6,500 IU of recombinant hCG) versus dual trigger (0.2 mg of triptorelin and 6,500 IU of recombinant hCG). Live-birth, clinical pregnancy, and implantation rates per cycle. A total of 376 patients with 378 completed cycles with embryo transfer were enrolled (hCG trigger/control group: n = 187; dual trigger/study group: n = 191). The dual trigger group demonstrated statistically significantly higher implantation (29.6% vs. 18.4%), clinical pregnancy (50.7% vs. 40.1%), and live-birth (41.3% vs. 30.4%) rates as compared with the hCG trigger group. There was no statistically significant difference in terms of patient demographics, cycle parameters, or embryo quality. Dual trigger of final oocyte maturation with a GnRH-agonist and a standard dosage of hCG in normal responders statistically significantly improves implantation, clinical pregnancy, and live-birth rates in GnRH-antagonist IVF cycles.

Effect of low dose hCG support on GnRH-agonist triggered IVF/ICSI cycles

To assess the reproductive outcome (pregnancy rate and OHSS rate) in high responders after triggering final oocyte maturation with a combination of GnRH-agonist and hCG (500 to 1,000 IU) as compared with those triggered with GnRH-agonist alone and those treated with long protocol. Controlled interventional study. In high responders, three groups of ovulation induction cycles were allocated as below: (30 cycles for each group) 1. GnRH-antagonist protocol, triggered with a combination of GnRH-agonist and hCG (500 to 1,000 IU, depending on the peak Estradiol level) for final oocyte maturation 2. GnRH-antagonist protocol, triggered with GnRH-agonist alone for final oocyte maturation. 3. Long protocol (triggered with 10,000 IU hCG for final oocyte maturation) The clinical pregnancy rate and the rate of ovarian hyperstimulation syndrome (OHSS) were compared among the three groups. No significant differences were seen regarding positive hCG/ET rate, clinical pregnancy rate, and rate of early pregnancy loss between group 1 (GnRH-agonist and hCG dual trigger) and group 3 (long protocol). A tendency of lower pregnancy rate was noted in group 2 (GnRH-agonist trigger alone). There was no OHSS in group 1 (GnRH-agonist and hCG dual trigger) and group 2 (GnRH-agonist trigger alone). For higher responders, dual trigger with a combination of GnRH-agonist and hCG is safe considering the complication of OHSS, and as effective as conventional long protocol regarding the pregnancy outcome.

A combination of GnRH agonist and low dose hCG for triggering ovulation in oocytes donors is associated with smaller ovarian size and lower estradiol levels following oocyte retrieval

To compare the effect of triggering ovulation with HCG, GnRh analog(GnRh-a) or a combination of GnRh-a and low-dose HCG on cycle outcome and ovarian size and estrodiol(E2) levels post-retrieval in oocyte donors(DE)

Pretreatment with gonadotropin-releasing hormone (GnRH) antagonists to prevent the flare-up effect of long-acting GnRH agonists: results of a pilot study

This study evaluated in vivo whether the flare-up effect of GnRH agonists can be suppressed through pretreatment with a GnRH antagonist. The classic flare-up effect caused by 3.8 mg goserelin acetate could not be suppressed through pretreatment using a single dosage of 3.24 mg cetrorelix acetate.

O-172: Human cumulus cells gene expression profile according to ovarian hyperstimulation protocols (hMG vs. recFSH)

The emergence of DNA chips offers the possibility to exhaustively catalogue the genes that are expressed in a given tissue at a given time. The individual characterization of gene activity at a genomic scale in cumulus cells of each patient will provide important insights to evaluate the effects of ovarian stimulation treatments under IVF conditions. We have studied the cumulus cells RNA expression profile in order to identify the molecular markers differentially regulated following recFSH and hMG stimulation protocols. Total RNA of cumulus cells collected from patients undergoing an intracytoplasmic sperm injection (ICSI) in our centre and stimulated with hMG (group G1) or recFSH (group G2) were extracted. The RNA expression profiles were evaluated on DNA microarray chips. Total genes expressed and genes differentially expressed between these two groups were quantified and analyzed. The patients referred for ICSI were stimulated with a combination of gonadotropin-releasing hormone agonist (GnRH-a) and recombinants recFSH or hMG. Cumulus cells were mechanically removed on the day of egg collection and frozen at -80°C in RLT buffer from the RNeasy kit (Qiagen) before RNA extraction. Pools of cumulus cells from a single patient were analyzed on Affymetrix™ HG-U133 plus 2.0 GeneChip oligonucleotide array chips made of 54675 probesets and representing roughly 35000 unique human known or predicted genes. The Affymetrix™ GeneChip Operating Software 1.2 (GCOS) was used to evaluate the signal intensity for each probeset (detection call ”present“ or ”absent“) and also to perform pair wise comparison between single gene expression raw values from cumulus cells of different stimulation protocoles. We determined genes exclusively or differentially expressed between the two groups. Differential expression was determined by selecting the probesets with the following constraints: ratios ≥ 3 with a significant change p-value (P ≤ 0.01), ”overexpressed“ probeset ”present“ with a signal value ≥ 20. A total of 11932 genes are present in both groups. 2341 and 1476 are exclusive to G1 and G2 respectively. However, only 64 and 260 genes represent a significant difference between the two groups. Most gene expression levels (n= 31811) do not change between the two groups. Among the genes that are expressed in the two groups, 518 are upregulated in G1 whereas 1742 are upregulated in G2. In G1, the LHCGR gene represents one of the strongest difference with a 6.7 fold increase in cumulus cells under hMG when compared with recFSH. In addition, other genes of interest include CD163 and PAPPA, upregulated with hMG, and GREM1 and CDW92 with recFSH. We identified human genes that are differentially expressed in patients that were undergoing distinct stimulation protocols. Understanding the relationships between ovarian stimulation and cumulus cell gene expression profile will permit to adapt stimulation protocols in order to elicit an adequate response benefiting the IVF outcome.

P-694: Gene expression profile of human immature and mature oocytes: A microarray approach

The understanding of the mechanisms regulating human oocyte maturation are still rudimentary. The aim of this study is to determine gene expression profiles of human nuclear mature and immature oocytes (germinal vesicle and metaphase I). We have identified transcripts differentially expressed between imature and mature oocytes and cumulus cells. The patients referred for cIVF or ICSI were stimulated with a combination of gonadotropin-releasing hormone agonist (GnRH-a) and recombinants FSH or Menopur. Germinal vesicle (GV, n=20), metaphase I (MI, n=20) and unfertilized MII (n=16) oocytes were collected 21 hours or 44 hours post insemination and frozen at -80°C in RLT buffer from the RNeasy kit (Qiagen) before RNA extraction. Pools of oocytes were analyzed by DNA microarrays on Affymetrix® HG-U133 plus 2.0 GeneChip oligonucleotide array chips made of 54675 probesets and representing roughly 35000 unique human known or predicted genes. The Affymetrix® GeneChip Operating Software 1.2 (GCOS) was used to evaluate the signal intensity for each probeset (detection call “present” or “absent”) and also to perform pair wise comparison between single gene expression raw values from oocytes of different stages. We determined genes exclusively expressed during one stage and a list of genes which expression progressively increased during oocyte maturation by selecting the probesets with the following ratios constraints: MI/GV > 1.73, MII/MI > 1.73 and MII/GV > 3, with a significant change p-value (P < 0.01) and where GV, MI and MII stands for signals values in these samples. Immature and mature oocytes express in average 8728 genes, but there is a clear decline from the GV and MI stages (n = 10 892 and 9682 respectively) to MII oocytes (n = 5 633). Nevertheless, within this contingent expression varies little between the GV, MI and MII samples (tight scatter plots = 0.63 and high correlation coefficients = 0.92). The most specific genes only found in one category were HKR3, ZNF165 and MAK for the GV stage, PTPN12 for the MI stage. Despite showing the lowest diversity of gene expression, MII oocytes displayed the most specific pattern among all oocytes by up-regulation of 444 genes. Some are specific to MII oocytes (SOX2OT, DMBX1/OTX3, TGFB3, BCL2L11 and CAS2) whereas others were gradually induced during maturation (HIC2, CDC25A, CCNJ, CCNG2, SOCS7 and CUL5). Very few genes were specifically over- or under-expressed in GV and MI cells. Interestingly, a marker like Epithin (ST14) is very specific of both GV and MI oocytes whereas it is absent from mature MII oocytes. Such gene represents a potential marker of human immature oocytes. We identified human genes that are specific to three oocyte developmental stages as well as genes which expression increase gradually with the oocyte maturation process. Understanding these factors’ function will greatly improve our command of in vitro fertilization procedures. These genes already give insights about vital regulation pathways and will provide candidate cytoplasmic and nuclear markers of human oocyte maturation.

Pulmonary endometriosis in a patient with unicornuate uterus and noncommunicating rudimentary horn

Objective: To report a rare case of a patient with catamenial hemoptysis, secondary infertility, and endometriosis associated with a unicornuate uterus and noncommunicating rudimentary horn. Design: Case report. Setting: University hospital. Patient(s): A 29-year-old woman who developed progressive catamenial hemoptysis and secondary infertility was evaluated at the University Hospital of Crete. Intervention(s): The complete history, laboratory data, laparoscopic findings, and chest magnetic resonance image of this patient were analyzed. A GnRH agonist, leuprolide acetate, was successfully administered. Main Outcome Measure(s): Diagnosis and appropriate treatment of pulmonary endometriosis in a patient with rudimentary uterine horn. Result(s): Treatment with a GnRH agonist achieved suppression of both menstruation and hemoptysis. After 6 months of normal menstrual activity, the symptoms reappeared. The patient was again treated with leuprolide acetate (3.75 mg/mo IM) for 6 months and remained asymptomatic. In fact, the patient became pregnant after cessation of therapy. Finally, the patient was treated successfully with removal of the rudimentary uterine horn during cesarean section. Three-year follow-up showed disappearance of the chest symptoms. Conclusion(s): Pulmonary endometriosis and unicornuate uteri are rare. To our knowledge, this is the first case of catamenial hemoptysis with a congenital müllerian anomaly. We describe successful management using a combination of GnRH agonist and surgical resection of the rudimentary uterine horn.

Estrogen mini-dose replacement during GnRH agonist therapy in central precocious puberty: a pilot study.

During GnRH agonist therapy of patients with central precocious puberty (CPP), growth is sometimes suppressed to subnormal velocity. The working hypotheses were that estrogen levels are suppressed by GnRH agonist therapy below normal prepubertal levels, that such suppression is responsible for the slow growth of girls with CPP during GnRH agonist therapy, and that a mini-dose of estrogen replacement will normalize growth. The present pilot study examined growth and bone maturation over 2 yr in 13 patients with CPP and compared therapy with a combination of GnRH agonist and 8 microg conjugated equine estrogen (group 1) to therapy with GnRH agonist alone (group 2). Both groups had adequate suppression of gonadotropins, and E2 levels were below detection levels of our assay throughout the study period. Group 2 patients decreased their growth velocity from 2.0 +/- 1.4 to -1.6 +/- 1.2 SD score compared with group 1, who maintained their growth velocity of 1.3 +/- 1.5 SD score and their height SD score for 2 yr (P < 0.01). In group 1 patients the ratio of the change in bone age/change in chronological age decreased from 1.2 +/- 0.7 to 0.75 +/- 0.3, and in group 2 patients it decreased to 0.6 +/- 0.3 and 0.4 +/- 0.2 (P < 0.05) during the first and second years of therapy, respectively. It is concluded on a pilot basis that estrogen suppression is responsible for the slow growth of girls with CPP during GnRH agonist therapy and that a mini-dose of estrogen replacement is safe and effective for at least 24 months in maintaining normal prepubertal growth without acceleration of bone maturation or pubertal development. The current pilot results do not suggest an indication or provide a justification for such therapy.