International Journal of

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Background: Polycystic ovarian syndrome (PCOS) patients are prone to premature LH surge and ovarian hyperstimulation syndrome (OHSS). Long GnRH analogue protocol and GnRH antagonist protocol are two methods utilized for induction ovulation in patients undergoing IVF/ICSI. Objective: The aim of this study was to compare the effects of GnRH agonists and antagonists in PCOS patients. Materials and Methods: A total of 60 PCOS patients under 35 years old were enrolled in this study. The patients have no history of thyroid disorder and hyperprolactinemia. All patients received OCP (LD) before starting the treatment. Then patients randomly divided into two groups. The agonist group underwent standard long GnRH analogue protocol. In antagonist group, HMG (150 IU/day) was started from third day of cycle. Then GnRH antagonist (0.25mg) was administered from 6th day after HMG initiation (LH?5 IU/ml) to the day of HCG injection. Follicular development monitored by vaginal ultra sonography and serum estradiol measurement. Results: There were no significant differences in age, duration of infertility, BMI, number of HMG ampules, number of follicles?18mm, serum estradiol level on 6th day of HMG initiation and HCG injection time, fertilization and pregnancy rate between two groups. However there were significant differences regarding duration of treatment, duration of HMG usage, LH level at the initiation of HMG, OHSS rate and number of Metaphase II oocytes between two groups (p<0.05). Conclusion: Usage of the GnRH antagonist may have more advantages such as the shorter duration of treatment and less gonadotrophin requirement. Furthermore, the incidence of OHSS can be reduced in GnRH antagonist comparing to agonist. For decreasing the risk of OHSS and abortion rate, we recommend long term use of OCP before starting the treatment

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Background: General concern is that the pregnancy rate is higher with GnRH-agonist as a protocol of pituitary suppression. GnRH-antagonist protocol provides a shorter period of administration and an easy flexible protocol.
Objective: In this study, the outcomes of GnRH agonist and antagonist in ICSI cycles are compared in normo responder patients.
Materials and Methods: In this randomized clinical trial, 300 normoresponders undergoing ICSI were randomly divided to GnRh agonist (n=150) and GnRh antagonist (n=150) groups. The main outcome measurements were chemical, clinical and ongoing pregnancy rates (PR).
Results: The mean duration of stimulation were 9.6±1.6 and 8.2±1.6 days in agonist and antagonist groups respectively (p=0.001). The mean number of MII oocyte retrieved in agonist and antagonist groups were 7.7±4.0 and 6.9±4.3 respectively (p=0.03). There was no significant difference between two groups regarding mean number of gonadotrophin ampoules, follicles, occytes, total embryos and good quality embryos, OHSS incidence, and abortion rate. Chemical pregnancy rate was 35.3% in agonist and 39.3% in antagonist group. Clinical pregnancy rate was 35.3% in agonist and 34% in antagonist group. Ongoing pregnancy rate was 45 (31.3%) in agonist and 44 (29.3%) in antagonist group. There was no significant difference between two groups in pregnancy rates.
Conclusion: In this study antagonist protocol was shown to be an easy, safe and friendly protocol in Iranian normoresponder patients, having similar outcomes with standard agonist protocol but shorter period of stimulation.

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Background: The differential efficacy between long GnRH agonist with antagonist can partly be due to the preexisting differences in the early antral follicles before ovarian stimulation.
Objective: To compare the effect of pretreatment by estradiol with GnRH antagonist on antral follicular size coordination and basal hormone levels in GNRH antagonist protocol.
Materials and Methods: On cycle day 3 (control/day 3), women underwent measurements of early antral follicles by ultrasound and serum FSH and ovarian hormones then were randomized to receive oral estradiol 4mg/day (n=15) or 3mg cetrorelix acetate (n=15) in luteal phase before subsequent antagonist protocol. Participants were re-evaluated as on control/day 3.
Results: There was a significant reduction of mean follicular sizes in each group after medical intervention (7.63±2.11 Vs. 4.30±0.92 in group A and 8.73±1.96 Vs. 4.13±1.11 in group B) (p=0.0001). The magnitude of follicular size reduction was significantly higher in group B (-4.60±2.04 Vs. -3.33±2.28) (0.027). There was a non significant attenuation of follicular size discrepancies in two groups. FSH and inhibin B levels in the day 3 of the next cycle in both groups were significantly decreased but did not have significant difference between two groups.
Conclusion: Both luteal E2 and premenstrual GnRH antagonist administration reduces the follicular sizes significantly and GnRH antagonist acts more potently than E2 in this way but attenuation of follicular size discrepancies in both treatment is not significant

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Background: GnRH agonist and antagonist were developed to control the premature release of LH surge. There is some difference between two protocols.
Objective: We compared the outcome of frozen-thawed embryo transfer in infertile women who used GnRH agonist or antagonist protocol for previous COH cycle and evaluation of any adverse effect of GnRH antagonist on oocyte and embryo.
Materials and Methods: The study group included all infertile women who referred to Yazd Research and Clinical Center for Infertility. Overall 20-35 years old women who were candidate for frozen-thawed embryo transfer with regard to inclusion and exclusion criteria were participated in the study. The patients based on previous control ovarian stimulation (COH) protocol divided in to two groups: GnRH agonist long protocol (n=165) and GnRH antagonist multiple dose protocol (n=165). Frozen-thawed embryos were transferred after endometrial preparation in both groups. Main outcome measures were: implantation, chemical and clinical pregnancy rate.
Results: The implantation and clinical pregnancy rate following cryopreserved embryo transfer in GnRH agonist group and antagonist group were 16.3% vs. 15.7% (p=0.806) and 38.1% (63/165) vs. 36.9% (61/165) (p=0.915) and chemical pregnancy rate was 44.8% (74/165) vs. 43.6% (72/165) (p=0.915) respectively.
Conclusion: There was no statistically difference between two groups in terms of implantation and pregnancy rate. Although pregnancy rate in fresh embryo transfer in antagonist cycles was lower than agonist groups, Therefore decrease in these parameters might be due to detrimental effect of GnRH antagonist on the endometrium, not embryo or oocyte.

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Background: Determination of oocyte fertilization and embryo quality are one of the most important purposes in ART cycles. Follicular fluid provides an important microenvironment for development of oocytes and some biochemical characteristics of the follicular fluid, such as pregnancy-associated plasma protein-A (PAPP-A), may play an important role in prediction of success rate of ART.
Objective: This study was performed to evaluate whether there was any difference in follicular fluid PAPP-A, fertilization, and embryo quality between GnRH agonist long protocol and flexible GnRH antagonist multiple-dose protocol in ART cycles.
Materials and Methods: A total of 100 women who were candidates for ART were enrolled the study and were divided into two groups, GnRH agonist (GnRHa) long protocol (n=51) and flexible GnRH antagonist (GnRHant) multiple-dose protocol (n=49). Follicular fluid sample was obtained from a single mature follicle and follicular fluid PAPP-A level, fertilization and embryo quality of the same oocyte were evaluated in both groups.
Results: There was no significant difference in the mean levels of follicular fluid PAPP-A between the GnRHa protocol and GnRHant protocol (3.5±1.4 vs. 3.8±1.9, respectively). The mean levels of follicular fluid PAPP-A in fertilized oocyte and good quality embryo were comparable in GnRHa and GnRHant protocols.
Conclusion: Our data indicated that no differences of follicular fluid PAPP-A levels were observed between cycles using GnRHa long protocol and those of using flexible GnRHant multiple-dose protocol.

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Background: Premature luteinization during in vitro fertilization was commonly happened before the introduction of GnRh analogues. High level of unwanted progesterone is associated with adverse pregnancy outcome and is thought to be induced by inappropriate LH elevation.
Objective: To evaluate the progesterone level on the day of Human Chorionic Gonadotropin (HCG) triggering in GnRh agonist and antagonist protocols, and its correlation with clinical pregnancy rate and miscarriage rate.
Materials and Methods: One hundred and seven women underwent intracytoplasmic sperm injection with long agonist protocol (n=46) or antagonist protocol (n=61). Blood sample was obtained from each patient for progesterone level measurement in HCG administration day, then patients were divided into two groups according to their serum progesterone levels on the HCG triggering day: progesterone level <1.2 ng/ml, and progesterone level ≥1.2 ng/ml. Clinical pregnancy and miscarriage rates were evaluated as main outcomes and biochemical pregnancy rate and implantation rate were considered as secondary outcomes.
Results: The increased prevalence rate of premature progesterone (progesterone level ≥1.2 ng/ml) in total patients was 13.1% (14/107) and in long agonist protocol group and antagonist protocol group was 15.2% (7/46) and 11.5% (7/61) respectively. Premature progesterone rise had no significant correlation with clinical pregnancy rate in total patients (p=0.174), agonist protocol (p=0.545), and antagonist protocol (p=0.129). Also premature progesterone rise had no significant association with miscarriage rate in total patients (p=0.077), agonist protocol group (p=0.383) and antagonist protocol group (p=0.087).
Conclusion: A significant rise in progesterone levels at the time of HCG triggering doesn’t lead to decrease in pregnancy rate and implantation rate and increase in miscarriage rate.

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Background: Gonadotropin-releasing hormone (GnRH) plays essential roles in embryo implantation, invasion of trophoblastic tissue, and steroid synthesis in the placenta.
Objective: The aim of this study was to evaluate the effect of GnRH antagonist administration on pregnancy outcomes in early implantation period.
Materials and Methods: In this retrospective study, 94 infertile women undergoing GnRH antagonist protocol who were at risk of ovarian hyperstimulation syndrome (OHSS) were included. Sixty-seven patients (group I) received Cetrorelix 0.25 mg/daily in the luteal phase for 3 days while in 27 participants (group II), it was not administered. Pregnancy outcomes were assessed based on chemical and clinical pregnancy rates.
Results: The pregnancy outcomes were not significantly different between two groups (p=0.224).
Conclusion: The present study proposed that luteal phase GnRH antagonist administration does not influence the chance of successful pregnancy outcomes.

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Background:Gonadotropin-releasing hormone (GnRH) antagonist protocol has been proposed as a potentially proper option for the patients with limited ovarian reserve. Nevertheless, there is no significant difference in terms of clinical pregnancy between the GnRH antagonist and agonist cycles. The use of aromatase inhibitors such as letrozole was suggested by some studies.
Objective: The object of this study was to evaluate the efficacy of letrozole co-treatment with GnRH-antagonist protocol in ovarian stimulation of poor responder patients undergoing intracytoplasmic sperm injection.
Materials and Methods: A double-blinded randomized control trial was conducted on 70 infertile women with poor ovarian response based on Bologna criteria in two groups: letrozole+GnRH-antagonist (LA) group and placebo+GnRH-antagonist (PA) group (n=35/each). The LA group involved at letrozole 2.5 mg daily over 5 days and recombinant human follicle stimulating hormone 225 IU/daily. The PA group received placebo over 5 days and recombinant human follicle stimulating hormone at the same starting day and dose, similar to LA group. GnRH-antagonist was introduced once one or more follicle reached ≥14 mm. The main outcome measures were the number of oocytes retrieved, fertilization rate, implantation rate, cycle cancellation rate, and clinical pregnancy rate.
Results: There were no significant differences in demographic characteristics between groups. There were no significant differences between groups regarding the number of oocytes retrieved (p=0.81), number of embryos transferred (p=0.82), fertilization rate (p=0.225), implantation rate (p=0.72), total cycle cancelation rate (p=0.08), and clinical pregnancy rate (p=0.12).
Conclusion: The use of letrozole in GnRH-antagonist cycles does not improve clinical outcomes in poor responder patients undergoing intracytoplasmic sperm injection.

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Background: Management of poor-responding patients is still major challenge inassisted reproductive techniques (ART). Delayed-start GnRH antagonist protocol isrecommended to these patients, but little is known in this regards.
Objective: The goal of this study was assessment of delayed-start GnRH antagonistprotocol in poor responders, and in vitro fertilization (IVF) outcomes.
Materials and Methods: This randomized clinical trial included sixty infertilewomen with Bologna criteria for ovarian poor responders who were candidate forIVF. In case group (n=30), delayed-start GnRH antagonist protocol administeredestrogen priming followed by early follicular-phase GnRH antagonist treatment for7 days before ovarian stimulation with gonadotropin. Control group (n=30) treatedwith estrogen priming antagonist protocol. Finally, endometrial thickness, the ratesof oocytes maturation, , embryo formation, and pregnancy were compared betweentwo groups.
Results: Rates of implantation, chemical, clinical, and ongoing pregnancy indelayed-start cycles were higher although was not statistically significant.Endometrial thickness was significantly higher in case group. There were nostatistically significant differences in the rates of oocyte maturation, embryoformation, and IVF outcomes between two groups.
Conclusion: Delayed-start GnRH antagonist protocol can be a new hope method totreatment poor ovarian responders.

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Background: Gonadotropin-releasing hormone agonists (GnRH-a) was increasingly used for triggering oocyte maturationfor the prevention of ovarian hyperstimulation syndrome. Studies suggest that GnRH-a might be used as a better trigger agent since it causes both Luteinizing hormone and follicle stimulating hormone release from a physiologic natural cycle.
Objective: The aim of this study was to evaluate the effect of dual-triggering in assisted reproductive technology outcomes.
Materials and Methods: 192 normal responder women aged ≤42 years and 18< Body Mass Index <30 kg/m2 enrolled in this single-blind randomized controlled trial. All participants received antagonist protocol. For final triggering, women randomly were divided into two groups. Group, I was triggered by 6500 IU human chorionic gonadotropin (hCG) alone, and group II by 6500 IU hCG plus 0.2 mg of triptorelin. The implantation, chemical, clinical and ongoing pregnancy, and abortion rates were measured.
Results: The mean of retrieved oocytes and obtained embryos were statistically higher in the dual-trigger group (group I), but the implantation and pregnancy rates were similar in two groups.
Conclusion: The results of our study did not confirm the favorable effect of dual-triggered oocyte maturation with a GnRH-a and a standard dosage of hCG as an effective strategy to optimize pregnancy outcome for normal responders in GnRH-antagonist cycles. We think that this new concept requires more studies before becoming a universal controlled ovarian hyperstimulation protocol in in vitro fertilization practice.

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Background: The use of embryo cryopreservation excludes the possible detrimental effects of ovarian stimulation on the endometrium, and higher reproductive outcomes following this policy have been reported. Moreover, gonadotropin-releasing hormone agonist trigger in gonadotropin-releasing hormone (GnRH) antagonist cycles as a substitute for standard human chorionic gonadotropin trigger, minimizes the risk of ovarian hyperstimulation syndrome (OHSS) in fresh as well as frozen embryo transfer cycles (FET).
Objective: To compare the reproductive outcomes and risk of OHSS in fresh vs frozen embryo transfer in high responder patients, undergoing in vitro fertilization triggered with a bolus of GnRH agonist.
Materials and Methods: In this randomized, multi-centre study, 121 women undergoing FET and 119 women undergoing fresh ET were investigated as regards clinical pregnancy as the primary outcome and the chemical pregnancy, live birth, OHSS development, and perinatal data as secondary outcomes.
Results: There were no significant differences between FET and fresh groups regarding chemical (46.4% vs. 40.2%, p=0.352), clinical (35.8% vs. 38.3%, p=0.699), and ongoing (30.3% vs. 32.7%, p=0.700) pregnancy rates, also live birth (30.3% vs. 29.9%, p=0.953), perinatal outcomes, and OHSS development (35.6% vs. 42.9%, p=0.337). No woman developed severe OHSS and no one required admission to hospital.
Conclusion: Our findings suggest that GnRHa trigger followed by fresh transfer with modified luteal phase support in terms of a small human chorionic gonadotropin bolus is a good strategy to secure good live birth rates and a low risk of clinically relevant OHSS development in in vitro fertilization patients at risk of OHSS.

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Background: Both oral contraceptive pills (OCPs) and estradiol valerate (E2) have been used to schedule a gonadotropin-releasing hormone antagonist in vitro fertilization (IVF) cycles. Since the suppression of follicle-stimulating hormone by OCPs can stay 5-7 days after stopping the pills, it seems that starting the gonadotropin-releasing hormone (GnRH) after 6 days of pre-treatment discontinuation may be important in IVF outcomes.
Objective: The aim of the present study was to determine the number of mature oocyte and pregnancy rate of three pretreatment methods for fresh embryo transfer cycles.
Materials and Methods: In this randomized controlled trial, two-hundred ten women (18-35 yr and less than 2 previous IVF attempts) undergoing IVF with the GnRH antagonist protocol were randomized to the OCP, E2, and no pretreatment arms. OCP group (n=53) received OCP (ethinyl estradiol30 μg and levonorgestrel150 μg), E2 group (n=63) received 4 mg/day oral E2 (17β‐E2) for 10 days from day 20 of the previous cycle and GnRH antagonist stimulation was started 6 days after the interruption of OCP and E2. The control group (n =70) did not receive any pretreatment.
Results: No significant difference was observed in the mean number of the mature oocyte, endometrial thickness, and embryo quality. The pregnancy rate in E2 group was higher than the two other groups (42.9% vs 39.6% and 34.3% in OCP and control group, respectively), but the difference was not statistically significant (p=0.59).
Conclusion: It seems OCP or E2 pretreatment could not improve the fresh IVF-embryo transfer outcomes

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Background: Because of the unexpected and often dramatic inhibition of luteinizing hormone (LH) secretion related with the usage of gonadotropin-releasing hormone (GnRH)-antagonist, there has been a probable need for exogenous LH supplementation. There is a basic and clinical evidences that show late development of follicle needs an LH but there is a threshold for LH requirements during folliculogenesis.
Objective: The purpose of this study was to evaluate the changes in serum LH and the identification of patients who benefit from the addition of LH. Materials and Methods: Seventy volunteers for antagonist protocol in IVF cycle were enrolled in this prospective cross-sectional study. The study was carried out in Reproductive Health Research Center, University of Medical Sciences between July 2016 and February 2016. Serum LH level was estimated 24 h before and after the first (GnRH) antagonist injection. The primary outcome was the serum level of LH and its change in the three groups and the secondary outcome was Egg and Embryo quality.
Result: LH changes above or below 50% had no effect on the number of follicle, the number of oocyte, Germinal vesicle oocyte, metaphase 1 oocyte, metaphase 2 oocyte, endometrial thickness, and chemical and clinical pregnancy.
Conclusion: We evaluated the changes of serum LH in the patients who were entered in the antagonist protocol. Our study showed no significant difference in LH levels 24 h before and after the injection of the antagonist between the three groups, and LH changes did not affect the outcome of pregnancy. 

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The authors have been informed of some errors that occurred in the published paper. The errors are listed as:

• In the M&M section in the main text, the word “performed” has been changed to “recruited”.
• Some references have been modified in the text.
• The type of randomization has been corrected as the random number table.
• The Mann-Whitney test was added in the statistical analysis section.
• The statistical test which has been used in Table I was added as a table subtitle.
• The study results were re-analyzed, and some p-values were modified in the text and table, especially in Table IV.

The authors wish to apologize for these errors.

The online version of the article has been updated on September 4, 2021 and can be found at http://journals.ssu.ac.ir/ijrmnew/article-1-813-en.html&sw=(http://doi.org/10.29252/ijrm.15.4.231).

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Background: Increased levels of kisspeptin are associated with hypothalamus-pituitary-ovary axis dysfunction. It may lead to the development of polycystic ovary syndrome (PCOS).
Objective: We aimed to investigate the effect of prenatal kisspeptin antagonist exposure on the development of PCOS in prenatally androgenized rats in adulthood.
Materials and Methods: In this experimental study, pregnant rats were injected with free testosterone (T, 5 mg/day) or T+P271 (kisspeptin antagonist) on the 20^th day of the pregnancy period (n = 5 in each group), while rats in the control group received solvent. Female offspring were examined in terms of anogenital distance (AGD), anovaginal distance (AVD), vaginal opening, serum total testosterone (TT) levels, ovarian follicles, and the regularity of estrous cycles in adulthood. AGD and AVD were measured using a vernier caliper. TT levels were measured using the enzyme-linked immunosorbent assay method. Ovaries were fixed in 10% formalin, tissue processing was done by a standard protocol, and then ovaries embedded in paraffin. 5 μm-thickness ovarian sections mounted on a glass slide, deparaffinized, and stained using Harris’s Hematoxylin and Eosin Y.
Results: AGD, AVD (p < 0.001), TT levels (p = 0.02), and the numbers of preantral and antral follicles (p < 0.001) in the ovaries were significantly decreased in prenatally T-P271-exposed rats compared to prenatally T-exposed rats. The age of vaginal opening was early in T-P271-exposed rats compared to prenatally T-exposed rats (p < 0.001). The number of corpora lutea was significantly increased in T-P271-exposed rats (p < 0.001). No cystic follicles were observed in the ovaries of prenatally T-P271-exposed rats. Prenatally T-P271-exposed rats had regular estrous cycles compared to prenatally T-exposed rats.
Conclusion: Prenatal exposure to kisspeptin antagonist can prevent PCOS development in prenatally androgenized rats in adulthood.