GnRHa Treatment Research Articles

Final height outcome of boys with idiopathic central precocious puberty treated with gonadotropin-releasing hormone analogue

Methods Twenty boys with ICPP were treated with GnRHa for (20. 0 ± 6. 1) months. At the beginning of therapy, mean chronological age and bone age was(11. 4 ± 1. 0) years and(13. 0 ± 0. 4) years, respectively. GnRHa was discontinued when the boys reached the chronological age and bone age of (13. 2 ± 1. 1) years and (13. 7 ± 0. 6) years, respectively. At the conclusion of the study, all the boys had been followed up for (3. 3 ± 1. 5) years and had achieved adult height. Comparisons were made among their predicted adult height(PAH), final adult height(FAH),and target height (THt). The long term outcome of final adult height in boys with ICPP was investigated after GnRHa treatment.

Sex-specific development of spatial orientation is independent of peripubertal gonadal steroids

Prenatal exposure to androgens has been shown to modulate brain development, resulting in changed behavioral attitudes, sexual orientation and cognitive functions, including processing of spatial information. Whether later changes in gonadotropic hormones during puberty induce further organizational effects within the brain is still insufficiently understood. The purpose of this study was to assess development of spatial orientation before and after the time of normal pubertal development, in an ovine model where half of the animals did not undergo typical reproductive maturation due to the pharmacological blockade of gonadotropin releasing hormone receptor (GnRHR) signaling. The study formed part of a larger trial and utilized 46 pairs of same sex Scottish Mule Texel Cross twins (22 female and 24 male). One twin remained untreated throughout (control) while the other received a subcutaneous GnRH agonist (GnRHa: Goserelin-Acetate) implant every fourth week. GnRHa treatment began at eight and 28 weeks of age, in males and females respectively, because the timing of the pubertal transition is sexually differentiated in sheep as it is in humans. Spatial orientation was assessed at three different time points: eight weeks of age, before puberty and treatment in both sexes; 28 weeks of age, after 20 weeks GnRHa treatment in males and before puberty and GnRHa treatment in females; and at 48 weeks of age, which is after the normal time of the pubertal transition in both sexes. Spatial orientation was tested in a spatial maze with traverse time as the main outcome measure. GnRHa treatment did not affect spatial maze performance as no significant differences in traverse time between treated and untreated animals were observed at any time-point. Adolescent females (48 weeks of age) traversed the maze significantly faster than adolescent males, whereas no sex differences in traverse time were seen at earlier developmental stages (eight and 28 weeks). Development of sex differences in spatial orientation was independent of exposure to pubertal hormones since puberty-blocked and control animals both showed the same pattern of spatial maze performance. This result demonstrates the prenatal nature of spatial orientation development. Furthermore, the unexpected finding that female animals outperformed males in the spatial orientation task, underscores the importance of the testing context in spatial orientation experiments.

Potential reasons for patients not pursuing fertility preservation after referral for consultation and counseling

It is recommended that fertility preservation should be addressed with every cancer patient of childbearing age and appropriate referral made to the fertility specialists. Most patients choose not to pursue any fertility preservation options even after extensive consultation and counseling with an REI. Our aim was to determine whether there were any obstacles or reasons for not pursuing fertility preservation before cancer therapy. Prospective cohort survey. We mailed surveys to patients who were referred to our oncofertility program from 2009 to 2011 but did not pursue any form of fertility preservation. The survey was to determine the potential reasons for not proceeding with any of the fertility preservation options. The reasons offered in the survey included lack of time for treatment due to need to start treatment immediately, spouse or family members were opposed, oncologist was opposed, ethical/religious reasons regarding freezing embryos and financial reasons. A total of 71 calls were received at the oncofertility hotline and 39 women (54.9%) were seen for consultation. Of the patients who were seen for consultation 11 patients (28.2%) opted for oocyte freezing and 10 patients (25.6%) opted for embryo freezing. Five patients (12.8%) opted for other treatments such as GnRHa treatment. Twenty-three women (58.9%) did not undergo any form of fertility preservation. We therefore sent the surveys to the 23 patients who did not proceed with any treatment. Preliminary results from some of the completed surveys show that financial reason is the major factor why most patients do not pursue treatment. However, a full analysis of all the completed returns will be presented at the meeting. Financial reasons may be the driving force behind patient's decisions not to pursue treatment. Better awareness of insurance companies regarding fertility preservation may increase insurance coverage for patients and allow more patients to pursue fertility preservation.

Long-term regression of experimental endometriosis in a rat model treated with local application of levonorgestrel-loaded biodegradable microspheres

A previous study demonstrated that local application of levonorgestrel-loaded polylactic acid microspheres (LNG microspheres) resulted in significant regression of endometriotic cysts in a rabbit model for 6 months without disturbing the metabolic parameters or ovarian function. In order to investigate the feasibility of local application of LNG microspheres as a long-term maintenance treatment for endometriosis, the suppressive effect of a single intra-cystic injection of LNG microspheres was studied for 1 year in a rat model. Twenty four rats with experimental endometriotic cysts were randomized to be treated with a single intra-cystic injection of LNG microspheres (n = 8); 6-month GnRH agonist (GnRHa, n = 8) or control (n = 8). Intra-cystic injection of LNG microspheres and GnRHa treatment caused comparable regression and atrophy in endometriotic cysts in the first 6 months. Compared with the control, the wet weight of the endometriotic cysts was significantly lower in both groups at Month 6 but by Month 12 only remained low in the LNG microspheres group (P < 0.01). The immunostaining of estrogen receptors (ERs) in both the epithelium and stroma and progesterone receptors (PRs) in the stroma was significantly weakened in the LNG microspheres group at Month 6 and was not fully restored at Month 12 (P < 0.01). Metabolic parameters and estrous cycle were not disturbed by local application of LNG microspheres. In a rat endometriosis model, the suppressive effect of a single intra-cystic injection of LNG microspheres was comparable to that of GnRHa, and was maintained for 1 year. The down-regulation of ERs and PRs might serve as possible mechanism of long-term effectiveness.

Development of psychophysiological motoric reactivity is influenced by peripubertal pharmacological inhibition of gonadotropin releasing hormone action – Results of an ovine model

This study reports the effects of peripubertal GnRH receptor inactivation on development of psychophysiological motoric reactivity (PMR; sometimes also called emotional reactivity), plasma cortisol concentrations and the relationship between plasma cortisol and PMR in male and female sheep. The study formed part of a larger trial and utilised 46 same sex twins. One twin remained untreated (control) while the other received a subcutaneous GnRH agonist (GnRHa Goserelin-Acetate) implant every 4th week, beginning at 8 and 28 weeks of age, in males and females, respectively (different, due to sex specific age of puberty). PMR, a measure of an animals' response to social isolation, was measured over a two minute period at 8, 28 and 48 weeks of age, using a three axis accelerometer. During the test period vocalisation rate was recorded. Cortisol was assayed in blood samples collected on a single day when animals were 40 weeks of age. PMR and vocalisation rate were significantly higher in females than males at all ages tested. At 28 weeks of age (20 weeks treatment) PMR was increased in treated males to the level seen in control females, by 48 weeks of age treated males' PMR was significantly less than controls. In females, 20 weeks of GnRHa treatment (28-48 weeks of age) was not associated with differences in PMR. Cortisol concentrations were significantly higher in females than males but were not affected by treatment. Plasma cortisol concentrations were positively correlated with PMR; this relationship being driven by the treated animals in both sexes. The results demonstrate that PMR is sexually dimorphic and cortisol dependent in sheep from at least 8 weeks of age. Importantly, they also demonstrate that long-term treatment of males with a GnRH agonist results in changes in age-dependent development of PMR.

Reproductive development, GnRHa-induced spawning and egg quality of wild meagre (Argyrosomus regius) acclimatised to captivity

The objective of the study was to acclimatise wild-caught meagre (Argyrosomus regius) to captivity to produce viable eggs for aquaculture production. Twelve meagre (3 males and 9 females, mean weight=20±7kg) were caught and transported to a land-based facility on 26 October 2006. During, March to June 2007, all three males were spermiating and five of the nine females were in vitellogenesis with mean maximum oocyte diameter ≥550μm. No spontaneous spawning was observed. Two hormone treatments, either a single injection of gonadotropin-releasing hormone agonist (GnRHa, 20μgkg(-1) for females and 10μgkg(-1) for males) or a slow-release implant loaded with the same GnRHa (50μgkg(-1) for females and 25μgkg(-1) for males), were used to induce spawning on three different dates on 26 March 2007, 4 May 2007 and 18 April 2008. From each spawning event, the following parameters were determined: fecundity, number of floating eggs, egg size, fertilisation and hatching success, unfed larval survival, and proximal composition and fatty acid profile of the eggs. In 2007, two females that were injected on 26 March and 4 May spawned a total of 5 times producing 9,019,300 floating eggs and a relative fecundity of 198,200 eggskg(-1) and two different females that were implanted on the same dates spawned 14 times producing 12,430,000 floating eggs and a relative fecundity of 276,200 eggskg(-1). In 2008, a pair that was implanted spawned five times producing a total of 10,211,900 floating eggs and a relative fecundity of 527,380 eggskg(-1). The latency period was 48-72h. Parameters were compared between hormone treatments, date of hormone induction and parents determined by microsatellites. Percentage hatch and egg size were 70±0.3% and 0.99±0.02mm, respectively, for GnRHa-implanted fish and were significantly higher (P<0.05) compared to 30±0.3% and 0.95±0.03mm, respectively, for injected fish. Few differences were observed in proximal composition and fatty acid profile and for all spawns mean (% dry weight) lipid content was 17.3±3.0%, carbohydrate was 4.4±1.9% and protein was 31.5±6.4% and the essential fatty acids: Arachidonic acid (ARA, 20:4n-6) ranged between 0.9 and 1% (of total fatty acids), eicosapentaenoic acid (EPA 20:5n-3) 7.7-10.4% and docosahexaenoic acid (DHA 22:6n-3), 28.6-35.4%. All good quality spawns were obtained in the second and/or third spawn after GnRHa treatment, whereas all bad quality spawns were obtained either on the first spawn or after the fifth spawn. Both spawning protocols gave commercially viable (1,000,000+) numbers of good quality eggs that could form the basis of a hatchery production.

Factors Affecting Growth Velocity during Gonadotropin-Releasing Hormone Agonist Treatment in Girls with Idiopathic Central Precocious Puberty

Results: GV during the first year of GnRHa treatment was 5.9 ±1.0 cm/yr and decreased significantly to 5.4±1.1 cm/yr during the second year of treatment (P = 0.005). GV during the third year (5.0±1.0 cm/yr) was not different from GV during the second year. During the second year of treatment, 8.2% and 36.7% of the girls had a GV < 4 cm/yr and < 5 cm/yr, respectively. Girls with relatively low GV during the second year of treatment (< 5 cm/yr) showed higher risk of advanced BA (≥ 11 yr) at 1 year (55.6% vs. 19.4%; odds ratio [OR], 5.2; P = 0.022). In multivariate logistic regression analysis, more advanced BA at 1 year (OR, 6.1; 95% confidence interval [CI], 1.57-23.87) and lower height SDS for BA at 1 year (OR, 0.24; 95% CI, 0.06-0.94) were associated with relatively decreased GV (< 5 cm/yr) during the second year of GnRHa treatment. Conclusion: GV during and after the second year of GnRHa treatment in girls with idiopathic CPP remains within the normal prepubertal range, and relatively low GV during GnRHa treatment is associated with more advanced BA and lower height SDS for BA.

Opposite Models of Expression of Androgen Receptor (AR) and Retinoic Acid Receptor-α (RAR-α) in the Onset of Male Germ Cell Development in Hormonally Manipulated Rats

ABSTRACTVitamin A together with testosterone (T) and FSH play an essential role in regulating spermatogenesis. Androgens and Vitamin A act through specific nuclear receptors: AR, retinoic acid receptors (RAR-α, β, γ). Present study aimed to investigate relationship between expression of RAR-α and AR in the initiation of spermatogenesis during puberty in conditions of neonatal hormonal manipulation.Experimental model of neonatal treatment with diethylstilbestrol (DES), GnRH-antagonist, T was used. Immunostaining for RAR-α and AR; 121-point counting were applied. Absolute nuclear volume (ANV) was estimated for Sertoli (SC), germ cells (GC) and their subtypes. At onset of puberty (d18) over expression of RAR-α was evident in SCs after DES-10 but not GnRHa treatment. Co-administration of T+DES restored the normal pattern of weak immunoreactivity. An opposite model of expression was seen for AR—loss of expression after DES but not GnRHa application and recovery by T-therapy. Similar reduction was found for spermatogonial ANV in both groups, whereas ANV of spermatocytes (Sc) is reduced in greater extent by DES (10x) than GnRHa (6x). Preleptotene (Pl), leptotene (L) and zygotene (Z) stages was also more affected by DES than GnRHa. Sertoli cell support toward spermatocytes (Sc/SC ratio) was more affected by DES (4x decrease) than GnRHa (2x) and the same tendency is found for Pl/SC and L-Z/SC.In conclusion, data suggest possible interplay between retinoid and androgen signaling in Sertoli cells in the differentiation of male germ cells. The anti-androgenic effect of estrogens on meiotic germ cells is probably mediated by augmentation of RAR- α expression.

Artificial fertilization of Senegalese sole ( Solea senegalensis): Hormone therapy administration methods, timing of ovulation and viability of eggs retained in the ovarian cavity

One method to overcome the difficulties involved in reproducing first generation Senegalese sole that are born in captivity (F1) is to obtain gametes by stripping and then using them for artificial fertilization. The aim of this paper was to determine an appropriate protocol for hormonal induction of ovulation and artificial fertilization. To induce ovulation, two different GnRHa administration methods, weekly injections versus a single GnRHa implant, were compared. The number of spawns per female and the relative fecundity were significantly higher in females that received the hormonal therapy through repeated injection (6.3 ± 0.8 spawns per female and 574.9 ± 67.2 × 10 3 eggs kg −1 BW) than those treated with a single implant (2.7 ± 0.6 spawns per female and 134.6 ± 40.9 × 10 3 eggs kg −1 BW). The lapsed time to ovulation and the changes in fertilization and hatching rates of eggs that were retained in the ovarian cavity after ovulation were determined after GnRHa treatment. At 16 °C, ovulation was detected at 41–44 h post-treatment in 70% of the females. The highest fertilization and hatching rates were obtained during the first 3 h after ovulation, and they significantly decreased thereafter. The results suggested that the GnRHa injection was the most appropriate administration method for the induction of ovulation. Ovulated eggs inside the female were viable for a short time, and therefore it is important to determine the time of ovulation after hormonal treatment.

Near-final height in patients with congenital adrenal hyperplasia treated with combined therapy using GH and GnRHa

Intrinsic limitations of glucocorticoid therapy in patients with congenital adrenal hyperplasia (CAH) determine frequent loss in final height. The association of secondary central precocious puberty and early epiphyseal fusion is also frequent. In these conditions, GnRHa treatment alone or in combination with GH has been indicated. This is a retrospective study, describing the estatural findings of CAH patients with significant decrease in height prediction, who were submitted to combined GH plus GnRHa therapy up to near-final height. We studied 13 patients, eight females and five males, eight with the classical and five with the nonclassical form of the disorder. Treatment with hydrocortisone (10-20 mg/m(2)/day) or prednisolone (3-6 mg/kg/day) was associated with GnRHa (3.75 mg/months) for 4.0 (1.5) years, and GH (0.05 mg/kg/day) for 3.6 (1.4) years. Stature standard deviation score for bone age improved significantly after GH treatment, becoming similar to target height at the end of the second year of GH treatment. We conclude that combined GH plus GnRHa therapy can be useful in a subset of CAH patients with significant reduction of predicted final height associated with poor hormonal control and central precocious puberty.

Comparison of the effects of different methods of mammalian and salmon GnRHa administration on spawning performance in wild-caught female carp ( Cyprinus carpio carpio) from the Caspian Sea

The effects of acute or sustained administration of mammalian and salmon GnRH agonists (mGnRHa and sGnRHa) were examined on the reproduction performance of wild-caught female carp from the Caspian Sea. Ripe broodfish were caught in the beginning of spawning season (early May) by seine from the southeast coastal waters of the Caspian Sea, Golestan province, Iran. To induce spawning fish were allocated into 8 groups and treated with 7% physiological saline (control), 4 mg kg − 1 carp pituitary (CP) extract (positive control), 20 μg kg − 1 mGnRHa or sGnRHa together with 40 mg kg − 1 Metoclopramide (acute treatments), a cholesterol pellet containing 20 μg kg − 1 mGnRHa or sGnRHa together with 40 mg kg − 1 Metoclopramide, and emulsified GnRHa in Freud's Incomplete Adjuvant (FIA) containing 20 μg kg − 1 mGnRHa or sGnRHa together with 40 mg kg − 1 Metoclopramide (mGnRHa-FIA and sGnRHa-FIA). After the final injection the fish were checked for spawning regularly. Ovulated fish were stripped manually and the eggs fertilized artificially. Significant differences were observed in the reproduction success, latency period and practical fecundity among treated groups ( p ≤ 0.05). Injection of CP or sGnRHa in acute or sustained methods increased significantly the reproduction success compared to acute injection, implanting of mGnRHa or controls ( p ≤ 0.05). In spite of the weaker effect of mGnRHa injection or cholesterol implant on induction of ovulation, treatment with mGnRHa-FIA showed similar effects as sGnRHa-FIA. Almost all sustained delivery systems of both GnRHa enhanced the practical fecundity of treated fish compared to CP and acute GnRHa injections. The latency periods of fish that received GnRHa-FIA and implant were near 24 and 50 h, respectively, which were significantly longer than that of fish given an acute GnRHa treatment or CP ( p ≤ 0.05). There was no significant difference in fertilization and hatching success of the produced progeny among treated groups ( p ≥ 0.05). The results showed that injection of GnRHa in FIA increased significantly the ovulation success of fish, but implanting fish with mGnRHa in a cholesterol pellet had variable effects with increasing fecundity but not higher ovulation success, compared to the CP treatment.

Induced ovulation of yellow catfish (Pelteobagrus fulvidraco) using a combination of a gonadotrop-releasing hormone analogue and domperidone

The effects of an intraperitoneal hormone injection of gonadotropin-releasing hormone agonist (D-Ala6, Pro9-NEt GnRHa) alone or in combination with a dopamine antagonist, domperidone (DOM), on ovulation induction in yellow catfish Pelteobagrus fulvidraco were tested. The hormone treatments were as follows: 6 mg kg−1 body weight (BW) of carp pituitary extract as a positive control, GnRHa 10, 20, 40 and 80 μg kg−1 BW and a combination of GnRHa and DOM as follows: 10 μg+5 mg, 20 μg+10 mg, 40 μg+20 mg and 80 μg+40 mg kg−1 BW. Physiological saline (0.7% NaCl) was used as a negative control. Significant differences in the ovulation ratio, latency period and ovulation index (OI) were observed among treatments (P<0.05). The combination of GnRHa and DOM at doses of 40 μg+20 mg kg−1 BW had higher values of the ovulation ratio and OI, and a shorter latency period compared with other treatments. The highest OI in GnRHa treatments was only 56.67%, suggesting a dopaminergic tone on gonadotropin secretion in this fish at the pre-ovulatory stage. Therefore, ovulation can be successfully induced in yellow catfish with 40 μg kg−1 GnRHa+20 mg kg−1 DOM without affecting the egg quality.

Overnight Luteinizing and Follicle Stimulating Hormone Profiles During GnRHa Treatment in Short Girls Born Small for Gestational Age

Since puberty starting at a height less than 140 cm might reduce adult height, postponement of puberty was studied in short pubertal girls born SGA. Data on overnight LH and FSH profiles during GnRHa treatment are very limited. To evaluate whether 3 months of GnRHa treatment results in sufficient suppression of pubertal LH and FSH profile patterns. To evaluate whether girls show sufficient pubertal suppression according to a consensus-based peak LH cut-off level of 3 IU/l during a GnRH agonist test. Twenty-one short pubertal girls born SGA. After baseline LH and FSH profiles, children received leuprorelide acetate depots of 3.75 mg subcutaneously, every 4 weeks. At baseline, amplitude and frequency of LH and FSH pulsatility were higher in girls with breast stage 3, compared to girls with breast stage 2. After 3 months of GnRHa treatment, all girls showed clinical arrest of puberty and their LH and FSH levels during overnight profiles had significantly decreased to prepubertal levels. In contrast, peak LH during the GnRH agonist test indicated insufficient pubertal suppression in 33% of girls. No differences in LH and FSH profiles were found between girls with a peak LH above or below 3 IU/l. After 3 months of GnRHa treatment, central puberty was adequately suppressed in all girls, as shown by the prepubertal LH and FSH profiles. The GnRH agonist falsely indicated insufficient pubertal suppression in 33% of these girls.

Fatores determinantes do ganho na altura em meninas com puberdade precoce central idiopática tratadas com análogo de GnRH

to evaluate predictive factors of response to GnRHa treatment in girls with idiopathic central precocious puberty. a retrospective cohort study was conducted involving 33 girls diagnosed with idiopathic central precocious puberty and treated with GnRHa. The following independent variables were assessed: age at the beginning of therapy and at the onset of symptoms, time elapsed since the appearance of pubertal characteristics and the beginning of treatment, bone age, bone age advance, duration of GnRHa treatment, actual height and Z-score, predicted height and Z-score and hormone measurements of FSH and LH after GnRH stimulation, which were correlated with gain in height as a dependent variable at treatment discontinuation, calculated by the difference between the predicted height at the end and beginning of treatment. For statistical analysis, Pearson's linear correlation was used, in addition to multiple linear regression analysis. the mean age at the beginning of treatment was 7.8+/-1.3 years, with a mean bone age of 10.1+/-1.6 years. Bone age advance was 2.3+/-1.1 years and was controlled during the treatment period. Gain in predicted height was 2.5+/-1.3cm. It was positively correlated with time elapsed since the beginning of symptoms and the beginning of treatment and with bone age advance, while negatively correlated with the Z-score of height at the beginning of treatment and predicted height at the beginning of treatment, and the latter was the main factor determining gain from treatment. girls who had the most significant compromise of predicted adult height, as detected by a larger deviation from the population (Z-score) and the most considerable advance in bone age, received benefit from GnRHa therapy, and they must not be excluded from the group to be treated.

Extended hypo-estrogenic treatment might enhance implantation in those who have failed to conceive after multiple embryo transfers

OBJECTIVE: It is well known that the pregnancy rate (PR) is higher in recipient women without ovarian function than those with ovarian function in oocyte donation programs, though the mechanism is not clear. We investigated whether extended hypo-estrogenic state with GnRHa treatment might be effective to enhance implantation in those who have failed to conceive after multiple embryo transfers.DESIGN: A prospective clinical study was carried out at a private IVF clinic between October 2006 and December 2007.MATERIALS AND METHODS: A total of 98 women, who had failed to conceive after three or more fresh and frozen thawed ETs (FET), entered this study. Thirty-three women who agreed to have this treatment were given GnRH agonist (Leuplin 1.88mg, Takeda, Japan) once a month for 1 to 3 months to induce at least one month duration of hypo-estrogenic state prior to FET. Sixty-five women who had FET during the same period were served as the control group and received OC for a month prior to FET. Endometrium was prepared with oral and patched estrogen (E) and progesterone (P) IM in all FET cycles. Only blastocysts were vitrified with cryoloop and transferred on the 5th day after P administration. Serum HCG was measured on the 14th day after P administration and pregnancy was determined with the presence of fetal heart beat.RESULTS: Thirty-three FET cycles were performed in the study group and 78 cycles were performed in control. Mean age of women was 36.9 in study group and 36.1 in control. The number of previous ET cycles was 4.1(3-6) and 4.1 (3-8), respectively. The number of patients with diagnosis of endometriosis and/or myoma was 5 and 9, respectively and there were no differences in their proportion of the groups. The mean number of blastocysts transferred was 1.4 in both groups and their grades at the time of vitrification were similar according to blastocyst grading score. The implantation rate and ongoing PR were 42.6% (20/47) and 51.5% (17/33) in study group, and 26.1% (29/111) and 28.2% (22/78) in control group, respectively. These differences were statistically significant (p<0.05).CONCLUSIONS: Although the mechanism is not clear, extended hypo-estrogeninc treatment with GnRH agonist may affect the sensitivity of E and P and/or enzyme expression, such as aromatase cytochrome p450 in endometrium and resulted in an increased implantation rate in those who had a history of multiple implantation failures. OBJECTIVE: It is well known that the pregnancy rate (PR) is higher in recipient women without ovarian function than those with ovarian function in oocyte donation programs, though the mechanism is not clear. We investigated whether extended hypo-estrogenic state with GnRHa treatment might be effective to enhance implantation in those who have failed to conceive after multiple embryo transfers. DESIGN: A prospective clinical study was carried out at a private IVF clinic between October 2006 and December 2007. MATERIALS AND METHODS: A total of 98 women, who had failed to conceive after three or more fresh and frozen thawed ETs (FET), entered this study. Thirty-three women who agreed to have this treatment were given GnRH agonist (Leuplin 1.88mg, Takeda, Japan) once a month for 1 to 3 months to induce at least one month duration of hypo-estrogenic state prior to FET. Sixty-five women who had FET during the same period were served as the control group and received OC for a month prior to FET. Endometrium was prepared with oral and patched estrogen (E) and progesterone (P) IM in all FET cycles. Only blastocysts were vitrified with cryoloop and transferred on the 5th day after P administration. Serum HCG was measured on the 14th day after P administration and pregnancy was determined with the presence of fetal heart beat. RESULTS: Thirty-three FET cycles were performed in the study group and 78 cycles were performed in control. Mean age of women was 36.9 in study group and 36.1 in control. The number of previous ET cycles was 4.1(3-6) and 4.1 (3-8), respectively. The number of patients with diagnosis of endometriosis and/or myoma was 5 and 9, respectively and there were no differences in their proportion of the groups. The mean number of blastocysts transferred was 1.4 in both groups and their grades at the time of vitrification were similar according to blastocyst grading score. The implantation rate and ongoing PR were 42.6% (20/47) and 51.5% (17/33) in study group, and 26.1% (29/111) and 28.2% (22/78) in control group, respectively. These differences were statistically significant (p<0.05). CONCLUSIONS: Although the mechanism is not clear, extended hypo-estrogeninc treatment with GnRH agonist may affect the sensitivity of E and P and/or enzyme expression, such as aromatase cytochrome p450 in endometrium and resulted in an increased implantation rate in those who had a history of multiple implantation failures.

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg−1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.

DEVELOPMENTAL PROGRAMMING: DO EXOGENOUS GONADOTROPINS NORMALIZE OVARIAN FUNCTION IN PRENATAL TESTOSTERONE-TREATED FEMALES?

Testosterone (T) treatment of fetal lambs from 30–90 d of gestation (T60; term 147 d) leads to hypergonadotropism, multifollicular ovaries and follicular persistence. The disrupted ovarian follicular dynamics may be the consequence of neuroendocrine defects resulting in altered gonadotropic drive or, alternatively, compromised ovarian sensitivity to gonadotropins. To test the hypothesis that reproductive dysfunction in T60 ewes is a consequence of abnormal gonadotropic drive, release of endogenous LH and biopotent FSH was blocked by administering gonadotropin-releasing hormone antagonist (GnRHA; acyline; 10 μg/Kg, 12-hourly) to control (C, n = 5) and T60 (n = 5) ewes for 9 days. On day 7 of GnRHA treatment, ewes were injected with LH (0.108 μg/kg) and native ovine FSH (0.173 μg/kg) at 2-h intervals for 36 h followed by hourly injections of LH (0.054 μg/kg) and a combination (1:4 ratio) of native FSH (0.043 μg/kg) and asialo FSH (FSH exposed to neuroaminidase; 0.173 μg/kg) for the next 12 h. Native FSH is less biopotent with a longer half life. Asialo FSH is short-lived but more biopotent. All ewes were then given 1500 IU of hCG to induce ovulation. Plasma estradiol (E) was measured at 4-h intervals during the LH/FSH drive. Transrectal ultrasonography was performed to monitor ovarian follicular dynamics at the start of GnRHA and daily beginning 1 d before and concluding 4 d after LH/FSH administration. Ultrasonography also was performed 10 d after hCG to assess corpus luteum (CL) development. Follicle number and diameter, basal and peak E concentrations and temporal changes in follicular size classes were analyzed by ANOVA, with repeated measures as needed. Total number of follicles tended to be greater (P=0.09) in T60 ewes before the start of GnRHA treatment. Six days after exposure to GnRHA, T60 ewes continued to have a greater number of follicles (P<0.05) due to increased number of <2 mm follicles (P<0.01). With advancing LH/FSH treatment, a progressive increase (P<0.01) in >4–8 mm size follicles (ovulatory range) were evident in both groups and there were no treatment differences. After hCG, follicles from both C and T60 females continued to grow (P<0.01) with some achieving >12–20 mm in size. The number of CL (C:4.0 ± 1.5; T60:3.8 ± 0.5) and luteal cysts (C:5.0 ± 1.1; T60:6.0 ± 3.3) did not differ between C and T60. Concentrations of E (pg/mL) were similar (C:9.1 ± 1.6; T60: 10.8 ± 1.1) at the start of FSH treatment. Peak circulating E (C:4.5 ± 1.1; T60: 7.1 ± 1.8) and area under the E curve (C:17.6 ± 3.4; T60: 31.8 ± 7.8) were similar after LH/FSH treatment. Based on these findings, we suggest that exogenous gonadotropin treatment following blockade of GnRH action with GnRHA equalized the follicular and circulating E responses of T60 to that of C. The superstimulation achieved with more biopotent, short-lived asialo FSH did not allow proper testing of the hypothesis. Development of follicles >12–20 mm that persisted even after hCG in both C and T60 females indicated that these follicles may have persisted because they failed to develop the attributes of a preovulatory follicle, perhaps due to the asialo FSH provided. Abnormal FSH drive may, therefore, be one mechanism by which follicular persistence may have occurred in prenatal T females. This study emphasizes the need for proper ovarian stimulation protocols to reduce the risk for multiple pregnancy and ovarian hyperstimulation syndrome. Supported by NIH-HD 41098 & P01 HD 4423. (platform)

Circulating ghrelin levels in girls with central precocious puberty are reduced during treatment with LHRH analog

Decreased levels of ghrelin have been measured in growing children during puberty. No data are available for girls with central precocious puberty (CPP). To explore ghrelin changes before, during, and after GnRH analog treatment in girls with CPP. A sample of 20 Caucasian girls (8.08 +/- 0.65 years of age) with CPP was recruited. Height and weight, bone age, LH, FSH, 17beta estradiol (E(2)), and ghrelin were measured before starting treatment with GnRH analog, 18 months after therapy began and again 6 months after therapy discontinuation. LH and E(2) serum levels decreased significantly during treatment (2.45 +/- 2.03 vs 0.67 +/- 0.49 UI/l, P < 0.01 and 28.17 +/- 9.7 vs 15 pmol/l, P < 0.01 respectively), returning to baseline levels after the discontinuation of therapy (4.75 +/- 1.66 UI/l and 29.23 +/- 6.99 pmol/l respectively). LH peaked following LHRH stimulation significantly (P < 0.01) decreased during treatment (24.45 +/- 14.17 vs 1.3 +/- 0.18 UI/l) and then increased after therapy discontinuation (12.58 +/- 6.09, P < 0.01). Ghrelin decreased significantly (P < 0.05) during treatment (1849 +/- 322 vs 1207 +/- 637 pg/ml), and increased, though not significantly (P = 0.09) after therapy withdrawal (1567 +/- 629 pg/ml). Contrary to what is expected in physiologic puberty, where ghrelin is progressively reduced, the prepubertal hormone milieau induced by GnRHa treatment in patients suffering from central precocious puberty (CPP) did not promote an increase in ghrelin circulating levels. Therefore, in CPP, ghrelin secretion seems to be independent from pubertal development per se. Concomitant estrogen suppression during treatment may play a potential role in the regulation of ghrelin secretion in these girls.

Prevention of severe menorrhagia in oncology patients with treatment‐induced thrombocytopenia by luteinizing hormone‐releasing hormone agonist and depo‐medroxyprogesterone acetate

Menorrhagia is a serious complication in young female oncology patients who suffer from severe thrombocytopenia during myelosuppressive treatment. To the authors' knowledge, little is known regarding the incidence of this complication or the effectiveness of possible therapies for its prevention. In this retrospective clinical study, after a thorough gynecologic evaluation, young female oncology patients with regular menstrual cycles undergoing myelosuppressive treatments received either depo-medroxyprogesterone acetate (DMPA), or D-tryptophan-6-luteinizing hormone-releasing hormone depot treatment (gonadotropin-releasing hormone agonist [GnRH-a]), or no treatment before the administration of myelosuppresive chemotherapy. Only patients who later developed severe thrombocytopenia (<25,000 platelets per muL) were included in the study. Daily blood counts, menorrhagia, nonvaginal bleeding episodes, and the need for blood products, gynecologic consultations, and other medical interventions were determined. Of 101 women with cancer who met the inclusion criteria, 42 patients received DMPA, 39 patients received GnRH-a, and 20 patients remained untreated. The mean duration (+/- standard deviation) of severe thrombocytopenia was 24.76 +/- 23.6 days. Four patients were not included because of significant gynecologic pathologies. General bleeding from nongynecologic sites was similar for all groups and was not associated with vaginal bleeding. Severe or moderate menorrhagia was documented in none of the 39 women who received GnRH-a, in 9 patients (21.4%) who received DMPA, and in 9 untreated patients (40%; P = .02). Fewer calls for urgent gynecologic consultations were documented in the GnRH-a group compared with the untreated group (P < .0001). Female patients undergoing myelosupressive therapy are at high risk of developing significant menorrhagia during prolonged, severe thrombocytopenia. Pretreatment gynecologic evaluation can detect significant pelvic pathologies. GnRH-a treatment effectively prevented menorrhagia, whereas DMPA administration was less effective.

Induction of spawning of captive-reared Senegal sole (Solea senegalensis) using different administration methods for gonadotropin-releasing hormone agonist

Captive-reared Senegal sole were treated with intramuscular injection or sustained-release implants of gonadotropin-releasing hormone agonist [D-Ala6, Pro9, NEt] GnRHa (GnRHa) to induce spermiation and spawning. Fish were treated with GnRHa during spring (April–May) or autumn (October), the natural spawning seasons known to occur in wild Senegal sole. During spring, females injected with a dose of 5μg GnRHa kg−1 three times a week, or treated with a single GnRHa-loaded implant (50μg kg−1) showed multiple ovulations and spawns within a period of approximately 30 days. Plasma levels of 17β-estradiol (E2) were elevated at 7days post-treatment, preceding the spawns with the highest fecundity, concomitant with a decrease in the levels of plasma testosterone (T). During autumn, however, females changed their responsiveness to GnRHa treatment, a dose of 5μg kg−1 being ineffective in inducing spawning, while a dose of 1μg kg−1 induced a few spawns. Accordingly, plasma E2 levels showed a small increase after injection of 1μg kg−1 GnRHa. In contrast to females, GnRHa injection or implantation in males was ineffective in enhancing spermiation or milt production, regardless of the season when the treatments were applied. However, GnRHa treatment in males increased plasma levels of T and 11-ketotestosterone (11-KT) during spring, while in autumn the levels of plasma androgens decreased after GnRHa injection. These results show that administration of GnRHa during spring either by injection or sustained-release implants induces multiple spawns in captive Senegal sole, although these treatments were ineffective in inducing or enhancing sperm production in males.