GnRH Group Research Articles

Use of repeat-dose GnRH agonist, hCG, and co-trigger for final follicular maturation have similar rates oocyte maturity

Use of GnRH agonists with or without hCG for final follicular maturation has become increasingly common in high responders as well as in patients with normal ovarian reserve. However, studies investigating the effect of trigger type on IVF outcomes have been inconsistent and the majority studies have looked at patients triggered with single-dose rather than repeat-dose GnRH. (1-3) The objective of this study was to determine if rate of oocyte maturity differs between patients triggered with repeat-dose GnRH agonist, hCG, and GnRH agonist + hCG co-trigger. All IVF stimulation cycles at a single academic center between December 2016 and February 2018 were included in the study. Cycles were categorized by type of trigger: repeat-dose GnRH agonist trigger (leuprolide acetate 4mg q 12 hours x 2 doses), hCG trigger (purified hCG 5,000-10,000IU), or co-trigger (leuprolide acetate 4mg q 12 hours x 2 doses + hCG 1,500-2,000IU). Retrieval was performed 36 hours after trigger injection. Cycles were evaluated for patient age, antral follicle count (AFC), days of gonadotropin stimulation, oocyte maturity rate (# MII oocytes/# follicles > 12 mm at time of trigger) and fertilization rate. Statistical analysis was performed using analysis of variance and multivariate logistic regression. A total of 132 women underwent 152 cycles during the study period. All cycles were GnRH antagonist protocol. Of the 152 IVF cycles included, hCG trigger was used in 93 cycles (61%), repeat-dose GnRH agonist was used in 43 cycles (28%), and GnRH agonist + hCG co-trigger was used in 16 cycles (11%). The groups did not differ by BMI or number of days of gonadotropin stimulation. Age was significantly higher and AFC lower in the hCG compared to the GnRH and co-trigger groups (Table 1). In a multivariate regression model, after controlling for age and AFC, no difference was observed between trigger type and maturity rate (p=0.669) or fertilization rate (p=0.181).Table 1IVF outcomes by trigger type: Repeat-dose GnRH agonist vs hCG vs GnRH agonist + hCG co-triggerhCG (n = 93)GnRH agonist (n = 43)Co-trigger (n=16)p-valueDays of stimulation9.9 (1.4)9.5 (0.9)9.4 (1.2)0.078Peak E2 (pg/mL)2095.9 (794)4932.4 (1145.9)3735.2 (530.1)<0.001Follicles > 12mm9.6 (1.4)9.5 (0.9)9.4 (1.2)0.078Oocytes retrieved11.8 (6.7)20.9 (6.2)20.6 (6.9)<0.001MII oocytes8.8 (5.3)15.8 (5.6)14.6 (5.4)<0.001Maturity rate (%)83.5 (19.4)83.7 (16.5)85.8 (19.8)0.9Fertilization rate (%)77.5 (22.4)82.3 (21.2)89.7 (11.8)0.079Data presented as mean +/- standard deviation, unless otherwise noted Open table in a new tab Data presented as mean +/- standard deviation, unless otherwise noted Cycles triggered with repeat-dose GnRH agonist, hCG, and GnRH agonist + hCG co-trigger have equivalent oocyte maturity and fertilization rates in our cohort. These findings are reassuring that choice of trigger does not appear to impact oocyte maturity, even after controlling for ovarian reserve and patient age. However, further investigation into the impact of trigger type, including optimal dosage of GnRH agonist, on embryo development, clinical outcomes, and live birth rate is warranted.

Timed artificial insemination plus heat II: gonadorelin injection in cows with low estrus expression scores increased pregnancy in progesterone/estradiol-based protocol

The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm2, GnRH= 2.37 cm2, HEAT group= 3.07 cm2, P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.

Effect of GnRH analogues for pituitary suppression on oocyte morphology in repeated ovarian stimulation cycles.

Objective:To compare the effect of pituitary suppression regimens on oocyte morphology in consecutive ICSI cycles of the same patients.Methods:Data was obtained from 200 matched consecutive intracytoplasmic sperm injection (ICSI) cycles performed in 100 couples undergoing the first cycle with the GnRH agonist and the following cycle with the GnRH antagonist regimen, from January 2010 to August 2016, in a private university-affiliated IVF centre. The effects of the pituitary suppression type on oocyte morphology were assessed by multivariate General Linear Models.Results:Mean interval between cycles was 185.32±192.85 days. Maternal age, body mass index, and total FSH dose administered were similar in both patients' cycles. Antagonist cycles presented lower incidence of dark cytoplasm (0.69±3.28% vs. 4.40±17.70%, p=0.047), Smooth endoplasmic reticulum (SER cluster (4.37±11.62% vs. 7.36±17.17%, p=0.046), and ZP defects (6.05±14.76% vs. 11.84±25.13%, p=0.049). Similar numbers of follicles retrieved oocytes, and mature oocytes were observed between the GnRH groups, as well as the fertilisation rate, number of obtained embryos, high-quality embryo rates, and the clinical outcomes.Conclusion:GnRH antagonist's inhibitory effect on the ovaries in consecutive ICSI cycles results in improved oocyte maturity and morphology, despite similar laboratory and clinical outcomes, compared to the GnRH agonist treatment.

Effectiveness of Different Hormonal Treatment Protocols on Treatment of Postpartum Anoestrus in Cross-Bred Dairy Cows

Postpartum anoestrus (PPA) is one of the major factors causing prolonged calving interval &amp; that leading to decreased annual milk production. Hormonal treatments can be effective tools for treatment of PPA when nutritional &amp; environmental management are good. So, objective of this study was to know the comparative effectiveness of different hormonal protocols on treatment of PPA in cross-bred dairy cows. Used protocols were: single dose of GnRH, Ovsynch with TRIU-B and single dose of PGF2α injection which were used in three different groups, each containing ten animals. GnRH and Ovsynch with TRIU-B were used in postpartum true anoestrus cattle with inactive ovaries while PGF2α was used in postpartum silent oestrus condition having palpable corpus luteum and the condition were identifi ed through rectal palpation and history record. Cows treated with Ovsynch+TRIU-B showed higher oestrus expression rate within one week (80%) than cows treated with GnRH (10%) and PGF2α (50%). Ovulation rate was also comparatively higher in Ovsynch+TRIU-B group (100%) than GnRH (90%) and PGF2α (70%). But the pregnancy rate, checked after 45 days of first AI through rectal palpation, was comparatively higher in GnRH group (40%) than in Ovsynch+TRIU-B (30%) and PGF2α (20%). Among treated cows, pregnancy rate was found to be signifi cantly higher (P=0.02) in cows with ≥100 days duration of postpartum anoestrus (53.84%) than in cows with 100 days duration (0%). In conclusion, ovsynch with intravaginal progesterone (p4) releasing device was more effective at inducing oestrus and synchronizing ovulation than the single GnRH injection. However, single GnRH injection was as effective as ovsynch with intravaginal p4 releasing device for pregnancy outcomes in anoestrous crossbred dairy cows in a commercial dairy farm where formulated concentrate ration is daily fed.

Effect of GnRH or hCG administration on Day 4 post insemination on reproductive performance in Merino sheep of North Patagonia

Different therapeutic strategies have been used with the objective of improve luteal function to reduce embryonic losses. The objective of this work was to study the effect of the administration of GnRH or hCG at Day 4 post fixed time artificial insemination (FTAI) on reproductive efficiency in Merino sheep during the breeding season in North Patagonia. Estrus of multiparous Merino ewes (n = 288) was synchronized by two injections of prostaglandins (PG; 125 μg, Cloprostenol), 14 days apart. Cervical FTAI was performed 53–56 h after the second PG with a dose of fresh semen (100 × 106 spermatozoa) from five Merino rams. In all ewes body condition score (BCS) was determined at FTAI. At 4 days post FTAI ewes were randomly assigned into three experimental groups: GnRH group (4 μg, Buserelin; n = 99), hCG group (300 IU, hCG; n = 92) and Control group (1 ml, saline solution; n = 97). Pregnancy and pregnancy losses were evaluated by ultrasonography on Days 33 and 90 post FTAI. Additionally, embryo crown-rump length (CRL) was measured by ultrasonography (n = 12 single-pregnant ewes by experimental group) at the first ultrasound. Date of birth, litter size and lamb weight were recorded (n = 111 pregnant ewes). Pregnancy rate on Days 33 and 90 post FTAI did not differ among treatment groups (P > 0.05). Pregnancy losses at Day 33 post FTAI were lower in the hCG group compared to the GnRH and Control groups (0, 3, 7.2%, respectively; P < 0.05). Pregnancy losses between Days 33 and 90 after FTAI were negligible (P > 0.05). The embryo CRL at Day 33 post FTAI was not increased by the hormonal treatments (P > 0.05). Moreover, it was lower in GnRH group compared to Control group (P < 0.05). Litter size tended to be greater in the GnRH group compared to the hCG and Control groups (P < 0.1). The birth weight of twin lambs tended to be higher in the GnRH group compared to the Control group (P < 0.1). The birth weight of single lambs was not affected by treatments (P > 0.05). Ram fertility and BCS of ewes at FTAI influenced the effect of hormonal treatments on reproductive parameters. In conclusion, administration of hCG or GnRH at Day 4 post FTAI does not improve pregnancy rate but treatment with hCG reduces pregnancy loss on Day 33 post FTAI. GnRH treatment improves litter size and twin lambs birth weight.

The GnRH analogue dephereline given in a fixed-time AI protocol improves ovulation and embryo survival in dairy cows

This study compares the fertility effects of inducing ovulation using the GnRH analogue, dephereline, versus natural GnRH at the end of a 5-day progesterone(P4)-based protocol for fixed-time artificial insemination (FTAI) in in heat-stressed and non-heat stressed lactating dairy cows. Cows were given GnRH (GnRH group, n = 369) or dephereline (DEPH group, n = 379) and were inseminated 14–20 h later. Dephereline treatment increased corpus luteum (CL) size on Day 7 post-AI compared with GnRH (P < .0001) while a one-mm increase in CL size was found to give rise to a 1.1-fold increase in the pregnancy rate at FTAI (P = .001). Based on odds ratios, the interaction between treatment and heat stress had a significant effect on the ovulation failure rate (P < .01). This meant that relative to non-heat-stressed GnRH-treated cows, ovulation failure was 2.9 times more likely in heat-stressed GnRH-treated cows (P = .001), 0.3 times less likely in non-heat-stressed DEPH-treated cows (P = .04) and was similar in heat-stressed DEPH-treated cows. Further, non-heat-stressed DEPH-treated cows were more likely to conceive by a factor of 1.6 than the remaining cows (P = .03). Finally, GnRH-treated multiparous cows were 9.9 times more likely to suffer pregnancy loss than the remaining cows (P = .03). Our results indicate that, compared to treatment with GnRH, dephereline reduced the risk of ovulation failure and consequently increased the pregnancy rate under heat stress conditions. In multiparous cows, dephereline treatment also reduced the negative age effect on pregnancy maintenance.

Evaluation of the effect of mating, intrauterine deposition of raw seminal plasma or seminal plasma purified β-NGF on endometrial vascularization in llamas

The aim of this study was to evaluate the endometrial vascularization area (EVA) of both uterine horns in llamas subjected to different intrauterine treatments resembling physiological conditions after a single mating. Llamas with a growing follicle (≥8 mm) were randomly assigned to: a) single mating with a fertile male (mating; positive control; n = 6); b) intramuscular administration of 50 μg of gonadorelin acetate plus an intrauterine infusion of 4 ml of PBS (GnRH; negative control; n = 4); c) intrauterine infusion of 4 ml of raw llama seminal plasma (SP; n = 4) or d) intrauterine infusion of 10 mg of β-NGF purified from llama semen diluted in 4 ml of PBS (spβ-NGF; n = 6). Females in GnRH, SP and NGF group received 50% of treatment volume into each horn by guiding an insemination pipet through the cervix. Ovaries were examined by ultrasonography every 12 h until Day2 (Day 0 = Day of treatment) to determine ovulation. Power-Doppler ultrasonography evaluation of EVA in a cross-section of the middle segment of each horn was conducted at 1 h before and 1, 3, 6, 12 and 24 h (intensive evaluation) and 2, 4, 6 and 8 days (long-term evaluation) after treatment administration. Serial EVA data was analyzed as a 2-by-2 factorial design for repeated measures using the MIXED procedure. The analysis included main effects of treatment (mating, SP, spβ-NGF or GnRH), uterine horn (left vs right), time, and their interactions. According to the 2 by 2 analysis there was no effect of uterine horn on EVA during the first 24 h and from Day 2 to Day 8 after treatment; therefore, data were grouped based on treatment type regardless of uterine horn for both periods of observation. Thus, EVA was affected by time (P < 0.04) and treatment by time interaction (P < 0.02) and tended (P = 0.07) to be influenced by type of treatment during the intensive evaluation period. Females on mating and spβ-NGF group showed a significant increase in EVA at 3 and 12 h after treatment compared to GnRH and SP groups. However, no effect of treatment, time or their interaction was observed during the long-term evaluation period. In spite of the limited number of animals used in this study, our results allow us to concluded that natural mating and intrauterine deposition of 10 mg of spβ-NGF induce a symmetrical increase in endometrial vascularization of both uterine horns during the first 24 h post treatment administration in llamas; however, this effect did not persist beyond that period.

Effect of Kisspeptin-10 on plasma luteinizing hormone concentrations and follicular dynamics during the luteal phase in cattle

Two experiments were carried out to determine the effect of Kisspeptin-10 on plasma LH concentrations and follicular dynamics during the luteal phase in cattle. We tested the hypothesis that a single treatment of Kisspeptin-10 will increase plasma LH concentration and the diameter of the dominant follicle, and induce ovulation during the luteal phase of the estrous cycle in cattle. In the Experiment 1, Hereford-cross heifers (n = 28, 14–16 mo) were given PGF2α im to induce luteolysis and ovulation. On Day 5 (Day 0 = ovulation), a new follicular wave was induced by ultrasound-guided follicular ablation. Heifers were treated on Day 10 (4 days after wave emergence) with 100 μg GnRH im (n = 9), 2 mL saline im (n = 7), 1 mg Kisspeptin-10 im (Kp im, n = 6) or 1 mg Kisspeptin-10 iv (Kp iv; n = 6). Blood samples were collected at −60, −15, 0, 5, 15min (0 min = time of injection) and every 15 min thereafter until 3 h. Transrectal ovarian ultrasonography was performed at 12 h intervals from Day 10–14. In Experiment 2, non-lactating beef cows on Day 5 were treated with 100 μg GnRH im (n = 9), saline im (n = 5), 10 mg of Kisspeptin-10 iv (Kp 10 mg; n = 5) or 15 mg of Kisspeptin-10 iv (Kp 15 mg; n = 5). Blood samples were collected at −15, 0, 15, 30, 60, 120, 180 min and twice daily ovarian ultrasonography was done from Day 5–10. In Expt 1, plasma LH concentrations increased for 1 h following Kp iv administration. The peak concentration occurred at 15 min and was higher in the Kp iv group than in the Kp im group (P = 0.01). The LH peak was 3.5-folds higher in the GnRH group than the Kp iv group (P < 0.0001). In Expt 2, GnRH induced higher (P < 0.001) plasma LH concentrations for all time-points than other groups. Kp 15 mg at peak (15min), 30 and 60 min induced higher (P < 0.0001) plasma LH concentrations than Kp 10 mg and saline. Kisspeptin-treated animals did not ovulate in either experiment while GnRH induced ovulation (n = 5/9 in Expt 1; 9/9 in Expt 2). The diameter of the dominant follicle was greater (P = 0.02) at 12–48 h after kisspeptin treatment (Kp groups combined) than the Saline group in Expt 2. In conclusion, Kisspeptin-10 increased plasma LH concentrations and follicle size, and although plasma LH concentrations were higher after iv than im administration, but at the doses used, Kisspeptin-10 did not induce ovulation during the luteal phase in cattle.

Effect of pre-treatment with gonadotropin-releasing hormone analogue GnRH-α on high-intensity focussed ultrasound ablation for diffuse adenomyosis: a preliminary study

Objectives: To evaluate the efficacy and safety of GnRH-α pre-treatment with HIFU ablation for diffuse adenomyosis.Methods: From January 2014 to December 2016, 61 patients were divided into two groups. Twenty-three patients with larger uteri received pre-treatment with GnRH-α and were then subjected to HIFU, and 38 patients underwent HIFU alone. The technical parameters included treatment time, sonication time, average sonication power, treatment intensity, total energy, non-perfusion volume (NPV) and NPV ratio. Intra-, post-procedural complaints, the relief rate of dysmenorrhoea and clinical effectiveness were followed up.Results: Although the lesion volume in the HIFU + GnRH group was larger than in the HIFU-only group, higher NPV, NPVR%, treatment intensity and total energy with shorter treatment and sonication times in the HIFU + GnRH group were obtained than that in the HIFU-only group. Significant differences were evident for NPV, NPVR%, average power, and total intensity energy (p < 0.05), but not for other parameters between the two groups (p > 0.05).Although no differences in the intra-, post-procedural VAS scores, in the adverse effects and in the relief rate or clinical effectiveness were apparent between the two groups (p > 0.05), the relief rate was better in the HIFU + GnRH group than that in the HIFU group from 6 to 12 months after treatment. Self-comparison of differences occurred in the two groups before and after treatment (p < 0.05) and were between the two groups after only 6 months (p < 0.05).Conclusion: The combination of HIFU with GnRH-α is more effective than HIFU alone for ablation of diffuse adenomyosis. Moreover, the GnRH-α pre-treatment with HIFU is safe.

Effect of GnRH analogue administration on Day 7 after natural mating on formation accessory corpus luteum, progesterone concentration and conception rate in llamas (Lama glama)

The objectives of the present study were to determine the effects of exogenous GnRH administered 7 days after breeding on the formation of an accessory corpus luteum (ACL), plasma progesterone (P4) concentrations and pregnancy rates. Adult females (n = 71) having a follicle ≥ 7 mm in diameter in the ovary were naturally mated (Day 0). On Day 7, ultrasonic examination was performed to confirm the occurrence of ovulation as evidenced by presence of an induced corpus luteum (ICL). Females with an ICL plus a dominant follicle ≥ 7 mm (n = 56) were treated with saline solution (SS, n = 29) or GnRH analogue (n = 27). On Day 14, the formation of an ACL was observed by ultrasonography. Blood samples were collected on Days 7 and 14 to quantify plasma P4 concentrations. On Day 14, 21 of 27 (77.8%) females in the GnRH group developed an ACL, whereas females in the SS group did not. Progesterone concentrations on Day 7 and 14 in those llamas diagnosed as pregnant on Day 30 were not different (P > 0.05) between groups. In addition, P4 concentration was similar for GnRH-treated females having two CL to those with a single CL. Pregnancy rates were similar (P > 0.05) between SS and GnRH groups (55.2% compared with 74.1% respectively) and the pregnancy rate for the GnRH group was not affected (P > 0.05) by the number of CL observed at Day 14 (66.6% and 75.6% for females with one and two CL respectively). In conclusion, GnRH administration on Day 7 after breeding leads to ACL formation; however, neither the plasma P4 concentration nor pregnancy rate was affected by having an ACL.

The effects of a single or double GnRH dose on pregnancy survival in high producing dairy cows carrying singletons or twins.

Pregnancy loss during the late embryonic and early fetal periods influences dairy herd economy. The objectives of this study were to (1) investigate the effects of a single or double GnRH dose administered at the time of pregnancy diagnosis (28–34 days post-AI) on the pregnancy survival of cows in their third lactation or further carrying live singletons or unilateral twins, and (2) examine the impacts of GnRH treatment on subsequent twin reduction in twin pregnancies. Cows carrying singletons (n = 1,054) or unilateral twins (n = 379) were assigned at the time of pregnancy diagnosis to the following groups: control (no treatment), GnRH (100 μg GnRH), and 2GnRH (200 μg GnRH). Pregnancy loss was recorded in 180 of the 1,433 cows (12.6%) at 58–64 days post-AI. Based on the odds ratios, there was a significant (P < 0.0001) interaction between the treatment group and twin pregnancy. This interaction implies that control cows carrying twins were 3.2 times more likely to suffer pregnancy loss than the other cows, whereas the GnRH and 2GnRH treatment groups cows carrying singletons or twins had pregnancy loss rates similar to the control cows carrying singletons. Twin reduction was observed in 35 twin pregnancies (9.2%). Cows in the GnRH and 2 GnRH groups were seven times more likely to show twin reduction than control cows. Our results indicate that GnRH administered at the time of pregnancy diagnosis had no beneficial effects in cows carrying singletons. In contrast, for twin pregnancies, the treatment increased the rate of pregnancy survival and was accompanied by an increase in the twin reduction rate.

88 Effect of Follicular Ablation and GnRH on Recipient Synchrony in Holstein Heifers

In Holstein heifer recipients that are synchronized with a single dose of prostaglandin F2α (PGF2α), only 65% (on average) are eligible to receive an IVF embryo 7 ± 1 days after oestrus. The high variability in time to onset of oestrus after PGF2α treatment and the small size of the corpus luteum (CL) on the day of embryo transfer (ET) may explain these results. Synchronizing ovulation might decrease the variability of the interval from treatment to ovulation and maximize the size of the CL. For this purpose, Holstein heifers were randomly allocated (using the lottery method of sampling) to 3 groups: (1) Control (n = 88), a single dose of PGF2α (500 μg of cloprostenol, Merck, Darmstadt, Germany; Day 0) and transrectal ultrasonography of the ovaries on Day 0 to determine the diameter of the largest follicle, on Days 2, 3, 4, and 5 to determine occurrence of ovulation, and on Day 10 to determine presence and diameter of the CL; (2) follicular aspiration (n = 15), ultrasound-guided follicular aspiration (FA) 4 days before PGF2α administration and ovarian evaluation as described above; and (3) FA + GnRH (n = 19), FA at Day –4, gonadotropin-releasing hormone (GnRH, 100 µg of gonadorelin acetate, Vetoquinol USA, Fort Worth, TX, USA) on Day 2, and ovarian examination as described in the control group. Data were analysed by one-way ANOVA and proportions by Chi-squared. The interval (days) from PGF2α treatment to ovulation was not different among groups (3.2 ± 0.4, 3.1 ± 0.2, 3.0 ± 0.1, respectively). However, the variability (residuals) in the FA + GnRH group was the smallest (P &lt; 0.05; high degree of synchrony) in the study (0.1 ± 0.1). Likewise, the FA group (0.5 ± 0.1) had a smaller interval (P &lt; 0.05) compared with the Control group (0.8 ± 0.1). Interestingly, 86% of heifers from FA + GnRH group ovulated in a single day (Day 3; P &lt; 0.05) compared with the FA and Control groups (55% and 53%, respectively). Considering the day of ovulation (only recipients that ovulated 2 to 4 days after PGF2α treatment) and CL diameter (&gt;20 mm; Nogueira et al. 2012 R. Bras. Zootec. 1, 2129-2133; Yoshida et al. 2012 Anim. Sci. J. 83, 207-212), heifers in the FA and FA + GnRH groups had higher proportion (P &lt; 0.05) of animals selected to be transferred at Day 10 compared with the Control group (93%, 95%, and 67%, respectively). In conclusion, FA induced a more synchronous ovulation and increased the number of recipients ready to be transferred at Day 10 (7 ± 1 days after ovulation). These effects were further improved with the administration of GnRH.

Hormonal therapeutic strategy on the induction of accessory corpora lutea in relation to follicle size and on the increase of progesterone in sheep

We determined the effect of GnRH or hCG treatment on day 4 post-time artificial insemination (FTAI) on the formation of accessory corpora lutea (acc-CL) and on the concentration of serum progesterone (P4) in sheep. Multiparous adult Merino ewes (n = 36) were synchronized for estrus using double injection of PGF2α agonist (125 μg Cloprostenol) with an interval of 14 days. At 53–56 h after the second PG application, FTAI was performed. On day 4 post FTAI, ewes were either treated with analogue of GnRH (4 μg buserelin; n = 12) or hCG (300 IU, hCG; n = 12) or saline solution (1 ml; Control; n = 12). Two laparoscopic ovarian examinations were performed on days 4 and 10 post FTAI. In the first observation, we determined the number of post ovulation corpora lutea (po-CL) and the site, number and diameter of follicles present in both ovaries. In the second laparoscopy, we observed the number of po-CL and acc-CL. The sizes of the follicles that generated the acc-CL were determined according to the position of the follicles observed in the first laparoscopy. Serum P4 concentration was determined on days 4, 7, 10, 13, 17 and 21 post FTAI by chemiluminescence. A similar follicular population in size and number was observed in the three experimental groups prior to the beginning of treatments (Follicles 2 mm: 6.4 ± 3.7, 3 mm: 3.0 ± 2.3, 4 mm: 1.1 ± 0.5, 5 mm: 1.4 ± 0.8; P ˃ 0.05). The formation of 1.0 ± 0.4 and 1.1 ± 0.3 acc-CL was observed in the GnRH and hCG groups, respectively (P ˃ 0.05), but was not observed in the Control group (P < 0.05). Follicle sizes from which acc-CL generated were 3, 4 and 5 mm and did not differ between hormonal treatments (P ˃ 0.05). The hCG group had higher mean concentrations of P4 on days 7, 10, 13 and 17 post FTAI compared with the GnRH group and the Control group (P < 0.05), while no differences were observed between these two latter groups (P > 0.05). Mean P4 concentrations in ewes treated with hCG showed no differences according to the size of the follicle from which acc-CL were generated (P ˃ 0.05). In conclusion, administration of hCG or GnRH on day 4 post FTAI induced the formation of one acc-CL from follicles of 3, 4 or 5 mm, indistinctly. However, serum P4 concentration increased significantly only in the hCG group. The serum P4 concentrations of acc-CL that originated from different follicle sizes did not differ.

Effect of resynchronization with GnRH or progesterone (P4) intravaginal device (CIDR) on Day 23 after timed artificial insemination on cumulative pregnancy and embryonic losses in CIDR-GnRH synchronized Nili-Ravi buffaloes

The objective of the present study was to determine the effect of resynchronization on Day 23 with either GnRH or P4 (controlled internal drug release device containing progesterone; CIDR) on pregnancy rate, cumulative pregnancy, and embryonic and fetal losses in CIDR-GnRH synchronized Nili-Ravi buffaloes. Buffaloes (n = 181) of mixed parity, lactating, 181 ± 73 days postpartum, a body condition score (BCS) of 3.2 ± 0.5 (scale of 1–5), and 450–600 kg weight were subjected to synchronization and resynchronization. All buffaloes received CIDR on Day −9.5. In addition, GnRH was injected 36 h after CIDR removal, and timed artificial insemination (TAI) was performed 18 h later (Day 0). On Day 23, buffaloes were randomly assigned to receive one of the following treatments: 1) CON (n = 63), 2) P4 (n = 55), and 3) GnRH (n = 63) for resynchronization (2nd AI). Pregnancy rate, and embryonic and fetal losses were monitored by serial ultrasonography on Days 30, 45, 60, and 90 after synchronization (1st TAI), respectively. The pregnancy rate in GnRH-treated buffaloes remained significantly and consistently higher (P < 0.05) than in the CON group at Days 30, 45, 60, and 90 after 1st TAI. Based on the pregnancy diagnosis, on Day 30 post 1st TAI, buffaloes that remained non-pregnant in the CON, P4, and GnRH groups received: 1) Artificial insemination on detected estrus (AIDE; n = 37), 2) CIDR-GnRH protocol (CIDR; n = 27), and 3) Ovsynch protocol (OVS; n = 23), respectively. The pregnancy rate in resynchronized buffaloes did not differ (P > 0.05) between the OVS and CIDR groups; whereas the, cumulative pregnancy rate in GnRH + OVS buffaloes (81%) after 1st and 2ndAI when determined on Day 64 was higher (P < 0.05) than that in CON + AIDE (59%) buffaloes. The embryonic losses were significantly lower (P < 0.05) in GnRH-treated (18%) buffaloes, than in CON (42%) buffaloes on Day 45 post 1st TAI. Fetal losses were fewer and did not differ (P > 0.05) due to treatments on Day 60 or 90 post 1st AI. In conclusion, 1) the pregnancy rate and cumulative pregnancy rate in GnRH-treated buffaloes were higher than in CON buffaloes on Day 64 after synchronization and resynchronization, and 2) embryonic and fetal losses were lower in GnRH-treated buffaloes than in CON buffaloes when determined from Day 31–90 post 1st TAI.

Nerve growth factor from seminal plasma origin (spβ-NGF) increases CL vascularization and level of mRNA expression of steroidogenic enzymes during the early stage of Corpus Luteum development in llamas

The objectives of the study were to determine the effect of seminal plasma β-NGF on Corpus Luteum morphology and function and level of mRNA expression of steroidogenic enzymes. Llamas were assigned (n = 12/per group) to receive an intramuscular dose of: (a) 1 ml phosphate buffered saline (PBS), (b) 5 μg gonadorelin acetate (GnRH), or (c) 1.0 mg of purified llama spβ-NGF. Ovaries were examined by transrectal B-mode ultrasonography from treatment to ovulation (Day 0 = treatment). B mode/Power Doppler ultrasonography and blood samples collection were performed at Days 4, 8 and 10 (n = 3 llamas per treatment group/per time point) to determine CL diameter, vascularization and plasma progesterone concentration respectively. Plasma progesterone concentration was analyzed in all llamas at Day 0. Then females were submitted to ovariectomy at Days 4, 8 and 10 (n = 3 llamas/treatment/time), CL was removed to determine vascular area, proportion of luteal cells and CYP11A1/P450scc and STAR expression by RT-PCR. Ovulation was similar between llamas treated with GnRH or spβ-NGF and CL diameter did not differ between GnRH or spβ-NGF groups by Day 4, 8 or 10. Vascularization area of the CL was higher (P < 0.01) in llamas from the spβ-NGF than GnRH-treated group by Day 4 and 8. Plasma progesterone concentration was higher (P < 0.05) in llamas from the spβ-NGF compared to females of GnRH group by Day 4 and 8. The proportion of small and large luteal cells did not differ between GnRH or spβ-NGF groups by Day 8. CYP11A1/P450scc was upregulated 3 folds at day 4 and 10 by spβ-NGF compared to GnRH. STAR transcription was 3 folds higher at day 4 in females treated with spβ-NGF. In conclusion, the luteotrophic effect of spβ-NGF could be related to an increase of vascularization and up regulation of CYP11A1/P450scc and STAR transcripts enhancing progesterone secretion.

Influence of two ovulation-inducing agents on the pituitary response and follicle blood flow in mares

The objective of the current study was to evaluate the effects of deslorelin and hCG, two ovulation-inducing therapies, on LH surge and follicle vascularity in mares. Thirty mares were either treated with 1.5 mg IM of deslorelin, 2,500 IU IV of hCG or 2 mL IM of NaCl 0.9% (GnRH, hCG and Saline groups, respectively). Power-flow Doppler examination and blood collection were performed every hour during the first 12 hours after treatment (H0) and every six hours between hours 12 (H12) and 30 (H30) after treatment. Moreover, endpoints were evaluated every hour through the last six hours before ovulation (OV-6 to OV-1). In GnRH group, plasma LH concentration progressively increased (P < 0.001) during the first 6 hours after treatment and remained high (P > 0.1) until OV-1. A significant increase in LH concentrations was first detected (P < 0.05) at 24 hours after treatment in hCG group, while no changes (P > 0.1) on LH levels were found during H0-H30 and between OV-6 and OV-1 in the Saline group. Independent of the treatment, significant variations on the percentage of the follicle wall with Doppler signals were not observed (P > 0.1) throughout the entire experiment. A weak correlation between the preovulatory follicle vascularity and the plasma LH concentration was found in GnRH, hCG and Saline groups (r = +0.29, +0.29 and −0.23, respectively; P ˂ 0.0001). These results described for the first time the immediate and continuous pituitary response to ovulation-inducing therapy with injectable deslorelin. Moreover, spontaneous and induced ovulations were not preceded by an increased follicle vascularity, which differs from previous reports in large animals.

Comparison of bone mineral loss by combined androgen block agonist versus GnRH in patients with prostate cancer: A 12 month-prospective observational study

The multi-centre, prospective, observational study was designed to examine the efficacy of continuous combined androgen block (CAB) vs. GnRH agonist monotherapy in terms of bone mineral density (BMD) change during 12 months post-androgen deprivation therapy (ADT) in Asian prostate cancer patients. Multiple regression analysis and estimated the 10-year probability of major fractures among the patients with Fracture Risk Assessment Tool were conducted to investigate the underlying factors affecting BMD. Paired t-test to evaluate the change of BMD from baseline to 12 month, and two sample t-test to examine the difference of BMD changes were used between two groups. BMD significantly decreased in both the CAB and GnRH groups, with no group wise differences. The proportion of osteopenia or osteoporosis was slightly increased after the 12-month post-ADT. Ten-year probability of hip fracture and major osteoporotic fracture was approximately 3% and 5%, respectively. In conclusion, a significant decrease of BMD by 12-month ADT was observed without any differences between the two groups, whereas ADT-related BMD loss did not induce detrimental effects on bone health in terms of increased bone fracture risk. This was the first prospective study on BMD changes as a predictor of fracture during ADT in an Asian population.

Efficacy of a neoadjuvant gonadotropin-releasing hormone antagonist plus low-dose estramustine phosphate in high-risk prostate cancer: a single-center study.

The optimal treatment for high-risk prostate cancer (Pca) remains to be established. We previously reported favorable biochemical recurrence-free survival (BRFS) for high-risk Pca patients treated with neoadjuvant therapy comprising a luteinizing hormone-releasing hormone agonist plus low-dose estramustine (LHRH+EMP) prior to radical prostatectomy (RP). In the present study, we evaluated the efficacy of neoadjuvant therapy comprising a gonadotropin-releasing hormone antagonist plus low-dose estramustine phosphate (GnRH+EMP) in patients with high-risk Pca. Between September 2005 and March 2016, we identified 406 high-risk Pca patients of whom 136 received neoadjuvant GnRH+EMP (GnRH group) and 270 received LHRH+EMP (LHRH group) before RP. We retrospectively evaluated the clinical and pathological covariates between the two groups. The endpoint was the rate of pathological T0 status. The rates of pathological T0 status were 11.0 and 8.9% in the GnRH group and LHRH group, respectively (P=0.490). The 2-year BRFS rates were 97.8% in the GnRH group and 87.8% in the LHRH group (P=0.027). Our findings suggest that neoadjuvant GnRH antagonist+EMP followed by RP may improve the pathological outcomes and reduce the risk of biochemical recurrence in patients with high-risk Pca. Further prospective studies to confirm these findings are warranted.

12 OVARIAN STRUCTURES, ESTRUS EXPRESSION, AND PREGNANCY RATE IN BEEF HEIFERS USING ESTRADIOL CYPIONATE OR GnRH AS OVULATION INDUCTORS IN TIMED AI PROTOCOL

The aim of this study was to evaluate the effect of presence of corpus luteum (CL) at the onset of timed AI (TAI) protocol, size of ovulatory follicle and oestrus expression on pregnancy rate using E2 cypionate (EC) or GnRH as ovulation inductor in Angus heifers. On day 0, heifers [n = 414; 317 ± 23 kg; 3.08 ± 0.21 body condition score (1 to 5 scale) and 22–24 months of age] received a 1-g progesterone intravaginal device (Primer®, Agener União, Brazil) and 2 mg of oestradiol benzoate, IM, (RIC-BE®, Agener União, Brazil). On day 8, the device was removed, were administered 500 μg of sodium cloprostenol, IM, (Estron®, Agener União, Brazil) and heifers were divided in 2 groups: EC [n = 213; 0.5mg EC, IM, (SincroCP®, Ouro Fino, Brazil) on day 8] and GnRH [n = 201; 25 μg of Licerelin, IM, (GestranPlus®, Agener União, Brazil) at TAI] TAI was performed 50 h after device removal. Heifers were tail painted (Tell Tail®, GEA, New Zealand) to control oestrus expression from device removal to TAI. Heifers were scanned (7.5-MHz linear transducer MediSono® P3V, USA) to determine presence of CL (Day 0), ovulatory follicle diameter (OFD; Day 10) and ovulation (Day 11). Pregnancy was diagnosed 30 days after TAI. Oestrus expression was confirmed when marking ink was removed on day 10. Data were analysed using logistic regression and ANOVA. Oestrus expression was higher (P &lt; 0.05) in EC (93.90%, 200/213) compared to GnRH group (67.66%, 136/201). The OFD differed (P &lt; 0.05) according to presence of CL on Day 0 (10.3 mm) or not (9.6 mm), but was similar in treatment groups. Ovulation rate did not differ in EC (85%, 17/20) and GnRH (95%, 19/20) groups. Pregnancy rates were similar (P &gt; 0.05) between EC (52.58%, 112/213) and GnRH (59.20%, 119/201) groups; however, in heifers that expressed oestrus, rates were higher (P &lt; 0.05) (EC = 56%, 112/200; GnRH = 71.3%, 97/136) than in those did not show (EC = 0%, 0/13; GnRH = 33.8%, 22/65). Presence of CL at onset of TAI protocol was associated with higher pregnancy (P &lt; 0.05) in GnRH group (68%, 68/100) compared with EC (54.46%, 61/112) group, but in heifers without CL, pregnancy were similar for GnRH (50.5%, 51/101) and EC (50.5%, 51/101) groups. Heifers were also classified in 3 groups according OFD: small (≤9.5 mm), medium (9.6–11.5 mm), and large (≥11.6 mm) to verify its effect on oestrus expression and pregnancy. Medium and large OFD were implicated in higher (P &lt; 0.05) oestrus expression [(EC = 100%, 51/51; 96.97%, 32/33) and (GnRH = 83.02%, 44/53; 97.44%, 38/39)] than small OFD [(EC = 85%, 51/60) and (GnRH = 23.8%, 15/63)]. Pregnancy was lower (P &lt; 0.05) in small OFD groups (EC = 28.3%, 17/60; GnRH = 25.4%, 16/63). In medium and large OFD, pregnancy was similar in EC (72.55%, 37/51; 54.55%, 18/33) groups, but differed (P &lt; 0.05) between GnRH large (82.05%, 32/39) and medium (62.26%, 33/53). Higher pregnancy was achieved in heifers with CL in the beginning of TAI protocol treated with GnRH. Size of ovulatory follicle was associated with higher oestrus behaviour and pregnancy rate in EC and GnRH treatments.

Effect of GnRH and hCG on progesterone concentration and ovarian and luteal blood flow in diestrous mares

The objective of the present study was to investigate the effect of reproductive hormones (GnRH, hCG, LH and progesterone) on the regulation of corpus luteum (CL) and ovarian blood flow. Diestrous mares received a single treatment of saline, 100μg gonadorelin (GnRH), or 1500IU hCG 10days after ovulation. Plasma LH and progesterone concentrations, resistance index (RI) for ovarian artery blood-flow, and percentage of corpus luteum (CL) with color-Doppler signals of blood flow were determined immediately before treatment (hour 0) and at hours 0.25, 0.5, 1, 1.5, 2, 3, 4, 5, and 6. In the GnRH group, LH increased (P<0.0001) between hours 0 and 0.25 and then progressively decreased; concentration of LH was not affected in the saline and hCG groups. Progesterone concentration was not different among groups. In the GnRH group, RI tended (P<0.07) to decrease between hours 0 and 1.5 and increased (P<0.01) between hours 1.5 and 4. In the hCG group, two transient RI decreases (P<0.05) occurred before hour 2. The percentage change from hour 0 in the percentage of CL with blood-flow signals was greater at hour 0.5 in the GnRH group than in the saline group and was intermediate in the hCG group. The similarity among groups in progesterone concentration indicated that changes in progesterone were not involved in the GnRH and hCG stimulation of ovarian vascular perfusion. Effects of treatment might have been mediated through LH; however, since hCG biological activity is primarily LH-like, the differences in timing and degree of ovarian and luteal blood flow changes after GnRH or hCG administration in the present study suggest that GnRH might have a direct effect on ovarian blood vessels and vascular control.