Human Chorionic Gonadotropin Treatment Research Articles

Gonadotropic Activity of Pituitary Extract of African Catfish, Clarias gariepinus, in Mammals using Immature Wistar Rats as Models.

The pituitary glands normally produce and stores gonadotropic hormones (GnH) that are responsible for ovulation and spermiation in animals. However, whether fish pituitary extracts can elicit same effects in treated animals need elucidation as a prelude to their practical usage in animals. The aim of this study was to investigate the oestrus-inducing potential of the pituitary gland extract of the Africa Catfish (Clarias gariepinus) in immature Wistar rats. The experiment involved the use of 18 immature female Wistar rats and 10 male catfish brood stocks with the use of six Wistar rats per groups as follows: Group A had human chorionic gonadotropin (hCG) treatment. Group B had only normal saline treatment as the control whereas Group C had the C. gariepinus pituitary extract administration to induce oestrus with treatments occurring twice six hours apart in each group. There was an obvious expression of visible signs of heat and the presence of uterine horn oedema with significant (p < 0.05) increase in reproductive tract weight and uterine width and length. However, only progesterone levels increased significantly (p < 0.05) in the hCG and the C. gariepinus pituitary extract treated groups compared to other assayed hormones. These results showed that C. gariepinus pituitary extract has the capacity to induce oestrus in Wistar rats because of its gonadotropic effects, which needs further investigations at higher doses and for longer exposure periods for possible oestrus induction and synchronization in higher mammals. Further favourable results could herald the possible patent of the catfish pituitary extract for either experimental or commercial use in mammals.

Effect of breeding season and age on follicular dynamics and hemodynamics in embryo donor mares subjected to luteolysis after embryo flushing.

Mares are the only companion animals simulating women in the large diameter of their follicles. Horses start reproduction at the age of three years, and some of them live for >30 years, so aging influences their reproductive capacity. Mares are sensitive to summer heat stress as they can sweat like humans. The current work aimed to study the effects of age (young versus senile), season (cold versus hot), and the hormonal treatments during embryo collection on the dominant and subordinate follicular dynamics and hemodynamics and circulating ovarian hormones in embryo donor mares ovulated twice spontaneously before inducing ovulation for flushing embryos. Spontaneous oestrous cycles were studied for young mares (<10 years; N = 6) or senile (>20 years; N = 5) during months of the cold season (November to April) and hot season (May to August). In young embryo donor mares, oestrous cycles after inducing ovulation and luteolysis were studied using Doppler ultrasound. Estradiol (E2), progesterone (P4), nitric oxide (NO), total cholesterol, and lactate dehydrogenase (LDH) were measured in blood serum. A decrease in the dominant follicle antrum diameter (p > 0.05) and LDH (p = 0.016) was observed after inducing luteolysis in young embryo donor mares. Both human chorionic gonadotropin (hCG) and PGF2α treatments increased dominant follicle area (p = 0.0001), antrum area (p = 0.001), perimeter (p = 0.001), granulosa area (p = 0.0001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001). The dominant follicle area, antrum area, perimeter, color area, granulosa area, LDH, cholesterol, NO, and E2 increased (p = 0.0001) during the oestrous cycles of the hot season, but the circulatory % (p = 0.0001) declined. Senile mares had lower dominant follicle area (p = 0.002), antrum area (p = 0.0001), granulosa area (p > 0.05), LDH (p = 0.001), cholesterol (p = 0.0001), NO (p = 0.0001), and E2 (p = 0.0001) but higher circulatory % (p = 0.0001) and color area % (p = 0.023). The dominant follicle possesses the largest diameter, area, perimeter, granulosa area, and color area but the lowest circulatory % during spontaneous oestrous cycles, after inducing ovulation, or luteolysis with significant effects of the day of the spontaneous oestrous cycles on their dynamics and hemodynamics. During hot months, mares treated with hCG ovulated 24 hours later and prostaglandin-induced luteolysis was followed by new ovulation five days later. Follicles ovulated during the hot months were larger than those ovulated during the cold months and both had nearly the same color area %. Senile mares ovulated follicles with a lower area and antrum area but a higher color area %, so senile mares can be used as embryo or oocyte donors during the hot season.

In view of ovarian steroidogenesis and luteal construction to explore the effects of Bushen Huoxue recipe in mice of ovarian hyperstimulation

Ethnopharmacological relevanceBushen Huoxue recipe (BSHXR) is a widely used prescription medicine for treating gynecological diseases. We have previously found that BSHXR can improve the pregnancy outcome of controlled ovarian hyperstimulation (COH) mice by modulating the abnormal high level of progesterone. While the pharmacological mechanism of such therapeutic effect is not clear. Aim of the studyWe aimed to investigate the effects of BSHXR on the ovarian steroidogenesis and luteal function in mice undergoing COH. Materials and methodsA COH mouse model was established via an intraperitoneal injection of 0.4 IU/g pregnant mare serum gonadotropin (PMSG) and 1 IU/g human chorionic gonadotropin (HCG). The histological features of ovaries were observed using hematoxylin-eosin staining. The expression levels of FSHR, LHCGR, and key molecules in ovarian steroidogenesis, including CYP11A1, CYP17A1, CYP19A1, HSD3B1, and StAR, were examined via immunohistochemical staining, western blotting, and RT-qPCR. CD31, VEGFA, and FGF2 levels were assessed to evaluate ovarian vascularization. The protein and mRNA levels of ovarian ERK1/2, p-ERK1/2, MEK1/2, and p-MEK1/2 were also detected using western blotting, RT-qPCR, or immunofluorescence staining. ResultsCOH mice had a significantly increased volume and weight of the ovary and number of corpora lutea. In particular, COH exhibited a long-term influence on ovarian FSHR and LHCGR expression, disrupting the levels of CYP11A1, HSD3B1, and CYP17A1, causing poorer luteal angiogenesis. Compared with normal mice, the expression levels of ovarian VEGFA and FGF2 in COH mice were considerably lower on Day 1 after PMSG. On concomitant HCG treatment, both VEGFA and FGF2 expression surged dramatically on ED1 and then declined on ED4 and ED8. Moreover, the expression pattern of MEK1/2-ERK1/2 was almost consistent with that of VEGFA and FGF2. After treatment, BSHXR increased ovarian LHCGR, FSHR, CYP11A1, HSD3B1, and CYP17A1 levels, boosted luteal vascularization, and restored MEK1/2-ERK1/2 signaling in COH mice. ConclusionBSHXR restored the abnormally high progesterone level by regulating the CYP11A1 and HSD3B1 expression as well as promoted luteal angiogenesis, which was related with LHCGR-MEK1/2-ERK1/2-VEGFA/FGF2 signaling pathway in the ovary. This effect prevented the fluctuation of sex hormones in COH mice and benefited the outcome of pregnancy.

Effects of reproductive hormones on reproductive biology of commonly consumed fish in Akwa Ibom State, Nigeria

Nutritionally proteinous food and feed are required for a balance healthy living. This study was designed to assess the efficacy of some reproductive hormones: human chorionic gonadotropin (HCG), Ovaprim (OVA), extract of carp pituitary (CPE) and deoxy- corticosterone acetate (DOCA) on some aspects of reproductive biology of gravid Heterobranchus longifilis and Clarias gariepinus. A 25 number each of gravid males and females H. longifilis and C. gariepinus were employed in this study. Five experimental set up were designed for the study. Each treatment group was dosed with the required amount of the four reproductive hormones while in the fifth treatment breeders were treated with normal saline. This served as the control experimental. All protocol followed standard procedure. Results showed the highest mean value of relative fecundity (148.177) recorded in H. longifilis treated with ovaprim. This was followed by HCG treated C. gariepinus (145.668). Egg fertility rate had the highest mean value of 61.4803 in C. gariepinus treated with ovaprim, followed by 55.583 mean value in H. longifilis with HCG treatment. Egg hatchability mean value of 70.9280 was the highest observed in H. longifilis with ovaprim treatment. The mean values of survival rate were high in both species with OVA and HCG treatment showing values of 61.86% and 61.00% respectively. Treatment of the species with these reproductive hormones yielded good outcomes, except in the case of DOCA. Thus, CPE, OVA and HCG are recommended for breeding of C. gariepinus and H. longifilis.

Long-acting recombinant human follicle-stimulating hormone (SAFA-FSH) enhances spermatogenesis.

Administration of follicle-stimulating hormone (FSH) has been recommended to stimulate spermatogenesis in infertile men with hypogonadotropic hypogonadism, whose sperm counts do not respond to human chorionic gonadotropin alone. However, FSH has a short serum half-life requiring frequent administration to maintain its therapeutic efficacy. To improve its pharmacokinetic properties, we developed a unique albumin-binder technology, termed "anti-serum albumin Fab-associated" (SAFA) technology. We tested the feasibility of applying SAFA technology to create long-acting FSH as a therapeutic candidate for patients with hypogonadotropic hypogonadism. SAFA-FSH was produced using a Chinese hamster ovary expression system. To confirm the biological function, the production of cyclic AMP and phosphorylation of ERK and CREB were measured in TM4-FSHR cells. The effect of gonadotropin-releasing hormone agonists on spermatogenesis in a hypogonadal rat model was investigated. In in vitro experiments, SAFA-FSH treatment increased the production of cyclic AMP and increased the phosphorylation of ERK and CREB in a dose-dependent manner. In animal experiments, sperm production was not restored by human chorionic gonadotropin treatment alone, but was restored after additional recombinant FSH treatment thrice per week or once every 5 days. Sperm production was restored even after additional SAFA-FSH treatment at intervals of once every 5 or 10 days. Long-acting FSH with bioactivity was successfully created using SAFA technology. These data support further development of SAFA-FSH in a clinical setting, potentially representing an important advancement in the treatment of patients with hypogonadotropic hypogonadism.

Effect of human chorionic gonadotropin treatment on the duration of oxytocin-induced prolonged corpus luteum function in mares

Oxytocin is used increasingly in mares to prolong corpus luteum (CL) function to suppress estrus. When mares develop prolonged CL function in response to oxytocin treatment, the CL generally remains functional for 60–90 days. However, a longer duration of CL function can make this method of estrus suppression even more clinically useful. We hypothesized that human chorionic gonadotropin (hCG) treatment during the period of oxytocin-induced prolonged CL function would extend the duration of CL function (i.e. beyond 90 days). Prolonged CL function was induced in mares (n = 14) by treating them with 60 units of intramuscular oxytocin once daily on days 7–14 after ovulation. Mares were then randomly assigned equally to a control group that received no additional treatment and an hCG-treated group that received 2,500 units of intramuscular hCG on days 30, 45, 60, 75, and 90 after ovulation. Jugular blood samples were collected for progesterone concentration determination on the day of ovulation and then 3 times (M, W, and F) weekly for 120 days. Duration of CL function was not different (p &gt; 0.05) and it was 78.0 ± 7.5 and 91.4 ± 20.4 days (mean ± standard deviation [SD]) in control and hCG-treated mares, respectively. Therefore, hCG treatment (during the period of oxytocin-induced prolonged CL function) did not extend CL function, hence alternative methods should be explored.

Effects of letrozole co-treatment on the cumulative live-birth rate among normal responders in gonadotropin-releasing hormone antagonist cycles.

Studies have shown that letrozole cotreatment can improve clinical outcomes in high and poor responders in GnRH-antagonist protocol. However, whether letrozole is also beneficial to normal responders is not known. To investigate the clinical value of letrozole cotreatment during ovarian stimulation in vitro fertilization for normal ovarian reserve patients who were treated with the GnRH antagonist protocol, we conducted a retrospective study that based data from 1 January to 31 December 2017 for all IVF-ICSI GnRH-antagonist protocols. A total of 252 women who aged <40 years, FSH <10 IU/L on day 3 and antral follicle counting (AFC) >6 were included in the analysis (96 in the letrozole group and 156 in the no-letrozole group). The cumulative live-birth rate was calculated as the first live birth achieved after all cycles having an embryo transfer (cycles using fresh embryos and frozen-thawed embryos) among both groups. The initial gonadotropin (Gn) dosage and total Gn dosage were significantly lower and the number of days of Gn treatment was significantly fewer in the letrozole group than the non-letrozole group (p < 0.05). There were also significant between-group differences in luteinizing hormone, estradiol, and progesterone concentrations; and the number of metaphase II oocytes on the day of human chorionic gonadotropin treatment (p < 0.05). There was a significant difference in the implantation rate between the two groups that the letrozole group higher than the non-letrozole group (39.79 vs. 27.96%, p = 0.006), but there was no significant difference in the cumulative live-birth rate. The combination of letrozole with a GnRH antagonist may have no effect on the clinical pregnancy rate or cumulative live-birth rate in patients with a normal ovarian reserve. However, letrozole may increase the rate of embryo implantation and may reduce the requirement for exogenous gonadotrophins and, consequently, the cost of an IVF treatment cycle. In addition, the decreased estradiol level in the ovarian simulation by letrozole supports letrozole can be a safe solution for fertility preservation in estrogen-related cancer patients.

Liver receptor homolog 1 (LRH-1) regulates follicle vasculature during ovulation in mice.

It is well-established that liver receptor homolog 1 (LRH-1/NR5A2) regulates the ovarian function and is required for ovulation and luteinization in mice. In the present experiment, we showed that LRH-1 is required to control vascular changes during ovulation, a novel mechanism of action of this orphan nuclear receptor. Liver receptor homolog 1 (LRH-1/NR5A2) is a key regulator of ovarian function, and recently, it has been suggested that it may regulate changes in follicular angiogenesis, an important event during the ovulatory process and luteal development. In the present experiment, the objective was to determine whether conditional depletion of LRH-1 in mice granulosa cells modified vascular changes during the periovulatory period and to explore the possible mechanisms of this modification. We generated mice (22- to 25-day-old) with specific depletion of LRH-1 in granulosa cells by crossing Lrh1 floxed (Lrh1 f/f) mice with mice expressing Cre-recombinase driven by the anti-Müllerian type II receptor (Amhr2-cre; conditional knockout or cKO mice). We showed that preovulatory follicles of LRH-1 cKO mice had a reduced number of endothelial cells in the theca cell layer at 8 h after human chorionic gonadotropin treatment compared with control (CON) mice. Additionally, mRNA and protein expression of leptin receptor (LEPR), a protein that stimulates angiogenesis in a vascular endothelial growth factor-A (VEGFA)-dependent manner, and teratocarcinoma-derived growth factor-1 (TDGF1), which may directly stimulate endothelial cell function, were reduced in LRH-1 cKO mice as compared to CON after the LH surge. These results showed that LRH-1 is necessary for the correct vascular changes that accompany ovulation in mice and that this effect may be regulated through VEGFA-dependent and VEGFA-independent pathways mediated by LEPR and TDGF1.

Evaluation of hormonal protocols for induction of synchronized estrus on reproductive indices in Kangal-Akkaraman ewes during the outbreeding season

The objective of this study was to assess the effect of human chorionic gonadotropin (hCG) and gonadotropin-releasing hormone (GnRH) treatment before sponge insertion and anti-Müllerian hormone (AMH) concentration in ewes which were either pregnant or non-pregnant during the previous breeding season. In the study, 375 (3–6-year-old) Kangal-Akkaraman ewes that conceived or did not conceive in the preceding breeding season, and 36 rams (3–6-year-old) were used. The sheep were randomly divided into 5 equal groups. Blood samples were taken before stimulation (day −4), the day stimulation was initiated (day 0), after stimulation (day 9), on the mating day (days 11–12). AMH concentrations were measured in the samples taken on − 4th day. As a conventional method, a sponge containing 20 mg of flugeston acetate (Chronogest® CR, MSD, Turkey) was placed intravaginally (day 0) into to study ewes. The ewes were intramuscularly administered 131.5 µg prostaglandin F2α (Estrumate, MSD, Turkey) and 600 IU equine chorionic gonadotropin (Chronogest® PMSG 6000, MSD, Turkey) simultaneously with sponge removal 9 days after insertion. No other treatment was applied to group 5 which served as control. Ewes in groups 1, 2, and 3 received an intramuscular injection of GnRH (4.2 µg of Buserelin acetate, Receptal, MSD, Turkey) at the time of insertion of progesterone sponge. GnRH or 600 IU hCG (Chorulon®, MSD, UK) was administered 4 days prior to sponge insertion for the ewes in the groups 2, and 3, respectively. As for the group 5 group, hCG (instead of GnRH) was administered to the ewes at the beginning of the progesterone treatment. Ewes in estrus were detected with a teaser ram and taken into a different pen with rams. Transrectal ultrasonography (Mindray DP50/Vet/US, rectal probe) was achieved during days the 30–35 following mating. Transabdominal ultrasonography was achieved during days 60–65 to detect late embryonic and early fetal death. The estrus rate was 71.19 % out of breeding season for all ewes. The pregnancy rate on day 30 post-mating was 32 % in the control group and was 31.00 %. No significant differences were observed between groups for pregnancy rates on day 30. The mean pregnancy rate in sheep that conceived and did not conceived in the preceding breeding season of the study was 32.03 % and 32.06 % respectively (P > 0.05). After treatments, AMH concentrations in non-pregnant were significantly higher than pregnant ewes (P < 0.005). In conclusion, although the AMH concentration is significantly higher in non-pregnant ewes, the AMH concentration may not be a suitable biomarker to predict induction success outside breeding season. Progesterone-based protocols may achieve an acceptable reproductive outcome in lactating ewes and the ones which experienced infertility.

Chronic Estrus Disrupts Uterine Gland Development and Homeostasis.

Female mice homozygous for an engineered Gnrhr E90K mutation have reduced gonadotropin-releasing hormone signaling, leading to infertility. Their ovaries have numerous antral follicles but no corpora lutea, indicating a block to ovulation. These mutants have high levels of circulating estradiol and low progesterone, indicating a state of persistent estrus. This mouse model provided a unique opportunity to examine the lack of cyclic levels of ovarian hormones on uterine gland biology. Although uterine gland development appeared similar to controls during prepubertal development, it was compromised during adolescence in the mutants. By age 20 weeks, uterine gland development was comparable to controls, but pathologies, including cribriform glandular structures, were observed. Induction of ovulations by periodic human chorionic gonadotropin treatment did not rescue postpubertal uterine gland development. Interestingly, progesterone receptor knockout mice, which lack progesterone signaling, also have defects in postpubertal uterine gland development. However, progesterone treatment did not rescue postpubertal uterine gland development. These studies indicate that chronically elevated levels of estradiol with low progesterone and therefore an absence of cyclic ovarian hormone secretion disrupts postpubertal uterine gland development and homeostasis.

Ovarian hyperstimulation syndrome in a bitch caused by recombinant human chorionic gonadotropin treatment of suspected luteal insufficiency – a case report

Two bitches with a history of hypoluteoidism were treated by recombinant human chorionic gonadotropin (r-hCG) in repeated doses during early dioestrus. The level of progesterone increased, but the therapy led to hyperstimulation of the ovaries which resulted in ovarian hyperstimulation syndrome (OHSS), with subsequent development of oestrogen toxicity. This is the first case documenting OHSS in a canine after administration of r-hCG. Although manifested during dioestrus in our case report, the occurrence of OHSS is associated with similar administration of r-hCG to women. The described use of r-hCG is not appropriate for luteal insufficiency treatment, but seems to have a place in assisted reproduction technology because of its surprising effects. Further research of luteotrophic factors in bitches is warranted because of these unpredictable results. Lower doses and frequency of administration need to be considered for a better outcome.

Anticonvulsant Effect of Asparagus racemosus Willd. in a Mouse Model of Catamenial Epilepsy.

Asparagus racemosus Willd. (Family Liliaceae), also known as female reproductive tonic, is traditionally used across the Sub-Himalayan region in Uttarakhand, India for treatment of epilepsy and disorders of female reproductive system. Therefore, in this study, we investigated the anticonvulsant effect of A. racemosus in a mouse model of catamenial epilepsy. We artificially increased progesterone and neurosteroid levels (a state of pseudo-pregnancy) in adult Swiss albino female mice by injecting pregnant mares' serum gonadotropin (PMSG) (5IUs.c.), followed by human chorionic gonadotropin (HCG) (5IUs.c.) after 46h. In the following 10days, A. racemosus treatment was given along with measurement of progesterone, estradiol, and corticosterone levels in the blood. Neurosteroid withdrawal was induced by finasteride (50mg/kg, i.p.) on treatment day 9. Twenty-four hours after finasteride administration (day 10 of treatment), seizure susceptibility was evaluated with the sub-convulsant pentylenetetrazole (PTZ) dose (40mg/kg i.p.). Four hours after PTZ, animals were assessed for depression like phenotypes followed by euthanasia and separation of brain parts (cortex and hippocampus). The results showed that PMSG and HCG treatment elevated progesterone and estradiol levels. Treatment with finasteride increased seizure susceptibility and depression due to decreased progesterone and elevated estrogen levels coupled with decreased monoamine and elevated corticosterone levels. A. racemosus treatment, on the other hand, significantly decreased seizure susceptibility and depression like behaviors, possibly because of increased progesterone, restored estradiol, corticosterone, and monoamine levels. We concluded that herbal formulations using A. racemosus root extracts may be used as monotherapy or adjuvant therapy along with available AEDs for the better and safe management of catamenial epilepsy as well as comorbid depression.

Role of testosterone to estradiol ratio in predicting the efficacy of recombinant human chorionic gonadotropin and testosterone treatment in male hypogonadism.

We aimed to investigate the role of testosterone to estradiol ratio in predicting the effectiveness of human chorionic gonadotropin and testosterone treatments in male hypogonadism. Thirty-six male patients with hypogonadotropic hypogonadism were included in the study. Seventeen (47.2%) patients received weekly recombinant human choriogonadotropin alpha (hCG) treatment (group-1) and 19 (52.8%) received testosterone replacement therapy (T treatment) every 21 days (group-2). Under these treatments, adequate frequency of morning erection (≥3/week), testosterone to estradiol ratio (T/E), and testicular volume changes were analyzed. The mean age of the patients was 28.5 ± 8.7 years. When the frequency of morning erection (≥3/week) was specified as adequate, the cut-off value for effective T/E ratio was found to be 12.0 (sensitivity 93.8%, specificity 90.0%). There was no significant difference between the treatment groups in terms of total testosterone levels, T/E ratio, or frequency of morning erections (≥3/week) (p > 0.05). However, there was a statistically significant difference between the groups in terms of median left-right testicular volume in favor of group-1 (p < 0,05). In patients with hypogonadism who are under treatment, elevated estradiol-induced erectile dysfunction symptoms may persist even if serum testosterone levels are normal. Testosterone to estradiol ratio can be used as a predictive value in the effective treatment of hypogonadotropic hypogonadism with hCG and T.

Optimization of Hormone for Artificial Breeding and Seed Production of Clarias batrachus (Linnaeus, 1758) Under Captive Condition

Aims: The study aimed to develop brood stock of Magur (Clarias batrachus), artificial breeding and seed production under captive condition using growth hormones in north-eastern Bangladesh.&#x0D; Study Design: Fifteen pairs of brood Clarias batrachus were used in this experiment. Brood fishes were reared up to maturation for spawning operation in the farm by providing artificial diet for 4-5 months before onset of breeding season. In the present experiment, the brood fishes were induced with three growth hormones viz., Ovupin (100 mg dompridone and 0.2 mg S-GnRHa) at the rate of 1 ml.kg-1 body weight, Flash (20 mg S-GnRHa, 10 mg dompridone IP and propylene glycol) at the rate of 1 ml.kg-1 body weight and Human Chorionic Gonadotropin (HCG) hormone at the rate of 2272 IU.kg-1 body weight; respectively. The broods were injected with single injection in all treatments.&#x0D; Place and Duration of Study: The present study was conducted on induced spawning of Magur (Clarias batrachus) at the Reliance Aqua Farm, Boilor, Trisal of Mymensingh district from March to August, 2016.&#x0D; Methodology: Sexually mature healthy, uninjured male and female fishes were collected from the broodstock ponds for induced breeding. The fishes were transferred in the clean and germ free conditioning tanks for about 6-7 hours. During conditioning, continuous splash of water was given by keeping the male and female separate. The fishes were then weighed individually and injected intramuscularly with Ovupin, Flash and HCG hormone respectively. Only females were injected at 45° angle of the caudal peduncle. About 23-24 hours of post-injection the testes from males were dissected and mixed with stripping eggs for fertilization.&#x0D; Results: The incubation period for fertilized eggs were 23-24 hours for all the treatments. Fertilization rates were 78.20±0.52, 93.09±0.61 and 81.60±0.97 % and hatching rates were 54.93±0.61, 75.77±0.47 and 64.41±0.71 % in Ovupin, Flash and HCG treatments, respectively.&#x0D; Conclusion: This study revealed higher fertilization (93.09 %) and hatching (75.77 %) rate was achieved with Flash hormone. Therefore, Flash hormone could be effective for the artificial breeding and seed production of Magur (Clarias batrachus).

Sperm production and quality in European eel (Anguilla anguilla) in relation to hormonal treatment.

Aquaculture production relies on controlled management of gametogenesis, especially in species where assisted reproduction is needed for obtaining gametes in captivity. The present study used human chorionic gonadotropin (hCG) treatments to induce and sustain spermatogenesis in European eel (Anguilla anguilla). The aim was to evaluate effects of strip-spawning timing (12 vs. 24hr) after weekly administration of hCG and the necessity of a primer dose (in addition to weekly hormonal treatment) prior to strip spawning (primer vs. no-primer) on sperm quality parameters. Sperm parameters included milt production (weight), density and sperm kinematics at Week 9, 11 and 13 after onset of treatment. Spermiation commenced in 11.5% of males in Week 5 and by Week 9, and all males produced milt. Male weight, milt production, sperm density and spermatocrit did not differ among hormonal treatments during the experimental period. Overall, male weight decreased from 106.3 to 93.0g, milt weight increased from 3.5 to 5.4g, sperm density counts decreased from 11.7×109 to 10.5×109 cells/ml, and spermatocrit decreased from 46.5% to 40.5%. Furthermore, spermatocrit was positively related to haemocytometer counts (R2 =.86, p<.001), providing a reliable indicator of sperm density. Differences in sperm kinematics were observed depending on strip-spawning timing after hormonal injection (12 vs. 24hr) but with no consistent pattern. These sperm quality parameters also did not consistently differ between the no-primer and primer treatments. Considering that each male may be stripped 4-5 times over the 2-3months spawning season, omitting the primer would reduce animal handling, material costs and labour intensity, while sustaining high-quality sperm production.

Effects of the formation and regression of accessory corpus lutea during pregnancy on plasma progesterone concentration and pregnancy status in cross-bred beef heifers

There was examination of effects of accessory corpus lutea (CLs) formation and regression during pregnancy on circulating progesterone (P4) concentrations and pregnancy maintenance in beef heifers. Heifers (Experiment 1, n = 75; Experiment 2, n = 496) were randomly assigned to either a human chorionic gonadotropin (hCG) treatment or untreated group 5 days post-estrus, followed by embryo transfer (ET) on Days 6–8 (Day 0 = Estrus). In Experiment 1, blood samples were collected from pregnant heifers on Days 33, 40, and 47 for conducting P4 assays. Plasma P4 concentrations were greater in hCG-treated heifers than in untreated heifers on Day 33. In hCG-treated heifers with accessory CL regression between Days 33 and 47, plasma P4 decreased to concentrations similar to those of untreated heifers after Day 40. In hCG-treated pregnant heifers in Experiment 2, CL regression by Day 50 of gestation was more frequent when CLs were contralateral (49.3 %) rather than ipsilateral (4.4 %, P < 0.001) to the original CL. The hCG treatment resulted in a greater pregnancy percentages on Days 30 (80.5 % and 68.6 %, P = 0.002) and 50 (76.2 % and 65.3 %, P = 0.007) compared with untreated heifers. There, however, were no differences in either pregnancy percentages on Days 30 and 50 or pregnancy losses between hCG-treated heifers with ipsilateral and contralateral accessory CLs. These results indicate accessory CL formation improves pregnancy percentages resulting from ET. Furthermore, plasma P4 decreases associated with accessory CL regression does not affect pregnancy loss in beef heifers.

Morphological changes in mouse ovary due to hormonal hypersecretion and matrix metalloproteinase -2 activity.

We analyzed whether aberrant gonadotropin secretion affects the morphological remodeling of murine ovarian tissues facilitated by activated matrix metalloproteinase (MMP) enzymes. Six mice were intraperitoneally injected with 5 IU of pregnant mare serum gonadotropin (PMSG) or human chorionic gonadotropin (HCG) every two days after estrus synchronization. Morphology and expression of various MMPs were assessed following the successful induction of hormonal secretion in these tissues. HCG treatment, but not PMSG treatment, resulted in the expanded production of granulose second follicular cells. In addition, the number of developing follicular cells in the HCG group increased compared with that in the PMSG group. Ovarian diameters were also very small in the PMSG group. Immunohistochemistry revealed decreased MMP-2 protein activity in the HCG group and increased MMP-2 activity in the PMSG group. Activity was particularly high in theca and granulose cells of the PMSG group, but only partial activity was observed in the theca cells of the HCG group. Vascular endothelial growth factor activity was increased in both the external and internal theca cell walls in the PMSG group while the HCG group showed high overall expression of this protein in the internal theca cells. These data indicate that follicular cell activity and remodeling of the ovaries differ based on the type of secretory hormone signals they receive. Inappropriate gonadotropin secretion may induce functional changes in the ovaries, and follicular remodeling may be facilitated by the activity of various MMPs.

Gestational trophoblastic disease in Switzerland: retrospective study of the impact of a regional reference centre.

The European Society of Medical Oncology (ESMO) recommends that countries should have reference centres to provide adequate diagnosis and treatment of gestational trophoblastic disease. A trophoblastic disease centre in the French-speaking part of Switzerland was inaugurated in 2009. The objectives of this study were to report the activity of the centre during the last 10 years and analyse gestational trophoblastic disease outcomes. This was a retrospective study with data collected from all cases of gestational trophoblastic disease referred to the centre from 2009 to 2018. All histological specimens as well as data for treatment and follow-up of gestational trophoblastic disease and neoplasia were reviewed. Clinical features, including age, prognostic score and International Federation of Gynecology and Obstetrics (FIGO) stages (in the case of gestational trophoblastic neoplasia), human chorionic gonadotropin (hCG) follow-up, treatment and outcome were reported. The centre registered 354 patients, and these patients presented 156 cases of partial hydatidiform moles, 163 cases of complete hydatidiform moles and 14 cases of gestational trophoblastic neoplasia. During follow-up, 35 gestational trophoblastic neoplasms were diagnosed after hCG persistence. After pathology review, the overall agreement rates between our centre and a participating provider hospital was 82%. Methotrexate was the first line of single-agent chemotherapy for most patients, with resistance rates of 23%. Multi-agent chemotherapy was used as first-line treatment for five patients. None of the patients followed up by the centre died from gestational trophoblastic disease. This study reflects the activity of the Swiss trophoblastic disease centre from the French-speaking part of Switzerland created in 2009, and its role as local and national reference centre, in terms of global health, for women with gestational trophoblastic disease.

Effect of adjunctive hormonal therapy on testicular descent and spermatogenic function among children with cryptorchidism: a systematic review and meta-analysis

To systematically evaluate the effect of adjunctive hormonal therapy on testicular descent and spermatogenic function among children with cryptorchidism. A comprehensive literature search of electronic databases up to February 21, 2019 was performed. Continuous data of fertility indices were evaluated using weighted mean difference (WMD) with 95% confidence interval (CI), while nominal data of fertility indices and the complete descent rates were analyzed by risk ratio (RR) with 95% CI. A fixed effect or random effect model was used. A total of 27 previous studies were included for the current analysis. Hormonal therapy increased complete testicular descent rate, reaching statistical significance (RR = 3.74; 95% CI, 2.78-5.04; P < 0.001). The success rate was 22.43%. A similar result was found in subgroup analysis of hormonal category and effect on unilateral or bilateral cryptorchidism. Studies reporting primary outcome as continuous data showed that cryptorchid males have significantly increased germ cell numbers per tubule (WMD = 0.10; 95% CI, 0.01-0.20, P = 0.032) after hormonal therapy. The nominal data of pooled studies showed no significant difference (RR = 1.62; 95% CI, 0.65-4.00, P = 0.298). In addition, a significant result was noted in the luteinizing hormone-releasing hormone (LHRH) therapy group but not in those undergoing human chorionic gonadotropin (hCG) treatment. Our findings have demonstrated that hormonal therapy can effectively increase the success rate of complete testicular descent, while some boys may benefit as regards improvement of the fertility index.

Selection of fewer dominant follicles in Trio carriers given GnRH antagonist and luteinizing hormone action replaced by nonpulsatile human chorionic gonadotropin†.

Studying selection of multiple dominant follicles (DFs) in monovulatory species can advance our understanding of mechanisms regulating selection of single or multiple DFs. Carriers of the bovine high fecundity Trio allele select multiple DFs, whereas half-sib noncarriers select a single DF. This study compared follicle selection during endogenous gonadotropin pulses versus during ablation of pulses with Acyline (GnRH antagonist) and luteinizing hormone (LH) action replaced with nonpulsatile human chorionic gonadotropin (hCG) treatment in Trio carriers (n=28) versus noncarriers (n=32). On Day 1.5 (Day 0=ovulation), heifers were randomized: (1) Control, untreated; (2) Acyline, two i.m. doses (Days 1.5 and D3) of 3μg/kg; (3) hCG, single i.m. dose of 50IU hCG on Day 1.5 followed by daily doses of 100IU; and (4) Acyline+hCG. Treatments with nonpulsatile hCG were designed to replace LH action in heifers treated with Acyline. Acyline treatment resulted in cessation of follicle growth on Day 3 with smaller (P<0.0001) maximum follicle diameter in Trio carriers (6.6±0.2mm) than noncarriers (8.7±0.4mm). Replacement of LH action (hCG) reestablished follicle diameter deviation and maximum diameter of DFs in both genotypes (8.9±0.3mm and 13.1±0.5mm; P<0.0001). Circulating follicle stimulating hormone (FSH) was greater in Acyline-treated than in controls. Finally, Acyline+hCG decreased (P<0.0001) the number of DFs from 2.7±0.2 to 1.3±0.2 in Trio carriers, with most heifers having only one DF. This demonstrates the necessity for LH in acquisition of dominance in Trio carriers (~6.5mm) and noncarriers (~8.5mm) and provides evidence for a role of GnRH-induced FSH/LH pulses in selection of multiple DFs in Trio carriers and possibly other physiologic situations with increased ovulation rate.