FSH Concentrations Research Articles

The function of the pituitary-testicular axis in dogs prior to and following surgical or chemical castration with the GnRH-agonist deslorelin.

Chemical castration, that is the reduction of circulating testosterone concentrations to castrate levels by administration of a GnRH-agonist implant, is a popular alternative to surgical castration in male dogs. Detailed information concerning the pituitary-testicular axis following administration of a GnRH-agonist implant is still scarce. Therefore, GnRH-stimulation tests were performed in male dogs, prior to and after surgical and chemical castration. This approach also allowed us to determine plasma concentrations of testosterone and oestradiol in intact male dogs for future reference and to directly compare the effects of surgical and chemical castration on the pituitary-testicular axis. In intact male dogs (n=42) of different breeds GnRH administration induced increased plasma LH, FSH, oestradiol and testosterone concentrations. After surgical castration basal and GnRH-induced plasma FSH and LH concentrations increased pronouncedly. Additionally, basal and GnRH-induced plasma oestradiol and testosterone concentrations decreased after surgical castration. After chemical castration, with a slow-release implant containing the GnRH-agonist deslorelin, plasma LH and FSH concentrations were lower than prior to castration and lower compared with the same interval after surgical castration. Consequently, plasma oestradiol and testosterone concentrations were lowered to values similar to those after surgical castration. GnRH administration to the chemically castrated male dogs induced a significant increase in the plasma concentrations of LH, but not of FSH. In conclusion, after administration of the deslorelin implant, the plasma concentrations of oestradiol and testosterone did not differ significantly from the surgically castrated animals. After GnRH-stimulation, none of the dogs went to pre-treatment testosterone levels. However, at the moment of assessment at 4,4 months (mean 133 days ± SEM 4 days), the pituitary gonadotrophs were responsive to GnRH in implanted dogs. The increase of LH, but not of FSH, following GnRH administration indicates a differential regulation of the release of these gonadotrophins, which needs to be considered when GnRH-stimulation tests are performed in implanted dogs.

Cryptorchidism: The dog as a study model.

Cryptorchidism (CO) or undescended testicle is an abnormality of male gonadal development that can generate long-term repercussions in men, such as infertility and germ cell neoplasia in situ (GCNIS). The origin of these alterations in humans is not completely clear, due to the absence of an animal model with similar testicular development as in humans with CO. This work intends to describe the testicular histological development of dogs with congenital CO, and determine whether the species could adequately serve as a study model for this pathology in humans. The study was carried out with 36 dogs, equally distributed in two groups: healthy control (CTRL) and CO groups. The contralateral testis to the undescended one in CO group of the animals was considered and analyzed. Each group was subdivided in three stages of development: (1) peripubertal stage (6–8 months), (2) young adult (9–48 months) and (3) senile (49–130 months). Histological development, the presence of cells with gonocyte morphology, cell proliferation, testicular lipoperoxidation and hormonal concentrations of testosterone, estradiol, FSH and LH were evaluated and described. In the cryptorchid testes, the first histological alterations appeared from the first stage of development and were maintained until the senile stage. A pronounced testicular lipoperoxidation occurred only in the second stage of development. The histological alterations due to CO were markedly evident in the young adult stage. Testosterone concentrations witnessed a decrease starting from in the second stage and kept on until the last stage. The contralateral testes of the CO animals showed alterations that positioned them between the control and CO testes. Testicular development of dogs with CO is similar to that of humans. The results of the study suggest that this species could serve as a suitable model for the study of CO in humans.

EFFECT OF DIFFERENT ESTRUS SYNCHRONIZATION ON SERUM E2, P4, FSH AND LH DURING DIFFERENT ESTRUS PERIODS AND PREGNANCY IN EWES

This research was designed to know the effect of different estrus synchronization protocols on Serum E, P, FSH, and LH during different estrus periods and pregnancy in Karadi and Arabi ewes. The study was carried out by using 30 ewes for each breed at 2-4 years. Ewes from each breed were equally and randomly distributed into three groups. The first group was treated with vaginal sponges saturated with (60 mg) of Medroxy progesterone acetate within a period of (14) fourteen days (T1). The second group was administrated with the MAP injected with a dose of 300 IU / head of the hCG (T2). The third group was treated with MAP treatment, and injected at a dose of 75 µg /head of GnRH (T3). The results of this analysis showed a significant increase in the estrogen concentration in the different reproductive stages; the significant level of estrogen in the estrus phase (60.96) pg/ml. The concentration of progesterone differed significantly in the different reproductive stages, with the highest concentration in the pregnancy period (16.33) pg/ml. In comparison, the FSH concentration differed significantly in the different reproductive stages with the highest concentration in the estrus period(3.243) mIU/ml. the highest significant level of serum LH is in the estrus period (5.230) mIU/ml. Thus, the study is concluded that hormones and phases may be effective in inducing follicular growth and ovulation and finally may increase pregnancy rate and production .

Bu-Shen-Ning-Xin decoction alleviates premature ovarian insufficiency (POI) by regulating autophagy of granule cells through activating PI3K/AKT/mTOR pathway

Purpose To explore the therapeutic effects of Bu-Shen-Ning-Xin decoction (BSNXD) on POI and the underlying mechanism. Methods VCD was used to induce the in vivo and in vitro POI model. HE staining was used to evaluate the pathological state of ovarian tissues. ELISA was used to detect the production of hormones in the serum and granule cells (GCs). An immunohistochemical assay was used to determine the expression of ATG7 and p-AKT in the ovarian tissues. The number of oocytes in POI rats was counted. The mitochondrial membrane potential (MMP) in oocytes and GCs was detected by flow cytometry. A Western blot assay was used to measure the expression of AKT, p-AKT, p-mTOR, mTOR, S6K, p-S6K, ULK1, p-ULK1, Beclin-1, Bcl-2, LC3-II, LC3-I, ATG7, and cleaved Caspase3. The numbers of autophagosomes were detected by transmission electron microscope and autophagic flux assay. The CCK-8 assay was used to detect the cell viability. Results Decreased primary follicles in the ovarian tissues, elevated concentration of FSH, and LH, suppressed concentration of E2 and AMH in the serum, reduced number of oocytes, and mitochondrial dysfunction in oocytes induced by VCD were significantly reversed by BSNXD. Activated autophagy state and inhibited PI3K/AKT/mTOR pathway stimulated by VCD in both ovarian tissues and GCs were dramatically reversed by BSNXD. The protective effect of BSNXD on VCD-treated GCs was abolished by LY294002, an inhibitor of the PI3K/AKT/mTOR pathway. Conclusion Our data revealed that BSNXD alleviated POI by regulating autophagy of granule cells through activating PI3K/AKT/mTOR pathway.

Influence of endometrioma size on ART outcomes

Research questionDoes endometrioma size affect the number of oocytes retrieved after ovarian stimulation in women with endometriosis-related infertility undergoing IVF/intracytoplasmic sperm injection (ICSI)? DesignCohort study of infertile women with unilateral or bilateral endometrioma(s) associated with deep infiltrating endometriosis, undergoing their first IVF/ICSI cycle between January 2014 and November 2021. A total of 326 women with an adequate imaging work-up with transvaginal ultrasound and/or magnetic resonance imaging performed by senior radiologists before the start of ovarian stimulation was included. Prognostic factors associated with the number of oocytes retrieved were analysed. IVF/ICSI outcomes were compared between five groups defined according to the largest endometrioma diameter (<2, 2 to <4, 4 to <6, 6 to <8 and ≥8 cm). ResultsFactors that significantly reduced the number of oocytes retrieved after adjustment by multiple linear regression were women's age (regression coefficient –0.18; 95% confidence interval [95% CI] –0.31 to–0.06; P = 0.005), smoking habit (–2.02; 95% CI –3.42 to -0.62; P = 0.005), day 3 FSH concentration (–0.20; 95% CI –0.39 to –0.02; P = 0.031) and a previous history of surgery for ovarian endometriosis (–1.32; 95% CI –2.63 to –0.02; P = 0.047). Antral follicle count and oestradiol concentration on the trigger day were positively correlated with the number of oocytes retrieved (0.14; 95% CI 0.08 to 0.19; P < 0.001 and 0.003; 95% CI 0.002 to 0.004; P < 0.001, respectively). The mean number of oocytes retrieved was not significantly different between the five groups (P = 0.413), nor were the cumulative live birth rate, the number of cancelled cycles and perinatal outcomes. ConclusionsNo significant difference in the number of oocytes retrieved was observed according to endometrioma size. This study suggests that ovarian stimulation can be of benefit to women irrespective of the endometrioma size.

Consumption of soy isoflavones during the prepubertal phase delays puberty and causes hypergonadotropic hypogonadism with disruption of hypothalamic-pituitary gonadotropins regulation in male rats

Isoflavones are phytoestrogens with recognized estrogenic activity but may also affect testosterone, corticosterone and thyroid hormone levels in experimental models. However, the molecular mechanisms involved in these alterations are still unclear. Isoflavones are present in soy-based infant formula, in breast milk after the consumption of soy by the mother and are widely used for the preparation of beverages consumed by toddlers and teenagers. In this sense, we proposed to investigate the effects of soy isoflavone exposure during the prepubertal period, a recognized window of sensitivity for endocrine disruption, over the hypothalamic-pituitary-testicular (HPT) axis. For this, 42 3-week-old male Wistar rats were exposed to 0.5, 5 or 50 mg of soy isoflavones/kg from postnatal day (PND) 23 to PND60. We evaluated body growth, age at puberty, serum concentrations of LH, FSH, testosterone and estradiol, and the expression of the transcripts (mRNA) of genes encoding key genes controlling the hypothalamic-pituitary-testicular (HPT) axis. In the hypothalamus, we observed an increase in Esr1 mRNA expression (0.5 and 5 mg). In the pituitary, we observed an increase in Gnrhr mRNA expression (50 mg), a reduction in Lhb mRNA expression (0.5 mg), and a reduction in Ar mRNA expression. In the testis, we observed an increase in Lhcgr mRNA expression (50 mg) and a reduction in Star mRNA expression (0.5 and 5 mg). The serum levels of LH (5 and 50 mg) and FSH (0.5 mg) were increased, while testosterone and estradiol were reduced. Puberty was delayed in all groups. Taken together, these results suggest that prepubertal consumption of relevant levels of soy isoflavones disrupts the HPT axis, causing hypergonadotropic hypogonadism and altered expression levels of key genes regulating the axis.

Ameliorate effects of resveratrol and l-carnitine on the testicular tissue and sex hormones level in busulfan induced azoospermia rats

Busulfan (Bus), is an alkylating agent widely used in chemotherapy which has been proven to possess toxic side effects on testicles. This study was carried out to compare the probable treatment effects of resveratrol (Res) or/and l-carnitine (Lca), as strong antioxidants, on the testicular tissue as well as on the level of sex hormones in busulfan-induced azoospermic rat models. A total of 78 adult male rats, were divided into six different experimental groups including: 1) Control; 2) Lca + Res; 3) BUS; 4) Bus + Lca; 5) BUS + Res and 6) Bus + Lca + Res. Busulfan was intraperitoneally administered in a single dose (10 mg/kg b.w), while resveratrol (20 mg/kg b.w/day) and l-carnitine (200 mg/kg b.w/day) were orally administered by gavage during 48 consecutive days to the rats. At the end of the experiment in all groups the level of LH, FSH, and testosterone were biochemically analyzed by ELISA and the testicular tissue evaluated histologically using stereological technique. Results showed that Lca or/and Res, increased the body and testis weight, the volume of the testis, interstitial tissue, germinal epithelium, and seminiferous tubule, the number of the different cells of germinal epithelium and the level of testosterone. On the other hand, Lca, Res and their combination decreased the concentration of LH and FSH compared to the group treated with Bus. In conclusion, these results suggested that l-carnitine or/and resveratrol treatment significantly attenuated busulfan -induced changes of the rat reproductive system led to the recovery of both testis and sperm parameters. However, co-administration of L-ca and Res was more effective than their individual treatment. This combination may alleviate the side effects of alkylating drugs, such as busulfan and may be beneficial for spermatogenesis.

Allium cameleon extract improved androgenesis and sexual behavior in male rat (Rattus norvegicus) via attenuation of oxidative damages increased sexual hormone and NO/cGMP cascade

Objective: Allium cameleon Cronquist (1981) (Liliaceae) is a bulb belonging to the garlic family, known as a medicinal plant used for its aphrodisiac activities in Cameroonian folk medicine. The present study was conducted to assess the aphrodisiac potentials of Allium cameleon aqueous extracts and investigate on the possible mechanisms by which this could be accomplished. Methods: The behavioral and biochemical tests were used to detect the aphrodisiac properties. The conceivable mechanisms of action of the aqueous extracts were investigated through determination of sexual hormones, nitric oxide, cGMP, and antioxidant activities in male rats treated with different plant extract concentrations. Results: Allium cameleon enhanced sexual excitement, improved semen quality, and the concentrations of testosterone, FSH and LH. Levels of endogenous antioxidant enzymes (CAT, SOD, and GSH) were improved. Penile nitric oxide and cGMP levels were boosted. Significant stimulation in sexual behavior, elevated spermatic production, raised of viability, morphology, count and sperm mobility were also observed. Treated animals exhibited elevated levels of endogenous antioxidant enzymes. Conclusion: These findings suggest that Allium cameleon possess aphrodisiac properties in rats that might be involve in the attenuation of oxidative damage, but also in the increase of sexual hormones production and NO/cGMP cascade.

Cell-free fat extract improves ovarian function and fertility in mice with premature ovarian insufficiency

BackgroundPremature ovarian insufficiency (POI) is a refractory disease that seriously affects the reproductive health of women and is increasing in incidence and prevalence globally. There is enormous demand to improve fertility in women with POI, while there is still lack of effective therapeutic methods in clinic. Cell-free fat extract (CEFFE) has been reported to contain thousands of active proteins which possess the ability to promote tissue repair in other diseases. In our study, we aimed to observe the efficacy and biosecurity of CEFFE on the repair of ovarian function and fertility of mice with POI and further explore the underlying mechanism.MethodsIn vivo, POI mice model, established by cyclophosphamide (CTX, 120 mg/kg) and busulfan (BUS, 12 mg/kg), was treated with CEFFE via the tail vein every two days for 2 weeks. Then, the weight of ovaries, estrous cycle and follicle count by H&E staining were measured. The content of AMH, E2 and FSH in serum was measured by Enzyme-linked immunosorbent assay. Fertility was evaluated by the number of oocytes retrieved, the development of embryos in vitro and the litter size. Biosecurity of parent mice and their pups were examined by body mass and visceral index. The proliferation and apoptosis of cells in ovaries were examined by immunohistochemistry and transmission electron microscopy. Furthermore, the mRNA-Seq of mouse ovarian granulosa cells was performed to explore underlying mechanism of CEFFE. In vitro, KGN cell line and human primary ovarian granulosa cells (hGCs) were treated with 250 μM CTX for 48 h with/without CEFFE. The proliferative ability of cells was detected by cell counting kit-8 assay (CCK-8) and EDU test; the apoptosis of cells was detected by TUNEL and flow cytometry.ResultsCEFFE recovered the content of AMH, E2 and FSH in serum, increased the number of follicles and the retrieved oocytes of POI mice (P < 0.05). CEFFE contributed to the development of embryos and improved the litter size of POI mice (P < 0.05). There was no side effect of CEFFE on parent mice and their pups. CEFFE contributed to the proliferation and inhibited the apoptosis of mouse granulosa cells in ovary, as well as in human ovarian granulosa cells (including KGN cell line and hGCs) (P < 0.05).ConclusionsThe treatment of CEFFE inhibited the apoptosis of granulosa cells and contributed to the recovery of ovarian function, as well as the fertility of mice with POI.

The probability and possible influence factors of intermittent ovarian function recovery in patients with premature ovarian insufficiency

Research questionWhat are the probability and underlying influence factors of intermittent ovarian function recovery in patients with idiopathic premature ovarian insufficiency (POI)? DesignThis was a retrospective cohort study conducted in tertiary hospitals recruiting 162 patients diagnosed with POI based on European Society of Human Reproduction and Embryology criteria from June 2015 to March 2022. The incidence of intermittent ovarian function recovery was evaluated, and the possible influence factors were investigated by univariate and multivariate analysis. ResultsAmong 162 POI patients, 48 (29.63%) presented intermittent ovarian function recovery, and 11 (6.79%) were natural pregnancies; 114 (70.37%) patients failed to show ovarian function recovery. No association was found between initial clinical features and intermittent ovarian function recovery. In contrast, the variables of FSH, LH, oestradiol, anti-Müllerian hormone (AMH), ovarian volume, passive smoking and weekly exercise time after diagnosis were correlated with intermittent ovarian function recovery in patients with POI and further analysis indicated that FSH concentration at diagnosis (odds ratio [OR] 0.964, 95% confidence interval [CI] 0.934–0.995, P = 0.023), passive smoking (OR 0.369, 95% CI 0.141–0.963, P = 0.042) and weekly exercise time after diagnosis (OR 5.592, 95% CI 1.83–17.088, P = 0.003) were influence factors of intermittent ovarian function recovery in POI patients. ConclusionsThe incidence of intermittent ovarian function recovery in patients with idiopathic POI was 29.63%, and the natural pregnancy rate was 6.79%. Lower FSH concentration at diagnosis, no passive smoking and a weekly exercise time ≥1.5 h after the diagnosis may be beneficial for intermittent ovarian function recovery in POI patients.

The Effect of Hydroalcoholic Calendula Officinalis Extract on Androgen-Induced Polycystic Ovary Syndrome Model in Female Rat

Background Polycystic ovary syndrome (PCOS) is the most common hormonal disorder in women of reproductive age, and the major cause of infertility. Today, using medicinal plants instead of chemical drugs could be an alternative treatment option for PCOS. The purpose of this study was to determine the effect of Calendula officinalis hydroalcoholic extract on PCOS in rats. Method 60 female adult rats were randomly divided into six groups, including control, sham, PCOS group, and treated PCOS groups receiving hydroalcoholic extract of Calendula officinalis with different dosages of 200, 500, and 1000 mg/kg. PCOS was induced by subcutaneous injection of DHEA 6 mg/100 g bw for 35 days. For two weeks, the extract was taken orally. The serum glucose, insulin, sex hormone levels, and oxidative status were measured at the end of the experiment. The ovaries were dissected for histomorphometric and pathological analysis. Results When compared to the control and sham groups, the PCOS group showed a significant increase in glucose, insulin, testosterone, and malondialdehyde (MDA) concentrations, cystic and atretic follicles, and thickness of the theca and tunica albuginea layers, and a significant decrease in LH concentration, total antioxidant capacity, corpus luteum, antral follicles, and oocyte diameter. The mean concentration of FSH, on the other hand, did not change significantly. A trend of improvement was found in the treated groups with high doses of Calendula officinalis extract. Conclusion In rats with PCOS and nonovulation, Calendula officinalis hydroalcoholic extract improved oxidative stress, restored folliculogenesis, and increased ovulation.

Effect of elevating luteinizing hormone action using low doses of human chorionic gonadotropin on double ovulation, follicle dynamics, and circulating follicle-stimulating hormone in lactating dairy cows

Double ovulation and twin pregnancy are undesirable traits in dairy cattle. Based on previous physiological observations, we tested the hypothesis that increased LH action [low-dose human chorionic gonadotropin (hCG)] before the expected time of diameter deviation would change circulating FSH concentrations, maximum size of the second largest (F2) and third largest (F3) follicles, and frequency of multiple ovulations in lactating dairy cows with minimal progesterone (P4) concentrations. In replicate 1, multiparous, nonbred lactating Holstein dairy cows (n = 18) had ovulation synchronized. On d 5 after ovulation, all cows had their corpus luteum regressed and were submitted to follicle (≥3 mm) aspiration 24 h later to induce emergence of a new follicular wave. Cows were then randomized to NoP4 (untreated) and NoP4+hCG (100 IU of hCG every 24 h for 4 d after follicle aspiration). Ultrasound evaluations and blood sample collections were performed every 12 h for 7 d after follicle aspiration. All cows were then treated with 200 μg of GnRH to induce ovulation. In replicate 2, cows (n = 16) were resubmitted to similar procedures (i.e., corpus luteum regression, follicle aspiration, randomization, ultrasound evaluations every 12 h, GnRH 7 d after aspiration). However, cows in replicate 2 received an intravaginal P4 device that had been previously used (∼18 d). Only cows with single (n = 15) and double (n = 16) ovulations were used in the analysis. No significant differences were detected for frequency of double ovulation, follicle sizes, and FSH concentrations across replicates (NoP4 vs. LowP4 and NoP4+hCG vs. LowP4+hCG), so data were combined. Double ovulation was 40% for control cows with no hCG (CONT) and 62.5% with hCG (hCG). Double ovulation increased as the maximum size of F2 increased: <9.5 mm and 9.5-11.5 mm (7.7%) and ≥11.5 mm (94.1%). The hCG group had more cows with F2 > 11.5 (69%) than with 9.5 ≥ F2 ≤ 11.5 (25%) and F2 < 9.5 (6%). In agreement, F2 and F3 maximum size were larger in the hCG group, but FSH concentrations were lower after F1 > 8.5 mm compared with CONT. In contrast, FSH concentrations were greater before deviation (F1 closest value to 8.5 mm) in cows with double ovulations than in those with single ovulations, regardless of hCG treatment. In addition, time from aspiration to deviation was shorter in cows with double rather than single ovulation and in cows treated with hCG as a result of faster F1, F2, and F3 growth rates before diameter deviation. In conclusion, greater FSH and follicle growth before deviation seems to be a primary driver of greater frequency of double ovulation in lactating cows with low circulating P4. Moreover, the increase in follicle growth before deviation and in the maximum size of F2 during hCG treatment suggests that increased LH may also have a role in stimulating double ovulation.

O-128 Letrozole co-treatment during ovarian stimulation in antagonist cycles augments follicle growth and exerts pronounced effects on endocrinology: a randomized controlled trial

Abstract Study question What are the endocrine and paracrine effects of letrozole (LZ) co-treatment during ovarian stimulation (OS) and is follicle growth and recruitment affected? Summary answer Letrozole induces marked changes in the follicular and luteal phase endocrinology causing potentiation of follicle growth and corpus luteum function without affecting follicle recruitment. What is known already Letrozole is a 3rd generation aromatase inhibitor that is well-established as an effective ovulatory agent, while its possible benefits in standard in vitro fertilization (IVF) protocols are less thoroughly investigated. Study design, size, duration Double-blinded, placebo-controlled, randomized study with LZ or placebo intervention during OS for IVF treatment, a preceding baseline natural cycle, and a succeeding follow-up visit, in total three consecutive cycles. Participants were enrolled between August 2016 and November 2018. Participants/materials, setting, methods Thirty-one healthy, normo-responding women underwent a natural baseline cycle and an antagonist stimulation cycle with either LZ 5 mg/day (n = 16) or placebo (n = 15) during the follicular phase. Every third day, monitoring of follicle count and -growth and blood analyses of 12 endocrine and paracrine parameters were performed. Follicle recruitment was correlated to endocrine parameters, and differences in follicle recruitment, follicle growth and hormone parameters between groups were assessed. Main results and the role of chance In the LZ group, the number of follicles &amp;gt; 16 mm at oocyte pick up was increased (+2.0, 95% CI [0.1;3.8], p = 0.036), while the total number of follicles were the same (+0.2, 95% CI [-5.8;6.1], p = 0.958). The FSH concentration was increased in the LZ group at stimulation day 5 (p &amp;lt; 0.05), though the total FSH consumption during OS was similar (difference: -20 IU, 95% CI [-175;136], p= 0.794). Letrozole co-treatment suppressed oestradiol (E2) concentrations in the follicular phase (area under the curve (AUC) -62%, p &amp;lt; 0.001) and luteal phase (AUC -31%, p = 0.046). We found increased follicular phase LH (AUC +20.5%, p = 0.033), androstenedione (AUC +22%, p = 0.016), testosterone (AUC +33%, p = 0.013), and 17-OH-progesterone (+132%, p = 0.027). Interestingly, the corpus luteum (CL) output in the LZ group was increased with elevated luteal phase progesterone (AUC +35%, p = 0.043), Inhibin A (AUC +67%, p = 0.011), androstenedione (AUC +22%, p = 0.006), and testosterone (AUC +22%, p = 0.012). Hormone parameters were similar between groups at the follow-up visit. There was no effect of LZ co-treatment on recruitment of the next follicle cohort (p = 0.821). Limitations, reasons for caution This study included a relatively small, selected group of healthy women with an expected normal ovarian function and reserve, and the effects of LZ may therefore be different in other patient groups. Wider implications of the findings We confirm previous findings concerning increased follicle growth, increased endogenous FSH and androgen production, and support the rationale for use of LZ co-treatment as a form of endogenous androgen priming sensitizing the follicle to FSH. Letrozole appears to improve the luteal phase with better stimulation of CL and P4 secretion. Trial registration number NCT02939898

P-677 Increasing the efficacy of FSH with a potentiating monoclonal antibody

Abstract Study question Can we potentiate FSH activity with antibodies to improve ovarian stimulation? Summary answer IGYXOS developed a potentiating monoclonal antibody that improves FSH potency and efficacy in vitro and in vivo as demonstrated in different animal models. What is known already FSH plays a key role in reproduction. For a controlled ovarian stimulation in IVF cycles, FSH-containing preparations are injected to patients in order to obtain mature and fertilizable oocytes. However, the efficacy of these treatments is frequently limited and sometimes several attempts are needed before reaching a pregnancy and a live birth. It was previously demonstrated that breeding animals treated with gonadotropins could develop anti-gonadotropin antibodies that enhance their bioactivity instead of inhibiting it. These females had a high kidding rate and sometimes were hyperprolific. Study design, size, duration IGYXOS developed a monoclonal antibody (mAb) directed against human FSH. The mAb CF12 was selected based on its ability to increase FSH activity in vitro and in vivo in female rat. Its potentiating activity was tested on different human FSH preparations. The potentiating effect of CF12 was further investigated in ewe treated with mAb alone or FSH alone. Participants/materials, setting, methods CF12 was tested in vitro on HEK 293 cells expressing the human FSH receptor and a cell reporter system for cAMP. In vivo, the mAb was tested according to the Steelman and Pohley protocol on different human FSH preparations. Its potentiating effect was also investigated in an ovulation induction protocol on mature ewes. Main results and the role of chance In HEK 293 cells expressing the human FSH receptor, CF12 combined to FSH improved both the potency (EC50) and the efficacy (Emax) of FSH by increasing the cyclic AMP production at all tested FSH concentrations. The potentiating activity was confirmed in immature female rats treated with hCG+FSH alone or combined to mAb, twice a day, during 3 days. The animals were sacrificed the fourth day of treatment and the weight of the ovaries was compared. Ovaries weight was significantly increased in the group treated with hCG/FSH+CF12 compared to hCG/FSH only. To further analyze the effect of CF12 on ovulation induction, it was investigated in ewe as a breeding animal model. Animals were first synchronized with a progestagen and then treated either with FSH alone or with CF12 alone. The number of ovulations was investigated by endoscopy by counting corpora lutea and the progesterone secretion was measured over the luteal cycle. All females treated with CF12 ovulated (100%) by contrast 40% ovulated with FSH treatment. The ovulation rate was 1.8 fold higher in group treated with CF12 compared to FSH group. Moreover, progesterone secretion during luteal phase was increased in animals treated with CF12 suggesting a better quality of corpora lutea. Limitations, reasons for caution After several proofs of concept obtained in different animal models, the compound is currently in the humanization phase before starting the CMC process and then preclinical studies. Wider implications of the findings The potentiating antibody CF12 is a new concept. It represents an interesting strategy in reproductive health, particularly to improve the IVF outcome. CF12 is currently developed for human application by our company and could be a game changer in the field of ovarian stimulation by enhancing the efficacy of FSH. Trial registration number not applicable

Experience, and gynaecological and reproductive health follow-up of young adult women who have undergone ovarian tissue cryopreservation

Research questionWhat are the experience, and gynaecological and reproductive health outcomes in young adult women who have undergone ovarian tissue cryopreservation (OTC)? DesignA retrospective observational study was conducted at a single institution between May 2019 and February 2021 including 87 women aged over 18 years undergoing OTC. Medical characteristics and questionnaire data collected more than 18 months after OTC were analysed. ResultsClose to 74% (n = 64/87) of women had a follow-up consultation and completed the questionnaire. Most women found the information provided on the OTC technique and the strategies proposed to restore fertility with ovarian tissue understandable and useful. The majority of patients thought that OTC had a positive impact on their well-being during disease treatment. Anti-Müllerian hormone serum concentration decreased significantly after treatment (P < 0.0001) and was significantly lower when patients received chemotherapy before OTC (P = 0.0039). The total cyclophosphamide equivalent dose was significantly higher in women with FSH concentrations above 25 IU/l after treatment (P = 0.0004). More than 70% of women who planned a pregnancy after the end of treatment succeeded, with a natural pregnancy rate close to 53%. Only nine patients (8.0%) underwent ovarian tissue transplantation for fertility restoration and six of them became pregnant and delivered at least once. ConclusionYoung adult women expressed a good satisfaction rate with OTC and that their experience had been beneficial. The usage rate of cryopreserved ovarian tissue remains low. The gynaecological and reproductive health follow-up consultation should be included in the supportive care provided following OTC.

Long-day photoperiods affect expression of OPN5 and the TSH-DIO2/DIO3 pathway in Magang goose ganders

This study sought to understand the regulation mechanism of OPN5 through the TSH-DIO2/DIO3 pathway mediated photoperiod on the breeding activity of short-day breeding birds. In this study, the reproductive activity of Magang goose was regulated by artificial light, and the reproductive activity of the ganders were determined according to the daily laying rate of female geese. The testicular development and the serum reproductive hormone concentrations of ganders were measured during the reproductive period (d 0), the reproductive degeneration period (d 13 and 27) and the resting period (d 45). The mRNA and protein expression patterns of OPN5, the HPG axis reproductive genes, and TSH-DIO2/DIO3 pathway related genes were examined. Results showed that the laying rate of geese and the gonadal indices (GSI) decreased gradually after the photoperiod increased. Histological observation found that the spermatogenic function of the testis was normal on d 0 and 13, while degeneration occurred by d 27 and 45. Serum testosterone, FSH, and LH concentration showed a slight increase on d 13, followed by a sharp decrease on d 27 and 45 (P < 0.01), while PRL concentrations were low on d 0 and 13, and increased rapidly on d 27 and 45 (P < 0.01).The expression pattern of GnRH, FSH, LH, and THRβ mRNA were similar, with high levels on d 0 and 13 and a decreasing trend on d 27 and 45 (P < 0.05 or P < 0.01); and GnRHR mRNA levels were higher on d 13 (P < 0.05), but then had decreased by d 27 and 45 (P < 0.01). The expression pattern of GnIH and GnIHR was similar, which was opposite to that of GnRHR. VIP, PRL, and PRLR increased gradually and peaked on d 45 (P < 0.01). The expression trend of TRH, TSHβ, and DIO2 was similar to that of GnRHR, and the expression abundance increased on d 13, and then decreased on d 27 and 45. GnRH protein expression was significantly higher than during the other 3 periods (P < 0.01) while the GnIH protein levels were extremely low on d 0, had gradually increased by d 13, and significantly increased by d 27 and 45 (P < 0.01). The protein expression trends of THR and DIO2 were similar to that of GNIH. DIO3 protein expression was low on d 0 and 13, and increased by d 27 and 45. These results suggest that when the photoperiod increased, the hypothalamus OPN5 gene and protein were upregulated and the pituitary TSHβ, TSHR, and hypothalamus THRβ, TRH, and DIO2 were downregulated, and thus the reproductive activity of geese was inhibited.

A Biphasic Pattern of Reproductive Hormones in Healthy Female Infants: The COPENHAGEN Minipuberty Study.

Minipuberty, a period of a transient activation of the hypothalamic-pituitary-gonadal (HPG) axis in both sexes, enables evaluation of gonadal function in infants suspected of hypogonadism. However, female minipuberty remains poorly elucidated. We aimed to establish continuous reference ranges for the most commonly used reproductive hormones and to evaluate the dynamics of the HPG axis in females aged 0 to 1 year. The COPENHAGEN Minipuberty Study (ClinicalTrials.gov ID: NCT02784184), a longitudinal, prospective cohort study. Healthy infants from Copenhagen. A total of 98 healthy, term female infants followed with 6 examinations including venipuncture during the first year of life. None. Serum concentrations of LH, FSH, inhibin B, anti-Müllerian hormone (AMH), estrone (E1), estradiol (E2), and SHBG were quantified using highly sensitive methods in 266 serum samples. Reference ranges were established for LH, FSH, inhibin B, AMH, E1, E2, and SHBG. Two peaks were observed in normalized mean curves for all hormones. The first peaks were timed around postnatal days 15 to 27 followed by a general nadir for all hormones around days 58 to 92. The second peaks occurred around days 107 to 125 for inhibin B, AMH, E1, E2, and SHBG and days 164 to 165 for LH and FSH. We present age-related, continuous reference ranges of the most commonly used reproductive hormones and present novel data revealing a biphasic and prolonged female minipuberty. NCT02784184.

Gonadotropin profile in experimentally induced hypothyroid and hyperthyroid cyclic female rats

The current study examined the association of thyroid disorders with reproductive dysfunction by determining its effect on gonadotropin secretion in cyclic female rats. Sixty cyclic females were assigned to three groups (20 each) and supplemented, for 30 days plus two consequent estrous cycles, with drinking water (control), methimazole in drinking water (0.02% w/v) (hypothyroid group), and thyroxine in drinking water (0.002% w/v) and gastric gavage of 200 μg/kg body weight (hyperthyroid group). At late proestrus, ten females from each group (for each cycle) were anesthetized and dissected. Blood samples were obtained to assess thyroid-stimulating hormone, free and total triiodothyronine, free and total thyroxin, follicle-stimulating hormone, luteinizing hormone, and prolactin concentrations. Ovarian and pituitary tissue samples were obtained for molecular analysis of ovarian thyroid receptor genes and pituitary TSH, FSHβ, and LHβ genes. In comparison with control, the Hypo group revealed increased serum concentrations of TSH and PRL and the expression levels of pituitary TSH and ovarian TRsTRs genes and significant decrease of FT3, TT3, FT4, TT4, FSH, and LHLH concentrations and the expression levels of pituitary FSHβ and LHβ genes. In contrast, the Hyper group showed increased serum FT3, TT3, FT4, TT4, and LHLH concentrations and the expression levels of pituitary LHβ and ovarian TRsTRs genes and decreased serum TSH FSH and PRL concentrations and pituitary FSHβ and TSH gene expression levels. It is concluded that thyroid dysfunction is associated with altered serum gonadotropin secretion and reproductive failure.

The Simulated Physiological Oocyte Maturation (SPOM) system in domestic animals: A systematic review

Simulated Physiological Oocyte Maturation (SPOM) mimics in vitro the physiological events of oocyte maturation. The system uses cAMP modulators in two steps (pre IVM and IVM) and has presented promising results that are arousing the curiosity of IVF programs in different animal species, generating several papers, adaptations, and controversies worldwide. This study systematically analyses the data in the literature on the use of SPOM and compares the outcomes to the original paper (Albuz et al. Hum. Rep., 25: 2999–3011 2010), classifying them into success or failure. The PubMed, Scopus, and Google Scholar databases were searched and 22 studies were included, from which data on 26 experiments were extracted and evaluated via descriptive statistical analysis. Only experiments that assessed the blastocyst rate (BR) were considered for the success parameter, i.e. success (increase in BR) or failure (either no difference or a reduction in BR). The experiments applied the SPOM system in the following species: cattle, sheep, goats, mice, mares and cats. Three experiments (3/26) could not be evaluated for success or failure, and of the remaining, 34.7% (8/23) succeeded in improving blastocyst production. More than two-thirds (69.2%, 18/26) of experiments were conducted in cattle; of those, 86.8% (13/15) used TCM-199 as the IVM media, and 22.2% did not use forskolin or IBMX modulators as indicated in the original study. Also, 27.7% (5/18) of the experiments in cattle used the same type and dose of FSH, and 22% (4/18) used the same protein source and concentration as indicated in the original study. All experiments conducted in mice (3) kept the parameters of the original study in terms of forskolin and IBMX doses and BSA and FSH concentrations, however, they removed cilostamide from IVM. Cilostamide was used during IVM in more than half (53.8%) of all experiments, but only in cattle and sheep. Considering oocyte and embryo assessments, six experiments assessed cAMP levels and most (5/6) of these observed an increase: in cattle (2), sheep (2), and mice (1). Ten experiments evaluated the effect of SPOM on nuclear maturation, and in 90% (9/10), the SPOM system was able to arrest meiosis (cattle, sheep and mice). Thirteen experiments evaluated the total cell number (cattle, mice and sheep), and six (6/13) showed an increase. Our findings clearly indicate difficulties in reproducing the SPOM system worldwide, demonstrating that the meiosis arrest is not sufficient to ensure successful SPOM application. They also suggest that the different supplements used in the IVM medium and/or their interaction with modulators for different durations may produce a significant bias that affects experimental success.

Fecundity profiles and physiological traits of Iraqi Awassi ewes supplemented mulberry leaves powder extract

The fecundity efficacy of ewes depends on reproductive hormones, ovarian activity so the current research was designed to improve the fecundity of Iraqi anestrous ewes by mulberry leaves (Morus alba) powder extract supplementation. 30th healthy non-pregnant ewes aged 2-3.5 year outbreeding season were divided into equal three groups, 1st group represent a (control) had fed the standard diet, second and third group represent T1 and T2 supplemented 15 and 30 mg/kg mulberry powder extract, respectively, for 40 day before sponge insertion. At day 28th of beginning supplementation the intra-vaginal sponge were inserted for 12 days. After sponges eliminated, estrous onset reported and blood collected before 20 and after 23 & 40 hrs of VSPR to estimate FSH, LH, progesterone and Estradiol concentration. Also, the fecundity, lambing and prolificacy rates had manifested. The results recorded a significant elevation in FSH, estradiol and LH while progesterone registered significant decreased in T1 &T2 comparing with C at 23 & 40 hr after VSPR. Also the fecundity, lambing rates were significantly higher in T1 &T2 in comparing with C. In concluded, that the mulberry leaves powder extract can augment the fecundity in anestrus Iraqi ewes by enhancing the fecundity profile and prolificacy rate.