Androgenic effects on ventricular repolarization: A translational study from the international pharmacovigilance database to iPSC-cardiomyocytes.

BACKGROUND: Male hypogonadism, arising from a range of etiologies including androgen-deprivation therapies (ADTs), has been reported as a risk factor for acquired long-QT syndrome (aLQTS) and torsades de pointes (TdP). A full description of the clinical features of aLQTS associated with ADT and of underlying mechanisms is lacking. METHODS: We searched the international pharmacovigilance database VigiBase for men (n=6 560 565 individual case safety reports) presenting with aLQTS, TdP, or sudden death associated with ADT. In cardiomyocytes derived from induced pluripotent stem cells from men, we studied electrophysiological effects of ADT and dihydrotestosterone. RESULTS: Among subjects receiving ADT in VigiBase, we identified 184 cases of aLQTS (n=168) and/or TdP (n=68; 11% fatal), and 99 with sudden death. Of the 10 ADT drugs examined, 7 had a disproportional association (reporting odds ratio=1.4-4.7; P<0.05) with aLQTS, TdP, or sudden death. The minimum and median times to sudden death were 0.25 and 92 days, respectively. The androgen receptor antagonist enzalutamide was associated with more deaths (5430/31 896 [17%]; P<0.0001) than other ADT used for prostate cancer (4208/52 089 [8.1%]). In induced pluripotent stem cells, acute and chronic enzalutamide (25µM) significantly prolonged action potential durations (action potential duration at 90% when paced at 0.5Hz; 429.7±27.1 (control) versus 982.4±33.2 (acute, P<0.001) and 1062.3±28.9ms (chronic; P<0.001), and generated afterdepolarizations and/or triggered activity in drug-treated cells (11/20 acutely and 8/15 chronically). Enzalutamide acutely and chronically inhibited delayed rectifier potassium current, and chronically enhanced late sodium current. Dihydrotestosterone (30nM) reversed enzalutamide electrophysiological effects on induced pluripotent stem cells. CONCLUSION: QT prolongation and TdP are a risk in men receiving enzalutamide and other ADTs. CLINICAL TRIAL REGISTRATION: URL: https://www.clinicaltrials.gov. Unique identifier: NCT03193138.

Evaluation of different markers of the ovarian reserve in patients presenting with premature ovarian failure.

Premature ovarian failure (POF) is a heterogeneous syndrome, possibly due to mutations of genes involved in the normal development of the ovary and/or the follicles. Based essentially on animal models, these mutations are associated with various ovarian histological phenotypes, from a complete absence of to a partial follicular maturation. The aims of our work were in one hand to determine if ovarian histology, compared to pelvic ultrasonography, would be helpful either in identifying which patients display an impaired follicular growth or in the orientation of the POF etiology; on the other hand, since developing follicles up to the antral stage are reported in POF and that Anti-Müllerian hormone (AMH) might be a good indicator of follicular presence, we decided to determine whether AMH should be a better marker to determine the presence of an ovarian reserve in POF patients. To try to answer to the first question, we studied first 166 patients suffering from POF with a normal karyotype. Vaginal ultrasonography (US) was performed in 134 patients and an ovarian biopsy was obtained in 67 women. The presence of follicles suggested at US was confirmed at histology in only 56% of the patients. Ovarian histology led to the distinction of two phenotypes (a) small-sized ovaries, deprived of follicles, and (b) normal-sized ovaries with partial follicular maturation. To confirm the value of ovarian biopsies, samples from 20 normal women have been studied, confirming that ovarian biopsy at random allow reliable assessment of follicular activity. To try to answer to the second question of our work, a cross sectional study analyzing serum AMH, ovarian histology and AMH immunoexpression in 48 POF patients, was performed. Serum AMH was significantly higher in women with more than 5 follicles at ovarian histology. Ovarian AMH immunostaining revealed a normal AMH expression in POF preantral follicles but a decrease expression at the early antral stages. In conclusion, ovarian histology appears to be a reliable tool to appreciate the follicular reserve, and helpful and complementary to clinical and hormonal phenotyping in order to orient the search for various genetic causes of POF syndrome. Finally, AMH levels in POF patients could identify women with persistent follicles.

Molecular pathology of the FSH receptor: new insights into FSH physiology.

Manipulations of mouse genome have helped to elucidate gonadotrophin function but important differences subsist between rodent and human reproduction. Studies of patients with mutations of gonadotrophins or gonadotrophin receptors genes allow understanding their physiological effects in humans. The correlation of the clinical phenotypes of patients with in vitro studies of the mutated receptor residual function and histological and immunohistological studies of the ovarian biopsies permits to understand which stages of follicular development are under FSH control. Total FSH receptor (FSHR) inactivation causes infertility with an early block of follicular maturation remarkably associated with abundant small follicles as in prepubertal ovaries and demonstrates the absolute requirement of FSH for follicular development starting from the primary stage. Partial FSHR inactivation, characterized by normal-sized ovaries, can sustain follicular development up to the early antral stages but incremental levels of FSH stimulation seem to be required for antral follicular growth before selection. These findings contrast with the traditional view of an initial gonadotrophin-independent follicular growth prior to the preantral-early antral stages. The presence of numerous reserve follicles in the ovaries of these patients may permit a future treatment of their infertility. The study of reduced FSHbeta or FSHR activity in genetically modified male mice models and in men suggests a minor impact of the FSHR on masculine fertility. Further studies on patients with a demonstrated total FSHbeta or FSHR inactivation are required to elucidate reported differences in spermatogenesis impairment. Finally, the studies of mutations of gonadotrophins and their receptors demonstrate differences in gonadotrophin function between genetically modified rodents and humans which suggest prudence in extrapolating observations in rodents to human reproduction. Ovarian hyperstimulation syndrome (OHSS) can infrequently arise spontaneously during pregnancy, but most often it is an iatrogenic complication of ovarian stimulation treatments with ovulation drugs for in vitro fertilization. The first genetic cause of familial recurrent spontaneous OHSS was identified as a broadening specificity of the FSHR for hCG due to naturally occurring heterozygous mutations located unexpectedly in the transmembrane domain of the FSHR. Broadening specificity of a G protein-coupled receptor is extremely rare. These observations led to the identification of the etiology of this previously unexplained syndrome and permitted to conceive novel models of FSHR activation. Susceptibility to iatrogenic OHSS or its clinical severity may be associated with FSHR polymorphisms with slightly different activities in vivo as suggested by several studies. The study of larger cohorts is needed to evaluate the clinical impact of these observations in the management of patients undergoing IVF protocols.

Luteinizing hormone pulsatility in patients with major ovarian hyperandrogenism.

Hyperandrogenism and ovulatory dysfunction are common in women with either polycystic ovary (PCOS) or ovarian virilizing tumor. However, contrasting with the numerous studies that have extensively described gonadotropin secretory abnormalities, principally increased LH pulse amplitude and frequency, few studies have concerned gonadotropin secretion in patients with ovarian virilizing tumors; low gonadotropin levels have occasionally been reported, but never extensively studied. The goal of the present study was to further evaluate the pulsatility of LH secretion in women with ovarian virilizing tumor compared with that of PCOS patients. Eighteen women with major hyperandrogenism (plasma testosterone level >1.2 ng/ml) were studied (5 women with ovarian virilizing tumor, 13 women with PCOS, and 10 control women). Mean plasma LH level, LH pulse number and amplitude were dramatically low in patients with ovarian tumors when compared to both PCOS (p<0.001) and controls (p<0.001). In case of major hyperandrogenism, LH pulse pattern differs markedly between women with ovarian virilizing tumor or PCOS, suggesting different mechanisms of hypothalamic or pituitary feedback.

[Semen characteristics and quality of the conceptus in fertilization in vitro]. / Caractéristiques du sperme et qualité du conceptus en fécondation in vitro.

OBJECTIVE: Assisted reproductive technology (ART) is associated with increased risks for the neonate, compared to natural fertility, mainly because of multiple pregnancies and increased maternal age. On the opposite, the impact of paternal factors has been scarcely studied, except for the relationship between surgically retrieved sperms and genetic abnormalities. PATIENTS AND METHODS: This study has been realized using the large French register on in vitro fertilization (FIVNAT) that collects information on 80% of French ART activity. For the study, all the pregnancies obtained from oocyte recoveries between January 1996 and December 2003, for which information on cycles could be linked to the pregnancy were included, i.e. 34223 pregnancies, resulting in 27025 deliveries and 33945 neonates. Sperm quality was defined either according to the semen origin (spouse's ejaculate, epididymis, testis, or donor), or according to the spermiogramme values for concentration, motility and morphology. The statistical analysis included the use of multivariate logistic models, with the main prognostic factors. RESULTS: The delivery, spontaneous abortion and ectopic pregnancy rates were not influenced by semen origin nor by quality, all the 95% confidence intervals including 1. The neonates conceived through surgically retrieved sperms were at a slightly increased risk of hypotrophy (weight under the 10% centile of reference curves) and of malformation (OR=1.18, 95CI: 1.03-1.36 and OR=1.30 [0,95-1,84], respectively). On the opposite, when analysis was restrained to ejaculated semen, no risk was demonstrated. DISCUSSION AND CONCLUSION: The semen quality has little impact on pregnancy issue. Only the semen origin seems to act on hypotrophy and malformation, but these results deserve to be further analyzed for confirmation.

Hormonal treatment of congenital adrenal hyperplasia due to 21-hydroxylase deficiency.

During childhood, the main aims of the medical treatment of congenital adrenal hyperplasia (CAH) secondary to 21-hydroxylase deficiency, are to prevent salt loss and virilization and to achieve normal stature and normal puberty. As such, there is a narrow therapeutic window through which the intended results can be achieved. In adulthood, the clinical management has received little attention, but recent studies have shown the relevance of long-term follow-up of these patients. Indeed, long-term evaluation of adult CAH patients enables the identification of multiple clinical, hormonal and metabolic abnormalities as bone mineral density alteration, overweight and disturbed reproductive functions. In women with classic CAH, low fertility rate is reported, and is probably the consequence of multiple factors, including neuroendocrine and hormonal factors, feminizing surgery, and psychological factors. Men with CAH may present hypogonadism either through the effect of adrenal rests or from suppression of gonadotropins resulting in infertility. These patients should therefore be carefully followed-up, from childhood through to adulthood, to avoid these complications and to ensure treatment compliance and tight control of the adrenal androgens, by multidisciplinary teams who have knowledge of CAH.

Characteristics of reproductive life and risk of breast cancer in a case-control study of young nulliparous women.

Between 1982 and 1985, a case-control study of nulliparous women, aged 25-45, was conducted to analyse the relationships between the risk of breast cancer and causes of nulliparity, including contraceptive methods. Fifty-one cases of breast cancer diagnosed less than 3 months before interview were matched with 95 controls on age at diagnosis, year of interview, and medical center. The causes of nulliparity related to female sterility or subfertility (mechanical or hormonal disorders) were not found to be associated with a significantly higher risk of breast cancer. The causes related to fertilization failure, i.e. no sexual partner, rare sexual intercourse (less than once per month), or partner with abnormal semen, were found to lead to an increased risk. Detailed analysis of contraceptive methods showed that the risk of breast cancer increased (p = 0.02) with a longer duration of use of barrier methods (withdrawal or condom). Conversely, the risk significantly decreased (p = 0.004) with a longer duration of use of non-barrier methods (oral contraceptives, IUD, cap, local spermicides, vaginal douche, safe period, or no method), i.e. methods allowing a direct exposure to human semen.

Oral contraceptive use and risk of benign breast disease in a French case-control study of young women.

We report the results of a French hospital-based case-control study designed to analyse the relation between the use of oral contraceptives (OC) and the risk of benign breast disease (BBD). The cases were 286 women, each less than 46 years old, with BBD histologically verified between 1982 and 1985. Controls were 382 patients, matched to cases on year of birth and month of interview, and who were hospitalized for a non-malignant disease other than BBD. Odds ratios were estimated by multivariate regression, taking into account level of education, place of residence, family history of breast cancer, age at menarche, number of children, age at first full-term pregnancy and Quetelet index. The risk of BBD was found to decrease significantly with a longer use of OC before the first full-term pregnancy (FFTP), but there was no association between the risk of BBD and the duration of OC use after FFTP. OC use before FFTP reduced the risk of non-proliferative disease, but did not significantly affect the risk of proliferative disease. These results did not depend on the amount of oestrogen (0.05 mg or more vs < 0.05 mg) contained in OC.

PIP: This article reports the findings of a French-based case-control study examining the relationship between risk of benign breast disease (BBD) and oral contraceptive (OC) use. Special emphasis was given to duration of OC use before and after the first full-term pregnancy (FFTP). Conditional multivariate logistic regression was the statistical process used to analyze the findings. 286 women with verified BBD were studied. 382 women, who were hospitalized for a non-BBD disease, were used as controls. Each case was matched with at least one control. 7 confounders were adjusted for. Confounders included family history of breast cancer, level of education, place of residence, age at menarche, number of children, age at FFTP, and Quetelet index. The risk of BBD decreased with longer OC use before the FFTP. There was no association observed between risk of developing BBD and the duration of OC use after the FFTP. OC use before the FFTP reduced the risk of non-proliferative disease, but did not significantly affect the risk of proliferative disease.

Conditions for choosing between drug-induced and surgical abortions.

In France, pregnant women with amenorrhoea of no more than 49 days intending to terminate pregnancy can choose between a surgical abortion via vacuum aspiration under local or general anesthesia and a drug method combining Mifepristone orally administered (RU 486 degrees), with a prostaglandin analogue. This prospective survey was conducted to study the conditions under which women choose their abortion method, and to evaluate the acceptability of each method after the abortion. The data gathered from 488 women were analyzed according to their initial decision, and then according to the method actually used. The majority (62%) chose RU 486. The women's choice was found to be linked to sociodemographic characteristics such as age, education, occupation, geographic origin, and certain attitudes towards pregnancy, as well as to the individual criteria of each method. The women who chose the drug protocol had most often already made their decision before going to the family planning center (68%), having been advised by their doctor (20%). They were slightly less satisfied with the abortion experience than they had expected: 12.4% were unsatisfied in the RU group and 3.6% in the aspiration group. They needed more rest and quiet afterwards (50%) than the other women. They were distinguished by their desire to verify the expulsion (63%). The length of pregnancy is therefore not the only criterion to be considered when recommending one or other of these methods: the women concerned have different requirements and should have several possibilities to choose from.

[Comparison of agonists versus antagonists for i.v.f. stimulation: the French FIVNAT survey 2001-2002]. / Comparaison agonistes / antagonistes dans les stimulations pour FIV: les données de FIVNAT 2001-2002.

In 2002, FIVNAT has continued its study on the use and the results of the ovarian stimulation protocols using GnRH antagonist for i.v.f. and i.c.s.i. The first part of this study demonstrates that the antagonist protocols were still used in more pejorative cases than the long agonist protocols but with a smaller difference compared to 2001. So far, the frequency of the antagonist protocols has jumped from 13% to 19% from 2001 to 2002. This increase was mainly at the expense of the long agonist protocols. In the overall population, the lower pregnancy rate per ovum pick-up with antagonists, demonstrated in 2001, has been confirmed. In the second part of the study, FIVNAT has selected a good prognosis group (women's age less than 35 years and attempts rank lower than 3). This group includes 17,532 cycles with agonists and 3229 cycles with antagonists. A comparison of all the biological and clinical data was made. Finally, the pregnancy rate per ovum pick-up was significantly reduced when the stimulation was done with an antagonist protocol (Anta) rather than with a long agonist protocol (Ago). In i.v.f. these rates were 20.8% (Anta) versus 25.8% (Ago) (P < 0.001). In i.c.s.i. they were 21.9% (Anta) versus 29.4% (Ago) (P < 0.001). Moreover, the absolute and relative differences were the same in 2001 and 2002, suggesting that the learning curve effect does not explain the differences.

[Biological effects of GnRH antagonists]. / Les antagonistes du GnRH: effets biologiques.

Numerous questions about the efficacy of GnRH antagonists, known for the prevention of undesirable LH surge, have been raised. Al-Inany in 2002 in a meta-analysis, IVF Germany Register and FIVNAT the French data base in 2003, have shown a decrease of the pregnancy rate after antagonists treatment. Among 20761 ART attempts for good prognosis women (<35 years, i.v.f. range 1 or 2) we have compared the clinical and biological parameters after agonists' long protocol versus antagonist treatment. The characteristics of the responses to the stimulation treatment were similar for both groups, whereas the duration of the stimulation and the doses of gonadotrophin used in the antagonist group was lower. The fertilization and embryo development rates were not modified. But we observed a decrease in the number of oocytes retrieved, of embryos obtained and of the pregnancy rate (P < 0.001). These results could be explained by endometrial modifications induced by antagonists but we cannot exclude an impact of oestradiol and LH levels. GnRH antagonists could be an inhibitor of the cell cycle by decreasing the synthesis of growth factors. The interaction of GnRH antagonists and GnRH receptors may compromise the mitotic programme of the cells and induce an alteration of folliculogenesis, embryo quality and implantation. More studies are necessary to understand these results. Using of GnRH antagonists involves specific patient information on the benefits and drawbacks of such treatment before ART.

Detection and management of late-onset 21-hydroxylase deficiency in women with hyperandrogenism.

Moderate forms of 21-hydroxylase deficiency (D21OH-NC), the so-called non-classical or late-onset forms are a frequently reported cause of hyperandrogenism in women [1-5]. The purpose of this collective and synthetic work was to provide the endocrinologist, gynecologist and dermatologist with consensual information so as to detect the maximum cases with acceptable cost-benefit ratio and to define the main lines of optimal patient management, given the data currently available in medical literature.

Serum anti-Müllerian hormone expression in women with premature ovarian failure.

BACKGROUND: Premature ovarian failure (POF) is generally irreversible. However, developing follicles up to the antral stage are reported in POF and anti-Müllerian hormone (AMH) might be a good indicator of follicular presence. This study analysed serum AMH, ovarian histology and AMH immunoexpression in POF patients. METHODS: A cross-sectional study of 48 POF patients in an Endocrinology Department setting. Patients had an ovarian biopsy simultaneously with serum AMH sampling and/or ovarian AMH immunostaining. RESULTS: Mean serum AMH was 1.04 +/- 1.66 ng/ml. Serum AMH was significantly higher in women with 15 or more follicles at ovarian histology (P = 0.001). Comparison of ovarian AMH immunostaining from POF patients and 10 normal controls revealed a normal AMH expression in POF pre-antral follicles, but a decreased expression at the early antral stages. Serum AMH was undetectable in 77% of the patients with 0-5 AMH immunopositive follicles and detectable in 100% of the patients with more than 15 AMH immunopositive follicles. CONCLUSIONS: AMH levels in POF patients could identify women with persistent follicles. The decrease of AMH immunoexpression in POF antral follicles could suggest a defect of antral development.

Establishing a national in vitro fertilization registry: methodological problems and analysis of success rates.

A national registry for in vitro fertilization (IVF) was established in France in January 1986 and has been in operation since that date. During 1987, data on 14,763 attempts were collected from 72 teams, representing 80 per cent of all French IVF activity. In this registry, a form is completed for every oocyte recovery attempt, containing information about the infertility diagnosis, the attempt itself and its result. This paper presents results for the years 1986 and 1987 and studies the relationship between pregnancy occurrence and some couple or cycle characteristics. Methodological aspects are discussed, concerning the possible sources of bias, and the analysis of results. Results obtained in this registry are compared with those from other similar registries.

[Experience of ectopic pregnancy]. / Vécu de la grossesse extra-utérine.

This survey relates to the experience and the management of women who have received a treatment for an ectopic pregnancy. It is based on 31 clinical interviews, conducted a month after the end of surgical or medical treatment. Among these women, 42% had had previous treatment for sterility. The main results show that 16% had never previously heard of ectopic pregnancy, and 29% do not see any reason why they had had one. For the patients, the diagnosis had not been made early enough: 45% feel that the consequences could have been serious if they had not taken the matter seriously themselves. Women clearly express some strong feeling against the health professionals (74%) who were responsible for their medical care, in spite of the attention they received, which reveals the psychological trauma they suffered. Although 55% of women felt depressed, for some of them the ectopic pregnancy seemed to operate as an exorcism from a previous more serious situation. They place their hope in a future pregnancy under close medical supervision. When it appears necessary, psychological support should be offered to these patients.

[Ovulation stimulation]. / Stimulation de l'ovulation.

In 1997, 2 new recombinant FSH were available. The analysis of the first 11,000 attempts IVF and ICSI received by FIVNAT for 1997 shows that recombinant FSH was used in about 20% of cases. Pregnancy rates were similar with urinary and recombinant FSH but the number of ampoulles was significantly lower with recombinant FSH.

In-vitro fertilization: influence of women's age on pregnancy rates.

An analysis of the influence of women's age on the results of in-vitro fertilization was performed, using 5590 attempts collected during the year 1986. Increasing age was found to be related to decreased success rates from 19.8% per attempt below the age of 25 years, to 9% per attempt at 40 years or more. A cut-off point was found between the ages of 36 and 37 years, using a mathematical model. The decrease was related to a reduction in oocyte production (4.3 +/- 2.8 at 25 years or less and 3.3 +/- 2.1 at 40 years or more, P less than 0.001), and to a reduced implantation rate, whatever the number of transferred embryos. These findings were not due to spouse's age, rank of attempt, infertility diagnosis or oocyte stimulation regimen, since the effect of age remained significant when a logistical model including these confounders was applied. Finally, a woman's age must be considered as a prognostic factor when IVF is proposed to infertile couples.