Abnormal Ovarian Function Research Articles

The physiology and pathophysiology of inhibin, activin and follistatin in female reproduction.

In the last 2 years, major advances have been made in the understanding of inhibin physiology. Discovery of an inhibin receptor and binding protein has expanded our knowledge of the mechanism whereby inhibin antagonizes activin action. Controlled experimental studies have clarified the regulation and physiology of inhibin A and inhibin B, providing evidence for their use as markers of ovarian function. Clinical studies continue to uphold the use of inhibin as a marker for ovarian cancer, but have not generally supported its use over standard prognostic markers in assisted reproductive technologies. Finally, ongoing work suggests alterations in inhibin and follistatin that may be linked to the pathophysiology of polycystic ovary syndrome. Thus, the mechanism of inhibin action and its role in normal and abnormal ovarian function continues to emerge.

Free Androgen Index and Leptin Are the Most Prominent Endocrine Predictors of Ovarian Response during Clomiphene Citrate Induction of Ovulation in Normogonadotropic Oligoamenorrheic Infertility1

We have previously demonstrated that obese hyperandrogenic amenorrheic women are less likely to ovulate after clomiphene citrate (CC) medication. The present study was designed to identify whether additional endocrine screening characteristics, all potentially involved in ovarian dysfunction in 182 normogonadotropic oligoamenorrheic infertile women, are associated with ovarian response, which may improve overall prediction of CC-resistant anovulation. Standardized endocrine screening took place before initiation of CC medication (50 mg/day; increasing doses up to 150 mg/day if required) from cycle days 3-7. Screening included serum assays for fasting insulin and glucose, insulin-like growth factor I (IGF-I), IGF-binding protein-1 (IGFBP-1), IGFBP-3, free IGF-I, inhibin B, leptin, and vascular endothelial growth factor. Forty-two women (22% of the total group) did not ovulate at the end of follow-up (a total number of 325 cycles were analyzed). Fasting serum insulin, insulin/glucose ratio, IGFBP-1, and leptin were all significantly different in univariate analyses (P < or = 0.02), comparing CC responders vs. nonresponders. Forward stepwise multivariate analyses in combination with factors reported earlier for prediction of patients remaining anovulatory after CC revealed a prediction model including 1) free androgen index (FAI = testosterone/sex hormone-binding globulin ratio), 2) cycle history (oligomenorrhea or amenorrhea), 3) leptin level, and 4) mean ovarian volume. These data suggest that decreased insulin sensitivity, hyperandrogenemia, and obesity, all associated with polycystic ovary syndrome, are prominent factors involved in ovarian dysfunction, preventing these ovaries from responding to stimulation by raised endogenous FSH levels due to CC medication. By using leptin instead of body mass index or waist to hip ratio, the previous model for prediction of patients remaining anovulatory after CC medication could be slightly improved (area under the curve from 0.82-0.85). This may indicate that leptin is more directly involved in ovarian dysfunction in these patients. The capability of insulin and IGFBP-1 to predict patients who remain anovulatory after CC disappears when FAI enters into the model due to a significant correlation between FAI and these endocrine parameters. This suggests that markers for insulin sensitivity (e.g. IGFBP-1 and insulin) are associated with abnormal ovarian function through its correlation with androgens, whereas leptin is directly involved in ovarian dysfunction.

Reproductive correlates of chronic fatigue syndrome

A case-control study was conducted to determine whether menstrual and gynecologic abnormalities precede the onset of chronic fatigue syndrome (CFS) in women with this disorder to a greater extent than that observed among healthy controls. We identified 150 women who met the 1988 Centers for Disease Control criteria for CFS from the Brigham and Women’s Hospital Cooperative CFS Research Center. A comparison group of 149 women being seen for nongynecologic conditions were selected from the waiting area of the Brigham and Women’s Hospital Internal Medicine outpatient department. Women with and without CFS completed self-administered questionnaires on menstrual, reproductive, and medical history. Women with CFS reported increased gynecologic complications and a lower incidence of premenstrual symptomatology. After adjustment for age, a somewhat greater number of cases compared with controls self-reported irregular cycles, periods of amenorrhea, and sporadic bleeding between menstrual periods. Factors suggestive of abnormal ovarian function—such as a history of polycystic ovarian syndrome, hirsutism, and ovarian cysts—were reported more often in CFS cases compared with controls. Frequent anovulatory cycles due to ovarian hyperandrogenism (PCOS) or hyperprolactinemia may increase risk for CFS through loss of the potential immunomodulatory effects of progesterone in the presence of continued estrogen production. We hypothesize that frequent anovulatory cycles due to PCOS and/or hyperprolactinemia may explain the increased reporting of gynecologic complications and the lower reported premenstrual symptomatology observed in women with CFS.

In UteroExposure to Low Doses of 2,3,7,8-Tetrachlorodibenzo-p-dioxin Alters Reproductive Development of Female Long Evans Hooded Rat Offspring

Prenatal administration of a single dose of 1 μg TCDD/kg induces malformations of the external genitalia and subfertility in female offspring (L. E. Gray, Jr., and J. S. Ostby (1995)Toxicol. Appl. Pharmacol.133,285–294). A cross-fostering study indicated thatin uterobut not lactational TCDD exposure (1 μg TCDD/kg on gestational Day 15) induces cleft phallus, vaginal thread formation, and reduced ovarian weight. TCDD treatment on the 15th day of pregnancy at 0, 0.05, 0.20, or 0.80 μg TCDD/kg delayed vaginal opening at 0.80 μg/kg in the progeny. A persistent vaginal thread was displayed by 27% of the progeny at 0.20 and 92% at 0.80 μg TCDD/kg. These effects did not appear to result from abnormal ovarian function during prepubertal development; neither serum estradiol levels nor ovarian estradiol production were reduced in 21- or 28-day-old progeny of dams exposed to 1 μg TCDD/kg. In addition, partial to complete clefting of the phallus was displayed in TCDD-treated rats (10% at 0.20 and 60% at 0.80 μg TCDD/kg) and these dosage levels also increased the length of the urethral slit, increased distance from the urethral opening to the tip of the phallus, and decreased distance from the urethral opening to the vaginal orifice. Although fertility rates were normal, time-to-pregnancy was delayed by treatment with 0.80 μg TCDD/kg. When necropsied at 20 months of age, females from the TCDD-dose groups displayed histopathological alterations of the reproductive tract. In summary, administration of TCDD at dosage levels of 0.2, 0.8, and 1.0 μg/kg produces morphological reproductive alterations in female rat offspring as a consequence ofin uteroexposure.

Identification of G alpha s messenger ribonucleic acid splice variants in human granulosa cells.

Granulosa cells are essential for follicular development and corpus luteum formation and their functions are regulated by gonadotrophins through G protein-coupled receptors. The dominant second messenger pathway involves the stimulation of cyclic AMP formation by G alpha s-linked receptors. In this paper we have investigated the expression of G alpha s mRNA splice variants in relation to expression of G alpha s protein isoforms in granulosa cells obtained from patients undergoing in vitro fertilization. We have carried out ribonuclease protection assays using cRNA riboprobes which are capable of detecting all G alpha s mRNA isoforms as well as quantifying total amounts of G alpha s mRNA. Granulosa cells express the message for G alpha s-Large and G alpha s-Small and the presence of two distinct protein products was confirmed by immunoblotting using the antibody RM/1. Moreover, the data show that a significant fraction of G alpha s-Large and G alpha s-Small mRNAs contain an extra CAG codon. This should generate proteins with an extra serine residue, resulting in G alpha s variants with the consensus sequence of a protein kinase C phosphorylation site. These results highlight the possible interaction between different signalling pathways in the control of cAMP production and the need to investigate the relationship between G alpha s variants and different adenylyl cyclase isozymes in patients with normal and abnormal ovarian function.

Endometriosis and polycystic ovary syndrome: enhanced stimulatory effect of peritoneal fluid on progesterone release from human granulosa-lutein cells

To compare the activity of peritoneal fluid (PF) from women with and without endometriosis or polycystic ovary syndrome (PCOS) on P release from cultured human granulosa-lutein cells. Case-control study. Medical school and hospital. Women with mild to moderate endometriosis or PCOS and controls undergoing laparoscopic sterilizations. Human granulosa-lutein cells, obtained from the IVF clinic, were cultured with increasing volumes of steroid-extracted PF samples with and without hCG. Progesterone concentrations in the media after 72 hours culture were measured by RIA. Peritoneal fluid stimulated P release in a dose-dependent manner and, at the highest dose, the response was enhanced significantly by PF from women with endometriosis and PCOS compared with control samples. The effects of PF plus hCG were not simply additive. There was a large potentiation of the response, which was greater in women with endometriosis and PCOS. Peritoneal fluid contains factors that stimulate P release and potentiate the response to hCG. The increased activity of PF associated with endometriosis and PCOS in part may have some significance in abnormal ovarian function associated with these syndromes.

Pregnancy rate in an oocyte donation program.

Oocyte donation programs offer an alternative treatment for infertile women with ovarian failure or abnormal ovarian function. Seventeen cycles of in vitro fertilization and embryo transfer with donated oocytes were performed in 13 women, with a mean age of 34.8 years. The hormonal replacement therapy consisted of a fixed dose of oral estradiol valerate, 6 mg daily, and intramuscular progesterone in oil, 100 mg daily. Estrogen and progesterone were continued for 10 more weeks after embryo transfer if pregnancy was established. After 13 embryo transfers, 8 pregnancies were obtained, for a pregnancy rate per transfer of 61.5%. Today seven pregnancies are progressing normally, including one set of twins. This results suggest that an oocyte donation program using a fixed and simple hormonal replacement therapy is an adequate treatment for these infertile couples.

Preliminary results suggesting exaggerated ovarian androgen production early in the course of polycystic ovary syndrome

Excess ovarian androgen production might be a cause of the polycystic ovary syndrome (PCO). Previous studies have evaluated adult women with long-standing abnormality of the hypothalamic-pituitary gonadal axis. Abnormal ovarian function in such patients could be a primary or even a secondary finding. For that reason, this study was designed to evaluate ovarian androgen production in symptomatic adolescent females. Simultaneous adrenal suppression, by using dexamethasone, and ovarian stimulation, by using gonadotropin-releasing hormone (GnRH), were achieved in 12 patients. Following stimulation, blood was serially obtained over 8 hr to measure gonadotropin, estrogen, and androgen responses. Based on the androgen response, patients could be divided into two groups. Group A (five) had a significant increase ( p < 0.01) in free testosterone, whereas group B (seven) had no increase in any androgen, including free testosterone (significantly different from group A, p = 0.01). All patients in group A had enlarged or cystic ovaries, whereas only one-quarter patients in group B had enlarged ovaries (significantly different from group A, p < 0.03). The pituitary and estrogenic response was similar in both groups. These preliminary data suggest that some patients with PCO (group A) have a primary abnormality in ovarian androgen production early in the course of their disease.

Effect of active immunization against androstenedione on ovarian function in dairy cows and prepubertal gilts

Two experiments were performed to assess the effects of immunization against androstenedione on ovarian function in cows during the first 100 days after calving and in gilts at the onset of puberty. Seven Holstein-Friesian cows were immunized against androstenedione-11α-human serum albumin (AND-HSA) at 3 weeks before and at 3 weekly intervals after calving and compared with ten cows immunized against HSA. Titres against androstenedione were significant but low and variable (1: 280-1: 4). Two cows with the highest titres exhibited abnormal ovarian function which could be attributable to treatment. One cow developed a large follicular cyst and nymphomania. The other cow exhibited mild super ovulation in her first cycle after calving and multiple ovulations during later cycles. Other cows in the AND-HSA group also had a tendency to develop larger than normal follicles and follicular cysts. Groups of twelve prepubertal gilts were immunized against either androstenedione 11α-hemisuccinyl bovine serum albumin (AND-BSA) or BSA at 100, 121 and 142 days of age. Titres against AND were significant but were low and variable (1: 2500-1: 10). There was no effect of immunization against AND on onset of ovulation, puberty or ovulation rate in the first or second ovulatory cycles.

Peripheral venous concentrations of oestrogens in postmenopausal women with ovarian cancer.

Serum concentrations of oestrone, oestradiol and oestriol were measured in 23 postmenopausal women, 12 with ovarian cancer and 11 with genital prolapse. Oestrone (387.6 pmol/l) and oestradiol (72.7 pmol/l) levels were higher in the cancer group than those in the control group (159.8 and 27.5 pmol/l respectively), while oestriol levels did not differ (434.5-270.8 pmol/l). The results indicate abnormal ovarian function in postmenopausal patients with ovarian cancer.

Abnormal Function of the Corpus Luteum in some Ewes with Phyto-Oestrogenic Infertility

Ewes with permanent phyto-estrogenic infertility show oestrus less regularly than normal ewes, and the present study examines the extent to which this results from abnormal ovarian function. Forty-nine affected ewes and 53 controls were run with rams fitted with marking crayons and harnesses, and crayon marks were recorded and laparoscopy performed at weekly intervals for 3 weeks. Fewer affected ewes showed oestrus accompanied by ovulation (28 v. 49, P less than 0.001), and four of these affected ewes had a second ovulation during the experiment. More of the ovulations observed in affected ewes were unaccompanied by behavioural oestrus than in controls (8 out of 38 v. 2 out of 50; P less than 0.05). Six affected ewes had no corpus luteum or oestrus, and five of these had adhesions over the genitalia. Hydrops uteri in five other affected ewes was accompanied by prolonged maintenance of the corpus luteum. Some other abnormalities were also observed. In a second study, plasma progesterone concentrations were measured twice daily in 12 affected ewes which were run with rams. Five ewes had oestrous cycles of abnormal duration (two of more than 23 days, two of 21 days, and one of 11 days), and these were accompanied by plasma progesterone patterns different from those of the ewes with an oestrous cycle duration of 16-18 days. It is concluded that the irregular oestrous cycles in affected ewes are due mainly to abnormal life span and progesterone secretion by the corpus luteum, which in turn largely result from changes in the uterus.

Radioimmunoassay of progesterone in saliva: application to the assessment of ovarian function.

We report a specific radioimmunoassay that has the required sensitivity (7 pg per assay tube) for determining progesterone concentrations in 400 microL of mixed saliva collected from normal women. The assay is precise: intra and inter-assay variation (CV) never exceeded 11.0 and 8.0%, respectively. The assay was used to determine progesterone in saliva samples collected daily for not less than 28 days by normal women and by patients having abnormal ovarian function. Four normal women provided matched saliva and plasma samples for accurate dating of the menstrual cycle by plasma progesterone, estradiol, lutropin, and follitropin. Nine further subjects collected saliva samples only, and from these data a provisional "normal range" was established. Progesterone concentrations in saliva during the follicular phase of the cycle were low (less than 100 pmol/L) but rose beginning on day 12 to reach peak values of 230-550 pmol/L on day 21. Thereafter, progesterone concentrations in saliva declined to values generally less than 170 pmol/L at the commencement of menses. Saliva samples from three patients attending an infertility clinic were also studied to assess ovarian function.

Histologic Study of Ovaries from Young Women with Endometrial Adenocarcinoma

A histologic study of 15 ovaries from patients in whom endometrial adenocarcinoma developed before the age of 40 years was performed. These ovaries were randomized with control groups of 21 age-matched normal ovaries and 25 ovaries from patients who had Stein-Leventhal syndrome. The study was done in an attempt to determine whether the ovaries from these young patients showed any histologic feature that would indicate abnormal ovarian function resulting in unopposed estrogenic stimulation of the endometrium. Histologic features assessed included type and size of cysts, presence and number of primary follicles, corpora lutea, corpora albicantia, and presence of stromal luteinization and sclerosis. Analysis of the results indicated that from a histologic point of view ovaries of patients less than 40 years old who have endometrial adenocarcinoma are more similar to normal ovaries than to polycystic ovaries. Only 11.1% of ovaries of the study group had features suggestive of polycystic ovarian disease.

Adenocarcinoma of the endometrium and abnormal ovarian function in young women

Twenty-four cases of endometrial adenocarcinoma in women under 40 years of age were reviewed in order to determine the relation to ovarian unopposed estrogenic stimulation. Such a relation should be strong in those cases in which the patients develop adenocarcinoma at a young age. The clinical and histologic characteristics of ovarian imbalance were compared with those of several control groups. The clinical control subjects included women over 50 years of age with adenocarcinoma of the endometrium matched for date of diagnosis and age-matched noncancer patients. The histologic control material consisted of sections previously interpreted as consistent with polycystic ovarian disease as well as histologically normal ovaries. These were randomized with similar specimens from our study group and were interpreted by a pathologist who was unaware of their respective origins. In our study group of young women with endometrial adenocarcinoma, 45.5 per cent were nulliparous and 52.4 per cent had grossly abnormal menses. Both percentages were significantly higher than in any of the control groups. Although this suggested ovarian abnormality, histologic analysis of available material did not reveal any significant difference in the microscopic appearances from normal ovaries. We concluded that the abnormality in steroid function may be of nonovarian origin.

Estrogen Dynamics and Hormonal Differences Associated with Lactational Performance of Cows Induced to Lactate

Five nonpregnant and nonlactating Holsteins (three cows and two heifers) were injected subcutaneously each 12 b of days 1 to 7 with estradiol-17β and progesterone (.1mg and .25 mg/day per kg bodyweight) dissolved in absolute ethanol to induce lactation. Estradiol-17β and -17α represented 81 to 89% of total estrogen in blood plasma from days 7 to 27, and -17α was 78 to 90% of total estrogen excreted in urine. Although conversion of absorbed estradiol-17β to free estrone occurred in blood plasma, conversion of estradiol-17β to free estradiol-17α was greater. Among estrogens, only estradiol-17β was higher in blood plasma after day 27 than in some untreated cows with abnormal ovarian function. The heifers, but not the cows, lactated from treatment. Compared with pretreatment (day 0) and end of treatment day 7 to 7.5, concentrations of prolactin in blood plasma were increased prior to first milking (day 12) of heifers as in other studies but were unchanged through day 38 in the cows. Epimers of estradiol, -17β in blood plasma and -17β and -17α in urine, were lower in heifers than in cows during day 13 and days 17 to 26 but not thereafter. However, failure to induce lactation in the cows appeared to be due to low secretion of prolactin.

The metabolism of steroid hormones in breast cancer: a reappraisal.

MacMahon (1), Lemon (2) and Smith and Smith (3) have emphasized that abnormal ovarian function or steroid metabolism not modified by pregnancy in early life, may well be implicated in human breast cancer etiology. Wynder (4) and Dao (5) have pointed toward an etiological role for estrogen metabolism and nutritionally-conditioned alteration of estrogen metabolism in breast cancer, as also have Stocks (6) and Hems (7). The study of steroid excretion patterns in women with breast cancer (or prior to the onset of the disease) is further recommended by Bulbrook’s observations in the women of Guernsey (8). Furthermore, there is some evidence (9) that at least part of the genetic predisposition of certain mouse strains is the result of genetically-determined ovarian function. These are but a few of the reports, published in the past decade, that pertain to steroid hormone metabolism in mammary cancer. However, before reappraising the data in this field it may be helpful to summarize what is now known of the fate of steroid hormones in the mammal.

The investigation of ovarian function by measurement of urinary oestrogen and pregnanediol excretion.

Urinary oestrogen and pregnanediol excretion was measued daily ("daily monitoring") for a complete cycle in 20 normally menstruating women, in one patient with an anovulatory cycle and for 28 days in a patient with secondary amenorrhoea. The measurements were also performed on urine specimens collected at weekly intervals for 4 to 6 weeks ("weekly tracking") from 506 patients with evidence of abnormal ovarian function. These included 9 patients with primary amenorrhoea, 132 patients with secondary amenorrheoa, 138 patients with oligomenorrhoea and 227 patients with evidence of ovarian dysfunction and cycle lengths of 25 to 42 days. The results were subjected to statistical analysis. In the normal cycles, ovulation could be identified on the criteria of a rising pregnanediol value reaching or exceeding 2-0 mg. per 24 hours for a period of 7 days or more. Valid conclusions on the overall mean oestrogen and pregnanediol values for a complete cycle could be made from the results of weekly tracking, irrespective of which day the tracking commenced. Correlations were obtained by comparing the mean and maximum urinary oestrogen values and the variability of the values with the evidence of ovarian function indicated by the clinical classifications of the patients, the duration of the disorders and the subsequent occurrence of uterine bleeding. Mean oestrogen values of 10 mug. per 24 hours or less were associated with lack of ovarian function. For values higher than this a discriminant function based on both the mean oestrogen value and the variability of the oestrogen values was useful in predicting onset of spontaneous menstruation. A single urine specimen collected 4 to 8 days before onset of menstruation showing a raised pregnanediol value of 2-0 mg. per 24 hours or more provided a valid test for ovulation in women with regular cycles, and a single urine specimen giving an oestrogen value of 10 mug per 24 hours or less gave a valid indication of absent ovarian function in women with amenorrhoea for two years or more. In all other circumstances serial sampling at weekly intervals provided a valid assessment of ovarian activity. Application of these principles allows the greatest amount of information on ovarian function to be obtained with the greatest economy of effort.

ニジマス卵の過熟現象 Ｉ “過熟卵”の形態ならびに出現時期

It has been frequently observed that during the stripping operation of the female rainbow trout, some eggs known as “over-ripened ova” contain oil globules which form into a lump. Furthermore, the yolk of such egg is characterized by a semi-transparency. It was confirmed from the present study that these eggs are not the products of abnormal ovarian function, but instead characterize a condition caused by an extended in situ existence within the body cavity following ovulation. This process of over-ripening could be divided into four stages according to the following description: normal ova immediately following ovulation (stage I) early stage of over-ripening (staeg II), followed by over-ripened ova (stage III), which then transform into the irregularly shaped ova with considerable decrease in their material content (stage IV). The progressive formation of over-ripened eggs, stage II, III and IV eggs could be generally recognized at about the 10th, 30th and 35th day following ovulation, respectively. During this process of over-ripening, partial collapse of the cortical alveoli and formation of a blastoderm-like structure permiated with oil globules occured. In the course of water-induced activation of eggs, the collapse of the cortical alveoli was restricted and bipolar differentiation was retarded in over-ripened eggs.

Normal and delayed ovulation in the human.

Better understanding of ovulation and ovarian function control has been initiated by better understanding of the etilogy of abnormal ovarian function and recognition of its role in reproductive wastage. Ovulation time an important factor in respect to normal ova production depends upon the function of all levels in the cortico-hypothalamic-pituitary-ovarian axis. Dysfunction at any level could lead to delayed ovulation. Evidence suggests that fertilization can occur at any time although the midcycle is the most fertile phase. Data indicate that fertilization of an overripe ovum beyond its full maturation may result in an increase of pregnancy abnormalities. Conversely fertilization of an unripe egg may have similar implications. Also affecting ovulation time is delayed endometrial development associated with shortened secretory phase. Postponed migration and implantation and subsequent delayed chorionic gonadotropin hormone production occur following fertilization of a belatedly shed ovum resulting in simulation of normal menstruation despite the fertilized ovums presence. Also discussed is corpus luteum defect and the above phenomena associated with various forms of reproductive wastage.

IN VITRO PRODUCTION OF ANDROGENS BY A VIRILIZING ADRENAL ADENOMA AND ASSOCIATED POLYCYSTIC OVARIES.

ABSTRACT A case of a virilizing adrenal adenoma in which bilateral Stein-Leventhal ovaries were found is presented. Despite the morphologic similarity, these ovaries did not possess the in vitro potential for increased androgen production demonstrated by polycystic ovaries removed from patients without apparent adrenal disease. These findings lend support to the thesis that, regardless of the organ source, increased androgen production leads to abnormal ovarian function reflected morphologically in follicle maturation arrest.