Reduced preovulatory granulosa cell steroidogenesis in women with endometriosis.

To examine the cause of altered follicular fluid steroid levels and lower in vitro fertilization rate observed in infertile women with minor endometriosis, we have compared the production of estradiol (aromatase activity) and progesterone of freshly isolated granulosa cells (3h. incubation) from such women and a control group with tubal or unexplained infertility, having IVF during unstimulated or gonadotropin-stimulated cycles. As previously observed, mature oocytes from women with endometriosis had a reduced fertilization and cleavage rate in vitro in unstimulated cycles (19/37[51%] vs. 69/94[73%], p < 0.05) and stimulated cycles (20/37[57%] vs. 32/39[82%], p < 0.01). Median [95%CI] basal aromatase activity was lower in endometriosis compared with control in unstimulated cycles (2.84[2.03-3.49] pmol E2/10(3) cells/3h, n = 31 vs. 3.63[2.72-3.49], n = 55, p = 0.057) and stimulated cycles (0.31[0.16-0.50], n = 14 vs. 0.99[0.70-1.52], n = 20, p < 0.001). Progesterone production followed a similar pattern in unstimulated (0.56[0.50-0.89] pmol/10(3) cells/3h, n = 29 vs. 1.23[0.69-1.54], n = 52,) and stimulated (0.37[0.20-0.73], n = 16 vs. 0.95[0.72-1.17], n = 21) cycles (p < 0.05). Addition of FSH, LH or hCG (30ng/mL) to the incubation medium enhanced progesterone production 2 to 3-fold, but had no effect on aromatase activity. Our results indicate a defect in granulosa cell steroidogenesis associated with endometriosis, which could affect oocyte function and explain the reduction in fertilizing capacity and subsequent competence of the corpus luteum, and the associated subfertility.

Female infertility.

This article has provided outcome-based evidence using easily understood graphic representation of cumulative pregnancy rates whenever possible for the methods used to investigate and treat female infertility. A scheme of basic routine investigations in specialist practice is developed and clear guidance provided on the choice of treatment for each couple.

Higher gonadotrophin surge-attenuating factor bioactivity is found in small follicles from superovulated women.

Ovine and rat pituitary bioassays for gonadotrophin surge-attenuating factor (GnSAF) were utilized to determine whether the level of GnSAF bioactivity in pooled human follicular fluid (hFF) from superovulated women varied according to follicle diameter (< or = 11 mm, 12-15 mm and 16-21 mm follicles examined using the ovine bioassay, or < or = 10 mm, 11-13 mm, 14-17 mm, 18-20 mm, 21-24 mm and > or = 25 mm follicles examined using the rat bioassay). When tested using dispersed ovine pituitary cells, GnSAF bioactivity, expressed in terms of the reduction in gonadotrophin-releasing hormone (GnRH)-induced LH secretion, was inversely related to follicle diameter (P < 0.01). In response to 5 microliters hFF/well from follicles of < or = 11, 12-15 and 16-21 mm diameter, GnRH-induced LH secretion was reduced to 40.5 +/- 6.9%, 65.2 +/- 6.6% and 83.7 +/- 7.9% of control respectively. A similar inverse relationship was observed when a second batch of hFF samples from different sized follicles was tested using rat pituitary cell monolayers. Expressing GnSAF bioactivity in terms of the dose required to suppress GnRH-induced LH secretion by rat pituitary cells to 50% of the maximal suppression observed (ED50), the three smallest follicle size pools contained the most GnSAF (ED50 values of 0.13, 2.79 and 5.36 microliters hFF/well from follicles of < or = 10, 11-13 and 14-17 mm respectively). The ED50 values for follicles of 18-20, 21-24 and > or = 25 mm were 8.81, 27.1 and 60.0 microliters hFF/well respectively. Thus hFF from follicles < or = 11 mm was over 450 times more potent than hFF from follicles > or = 25 mm in suppressing GnRH-induced LH release. The ED50 values for inhibin bioactivity (measured as the suppression of basal FSH secretion from rat pituitary monolayers) were much less variable than those for GnSAF bioactivity (between 0.85 and 0.13 microliters hFF/well). Inhibin immunoreactivity, measured by a two-site immunoradiometric assay, followed the same pattern as inhibin bioactivity with lowest concentrations in the smallest follicles (41.96 ng/ml) and highest concentrations in the three largest follicle size groups (56.48-64.48 ng/ml). The specific effects of inhibin on GnRH-induced LH and basal FSH release in these pituitary bioassays were determined by incubating culture dishes with pure recombinant human inhibin at doses of 0.025-25 ng/well. In both the sheep and rat pituitary monolayers, basal FSH was suppressed (ED50 = 0.02 and 0.16 ng/well respectively).(ABSTRACT TRUNCATED AT 400 WORDS)

Intraepithelial vaginal neoplasia following immunosuppressive therapy treated with topical 5-Fu.

A case is reported of the successful treatment of multifocal intraepithelial neoplasia of the vagina by topical application of a cytotoxic agent, 5-fluorouracil. These lesions followed prolonged immunosuppressive therapy after renal transplantation. Intraepithelial neoplasia was present also in the cervix and was treated by conization.

Use of follicle-stimulating hormone alone (urofollitropin) to stimulate the ovaries for assisted conception after pituitary desensitization.

OBJECTIVE: Analyse outcome of FSH therapy alone after pituitary desensitization for assisted conception to compare with published results of conventional hMG. DESIGN: Descriptive study of complete series of patients undergoing IVF-ET or GIFT treatment using the chosen drug protocol without exception. SETTING: University private IVF-ET clinic within a comprehensive fertility service. PATIENTS: All couples (n = 773), including 10% > or = 40 years old, treated by IVF-ET or GIFT (1,097 cycles started and 1,012 attempted egg recoveries) for mainly tubal pelvic infective damage (36% of cycles), endometriosis (16%), sperm disorders (14%) and prolonged unexplained infertility (34%) during 3 calendar years, 1990 to 1992. INTERVENTIONS: Ovarian stimulation using FSH alone (urofollitropin, Metrodin; Serono Laboratories Limited, Welwyn Garden City, United Kingdom) after pituitary desensitization using buserelin acetate nasal spray (Suprefact; Hoechst, Hounslow, United Kingdom) from the previous midluteal phase, monitored by ultrasonography and serum E2 measurements, followed by standard IVF-ET or GIFT treatment methods limited to the transfer of no more than three embryos or eggs. OUTCOME MEASURES: Rates per cycle started of cancellation of egg recovery, failure of egg recovery, clinical pregnancy (ultrasound detection of sac), livebirths, and cumulative pregnancy rates (PR) and birth rates. RESULTS: In women < 40 years old and men with favorable sperm (77% of couples and 84% of cycles) the cycle cancellation rate of egg recovery was 7%; attempted egg recovery was successful in every case. For IVF-ET the clinical PR per started cycle and the livebirth rate were 27% and 23%, respectively, and for GIFT 39% and 33%, respectively. The four-cycle cumulative PR by either treatment was 77% and livebirth rate 68%. In women > 40 years old, the cycle PR and birth rate were 14% and 8%, respectively. In cases of sperm disorder the rates in women < 40 years old were 17% and 14%, respectively, and > 40 years old were 18% and 0, respectively. CONCLUSIONS: By comparison with the best worldwide results of assisted conception employing pituitary desensitization, the findings demonstrate that FSH alone to stimulate the ovaries is fully effective and highly successful, and supplementation with LH is not needed.

The age-related decline in female fecundity: a quantitative controlled study of implanting capacity and survival of individual embryos after in vitro fertilization.

OBJECTIVE: To determine strictly comparable rates per embryo of implantation and birth of a baby related to the woman's age, which would be representative of natural fertility at least in relative terms. DESIGN: Comparative study of IVF-ET results controlling for confounding variables including cause and duration of infertility, history of previous pregnancy, hormonal treatment, rank cycle of treatment, and numbers of embryos transferred and available. SETTING: University comprehensive fertility service. PATIENTS: All couples (n = 561) in their first cycle of treatment reaching oocyte collection, women with normal uterus and ovulatory cycles, and men with normal sperm. INTERVENTIONS: Standardized methods of pituitary desensitization, ovarian stimulation, and IVF-ET, and maximum of three embryos transferred. MAIN OUTCOME MEASURES: Oocytes, pregnancies, and live births per cycle; fertilization and cleavage rates; embryo implantation and live baby rates. RESULTS: The numbers of oocytes and consequent embryos declined with age but fertilization and cleavage rates rose slightly. Embryo implantation rates were reduced when no more than three embryos were available (9.3 percent), especially in women aged 35 to 39 years (6.2 percent) or older compared with four or more embryos (17.1 percent) but were equally low in all women over 40 years even with more embryos (6.1 percent). In the age bands 25 to 29, 30 to 34, 35 to 39, and 40 to 44 years, the rates per embryo of implantation were 18.2 percent, 16.1 percent, 15.3 percent, and 6.1 percent, respectively, and of a live baby were 15.7 percent, 12.1 percent, 12.0 percent, and 3.5 percent. CONCLUSIONS: Embryo implanting ability and survival decline gradually after 30 years of age, but by more than two thirds after 40 years and in younger women with reduced ovarian capacity.

Onset of the preovulatory luteinizing hormone surge: diurnal timing and critical follicular prerequisites.

OBJECTIVE: To determine the diurnal variation in the onset of the preovulatory LH surge in women. DESIGN: Prospective open cohort study. SETTING: University hospital research program. PATIENT(S): Thirty-five women with infertility resulting from tubal damage that was associated with minor endometriosis or with infertility of prolonged unexplained etiology. INTERVENTION(S): Women underwent transvaginal ultrasonography and serum E2 estimation daily during monitored cycles before unstimulated natural cycle IVF: exogenous gonadotropins were not administered. MAIN OUTCOME MEASURE(S): Serum E2 concentration, follicle diameter, and endometrial thickness. RESULTS: Of 169 cycles. 155 progressed to an ovulatory LH surge, of which 146 occurred within 8 hours of assessment of the outcome measures. The relationship between follicle diameter and E2 was weak, but an abnormal value for one always was countered by a normal value for the other. CONCLUSIONS: Most women begin the preovulatory LH surge between midnight and 8:00 A.M., but with no particular variation by day of the week. The relationship between follicle size and serum E2 is not sufficiently strong to predict the LH surge confidently on the basis of only one variable, but the LH surge is unlikely to occur before either the follicle diameter has reached 15 mm and/or the serum E2 level has reached 600 pmol/L.

Delayed conception and active and passive smoking. The Avon Longitudinal Study of Pregnancy and Childhood Study Team.

OBJECTIVE: To determine whether passive as well as active smoking by women or smoking by men is associated with delayed conception, after adjustment for confounding factors. DESIGN: Population study of couples expecting a baby. Logistic regression was performed to identify factors associated with delayed conception. SETTING: The Avon Health Authority area, United Kingdom. PATIENT(S): All couples expected to deliver between April 1991 and December 1992. INTERVENTION(S): Questionnaires administered early in pregnancy. MAIN OUTCOME MEASURE(S): Time taken to conceive, categorized as <6 months, 6-11 months, 1-3 years, and >3 years. RESULT(S): After correction for confounding factors, delayed conception was statistically significantly associated with both active smoking by the woman (odds ratio [OR] 1.23 [95% CI 0.98-1.49] for > 6 months and 1.54 [95% CI 1.19-2.01] for >12 months) and her exposure to passive smoking (OR 1.17 [95% CI 1.02-1.37] and 1.14 [95% CI 0.92-1.42]) compared with women with no exposure to tobacco smoke (referent). Heavy smoking by men was independently associated with delayed conception. In active smokers, the effect increased with the number of cigarettes. CONCLUSION(S): Smoking by men and passive and active smoking by women are associated with delayed conception.

Relationship between midcycle luteinizing hormone surge quality and oocyte fertilization.

OBJECTIVE: To determine whether alterations in preovulatory follicular fluid (FF) levels of LH, FSH, and steroids are associated with the probability of fertilization. DESIGN: Retrospective analysis of prospective study results. SETTING: Reproductive medicine clinic of a university teaching hospital. PATIENT(S): Infertile women, with unstimulated, apparently regular cycles in an IVF research program. INTERVENTION(S): Measurement of preovulatory FF levels of LH, FSH, E2, and P and serum LH levels by fluoroimmunometry. MAIN OUTCOME MEASURE: Oocyte fertilization. RESULT(S): There were 84 transferable embryos (rate of normal fertilization and cleavage, 67%), and 41 oocytes (33%) failed to fertilize. Analysis of the matched FF showed that the median concentration of FF LH was significantly higher for cleaving embryos than for unfertilized oocytes (14.6 vs. 10.4 IU/L). Serum LH concentrations were similarly higher in cycles with cleaving embryos. There were no statistically significant differences in FF concentrations of FSH, E2, or P in the two groups. CONCLUSION: Reduced preovulatory FF LH levels are associated with impaired fertilization of oocytes in vitro, despite normal FF FSH and steroid levels.

The value of a single serum progesterone measurement in the midluteal phase as a criterion of a potentially fertile cycle ("ovulation") derived form treated and untreated conception cycles.

A single midluteal serum progesterone concentration was obtained in 212 untreated cycles in 113 infertile patients, including 138 cycles in 72 patients in whom tubal, seminal, and cervical causes of infertility had been excluded. There were 16 conception cycles in the latter group. In an extended study a total of 21 untreated singleton conception cycles have been observed with a mean progesterone value of 40.7 nmol/l (12.8 ng/ml), 95% confidence limits of 28 to 53 nmol/l (8.8 to 16.7 ng/ml), and a range of 27 to 53 nmol/l (8.5 to 16.7 ng/ml), which extended significantly above as well as below the conception range, indicating that there is an optimal range for fertility with both an upper and a lower limit. The lower limit is of greater practical importance; and, partly to allow for assay variation, we suggest it should be taken as 30 nmol/l (9.4 ng/ml). It provided a clinically reliable criterion of potential fertility ("ovulation") in related studies. Our findings in treated conception cycles suggest that a higher value may be needed after treatment with clomiphene or gonadotropins because of the contribution from other stimulated follicles.

Post-pill amenorrhea: a causal study.

The distribution of distinguishable disorders in 102 patients with post-pill amenorrhea has been compared with that in 166 other patients with secondary amenorrhea, including 123 patients who had never used oral conception. In the post-pill group there was a 13% to 21% excess of patients who had a functional disorder without any weight loss or psychological disturbance to account for it and who typically were estrogenized and demonstrated intact feedback systems by their ovulatory response to clomiphene (the so-called cycle initiation defect). This excess might be explained by biased selection of oral contraception, but it seems wiser to assume a causal relationship, which amounts to a true risk of post-pill amenorrhea in less than 1 per 1000 users. No risk was revealed of primary ovarian failure, hyperprolactinemia, or pituitary tumor.

Periovulatory human oocytes, cumulus cells, and ovarian leukocytes express type 1 but not type 2 11beta-hydroxysteroid dehydrogenase RNA.

OBJECTIVE: To further elucidate cortisol metabolism in the follicular microenvironment at the time of oocyte retrieval, the presence of 11beta-hydroxysteroid dehydrogenase (HSD) messenger (m)RNA transcripts in oocytes; cumulus cells; granulosa cells; and CD45(+), CD15(+) leukocytes was assessed semiquantitatively. DESIGN: Controlled study using semiquantitative assessment of 11beta-HSD mRNA. SETTING: University IVF center. PATIENT(S): Twenty-six patients undergoing controlled ovarian hyperstimulation for assisted conception. INTERVENTION(S): Metaphase II oocytes; cumulus cells; granulosa cells, and CD45(+), CD15(+) leukocytes from individual follicular fluid aspirates. MAIN OUTCOME MEASURES: Semiquantitative analysis of PCR products after total RNA extraction and complementary DNA synthesis. RESULT(S): Periovulatory human oocytes; cumulus cells; CD45(+), CD15(+) leukocytes; and granulosa cells consistently express type 1 but not type 2 11beta-HSD mRNA. Expression of mRNA is greatest in cumulus cells. Type 1 11beta-HSD mRNA expression varies considerably in all cell types and among individual follicles and patients. CONCLUSION(S): These studies of mRNA expression suggest that the enzymes present both in and around the periovulatory oocyte will favor a high-cortisol environment.

Urinary total oestrogen values determined by an automated method in normal and abnormal pregnancies.

Urinary total oestrogen values in late pregnancy have been determined by an automated, fluorimetric method based on the Kober reaction and Ittrich extraction. The normal range was obtained from 360 samples from 56 patients. The mean value at 40 weeks gestation was 52.6 mg/24 h and the 95 per cent probability bounds were 29.1 and 95.3 mg/24 h, obtained after initial calculations on logarithm values. "Small for dates" foetuses in good condition at birth were associated with oestrogen values generally below the normal mean. They were usually well discriminated from foetuses that died in the uterus. These were always associated with oestrogen values below or approaching 10 mg/24 h before death except when growth had been normal and death was due to mechanical complications at delivery.

Serum prolactin concentrations related to copper or inert intrauterine devices (IUDs) in women.

A report that serum prolactin concentrations were almost doubled by the use of copper IUDs led us to study the effect of both copper and inert IUDs. We studied 105 normal women in whom all other prolactinergic factors had been excluded; 25 of them were using copper IUDs, 25 were using inert IUDs and there were 55 controls. No differences were found in the mean prolactin levels between these groups (4.3, 4.5 and 4.3 microgram/l, respectively). We conclude that copper and inert IUDs do not affect prolactin secretion.

PIP: The results of a cross-sectional study designed to determine the effects of IUD material (i.e., copper or inert) on prolactin levels in serum are presented. A consecutive series of 105 healthy volunteers were studied. Prolactin was measured by radioimmunoassay. The serum prolactin determinations obtained are shown figuratively; there were no significant differences between the geometric mean values in women (n=25) with copper (4.3 mcgm/liter) or inert IUDs (n=25) (4.5 mcgm liter) or controls (4.3 mcgm); nor were differences discernable between the combined IUD groups (4.4) compared with controls (P .86 in all cases). Copper and inert IUDs do not affect prolactin secretion.

Effectiveness of infertility treatments: choice and comparative analysis.

Choice of infertility treatments usually depends on a balance of the chances of conceiving with or without treatment, and with more or less complex and costly treatments, and on other factors such as duration of infertility and the woman's age. Pregnancies often occur independent of treatment and prospective controlled trials are needed. Comparability between reported results requires pregnancy and birth rates to be calculated in a time-specific or cycle-specific way. Cumulative rates are preferable to account for the usual tendency of fecundity to fall progressively. This review focuses on such published data in order to assess the relative effectiveness of treatments, both conventional and assisted conception methods. The main conclusions are: (1) The only treatments that can achieve a normal chance of pregnancy are the ovulation induction methods in cases of oligomenorrhea/amenorrhea, and the assisted conception methods for other female causes and unexplained infertility. (2) Tubal/pelvic infective damage and endometriosis require new severity classifications which are sensitive to functional potential before and after surgery, and in vitro fertilization (IVF) would often be indicated as the primary choice. (3) Duration of unexplained infertility determines the need and therefore benefit of any of the treatments used, of which the assisted conception methods are by far the most effective. (4) Interpretation of reported results of treatments for 'male factor' infertility is critically affected by the diagnostic accuracy of defining sperm dysfunction. (5) In cases of well-defined sperm dysfunction there is little or no therapeutic benefit to the chance of natural conception, nor by intrauterine insemination; there is moderate success by IVF, but no proven benefit over standard IVF by any micromanipulative method except probably intracytoplasmic sperm injection.

Expectations of assisted conception for infertility.

OBJECTIVE: To provide reliable prognostic information for couples seeking assisted conception. DESIGN: Analysis of four years' practice (1988-91). SETTING: Private university service linked with NHS reproductive medicine services. PATIENTS: 804 couples with various causes of subfertility, median duration five years, median age of women 34 years. INTERVENTIONS: 1280 completed cycles: 950 in vitro fertilisation, 144 gamete intrafallopian transfer, and 186 intrauterine insemination and superovulation. MAIN OUTCOME MEASURES: Pregnancy and birth rates per cycle and cumulative pregnancy and take home baby rates per couple. RESULTS: In women under 40 years and men with normal sperm, whatever the cause of infertility, results with in vitro fertilisation improved steadily reaching a pregnancy rate per cycle of 30% (95% confidence interval 26% to 35%) during 1990-1 and birth rate per cycle of 29% (23% to 35%) in 1990. Pregnancy and birth rates for gamete intrafallopian transfer were 36% (28% to 44%) and 26% (17% to 37%) and for intrauterine insemination 18% (12% to 24%) and 16% (10% to 22%). After six cycles cumulative probability of pregnancy was 82% and cumulative take home baby rate 70%. Considering only in vitro fertilisation and gamete intrafallopian transfer after four cycles the pregnancy rate was 78% (66% to 91%). CONCLUSIONS: Conception is less likely in women over 40 and men with sperm dysfunction. For other couples the prognosis for a live birth is at least as good as for fertile couples if they persist with treatment.