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Plasma steroid and gonadotrophin profiles in ovulatory but infertile women.

A group of twelve patients with a history of infertility lasting at least 2 years despite evidence of regular ovulatory cycles, were selected from an Infertility Clinic after thorough investigation had revealed no other obvious cause for their infertility. Daily plasma gonadotrophin and steroid concentrations were measured in twenty cycles from these women and the results compared with the hormonal profiles obtained from a group of eleven control 'normal' women. No significant differences were found in the concentrations of LH, FSH or oestradiol although plasma progesterone concentrations were significantly lower in the infertile women, particularly during the first half of the luteal phase. In the absence of any other demonstrable defect, it was assumed that the decreased progesterone secretion was directly or indirectly implicated in the pathogenesis of their infertility, and a number of therapeutic regimes (progesterone supplementation of the luteal phase or additional stimulation by either anti-oestrogens or exogenous gonadotrophins) were assessed for their efficacy in correcting this defect. Although plasma progesterone concentrations after most of these regimes were significantly increased, none of the patients in this series conceived.

Adult↗

The role of steroid signals in the maturation of mammalian oocytes.

The resumption of meiosis in mammalian oocytes is associated with sequential changes in follicular steroidogenesis, the principal features of which are an initial stimulation and then suppression of the secretion of oestrogens and androgens, followed by a steady increase in the secretion of progesterone. The addition of steroid enzyme inhibitors to isolated follicles in vitro alters the normal profile of steroids secreted during maturation and induces intracellular changes in the oocytes which are expressed as nuclear abnormalities at fertilization. Different abnormalities are induced by selectively modifying steroid biosynthesis during maturation, the maturing oocyte being more sensitive to an imbalance in the steroid profile rather than the total inhibition of steroid secretion. The cause of the developmental aberrations may either result from or be associated with the abnormal patterns of proteins synthesized by oocytes denied the support of the correct balance or sequence of steroids during maturation.

Androgens↗

DNA methylation profiles of female steroid hormone-driven human malignancies.

Tumor DNA contains valuable clues about the origin and pathogenesis of human cancers. Alterations in DNA methylation can lead to silencing of genes associated with distinct tumorigenic pathways. These pathway-specific DNA methylation changes help define tumor-specific DNA methylation profiles that can be used to further our understanding of tumor development, as well as provide tools for molecular diagnosis and early detection of cancer. Female sex hormones have been implicated in the etiology of several of the women's cancers including breast, endometrial, ovarian, and proximal colon cancers. We have reviewed the DNA methylation profiles of these cancers to determine whether the hormonal regulation of these cancers results in specific DNA methylation alterations. Although subsets of tumors in each of these four types of cancers were found to share some DNA methylation alterations, we did not find evidence for global hormone-specific DNA methylation alterations, suggesting that female sex hormones may participate in different tumorigenic pathways that are associated with distinct DNA methylation-based molecular signatures. One such pathway may include MLH1 methylation in the context of the CpG island methylator phenotype.

Adaptor Proteins, Signal Transducing↗

Effects of cyproterone on the steroid biosynthesis in the human ovary in vitro.

To examine the direct effects of cyproterone on the ovarian biosynthesis of steroids, tissue slices obtained from two cyclically functional human ovaries were incubated with [4-14C]pregnenolone as precursor. The steroid production was determined by measuring the concentration of eleven 14C-labelled steroids in the medium at the end of a 3-h incubation. Under control conditions, the in vitro-synthesis of 17 alpha-hydroxypregnenolone, dehydroepiandrosterone, androstenediol (5-ene-3 beta-hydroxy-steroids) and androstenedione represents the characteristic profile of steroids of the ovary from the follicular phase. The addition of cyproterone to the incubation medium inhibited the biosynthesis of androstenedione, whereas 17 alpha-hydroxypregnenolone and dehydroepiandrosterone increased. The in vitro-synthesis of progesterone, 17 alpha-hydroxyprogesterone, oestrone and oestradiol-17 beta represents the characteristic profile of steroids of the luteal ovary. The biosynthesis of progesterone, 17 alpha-hydroxyprogesterone and oestradiol-17 beta is inhibited by cyproterone. The principal manifestation of the effects of cyproterone on the two different profiles of steroids is an apparent inhibition of the 3 beta-hydroxysteroiddehydrogenase-delta 5-4-isomerase activity. It is noteworthy, that the effects of the "pure antiandrogen" cyproterone are similar to those effects previously published for progestagens (chlormadinone acetate, norethisterone acetate and D-norgestrel). A simplified concept of the direct effects of cyproterone and progestagens on the steroidogenesis in the human ovary is proposed. The results indicate, that cyproterone, in addition to its well-known androgen receptor blocking effect, act directly on the steroid biosynthesis in the human ovary in vitro.

Androgens↗

[Sex steroids versus lipid profile and the degree of coronary artery stenosis in men with angiographically documented coronary artery disease].

The aim of the study was to assess the correlations between the levels of sex hormones and blood lipid profile as well as indexes of coronary artery stenosis in men with angiographically documented coronary artery disease. 111 men, aged 36-73 yrs (av. 55) were studied. In all the patients levels of testosterone (T), dehydroepiandrosterone sulfate (DHEA-S), estradiol (E), SHBG, LH and FSH were measured. The level of bioavailable testosterone (BT) was calculated knowing SHBG level. Total cholesterol (TCh), HDL-cholesterol, LDL-cholesterol and triglycerides (TG) levels were estimated as well as the degree of coronary artery stenosis was estimated by means of modified indexes. For statistics R-Spearman test was used. Summing coronary stenosis index correlated significantly with T-Ch and LDL-Ch levels. Positive correlation was found between blood level of E and TCh as well as between E and LDL-Ch. BT correlated partially with LDL-Ch level. No correlations were found between the levels of T, SHBG, DHEA-S, FSH, LH and lipid profile. The level of DHEA-s revealed negative correlation with age, while the level of SHBG increased with ageing leading to the decrease of the value of BT but not total T. None of studied hormones correlated with coronary indexes. Our results suggest that estradiol and BT may promote the formation of atherogenic lipid profile leading to atherosclerosis in men.

Adult↗

Calbindin-D28k immunoreactivity is a marker for a subdivision of the sexually dimorphic nucleus of the preoptic area of the rat: developmental profile and gonadal steroid modulation.

Calbindin-D28k (calbindin) is a 28 kilodalton calcium binding protein which potentially plays a role in neuroprotection. We report here the normal development and gonadal steroid modulation of a sexually dimorphic group of calbindin immunoreactive cells within the sexually dimorphic nucleus of the preoptic area (SDN) which we call the calbindin-immunoreactive SDN or CALB-SDN. Beginning on PN2, a faintly immunoreactive CALB-SDN is present, however, the volume is not sexually dimorphic. On PN4, the staining of the CALB-SDN appears more robust but the volume is still not sexually dimorphic. By PN8 and extending through PN12 and PN26, the latest age analysed, the volume of the CALB-SDN is larger in males by two- to fourfold. Cresyl violet counterstain reveals a similar developmental profile of the SDN as well as clusters of darkly staining calbindin immunonegative cells which lie around the CALB-SDN. Castration of males on PN0 decreases the volume of the CALB-SDN by PN12 and administration on the day of birth and PN1 of either testosterone propionate or oestradiol benzoate, but not dihydrotestosterone propionate to females increases the volume of the CALB-SDN by PN12. By demonstrating the sexual dimorphism and gonadal steroid modulation of the CALB-SDN, we hereby establish that calbindin is a specific marker of a subdivision of the SDN and can be used as such in future studies.

Aging↗

Infertility in women with apparently ovulatory cycles. I. Comparison of their plasma sex steroid and gonadotrophin profiles with those in the normal cycle.

Serial daily plasma profiles of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone, 17alpha-hydroxyprogesterone and oestradiol 17beta throughout single menstrual cycles from seven patients with a history of infertility but with normal menstrual rhythm and no apparent ovarian dysfunction were compared with those from apparently normally menstruating women. This comparison showed that the infertile group produced poor corpora lutea after poor follicular growth. The poor follicular growth may be related to depressed pituitary release of FSH in response to a negative feedback effect of elevated progesterone levels.

Anovulation↗

Suspending agent effects on steroid suspension dissolution profiles.

Dissolution profiles and particle-size analyses were determined for two lots of prednisolone acetate. The effects of common suspending agents on dissolution and particle-size distributions of these suspensions also were investigated. Lot-to-lot variation in the prednisolone acetate dissolution rate was observed and was apparently related to the percentage of fine particles within the distribution. Carboxymethylcellulose sodium inhibition of prednisolone acetate dissolution occurred with only one lot of raw material and seemed to be related to aggregation of the fine particles. Hydroxypropyl methylcellulose inhibited both prednisolone acetate lots and was observed with or without small particle aggregation. The dissolution variations observed have important implications in suspension formulation.

Excipients↗

Infertility in women with apparently ovulatory cycles. II. The effects of clomiphene treatment on the profiles of gonadotrophin and sex steroid hormones in peripheral plasma.

Six infertile patients had been studied previously (Dodson et al., 1975b) and were shown to produce inefficient corpora lutea which appeared to be predetermined by ovulation of "poorly grown" follicles. In a second cycle these infertile patients were treated with 50 mg of clomiphene per day for five days. The resulting plasma sex steroid and gonadotrophin profiles were compared with those found before treatment and with the profiles in normal patients (Dodson et al., 1975a). Treatment with clomiphene appeared to produce increased follicular growth and more active corpora lutea.

Clomiphene↗

Tissue-specific transcription profiles of sex steroid biosynthesis enzymes and the androgen receptor.

17beta-hydroxysteroid dehydrogenase (17beta-HSD) and 5alpha-reductase isoenzymes play a crucial role in the formation and metabolism of sex steroids. Not only the key androgens testosterone and dihydrotestosterone but also their precursors are potent activators of the androgen receptor and are, therefore, likely to act as determinants of male sexual differentiation and maturation in a differentially regulated way. The aim of the present study was to relatively quantify the expression of the mRNA of 17beta-HSD isoenzymes, namely, type 1, 2, 3, 4, 5, 7, and 10, together with the 5alpha-reductase type 1 and 2, and the androgen receptor in normal human males and females. RNA was isolated from peripheral blood cells of both sexes and from genital skin fibroblasts (GSFs) of two different localizations (foreskin and scrotal skin) obtained from phenotypically normal males. mRNA expression was semi-quantified by quantitative reverse-transcriptase polymerase chain reaction with the LightCycler Instrument (Roche). The examined enzymes show statistically significant differences in their transcription pattern between the blood and the GSF RNA samples. Within the GSF samples, there are also significant variations between the two examined localizations in the transcription of 17beta-HSD type 1, 2, 4, and 5 as well as for the androgen receptor. We found large interindividual variation of enzyme transcription patterns in all investigated tissues. In peripheral blood cells, no sex-specific differences were seen. We conclude that sex steroid enzymes are expressed not only in genital primary target tissues but also in peripheral blood. The expression in different target tissues may contribute to both the individual sexual and tissue-specific phenotype in humans.

17-Hydroxysteroid Dehydrogenases↗

Biological profile of the steroid 5 alpha-stigmastane-3 beta, 5, 6 beta-triol-3-monobenzoate.

The steroid 5 alpha-stigmastane-3 beta, 5, 6 beta-triol-3-monobenzoate at a dose of 40 mg/kg/day showed significant antifertility activity when administered orally in a single or in consecutive doses during pre-implantation stage of pregnancy in mice. In the rabbit interceptive activity was obtained when the test steroid was administered on Day 8 of pregnancy in a single dose level. The compound failed to show any distinct estrogenic or androgenic property following bioassay method. But in McPhial test, the compound showed some progestagenic activity.

Animals↗

Seasonal profiles of sex steroids in blood plasma and ovarian tissue of Clarias batrachus.

Sex steroids were measured in plasma and ovaries of the catfish, Clarias batrachus, mainly over the course of their annual reproductive cycle. Plasma testosterone levels increased in a stepwise fashion, the first significant increase occurring in the previtellogenic phase (March) and the second in the early vitellogenic phase (May). The levels remained elevated till the early postvitellogenic phase (July) and reached basal levels in the regressed phase (September). Estradiol 17 beta and estrone were at low levels in the previtellogenic phase, increased rapidly in the vitellogenic phase, and attained their peaks in the late vitellogenic phase. Ovarian testosterone levels were very low without any significant change from the late previtellogenic to the postvitellogenic phases (April to August). Estradiol-17 beta and estrone levels in the ovary increased significantly in the previtellogenic phase (April), reached a peak in the vitellogenic phase (June), and declined in the postvitellogenic period (July). Ovarian levels of sex steroids were always lower than plasma levels, indicating low storage and rapid secretion. Gonadosomatic index (GSI) was low in the regressed and previtellogenic phases and increased in the vitellogenic and postvitellogenic phases, with a peak in July. Plasma estradiol-17 beta levels could be correlated with GSI from May to July, and plasma estrone and testosterone levels with GSI in early June and July, respectively.

Animals↗

Analysis of profiles of unconjugated steroids in rat testicular tissue by gas chromatography-mass spectrometry.

A gas chromatographic-mass spectrometric (GC-MS) method for analysis of unconjugated steroids in a rat testis is described. A combined solvent-solid extraction procedure, utilizing Lipidex 1000 and Sep-Pak C18, gives a 25-fold purified extract. Steroids in this extract are fractionated by straight phase high-performance liquid chromatography (HPLC) on a LiChrosorb DIOL column in n-hexane-2-propanol, 92:8 (v/v). Four fractions are collected and the steroids are converted to tert-butyldimethylsilyl (TBDMS), 3-enol-TBDMS, and mixed TBDMS-trimethylsilyl (TMS) derivatives using TBDMS- and TMS-imidazole with sodium formate as catalyst under conditions suitable for the steroids present in the respective fractions. The derivatives are purified by reversed phase HPLC in 100% methanol and are analyzed by GC-MS, using selected ion monitoring of the major ions of high mass. For quantification, a mixture of known amounts of ten 14C-labelled steroids, [3H]estradiol and [2H3]estradiol are added to the testis homogenate. The mean concentrations (ng/g wet wt) of the twelve steroids determined were: 4-androstene-3, 17-dione, 4.0; testosterone, 127; 17 beta-hydroxy-5 alpha-androstan-3-one, 4.5; 5 alpha-androstane-3 alpha, 17 beta-diol, 5.7; 5 alpha-androstane-3 beta, 17 beta-diol, 1.5; progesterone, 5.5; 17 alpha-hydroxyprogesterone, 14.4; 3 beta-hydroxy-5-androsten-17-one, 0.07; 5-androstene-3 beta, 17 beta-diol, 0.25; 3 beta-hydroxy-5-pregnen-20-one, 10.3; 3 beta, 17 beta-dihydroxy-5-pregnen-20-one, 0.95; and estradiol, 0.025. Variations between animals were large whereas testes from the same animal in most cases had similar steroid concentrations.

Animals↗

Androgen metabolism assessment by routine gas chromatography/mass spectrometry profiling of plasma steroids: Part 1, Unconjugated steroids.

Using gas chromatography/mass spectrometry we have developed a method for the simultaneous determination of six plasma steroids: testosterone, 4-androstenedione, 17 alpha-hydroxyprogesterone, 5 alpha-androstane-3 alpha,17 beta-diol, 5 alpha-dihydrotestosterone, and dehydroepiandrosterone. For each analyte, a deuterium-labeled internal standard was used for quantification. Due to the high isotopic purity of our standards, no complex corrections for isotope contributions were necessary. The procedure provides a sensitive and specific technique with good accuracy and precision.

17-alpha-Hydroxyprogesterone↗

Alteration of urinary profiles of endogenous steroids and polyunsaturated fatty acids in thyroid cancer.

To explore the possible involvement of a relationship between steroids and polyunsaturated fatty acids (PUFAs) in thyroid cancer, we studied the concentration levels of 21 endogenous urinary androgens and glucocorticoids, and 6 urinary PUFAs in female patients with thyroid cancer (n=29, 49.4+/-12.6 years) and in normal female subjects (n=20, 48.3+/-11.9 years). Using gas chromatography-mass spectrometry in selected ion-monitoring mode, we performed quantification of androgens, glucocorticoids, and PUFAs. In the case of the urinary androgens and the glucocorticoids, the concentration of androgens was not altered between patients with thyroid cancer and normal controls. However, the concentration of glucocorticoid significantly decreased in patients with thyroid cancer compared to that of the normal controls. Also, in case of urinary PUFAs, the precursor of cholesterol, the concentration of linoleic acid, arachidonic acid, and docosahexaenoic acid (DHA) were decreased in patient with thyroid cancer (P<0.05). In this results, it can be concluded that the glucocorticoids milieu induced from the decreasing urinary concentration of DHA may exert an important influence upon thyroid cancer. Consequently, the change of urinary glucocorticoids concentration may play an important role in thyroid cancer.

Chromatography, Gas↗

Altered profile of endogenous steroids in the urine of patients with prolactinoma.

OBJECTIVE: The aim of this study was to investigate the possible effect of prolactin on the metabolism of androgens and estrogens in patients with prolactinoma. To accomplish this, prolactin, urinary androgen, and estrogen metabolite levels were determined. In order to indirectly evaluate the possible involvement of enzymes, the concentration ratios of precursor metabolite to product metabolite were also compared with controls. METHODS: Urine samples were obtained from 27 female patients with prolactinoma (macro, micro, and idiopathic) and from 31 age-matched normal female subjects. Urinary metabolites of 21 androgens and corticoids and 20 estrogens were analyzed by a gas chromatography-mass spectrometry system. RESULTS: In patients with prolactinoma, urinary 17-ketosteroids, and all estrogen metabolite concentrations were elevated. The ratios of delta 5/delta 4-steroids and 5 beta/5 alpha-hydrogensteroids were higher in the patients with prolactinoma than in normal female controls. In addition, no significant differences between patients and controls were observed in the precursor metabolite to product metabolite ratios relating to estrogen metabolism. CONCLUSION: Our data suggests that enhanced PRL levels have a direct effect on urinary steroid secretion and metabolism, probably due to lowered activities of 5 alpha-reductase and 3 beta-hydroxysteroid dehydrogenase in the patients with prolactinoma.

Adult↗

The effect of transdermal oestradiol on bleeding pattern, hormonal profiles and sex steroid receptor distribution in the endometrium of Norplant users.

The most common side-effect and reason for discontinuation with Norplant use is bleeding disturbance. The aim of this study was to investigate whether the 6 week application of a patch which released 100 micrograms/day oestradiol would reduce the number of abnormal bleeding days or eliminate the problem. Another objective was to find out the correlation between the bleeding pattern and endometrial concentrations of oestrogen receptor (ER) and progesterone receptor (PR). Of 98 Norplant users, 34 patients had normal bleeding patterns and 64 patients had abnormal bleeding patterns. An oestradiol patch or a placebo patch were randomly used to treat 33 and 31 women with abnormal bleeding respectively. There was a clinical improvement in the oestradiol group compared with the placebo group, although this was not statistically significant. There were no correlations between PR and ER concentration and the serum oestradiol, progesterone, levonorgestrel and sex hormone-binding globulin concentrations. Significantly increased mean immunostaining scores of stromal PR were observed in those Norplant users whose endometrium had an atrophic histological appearance. The serum oestradiol concentration did not show a significant change after treatment with the oestradiol patch compared with the placebo patch.

Administration, Cutaneous↗