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Embryonic treatment with xenobiotics disrupts steroid hormone profiles in hatchling red-eared slider turtles (Trachemys scripta elegans).

Many compounds in the environment capable of acting as endocrine disruptors have been assayed for their developmental effects on morphogenesis; however, few studies have addressed how such xenobiotics affect physiology. In the current study we examine the effects of three endocrine-disrupting compounds, chlordane, trans-nonachlor, and the polychlorinated biphenyl (PCB) mixture Aroclor 1242, on the steroid hormone concentrations of red-eared slider turtle (Trachemys scripta elegans) hatchlings treated in ovo. Basal steroid concentrations and steroid concentrations in response to follicle-stimulating hormone were examined in both male and female turtles treated with each of the three compounds. Treated male turtles exposed to Aroclor 1242 or chlordane exhibited significantly lower testosterone concentrations than controls, whereas chlordane-treated females had significantly lower progesterone, testosterone, and 5[alpha]-dihydrotestosterone concentrations relative to controls. The effects of these endocrine disruptors extend beyond embryonic development, altering sex-steroid physiology in exposed animals.

Androgens↗

Normal and elevated 3 alpha-androstanediol glucuronide concentrations in women with various causes of hirsutism and its correlation with degree of hirsutism and androgen levels.

We investigated peripheral androgen metabolic activity in 54 hirsute females (HF) by evaluating the serum 3 alpha-androstanediol glucuronide (3AG) concentration, hirsutism score (HS), and etiology of hirsutism. Based on basal and ACTH-stimulated steroid profiles (1 h post-Cortrosyn, 0.25 mg, i.v. bolus), the causes of hirsutism were determined to be increased adrenal androgen production (greater than 2 SD above normal mean), increased ovarian testosterone (T) production (greater than 2 SD above normal mean basal T of ovarian source only), or idiopathic cause (normal steroid profile). Serum 3AG levels in each group of HF were significantly higher (P less than 0.01-0.001) than those in normal females [normal: 2.9 +/- 0.94 nmol/L (n = 28); HF: increased adrenal androgen production of undefined cause, 7.7 +/- 7.5 nmol/L (n = 14); 21-hydroxylase deficiency, 7.6 +/- 7.4 nmol/L (n = 5); increased ovarian T production 5.5 +/- 3.5 nmol/L (n = 18); idiopathic cause, 5.8 +/- 4.8 nmol/L (n = 17)]. However, normal 3AG levels (less than 5.2 nmol/L) were present in 50-67% of HF in each group. Collectively, 3AG levels in HF correlated significantly (P less than 0.01) with dehydroepiandrosterone (DHEA; r = 0.41) and DHEA sulfate (DS; r = 0.44), while the correlation with androstenedione (r = 0.15) or T (r = 0.19) was not significant. Serum 3AG and adrenal androgen levels decreased in all subjects after dexamethasone treatment (0.5-1 mg at hour of sleep; 2 mg/day for 3-5 days). The correlation between 3AG and HS was significant (r = 0.6-0.74; P less than 0.01-0.001) only in HF with increased adrenal androgen secretion and idiopathic cause, and was not significant (r = 0.42) in HF with increased ovarian T secretion. There was no significant correlation between androgen levels and HS. We conclude that the serum 3AG level was not consistently elevated in HF and did not differ significantly between the various causes. Significant correlations between 3AG and DHEA/DS levels, and the simultaneous decrease in 3AG and adrenal androgens after dexamethasone administration in HF suggest that adrenal androgens contribute significantly to 3AG production. The significant correlation between 3AG and HS in HF with increased adrenal androgen secretion and idiopathic cause indirectly suggests an adrenal androgen contribution to both 3AG production and hirsutism in these HF. The insignificant correlation between 3AG and HS in HF with increased ovarian T secretion may result from a confounding effect of ovarian T on hirsutism.

Adolescent↗

Steroid hormone profiles in women treated with aminoglutethimide for metastatic carcinoma of the breast.

Recent evidence suggests that aminoglutethimide (AG), a known inhibitor of adrenal steroidogenesis, is a potent blocker of aromatase and thus of estrogen production. These properties of AG have been exploited clinically to reduce the biosynthesis of adrenal estrogen precursors and extraglandular estrogen production in postmenopausal women with metastatic breast carcinoma. In this study, we have explored the effects of AG on a variety of steroids, including delta 5-C19 and -C21 compounds and delta 4-C19 and -C21 steroids as well as plasma and urinary estrogens in a series of postmenopausal women with breast cancer treated for 2 to 26 weeks. Plasma concentrations of delta 5-C21 and -C19 compounds were reduced 3- to 5-fold during AG therapy and remained suppressed over the duration of the study. By contrast, the delta 4-steroids such as progesterone, androstenedione, and 17 alpha- hydroxyprogesterone rose 2- to 10-fold during the initial 2 weeks of AG treatment and then fell back to starting levels or were suppressed. Plasma levels of the potent androgens testosterone and dihydrotestosterone were relatively preserved during AG therapy. The possible contribution of the postmenopausal ovary to the above hormone levels during AG therapy was examined by comparing steroid values from surgically castrated and spontaneously menopausal women. No statistically significant differences between the two groups were observed. In response to AG therapy, plasma levels of estrone and estrone sulfate were decreased 61 to 72%, and urinary estrone similarly fell 85% over the 12-week period. Estradiol concentrations in urine and plasma were similarly reduced 40 to 66% from basal values over this same period.

Adrenal Glands↗

C19 and C21 5 beta/5 alpha metabolite ratios in subjects treated with the 5 alpha-reductase inhibitor finasteride: comparison of male pseudohermaphrodites with inherited 5 alpha-reductase deficiency.

Male subjects administered the 5 alpha-reductase inhibitor finasteride were studied to determine its effect on C19 and C21 5 alpha-metabolism. Plasma testosterone (T) and 5 alpha-dihydrotestosterone (DHT) were measured and T/DHT ratios determined at doses of 0.2-80 mg. Urinary etiocholanolone (5 beta)/androsterone (5 alpha) ratios and 5 beta/5 alpha metabolite ratios of cortisol, 11 beta-hydroxyandrostenedione, and corticosterone were also measured. The steroid profile was compared to male pseudohermaphrodites with inherited 5 alpha-reductase deficiency who have a global defect in C19 and C21 5 alpha-metabolism. The mean plasma DHT levels were decreased at all doses, resulting in elevated T/DHT ratios. The mean urinary etiocholanolone/androsterone, 11 beta-hydroxyetiocholanolone/11 beta-hydroxyandrosterone, tetrahydrocortisol/allotetrahydrocortisol, and tetrahydrocorticosterone/allotetrahydrocorticosterone ratios were elevated compared to pretreatment levels and placebo control values. The mean ratios appeared to be dose dependent for plasma T/DHT, urinary etiocholanolone/androsterone tetrahydrocorticosterone/allotetrahydrocorticosterone ratios. The mean 11 beta-hydroxyetiocholanolone-hydroxyandrosterone ratio was maximally elevated at the lowest doses. The results indicate that finasteride has a broad steroid spectrum inhibiting C19 and C21 5 alpha-steroid metabolism and affecting hepatic and peripheral 5 alpha-metabolism. These results suggest that a single gene codes for a single 5 alpha-reductase enzyme with affinity for multiple steroid substrates. The steroid profile is strikingly similar to that of male pseudohermaphrodites with inherited 5 alpha-reductase deficiency.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Hormonal diagnosis of 21-hydroxylase deficiency in plasma and urine of neonates using benchtop gas chromatography-mass spectrometry.

We aimed at measuring the first plasma concentrations of 17-hydroxyprogesterone (17OH-P) determined by benchtop isotope dilution/gas chromatography-mass spectrometry (ID/GC-MS) in term neonates with or without 21-hydroxylase deficiency. Plasma samples from normal cord blood specimens (n=30), unaffected neonates (n=38) and neonatal patients with classical 21-hydroxylase deficiency (eight salt-wasters, three simple virilizers) were analyzed. Steroid profiling of random urinary specimens by GC-MS served as a confirmatory test for 21-hydroxylase deficiency. 17OH-P (nmol/l) in cord blood plasma lay between 11.66 and 75.92 (median 24.74). It declined shortly after birth. In the first 8 days of life, the time that screening for 21-hydroxylase deficiency is performed, 17OH-P ranged between undetected levels and an upper limit of 22.87 (median 4.11). Thereafter (days 9-28) its concentrations lay between 2.18 and 20.30 (median 6.22). Except one simple virilizer, all other patients with 21-hydroxylase deficiency had clearly elevated plasma 17OH-P at the time that screening for 21-hydroxylase deficiency would be performed. We suggest ID/GC-MS, which provides the highest specificity in steroid analysis, for checking suspicious concentrations of 17OH-P in neonates and underscore the potential of urinary steroid profiling by GC-MS as a rapid, non-invasive and non-selective confirmatory test for congenital adrenal hyperplasia.

17-alpha-Hydroxyprogesterone↗

Limited value of serum steroid measurements in identification of mild form of 21-hydroxylase deficiency.

BACKGROUND: ACTH stimulation test is widely used as a basic diagnostic method for non-classical congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21-OHD). However, the interpretation of this test has not been definitely established. To determine the cut-off values of basal and post-ACTH serum 17-OHP concentrations, data of patients with suspected 21-OHD has been analysed. PATIENTS AND METHODS: Two hundred and eighty-seven patients with postnatal/peripubertal virilization were investigated. Serum steroid concentrations were measured by RIA, urinary steroid profile was determined by capillary gas chromatography and mutation analysis of CYP21 gene was performed by allele specific PCR. 21-OHD was diagnosed by elevated serum 17-OHP concentrations, high level of the urinary 17-OHP metabolites and/or homozygosity for CYP21 mutations. RESULTS: Twenty-one patients of the total of 287 subjects (7.3 %) were identified as having 21-OHD. The numbers of 21-OHD patients compared to total numbers of patients with different ranges of serum 17-OHP were as follows: basal values below 3.5 ng/ml (mean + 1 SD) 0/225; between 3.5 - 6.6 ng/ml 3/41; above 6.6 ng/ml (mean + 2 SD) 18/21. Post-ACTH values below 6.4 ng/ml (mean + 1 SD) 0/226, between 6.4 - 10.3 ng/ml 0/35, above 10.3 ng/ml (mean + 2 SD) 21/26. CONCLUSION: There are patients with inappropriate peripubertal virilization who have slightly elevated 17-OHP concentrations. In this subgroup of patients more sensitive and specific methods are needed to establish the diagnosis of 21-OHD. Therefore we suggest performing an ACTH stimulation test in patients with a morning 17-OHP level above 3.5 ng/ml. Furthermore, urinary steroid profile and/or CYP21 gene analysis are needed in patients with a stimulated 17-OHP value between 10 and 30 ng/ml. These tests will distinguish between patients with non-classical 21-OHD and patients with other disorders.

Adolescent↗

Papillary hidradenoma: immunohistochemical analysis of steroid receptor profile with a focus on apocrine differentiation.

AIM: To make a quantitative evaluation by image analysis of oestrogen receptors, progesterone receptors, and androgen receptors in papillary hidradenomas and anogenital sweat glands. METHODS: 20 papillary hidradenomas and the anogenital sweat glands detected in surgical specimens selected from 10 vulvectomies for squamous carcinoma, eight haemorrhoidectomies, and one anal polypectomy, all from female patients, were investigated by the avidinstreptavidin peroxidase testing system. RESULTS: 90% of papillary hidradenomas and almost all the anogenital sweat glands showed immunoreactivity for oestrogen receptor and, more weakly, for progesterone receptor, with immunolabelled nuclear area ranging from 10% to 90%. Conversely conventional sweat glands did not show any nuclear staining. Overexpression of androgen receptors occurred in 20% of papillary hidradenomas, with nuclear staining strictly bordering papillary epithelium with apocrine differentiation. There was no immunoreactivity for androgen receptors in anogenital sweat glands. CONCLUSIONS: Oestrogen and progesterone receptors seem to represent reliable markers for differentiating between anogenital sweat glands and conventional sweat glands, and a further link to explain why papillary hidradenomas occur almost exclusively in the female anogenital region. Positivity for oestrogen/progesterone receptors suggests that epithelia either of anogenital sweat glands or of papillary hidradenomas are controlled by ovarian steroid hormones. Androgen receptor nuclear staining of the epithelium with apocrine differentiation in vulvar papillary hidradenoma strengthens its homology with breast duct papilloma.

Adenoma, Sweat Gland↗

Animal models for hormone-dependent human breast cancer. Relationship between steroid receptor profiles in canine and feline mammary tumors and survival rate.

The present study shows that canine and feline mammary tumors, like human breast tumors, can be polyreceptive, i.e., they can contain estrogen (ER), progestin (PR), androgen, glucocorticoid, and/or mineralocorticoid cytosol receptors. Furthermore, a follow-up of 45 bitches with mammary carcinoma has indicated that the survival rate is significantly higher in animals with receptor-rich (ER and/or PR) tumors. This indicates that these canine mammary tumors should be evaluated further for their suitability as an animal model for hormone-dependent human breast carcinoma.

Animals↗

Steroid hormone profile during gestation and parturition of the guppy (Poecilia reticulata).

The guppy is an ovoviviparous teleost with fertilization and gestation occurring intrafollicularly, and ovulation and hatching at around parturition. The levels of estradiol-17 beta (E), testosterone (T), cortisol (F), 17 alpha-hydroxyprogesterone (17-OHP), 17 alpha-20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P), and progesterone (P) in the guppy at different stages of gestation were measured in 60 microliters of serum with a highly specific, sensitive, precise, and accurate procedure involving high-performance liquid chromatography and radioimmunoassay. Levels of E and T increased during vitellogenic growth of oocytes and declined sharply after fertilization, increasing again gradually during late gestation to reach prefertilization levels at periparturition. F levels decreased significantly during fertilization followed by an increase during gestation and a sharp decline again at periparturition. The level of 17-OHP was significantly lower at periparturition than at other stages, while that of 17,20 beta-P was significantly higher in periparturition than in late gestation. There was no significant change in the levels of P at all stages of gestation, suggesting that, unlike mammals, P may not be involved in maintaining gestation in this fish. Changes in the levels of E and T seem to be related to oocyte growth rather than gestation. The role of F in gestation is uncertain since the effect of stress could not be discounted or defined in this study. Thus, on the present results, the role of any particular steroid in maintaining gestation of the guppy is not obvious. The elevated levels of E at term may facilitate parturition in addition to its role in vitellogenesis during oocyte growth.

Animals↗

Malignant interstitial cell carcinoma of the testis: report of two cases with steroid synthetic profiles, response to therapy, and review of the literature.

Two cases of malignant interstitial cell carcinoma of the testis are reported. The first patient had no evidence of a virilizing syndrome. Basal plasma testosterone (T) was decreased, whereas plasma luteinizing hormone, estrone (E1), and androstenedione were elevated. These findings were diagnostic of a defect in T secretion as a result of a partial 17-hydroxysteroid dehydrogenase deficiency as seen in male pseudohermaphroditism. In the second patient, showing gynecomastia and atrophic testis, endocrine studies revealed high plasma T and estradiol (E2); all measured delta 4 and delta 5 precursors of T were elevated resembling the pattern seen in virilizing adrenal carcinomas. Both patients were treated with radiotherapy without demonstrable effect. Chemotherapy--consisting of a combination of cis-platinum, vinblastine, and bleomycin; then cyclophosphamide, doxorubicin, and vincristine; and finally o,p'-DDD--was unsuccessful in reducing tumor bulk. Since malignant Leydig cell carcinomas are rare, this paper reviews the literature and makes recommendations concerning treatment.

17-Ketosteroids↗

Oral testosterone administration detected by testosterone glucuronidation measured in blood spots dried on filter paper.

BACKGROUND: Blood sampling is not a common practice for sports drug testing. Our aim was to investigate whether dried blood spots on filter paper could be an alternative to plasma samples for monitoring steroid profiles in dope testing. METHODS: We collected dried blood spots and plasma from six healthy Caucasian subjects after an oral 120-mg dose of testosterone undecanoate (TU). Nonconjugated testosterone, testosterone glucuronide (TG), androsterone glucuronide (AG), and etiocholanolone glucuronide (EtG) were measured by gas chromatography-mass spectrometry in both matrices. 17alpha-Hydroxyprogesterone (17alphaOHP) and luteinizing hormone (LH) also were measured in the plasma samples. For comparison, similar measurements were done on samples obtained from the same subjects given 25 mg of testosterone propionate (TP) plus 110 mg of testosterone enanthate (TE) intramuscularly after a wash-out period. RESULTS: After oral TU intake, TG, AG, and EtG increased sharply, whereas nonconjugated testosterone did not change significantly. Results on dried blood spots correlated well with those on plasma. The TG/testosterone ratio in blood or plasma was verified to be a sensitive and specific marker (significantly increased for up to 8 h after intake; P <0.05) for oral TU intake but not for intramuscular administration of TP plus TE. Little suppression of plasma LH and 17alphaOHP was observed after a single oral dose of TU. One subject did not show a significant increase of blood TG after oral TU intake. CONCLUSIONS: The measurement of glucuronide conjugates in blood and plasma samples is relevant for sports drug testing when analyzing the steroid profile. Dried blood spots collected on filter paper are a suitable alternative to plasma for detecting testosterone abuse.

Administration, Oral↗

Effects of dietary fat and season on steroid hormonal profiles before parturition and on hormonal, cholesterol, triglycerides, follicular patterns, and postpartum reproduction in Brahman cows.

Spring-calving Brahman cows (S) artificially inseminated to Brahman, Angus, or Tuli sires and fall-calving Brahman cows (F) naturally bred to Brahman were allotted randomly to receive 3.74% (LF; n = 9 S and 6 F), 5.20% (MF; n = 8 S and 6 F), or 6.55% dietary fat (HF; n = 8 S). Diets were formulated to contain differing fatty acid concentrations and to be isocaloric and isonitrogenous. Cows were bled and fed twice daily from 2 wk before expected calving date through d 21 after calving. Ultrasonography was performed on d 14 and 21 after calving. From d 21 to 90 after calving a sterile bull equipped with a chin-ball marker was placed with the cows to aid in estrus detection. In both seasons progesterone decreased (P < .01) and estradiol-17 beta increased (P < .01) as parturition approached. Cows receiving MF and HF had increased (P < .01) total numbers of follicles compared to LF cows, and cows receiving MF had larger (P < .01) follicles. During the spring, cows receiving HF and cows bred to Brahman or Tuli sires had longer (P < .01) gestation lengths. Progesterone concentrations before calving were affected (P < .01) by treatment x sire and estradiol-17 beta by a time x treatment interaction (P < .01). Cholesterol after calving was higher (P < .01) in HF cows than in LF or MF cows. In the fall, LF cows had heavier (P < .01) calves than cows receiving MF. Birth weight was also affected (P < .01) by treatment x sex of calf. Progesterone was affected (P < .01) by treatment x sex of calf. Estradiol-17 beta was affected (P < .01) by sex of calf and treatment x sex of calf. Across seasons, by d 90 after calving, 9 of 15 (60%) LF and 11 of 15 (73.3%) MF cows showed estrual behavior. Cows in the spring had increased (P < .01) numbers and larger follicles compared to the fall. In conclusion, dietary fat may influence steroid hormone concentrations before calving, calf birth weight and postpartum follicular populations; furthermore, follicular populations may also be influenced by season.

Animals↗

Assessment of urinary gonadotropin and steroid hormone profiles of female cynomolgus monkeys after treatment with Pueraria mirifica.

This study investigated the changes in the urinary hormone levels of female monkeys (Macaca fascicularis) after single-dose and long-term treatments with Pueraria mirifica (PM). The monkeys were separated into 3 groups (n=3) and orally treated with 10, 100, or 1,000 mg of PM in each group. Two series of experiments were performed. In the first series of experiments, the monkeys were orally treated with a single dose of PM. The experimental schedule was divided into a one menstrual cycle pretreatment period and a two menstrual cycle post-treatment period. In the second series of experiments, the monkeys were orally treated daily with PM for 90 days. The experiment schedule was divided into a one menstrual cycle pretreatment period, a three menstrual cycle treatment period, and a two menstrual cycle post-treatment period. Urinary samples were collected daily and assayed for the FSH, LH, estradiol, and progesterone levels. The results showed that there were no changes in the FSH, LH, estradiol, and progesterone levels after treatment with a single dose of 10, 100, or 1,000 mg of PM or after daily treatment with 10 mg of PM for 90 days compared with the levels observed during the pretreatment period. Daily treatment with 100 mg and 1,000 mg of PM for 90 days only produced a clear reduction in the urinary FSH levels. This suggests that changes of urinary FSH levels can be considered an indicator for study of estrogenic effects on hormonal levels in female monkeys.

Animals↗

Steroid receptor profile and receptor stability in subfractions of human prostatic tissues. Critical aspects on microassays.

Androgen (AR), progesterone (PR), and estrogen (ER) receptor contents in cytosol and salt-extractable nuclear subcompartments from 6 normal, 39 benign hyperplastic (BPH), and 7 malignant prostatic tissue specimens were analyzed by radioligand-binding assay techniques. In addition, the temperature stability of AR and PR was measured in another three BPH specimens. Five punch-needle biopsy samples from prostate cancers were also analyzed for AR and PR content. All receptor data were calculated from saturation analyses. The highest AR content was found in the cytosol and nucleic from malignant prostatic tissues. The highest PR concentrations were found in BPH cytosol, whereas nuclei of all types of tissues were negative with regard to this receptor. Markedly lower concentrations of ER were found in cytosol and nuclei from BPH as compared with malignant and normal tissues. PR was the most temperature-stable receptor; a marked receptor loss at room temperature was not registered until after 12 h. AR was stable for 4-5 h in cytosol and for 8-9 h in nuclei. Needle-biopsy specimens from prostate cancer showed highly variable and confusing results for AR and PR content, indicating that microassay studies using biochemical techniques on small tissue samples are unreliable and should not be recommended.

Biopsy↗

Analysis of metabolic profiles of steroids in faeces of healthy subjects undergoing chenodeoxycholic acid treatment by liquid-gel chromatography and gas-liquid chromatography-mass spectrometry.

The multicomponent analysis of faecal steroids is described. Steroids were removed from faeces by solvent stripping in a Soxhlet apparatus and the resulting extracts were fractionated by diethylaminohydroxypropyl Sephadex column chromatography into neutral sterols, free bile acids, glycine conjugated bile acids, taurine conjugated bile acids and sulphated steroids. In this study the method has been applied for faecal steroid analyses of healthy subjects undergoing chenodeoxycholic acid therapy. Chenodeoxycholic acid administration causes a considerable increase in the concentration of faecal lithocholic acid which is a known comutagenic bile acid. Furthermore it has been shown that conjugated bile acids can account for between 10 and 20% of the faecal bile acid pool. The method described is convenient and may be useful for epidemiological studies which require a large number of faecal samples to be analysed.

Bile Acids and Salts↗

Thermospray HPLC/MS: a new mass spectrometric technique for the profiling of steroids.

The analysis of various steroid classes by thermospray HPLC-MS using solvent systems containing 0.1 M ammonium acetate has been described. For simple unconjugated 3-oxo-4-ene steroids the positive ion spectra are dominated by a parent ion M + H+ and with increasing numbers of hydroxyl group intense ions formed by sequential losses of water (M + H- n18)+ become important. Steroids with dihydroxyacetone side-chains readily lose these side-chains and the resulting (M + H-60)+ fragment is the base peak in their spectra. The (M + H-60)+ ion is not important for most steroids with glycerol-type side-chains. Although competition between thermal degradation and vaporization was observed at lower concentrations, the effect was minimized after optimizing conditions and the protonated molecular ion was easily detected when as little as 1-10 pmol of material were injected on-column. Steroid glucuronides when analyzed in the negative ion mode give simple spectra with base peak and parent ion (M-H)-. Lack of fragmentation permits facile and sensitive measurement of individual glucoronides by selected-ion-monitoring. Extensive fragmentation is seen in the positive ion mode with sequential losses of H2O from the molecular ions (M + NH4)+ and from the aglycone fragment ion. For simple unconjugated steroids the sensitivity of HPLC-MS in selected-ion-monitoring mode can be excellent. When the protonated molecular ion of testosterone was monitored the signal/noise ratio for 30 pg testosterone was about 10.

Chromatography, High Pressure Liquid↗