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PubMed2000

Steroid profiles determined by capillary electrochromatography, laser-induced fluorescence detection and electrospray-mass spectrometry.

Macroporous, monolithic capillary electrochromatography (CEC) columns, featuring a hydrophobic stationary phase, have been applied to the separations of steroids with good column efficiency. Using isocratic and gradient elution runs, mixtures of neutral or conjugated steroids could be resolved. While dansylated ketosteroids were detectable through laser-induced fluorescence at attomole levels, the CEC columns coupled to electrospray-ion-trap mass spectrometry featured femtomole detection limits.

Electrophoresis, Capillary↗
PubMed1996

Effects of different types of dietary fiber preparations isolated from bamboo shoots, edible burdock, apple and corn on fecal steroid profiles of rats.

This study was conducted to evaluate the effects of different types of dietary fibers (DF) under the conditions with or without cholesterol (Chol) loading on the amount and composition of steroids in rat feces. Rats were fed Chol-unsupplied diets containing 10% lard and 5% DF preparation isolated from four kinds of food, bamboo shoots, edible burdock, apple and corn, for three weeks. The respective diets were supplemented with 0.5% Chol and then given to the rats for a further two weeks. The excretion of total bile acid (BA) and several major BAs increased significantly in the apple group with or without Chol loading when compared with that in the cellulose (CP) or other DF groups. The tendency in the apple group was more noticeable when the diet was supplemented with Chol. This is presumably a major reason for the tendency of decrease in serum and liver Chol concentrations in the apple group. The ratio of secondary BAs to total BA in the feces was significantly low in the apple group. Although the lithocholic acid (LCA)/deoxycholic acid (DCA) ratio, a risk index for colorectal cancer, was significantly lower in the bamboo, burdock and apple groups than in the CP or corn groups when given the diet without Chol, the differences disappeared with the addition of Chol. The proportion of coprostanol, a secondary metabolite of Chol, was smaller in the former three groups than in the CP or corn groups. These results suggest that the intake of some DF by host animals works beneficially for the microbial conversion of BA and Chol in the large intestine but that the addition of Chol acts to cancel such beneficial effects.

Animal Feed↗
PubMed1996

Comparison of salivary steroid profiles in naturally occurring conception and non-conception cycles.

Oestradiol and progesterone profiles from naturally occurring conception and exposed non-conception cycles were compared to assess the impact of natural variation in concentrations of ovarian steroid hormones on female fecundity. In a prospective, longitudinal study, 24 women collected saliva samples twice daily and recorded intercourse for up to 1 year or until a pregnancy was clinically confirmed. Oestradiol and progesterone concentrations were measured by a salivary radioimmunoassay. Average mid-follicular oestradiol concentrations were significantly higher in conception than in non-conception cycles (12.6 +/- 1.7 versus 8.5 +/- 0.6 pmol/l, P < 0.01). A separate analysis, including only cycles from those women who contributed both conception and non-conception cycles, demonstrated an even more pronounced difference in mid-follicular oestradiol concentrations, not just for conception and non-conception cycles as groups (14.5 +/- 2.3 versus 6.5 +/- 0.7 pmol/l, P < 0.001), but also between the conception and average non-conception concentrations of individual women. Among these women, relative mid-follicular oestradiol concentration was highly correlated with the probability of successful conception. In addition, relative body weight was significantly positively correlated with mid-follicular oestradiol concentration. These findings indicate that variation in follicular development, reflected in variation in follicular oestradiol concentrations, is an important indicator of fecundity.

Adult↗
PubMed1999

Profiling steroid hormones in amniotic fluid of midpregnancy by routine stable isotope dilution/gas chromatography-mass spectrometry: reference values and concentrations in fetuses at risk for 21-hydroxylase deficiency.

Using routine stable isotope dilution/gas chromatography-mass spectrometry, 17-hydroxyprogesterone, androstenedione, testosterone, dehydroepiandrosterone, androstanediol, and 5alpha-dihydrotestosterone have been profiled in amniotic fluid of midgestation in 77 normal fetuses and 38 untreated or dexamethasone-treated fetuses at risk for 21-hydroxylase deficiency. Dexamethasone was suspended 5-7 days before amniocentesis. In normal fetuses, amniotic fluid concentrations (median, range; nanograms per mL) of 17-hydroxyprogesterone did not reveal a sex difference (1.48, 0.21-4.96), whereas those of androstenedione were lower in females (0.53, 0.00-2.71) than in males (0.93, 0.29-1.98). Testosterone levels were higher in males (0.24, 0.00-0.50) than in females (0.00, 0.00-0.27). No sex difference was found for dehydroepiandrosterone (0.47, 0.19-1.77). Levels of androstanediol and 5alpha-dihydrotestosterone were below the detection limit of our method in most cases. Regarding prenatal diagnosis of 21-hydroxylase deficiency, 17-hydroxyprogesterone and androstenedione presented the diagnostically most valuable steroids and were of equal diagnostic potential. They permitted successful diagnosis in 36 of 37 fetuses at risk: 12 were untreated and unaffected, 13 were treated and unaffected, 4 were untreated and affected (3 salt wasters and 1 simple virilizer), and 8 were treated and affected (5 salt wasters and 3 simple virilizers). In the latter group, one simple virilizer revealed normal steroid concentrations. Isotope dilution/gas chromatography-mass spectrometry, providing the highest specificity in steroid analysis, is proposed for routine use in clinical steroid analysis whenever maximal reliability is requested. Our study provides the first mass spectrometric reference data on amniotic fluid steroid concentrations and underscores the high accuracy of prenatal hormonal diagnosis of 21-hydroxylase deficiency.

Adrenal Hyperplasia, Congenital↗
PubMed2005

Urine steroid profile of judo competitors affected by acute physical exercises.

A heterogeneous group of 10 male and 15 female judo players are utilized in this study. The subjects complete a standardized maximal treadmill exercise test. Urine samples are collected at the pre- and postexercise stages. The urine steroids are measured using a gas chromatography-mass spectrometry instrument. In rest and after exercise, significantly higher testosterone and epitestosterone concentrations in males (p < 0.01) are found. The etiocholanolone-dehydroepiandrosterone (DHEA) ratio is significantly lower in males than females (p < 0.05). In both males and females, etiocholanolone concentration significantly decreases with the effect of exercise (p < 0.05). 11-OH etiocholanolone concentration also significantly decreases, but only in females (p < 0.05). Positive correlation is found between the changes of the etiocholanolone and epitestosterone concentration caused by exercise.

Androstanols↗
PubMed2004

Gonad development and plasma steroid profiles by HRGC/HRMS during one reproductive cycle in reference and leachate-exposed female perch (Perca fluviatilis).

Endocrine processes were studied in reference female perch (Perca fluviatilis) from Lake Djursjön and in leachate-exposed females from Lake Molnbyggen during one reproductive cycle. A high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) technique enabled the analysis of circulating sex steroids [progesterone (P), 17alpha-hydroxyprogesterone (17alpha-OHP), androstenedione (A), testosterone (T), estrone (E1), and 17beta-estradiol (E2)], the maturation-inducing hormone (MIH) 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-P), and glucocorticoids [11-deoxycortisol (11-DC) and cortisol]. Body and organ measurements, ratio of sexually mature (SM) females, and frequencies of body lesions, were also recorded. High frequencies of body lesions and a low ratio of SM females were generally found in Molnbyggen. These females also had lower gonadosomatic index (GSI) and liver-somatic index (LSI) in the early stages of vitellogenesis, indicating either a delayed onset of oogenesis or an impaired vitellogenesis. Non-reproducing females from both lakes showed a similar LSI cycle as SM females. Increasing T and A levels in SM females from Djursjön coincided with the GSI peak at the time of spawning. These two androgens were on several occasions lower in SM females from Molnbyggen, possibly the result of a disrupted 17,20-lyase activity of the P450 17alpha enzyme because only minor effects were observed on circulating P and 17alpha-OHP levels. E2 and E1 levels increased during vitellogenesis in females from both lakes, with E1 levels more closely resembling the peaks of A and GSI. Peak levels of 17alpha,20beta-P at the time of spawning confirmed its function as the MIH in perch, while the simultaneous peak of 11-DC suggests that it also may be involved in final maturation of the oocytes.

Animals↗
PubMed1993

Is there an altered steroid profile in patients with endometrial carcinoma?

BACKGROUND: Besides the well-known association between endometrial carcinoma (EC) and unopposed estrogen, androgens also may play a role in this respect; however, previous studies on endogenous estrogens and androgens in patients with EC and control subjects have yielded mostly divergent results, probably because of the use of poorly defined control groups. METHODS: Circulating steroid and pituitary hormones and sex hormone-binding globulin were measured in patients with EC and matched control groups of either patients with nonmalignant postmenopausal bleeding (hospital control subjects) and healthy, symptom-free women (nonhospital control subjects). RESULTS: Patients with EC had higher serum levels of adrenal C21 and C19 steroids, estrogens, and biologically active testosterone than nonhospital control subjects, whereas the hospital control subjects constituted an intermediate group in this respect. CONCLUSIONS: The results clearly indicate an altered steroid homeostasis, probably reflecting an increased "adrenal drive" in patients with EC. The results also stress the necessity of using strictly defined, healthy, symptom-free control subjects instead of the frequently used hospital control subjects when studying more subtile endocrine aberrations.

Adenocarcinoma↗
PubMed1997

[Sex steroid profiles and ovarian activities of the female panda Yan Yan in the Berlin Zoo].

The nine years old giant panda YAN YAN was received in April 1995 on loan for 5 years to the Berlin Zoo. Urine samples were collected daily or every second day from April 1995 to June 1996 in order to follow up sex hormone secretions and ovarian activities. Conjugated steroids were hydrolysed, extracted and measured with two enzyme immuno assays (EIA) being specific for either total oestrogenes or pregnandiol. The evaluation of the hormone secretion pattern yielded the following results: There is a significant synchronous cross correlation between estrogen and progestin metabolites secretion indicating its simultaneous synthesis. In addition, we found a regular increase and decrease of both hormones with a 13 days interval. This secretory pattern indicates repeating development and atresia of follicle cohorts with a cycle length of about 13 days. Only a single period of slightly elevated oestrogen synthesis was monitored in Feb. 96 without any signs of oestrus. Obviously the stimulation of ovarian function was insufficient for complete ovulation and corpus luteum formation.

Animals↗
PubMed1997

[Diagnosis of the adrenogenital syndrome caused by 11beta-hydroxylase deficiency using gas chromatographic-mass spectrometric analysis of the urinary steroid profile].

PROBLEM AND OBJECTIVE: In 5-7% of all cases of congenital adrenogenital hyperplasia (AGH) there is 11 beta-hydroxylase deficiency (11 beta-HM). Its clinical picture is characterised by hyperandrogenism and, in some cases, arterial hypertension. The diagnosis of the enzyme deficiency depends on a reliable method of analysing the hormone in plasma and urine. As little is known and data often contradictory about the pattern of urinary steroid excretion in 11 beta-HM, these steroid metabolites were measured by a highly specific method. PATIENT AND METHOD: The pattern of urinary excretion of steroids was determined by gas chromatography and mass spectrometry (GC/MS) in 16 children and adults (11 males, 5 females: mean age 9(8)/12 [2/12-20(3)/12] years) with 11 beta-HM. RESULTS: In all patients there was greatly increased excretion of tetrahydrated (TH) and hexahydrated (HH) metabolites of 11-desoxycortisol (S) and desoxycorticosterone (DOC). The excretion of THS and THDOC was extremely increased in all patients. The metabolites 5 alpha-THS as well as 20 alpha- and 20 beta-isomers of HHS, not normal found in healthy persons, were present in 15 patients (94%), while the 20 alpha- and 20 beta-isomers of 5 alpha-HHS were demonstrated in 14 (88%). For the first time, 20 alpha- and 20 beta-isomers of 5 alpha-HHS were shown to be typical urinary steroid metabolites in 11-HM. The excretion of cortisol metabolites is typically decreased in 11 beta-HM. No corticosterone metabolites were found. CONCLUSION: The urinary steroid excretion pattern, measured by GC/MS, is a noninvasive, highly specific and nonselective method in the differential diagnosis of abnormal steroid metabolism.

Adolescent↗
PubMed2005

Machine learning approaches for phenotype-genotype mapping: predicting heterozygous mutations in the CYP21B gene from steroid profiles.

OBJECTIVE: Non-linear relations between multiple biochemical parameters are the basis for the diagnosis of many diseases. Traditional linear analytical methods are not reliable predictors. Novel nonlinear techniques are increasingly used to improve the diagnostic accuracy of automated data interpretation. This has been exemplified in particular for the classification and diagnostic prediction of cancers based on expression profiling data. Our objective was to predict the genotype from complex biochemical data by comparing the performance of experienced clinicians to traditional linear analysis, and to novel non-linear analytical methods. DESIGN AND METHODS: As a model, we used a well-defined set of interconnected data consisting of unstimulated serum levels of steroid intermediates assessed in 54 subjects heterozygous for a mutation of the 21-hydroxylase gene (CYP21B) and in 43 healthy controls. RESULTS: The genetic alteration was predicted from the pattern of steroid levels with an accuracy of 39% by clinicians and of 64% by linear analysis. In contrast, non-linear analysis, such as self-organizing artificial neural networks, support vector machines, and nearest neighbour classifiers, allowed for higher accuracy up to 83%. CONCLUSIONS: The successful application of these non-linear adaptive methods to capture specific biochemical problems may have generalized implications for biochemical testing in many areas. Nonlinear analytical techniques such as neural networks, support vector machines, and nearest neighbour classifiers may serve as an important adjunct to the decision process of a human investigator not 'trained' in a specific complex clinical or laboratory setting and may aid them to classify the problem more directly.

Adult↗
PubMed1986

Effects of chronic ethanol ingestion on steroid profiles in the rat testis.

Testosterone, seven of its potential precursors, three of its metabolites and estradiol were analyzed in testes from rats given ethanol for 23 days in a nutritionally adequate liquid diet. The results were compared to those obtained with pair-fed control rats. The concentrations of pregnenolone, progesterone, 17-hydroxyprogesterone, androstenedione and testosterone were markedly lowered in four of the five rats given ethanol. The concentrations of the other 3 beta-hydroxy-delta 5 steroids and estradiol were unchanged, resulting in significantly increased ratios between 17-hydroxypregnenolone and 17-hydroxyprogesterone (P less than 0.025) and between androstenediol and testosterone (P less than 0.025) in the ethanol-treated rats. The results indicate that chronic ethanol administration reduces formation of testosterone by affecting a step prior to pregnenolone. There may also be an effect on the conversion of some 3 beta-hydroxy-delta 5 to the corresponding 3-oxo-delta 4 steroids. The levels of testosterone and three other steroids in testes of rats given the liquid diet were significantly lower than those in testes of animals fed a standard rat chow. This indicates a dietary influence on testicular steroid concentrations.

Alcoholism↗
PubMedDate not supplied

Morning steroid profile in children with congenital adrenal hyperplasia under different hydrocortisone schedules.

We studied 13 children with 21-hydroxyalse deficiency to explore the immediate potential suppressive effect of hydrocortisone dose schedule on the adrenal cortex. They were given 20 mg/m2 daily in a controlled trial. After random administration of a greater dose in the morning (7 patients) or at night (6 patients), we measured plasma levels of 17-hydroxyprogesterone, testosterone, and androstenedione at times-24, 0, 2, 4, and 6h. Considerable fluctuation of the steroid levels, unrelated to the drug intake, was observed. There was no statistically significant differences between the "morning dose" and "night dose" groups for any steroid. We conclude that; (i) the greater night dose did not avoid the 17-hydroxyprogesterone morning peaks, and (ii) the variation in plasma steroid levels is so marked that a single morning sample is unreliable to reflect the degree of adrenal suppression.

17-alpha-Hydroxyprogesterone↗