Steroid hormone accumulation in human breast cyst fluid.

Elevated concentrations of peptide hormones have been described previously in human breast fluid. In the current study, the levels of cortisol, progesterone, testosterone, dihydrotestosterone, androsterone, androsterone sulfate, dehydroisoandrosterone, dehydroisoandrosterone sulfate, estradiol, estrone, estradiol sulfate, and estrone sulfate were measured. The levels of the four 17-ketosteroids and the two estrogen sulfates were markedly elevated over the plasma level, while that of the other compounds was the same or only slightly higher than the plasma levels of the same compounds.

PIP: As part of a program to explore whether a relationship existed between the compounds present in breast cyst fluid and the risk for breast cancer, concentrations of a wide variety of hormones, enzymes, tumor-associated antigens, and ions in breast cyst fluid were determined. In this report, the concentrations of steroid hormones in cyst fluid samples from the same population used for previously published studies of peptide hormone concentrations were measured. This study determined the levels of cortisol, progesterone, testosterone, dihydrotestosterone, androsterone, androsterone sulfate, dehydroisoandrosterone, dehydroisoandrosterone sulfate, estradiol, estrone, estradiol sulfate, and estrone sulfate. In previous studies, levels of peptide hormones were shown to be elevated in breast cyst fluid, and in this study, the levels of the 4 17-ketosteroids and the 2 estrogen sulfates were markedly elevated in the breast cyst fluid over the plasma level. The concentrations of the other compounds were either the same or only slightly higher than plasma levels of the same compounds. To date, however, no correlation has been obtained between the occurrence of breast cancer and levels of any of these assayed hormones.

Serotonergic responsivity in male young adults with autistic disorder. Results of a pilot study.

Altered serotonergic function has been postulated to exist in autistic disorder. Central serotonergic responsivity was assessed with a neuroendocrine challenge test in seven male young adults with autistic disorder and in seven age- and gender-matched healthy controls. Binding indexes and physiologic responsivity of the platelet serotonin-2 (5-HT2) receptor complex were also measured, as was whole-blood serotonin content. Compared with controls, autistic subjects had substantially blunted prolactin release in response to a 60-mg oral dose of fenfluramine hydrochloride, an indirect serotonin agonist [corrected]. Furthermore, the magnitude of serotonin-amplified platelet aggregation, mediated by the platelet 5-HT2 receptor complex, was reduced in the autistic group, as was the mean number of platelet 5-HT2 receptor sites. Among autistic subjects, fenfluramine-induced prolactin release correlated positively with the serotonin-amplified platelet aggregation response and negatively with whole-blood serotonin content. The results of the present study are compatible with the hypothesis that central serotonergic responsivity is decreased in male autistic young adults. Correlations between central and peripheral serotonergic measures in autistic subjects suggest that systemic alterations in serotonergic function may occur in autism.

Twenty-four-hour cortisol profiles demonstrate exaggerated nocturnal rise in diabetic children.

Plasma cortisol was measured every 20 min for 24 h in 12 normal and 8 insulin-dependent, nonketotic, diabetic children treated with one daily injection of insulin. Plasma glucose was also measured every 20 min in the diabetic children. The diurnal pattern of cortisol secretion was identical. The mean 24-h plasma cortisol was similar in both groups, but significantly elevated in the diabetic children between 0200 and 0920 h. Peak cortisol levels were higher in the diabetic than in the normal children. No correlation was found between average plasma glucose and average plasma cortisol, or between the nocturnal change in plasma glucose and average nocturnal plasma cortisol in the diabetic subjects. These studies demonstrate an exaggeration of the normal nocturnal rise in plasma cortisol in diabetic children not related to the levels of plasma glucose.

6 beta-Hydroxycortisol: a noninvasive indicator of enzyme induction.

We measured urinary 6 beta-hydroxycortisol (6 beta OHF) excretion in normal children and in children receiving anticonvulsant therapy with phenobarbital or diphenylhydantoin, 6 beta OHF excretion increased 4- to 7-fold during anticonvulsant therapy. The marked increase in the ratios of 5 beta OHF to 17-hydroxycorticosteroid and 6 beta OHF to free cortisol suggests that the measured increase in urinary 6 beta OHF may serve as an index of induction of the microsomal mixed function oxidase system. This noninvasive approach allows rapid assessment of the effects of drugs and foreign compounds on microsomal hydroxylation of cortisol. This convenient probe can be applied, with particular ease, to further studies in children in whom characterization of the effects of drugs and xenobiotic compounds on hepatic hydroxylation is of interest.

Androsterone sulfate concentrations in plasma in hypo- and hyperthyroidism.

Androsterone sulfate concentrations have been measured in the plasma of 16 hypothyroid women and 14 hyperthyroid women by a gas-liquid chromatographic technique and in 23 hypothyroid and 18 hyperthyroid women by radioimmunoassay. In obth studies androsterone sulfate concentrations were significantly higher in the hyperthyroid groups. In one hypothyroid subject, blood was obtained at 20-minute intervals over 24 hours before and after the administration of triiodothyronine; plasma androsterone sulfate, initially 25 mug per dl, increased three-fold as a consequence of treatment.

The metabolism and 24-hour plasma concentrations of androsterone in man.

A radioimmunoassya for free androsterone permitting facile analysis of plasma taken at 20 min intervals over 24 h has revealed a curve of varying concentrations which formed a pattern similar to curves of cortisol and dehydroisoandrosterone, boht bein primary adrenal secretory products and the latter being the major precursor of androsterone. The mean of the average 24 h concentrationof androsterone in men was 55 +/- 14 ng/dl (n = 8). Such temporally related behavior requires rapid production and removal of both 17-ketosteroids, processes which have been confirmed by tracer studies where it has been shown that in a two compartment system, androsterone has a t1/2 of 25 +/- 9 min for the fast component and metabolic clearance rate of 4050 +/- 1315 L per day (n = 10). The data are in accord with an approximate production rate of androsterone which is consistent with values reported for its urinary excretion.

The mean 1300-1600 h plasma cortisol concentration as a diagnostic test for hypercortisolism.

The 24-h mean plasma cortisol concentration was compared with the mean plasma cortisol concentrations during short subperiods of the day in 88 normal subjects and 223 patients with a very wide range of mean 24-h cortisol levels. The correlation between the 1300-1600 h mean plasma cortisol concentration and the mean 24-h plasma cortisol concentration was extremely high in all groups. Mean cortisol concentrations during this short subperiod powerfully discriminate cortisol hypersecretors (patients with Cushing's syndrome, anorexia nervosa, or prostate cancer) from normal controls. Hence, it is suggested that the mean or integrated 1300-1600 h plasma cortisol concentration can be used as a reliable afternoon cortisol test for the presence of hypercortisolism.

Determination of plasma dexamethasone in the mother and the newborn after administration of the hormone in a clinical trial.

A RIA method is described for the measurement of dexamethasone in maternal, cord, and neonate plasma in a collaborative multicenter clinical trial to evaluate the efficacy of antenatal steroid therapy in the prevention of respiratory distress syndrome. The antiserum raised against dexamethasone-3-carboxymethyloxime-BSA conjugate was highly specific in that the endogenous steroids and 11-dehydrodexamethasone, a metabolite of dexamethasone, had a cross-reaction of less than 2.0%. Both pregnancy and cord plasma had to be purified by either gel filtration and/or paper chromatography. The overall recoveries of dexamethasone were 75.3 +/- 6.7% and 48.9 +/-6.7% for maternal and cord plasma samples, respectively. The intra- and interassay coefficients of variance for maternal plasma were 8.4% ad 9.1%, respectively, and for cord plasma were 11.9% and 10.9%, respectively. There was a good correlation between the dexamethasone values obtained by RIA when compared with those obtained by performance liquid chromatography in some cord plasma specimens. The recoveries of added dexamethasone and its metabolite by high performance liquid chromatography were also found to be good. The average maternal plasma dexamethasone level was 37 ng/ml 2 h after im injections of 5 mg dexamethasone phosphate every 12 h. The half-life of dexamethasone measured after the discontinuation of the drug was 216 min. Dexamethasone appeared too be cleared very rapidly from the circulation of the fetus and neonate. Because 11-dehydrodexamethasone was not present in significant amounts in the neonate, it was concluded that other factors in addition to the conversion of dexamethasone to its 11 dehydro metabolite were responsible for the rapid clearance of dexamethasone from fetal and neonatal circulation.

The effect of medroxyprogesterone acetate on the pituitary-adrenal axis.

Twelve cancer patients and one patient with diabetes mellitus were treated with medroxyprogesterone acetate (MPA) by intramuscular injection in a total weekly dose of 400, 700, or 1200 mg. The treatment reduced the plasma cortisol concentration by 76% in the AM hours (21 leads to 5.0 mug/dl) and by 75% in the PM hours (12.8 leads to 3.2 mug/dl). Cortisol production rate decreased by 67% (19 leads to 6.2 mg/24 hrs). The 24 hour profile of plasma cortisol concentration measured in 3 patients showed zero secretion over this period. Low plasma ACTH values prevailed during treatment, and a blunted response to maximal ACTH stimulation was found. No evidence of adrenal insufficiency was observed in any patient, even though in some patients the plasma cortisol concentration remained at zero for many weeks. MPA has cortisol-like effects and the suppression of adrenal function is probably mediated by a negative feedback action on the hypothalamus or pituitary.

PIP: The effects of medroxyprogesterone acetate (MPA), administered im in a total weekly dose of 400, 700, or 1200 mg, on the pituitary-adrenal axis were studied in 12 cancer patients and 1 patient with diabetes mellitus. MPA reduced mean A.M. plasma cortisol concentrations by 76% and mean P.M. cortisol concentrations by 75%. The 24-hour production rate of cortisol was reduced by 67%. 41% of the individual A.M. cortisol determinations and 61% of the individual P.M. determinations were not significantly (p greater than .001) different from 0. Plasma ACTH levels were low throughout treatment, and the response to maximal ACTH stimulation was not pronounced. Evidence of adrenal insufficiency was not observed in any of the patients. The results demonstrate the corticosteroid replacement properties of MPA, and it is suggested that the suppression of adrenal function is most likely mediated by a negative feedback action on the hypothalamus or pituitary.

Effect of flutamide on cortisol metabolism.

The effect of flutamide on cortisol metabolism was studied in eight patients with prostate cancer. Flutamide markedly decreased the formation of 3 alpha, 17,21-trihydroxypregnane-11,20-dione (THF), and the 11-oxy-17-ketosteroid metabolites by 72%, 50%, and 46% respectively; however, 3 alpha, 11 beta, 17,21-tetrahydroxy-5 alpha- pregnan-20-one was increased by 46%. The 24-h mean plasma cortisol concentration was not altered. The cortisol production rate decreased by an average of 53% (from 32.7 to 15.5 mg/24 h). The effect of the drug on plasma cortisol kinetics was studied in three patients. This showed that flutamide increased the t1/2 (from 80 to 108 min) but decreased the distribution volume (from 17.8 to 13.8 liters) and the MCR (from 222 to 130 liters/24 h). The changes in THE and THF formation and in the t1/2 and MCR of [C]cortisol are similar to the effects observed in patients with intrahepatic cholestasis. It is suggested that in the case of flutamide these changes were also due to a cholestasis-producing effect of the drug on the liver. As the clinical response to the drug did not correlate with the cortisol metabolic changes, its therapeutic effect was probably not mediated by its effects on cortisol metabolism.

The production rate of cortisol declines during recovery from anorexia nervosa.

The plasma concentration of cortisol is elevated in many patients with anorexia nervosa. It has remained unclear whether this elevation of plasma cortisol level is due only to a slowing of the rate of cortisol metabolism or whether there is, as well, an increase in adrenal secretory activity in anorexia nervosa. We studied adrenocortical activity in 9 female patients and one male patient with anorexia nervosa before and during recovery. The 24-h mean level of plasma cortisol and the rate of urinary free cortisol excretion decreased during recovery, from 11.4 to 7.4 micrograms/dl and from 225 to 116 micrograms/day, respectively (P less than 0.005 and P less than 0.10, paired t test). These changes were associated with a significant decline in the rate of cortisol production from 24.3 to 17.9 mg/day as measured by radioisotope dilution (P less than 0.005). These results suggest that adrenal secretory activity is increased in anorexia nervosa and that the elevation of plasma cortisol level observed in this syndrome reflects not only a slowing of cortisol metabolism but also a rise in cortisol production.

Age variation of the 24-hour mean plasma concentrations of androgens, estrogens, and gonadotropins in normal adult men.

The 24-h mean plasma concentrations of androgens (dihydrotestosterone and total and free testosterone), estrogens (estrone and estradiol), and gonadotropins (LH and FSH) were measured in 35 healthy men, aged 21-85 yr, who were rigorously screened to exclude factors known or suspected to alter endocrine function. The plasma total testosterone concentration showed a slow continuous decline with age, decreasing about 35% between 21 and 85 yr of age; the free testosterone level was closely correlated with that of total testosterone over the entire observed concentration range. The concentrations of dihydrotestosterone, estrone, estradiol, and LH were age invariant. The concentration of FSH showed a continuous linear increase with age; the level at age 85 was about 2.5 times the level at age 21. The following conclusions were drawn. 1) Testosterone secretion appears to decline slowly and continuously throughout adult life in men. 2) Measurement of the plasma free testosterone level adds no independent information in healthy men, since its level is closely correlated with that of total testosterone at all concentrations. 3) The continuous rise with age in FSH concentration while LH is age invariant cannot be explained by changes in testosterone or estrogen production, but might be due to a decline of inhibin production with age.

Obese young men have elevated plasma estrogen levels but obese premenopausal women do not.

The 24 hr mean plasma concentrations of estrone (E2) and estradiol (E2) were measured in 18 healthy, regularly cycling obese women; 16 healthy, regularly cycling nonobese women; 18 healthy obese men; and 33 healthy nonobese men. The obese men showed significant elevations of both E1 (67 pg/ml versus 49 pg/ml control; P less than 0.005) and E2 (37 pg/ml versus 28 pg/ml; P less than 0.005), but the obese women showed no significant elevation of either E1 or E2. The most likely explanation for the absence of significant hyperestrogenemia in the obese women despite evidence that such women have increased androstenedione-to-estrone conversion is that the latter source of estrogen is too small in comparison with estradiol secretion to cause a statistically detectable increment in plasma estrogen levels.

Sex difference in the influence of obesity on the 24 hr mean plasma concentration of cortisol.

The 24 hr mean plasma cortisol concentration was measured in 65 healthy women ranging from 21% below to 218% above desirable weight and in 47 healthy men ranging from 5% below to 330% above desirable weight. In the women, there was a clear-cut inverse linear correlation between the plasma cortisol concentration and the percent deviation from desirable weight (y = 7.5 -- 0.3 x; r = -0.49; p less than 0.001); the relation of free to total cortisol concentration was weight-invariant; the MCR of cortisol in the most obese women was much higher than that of nonobese women (340 +/- 76 versus 211 +/- 31 liters/gm urinary creatinine; p less than 0.01). In the men, the plasma cortisol level and MCR were weight-invariant. To account for the finding in women of a linear correlation of the decrement in plasma cortisol level with the percent deviation from desirable weight (which in turn is nearly perfectly correlated with the total body fat content), we postulate that a given weight of adipose tissue in women takes up a constant amount of cortisol; this in turn suggests that their adipose tissue contains a saturable binding system such as corticosteroid receptor. By the same logic, the weight-invariance of plasma cortisol and MCR in men suggests the absence of significant amounts of corticosteroid receptor in their adipose tissue. The finding that the increased cortisol MCR of obese women results in decreased plasma cortisol levels rather than an increase in cortisol production (the latter, corrected for muscle mass, is normal in obesity: Strain et al, Metabolism 29:980, 1980) suggests a defect in their cortisol ACTH feedback system. Such a defect, presumably hypothalamic, is not unexpected in the light of reports of defective hypothalamic control of prolactin and growth hormone secretion in obesity.

Mild Hypogonadotropic hypogonadism in obese men.

To evaluate the pituitary-gonadal axis of obese men, we compared the 24-hour mean plasma concentrations of total and free testosterone and of dihydrotestosterone, FSH, and LH in 21 healthy obese men, aged 18-50, and 24 age-matched healthy nonobese men. In the obese men, we also measured the volume of ejaculate and the number and motility of sperm, and investigated libido by psychiatric interview, and potency by history and by measurement of nocturnal penile tumescence. As a group, the obese men had less than two-thirds the normal mean plasma levels of total testosterone, free testosterone, and FSH; the difference from normal was highly significant for all three. 24 hr LH levels were normal, which is inappropriately low in view of the subnormal testosterone levels. 24 hr mean levels of dihydrotestosterone and spermatogenesis, libido, and potency were essentially normal. Taken together, the findings represent a state of mild hypogonadotropic hypogonadism, which thus appears to be characteristic of obese men. This abnormality probably results from partial suppression of the pituitary by the elevated plasma estrogen levels we and others find in these men.

Profile of 24-hour plasma glucose and insulin concentrations. Their variation with two methods of total parenteral nutrition administration.

The concentration of glucose and insulin in the plasma during total parenteral nutrition (TPN) was measured every 20 minutes during a 24-hour period in three subjects with granulomatous enterocolitis. During a gravity drip, parallel variations in rate of flow and plasma glucose and insulin concentrations were seen, and both hypoglycemia and hyperglycemia occurred. During a constant (pumped) infusion, plasma glucose and insulin concentrations were stable and normal. Glucose excretion in the urine was negligible and adequate gains in weight occurred, suggesting utilization of the administered nutrients. Hypertonic solutions for TPN should be infused at a constant rate either by careful attention to the flow rate during changes of intravenous bottles and tubing or by using a constant-infusion pump. Infusion of TPN solutions at a constant rate minimizes the risk of hypoglycemia or hyperglycemia.

Cortisol measurements in patients receiving oral corticosteroid replacement treatment.

Eight women receiving corticosteroid replacement in the form of 50 mg cortisone acetate or 40 or 50 mg cortisol orally daily were studied. The cortisol "urinary productions rate" and "blood production rate" measurements suggested that the steroid was rapidly metabolized in the gut or in the first passage through the liver. The 24-hour mean plasma cortisol concentration obtained from blood samples drawn every 20 minutes over a 24-hour period and the "blood production rate" were close to the values in normal women. However, since the normal 24-hour plasma cortisol concentration profile could not easily be reproduced and this may be relevant for optimal physiologic function, the patient's clinical status will continue to be the main guide to the choice of the appropriate replacement dose.

Radioimmunoassay of androsterone and androsterone-3-sulfate in plasma.

Details of a sensitive and specific radioimmunoassay for androsterone (1) and androsterone sulfate in plasma have been presented. Benzene extracts of plasma were chromatographed on alumina to isolate the androsterone fraction either (a) directly after extraction (A) or (b) after solvolysis (AS). Following treatment with rabbit anti-A-17-BSA, antibody bound steriod was precipitated by ammonium sulfate. Androsterone concentrations in normal male plasma averaged 57 +/- 24 (S.D.) ng/dl, range 35-135 ng/dl and for normal women, 44 +/- 21 (S.D.) ng/dl, range 18-98 ng/dl. Androsterone sulfate concentrations were: males 55 +/- 28 mug/dl (range 10-114 mug/dl); premenopausal females 52+/- 31 mug/dl (range 16-318 mug/dl).

Preparation and antigenic properties of androsterone-7-BSA conjugate.

The 7-carboxymethoximino derivative of androsterone (1) has been prepared from dehydroisoandrosterone-17-ethyleneketal by a sequence involving inversion at C-3, introduction of a carbonyl at C-7, and reduction of the double bond at C-5. The substance was condensed with BSA by the carbodiimide procedure to afford a conjugate which produced anti-androsterone antiserum in innoculated rabbits. The antiserum is sufficiently active to be useful in radioimmunoassay procedures.

Elevated 24-hour mean plasma concentration of FSH in men with cirrhosis of the liver.

Twenty-four-hour mean plasma concentrations of estradiol, estrone, testosterone, LH and FSH were measured in 6 men with typical alcoholic liver cirrhosis and in 17 age-matched controls. Plasma estradiol levels were markedly elevated in the cirrhotic patients, as has been reported before. Plasma levels of estrone, testosterone and LH did not differ significantly from normal. The most striking finding was marked elevation of the plasma FSH level: the mean value in the cirrhotic group was 19.5 mIU/ml (95% confidence limits, 13-29), more than twice the mean control value, 9.7 mIU/ml (95% confidence limits, 7.0-13) (p less than 0.0005). This consistent elevation of FSH in cirrhotic men may be due to a deficient testicular secretion of inhibin as a result of alcohol toxicity to the testes.