Free 17-hydroxycorticosteroids Research Articles

Clinical and Genetic Heterogeneity, Overlap with Other Tumor Syndromes, and Atypical Glucocorticoid Hormone Secretion in Adrenocorticotropin-Independent Macronodular Adrenal Hyperplasia Compared with Other Adrenocortical Tumors

ACTH-independent macronodular adrenal hyperplasia (AIMAH) is often associated with subclinical cortisol secretion or atypical Cushing's syndrome (CS). We characterized a large series of patients of AIMAH and compared them with patients with other adrenocortical tumors. We recruited 82 subjects with: 1) AIMAH (n = 16); 2) adrenocortical cortisol-producing adenoma with CS (n = 15); 3) aldosterone-producing adenoma (n = 19); and 4) single adenomas with clinically nonsignificant cortisol secretion (n = 32). Urinary free cortisol (UFC) and 17-hydroxycorticosteroid (17OHS) were collected at baseline and during dexamethasone testing; aberrant receptor responses was also sought by clinical testing and confirmed molecularly. Peripheral and/or tumor DNA was sequenced for candidate genes. AIMAH patients had the highest 17OHS excretion, even when UFCs were within or close to the normal range. Aberrant receptor expression was highly prevalent. Histology showed at least two subtypes of AIMAH. For three patients with AIMAH, there was family history of CS; germline mutations were identified in three other patients in the genes for menin (one), fumarate hydratase (one), and adenomatosis polyposis coli (APC) (one); a PDE11A gene variant was found in another. One patient had a GNAS mutation in adrenal nodules only. There were no mutations in any of the tested genes in the patients of the other groups. AIMAH is a clinically and genetically heterogeneous disorder that can be associated with various genetic defects and aberrant hormone receptors. It is frequently associated with atypical CS and increased 17OHS; UFCs and other measures of adrenocortical activity can be misleadingly normal.

Comprehensive study of urinary cortisol metabolites in hyperthyroid and hypothyroid patients

To further analyse the significance and mutual relationship of thyroid function-linked alterations in cortisol metabolism that have been separately and variously reported. Twenty-four-hour urine samples from 21 patients with hyperthyroidism (Graves' disease), 16 patients with hypothyroidism (Hashimoto's thyroiditis), 21 healthy age- and sex-matched controls for hyperthyroidism, and 16 healthy age- and sex-matched controls for hypothyroidism were evaluated for 6beta-hydroxycortisol (6beta-OHF), tetrahydrocortisol (THF), tetrahydrocortisone (THE), allo-tetrahydrocortisol (allo-THF), urinary free cortisol (UFF), urinary free cortisone (UFE) and 17-hydroxycorticosteroid (17-OHCS). Urinary 17-OHCS, THE and allo-THF levels increased considerably in hyperthyroid patients compared to the controls, while UFF and THF showed no difference between the two groups. Urinary 6beta-OHF was significantly lower in the hyperthyroid patients than in the controls. Both the urinary allo-THF + THF/THE and the UFF/UFE ratios were significantly lower in the hyperthyroid patients than in the controls, whereas only the former was significantly higher in the hypothyroid patients than in the controls. The urinary allo-THF/THF ratio was significantly higher in the hyperthyroid patients and significantly lower in the hypothyroid patients than in the controls. In an analysis of pooled subjects including all groups (n = 64), free T4 levels correlated negatively (P < 0.0001) with the urinary allo-THF + THF/THE ratio but not with the UFF/UFE ratio. The serum levels of free T4 correlated positively (P < 0.0001) with the urinary allo-THF/THF ratio. The thyroid hormones seem to affect the total 11beta-HSD activity (allo-THF + THF/THE) more strongly than the renal 11beta-HSD2 activity (UFF/UFE). 5alpha-reductase activity (allo-THF/THF) is also enhanced in hyperthyroidism, while the reduction of urinary 6beta-OHF in hyperthyroidism might be a secondary effect of the altered activity of the total 11beta-HSD and 5alpha-reductase.

High fluid intake increases urine free cortisol excretion in normal subjects.

To test the hypothesis that increased fluid intake increases the urine free cortisol, we prescribed 5 liters of fluid intake per day or a normal fluid intake according to a randomized cross-over design in six normal volunteers. Each period lasted 5 days, with a 2-day washout period of normal fluid intake between the two periods. Urine free cortisol, 17-hydroxycorticosteroids, and creatinine were measured daily during each study period, and the average value over each 5-day period was calculated for each subject. High fluid intake caused a significant increase in the mean urine free cortisol [126 +/- 33 (SD) vs. 77 +/- 18 micrograms/day, P < 0.005]. The frequency of urine free cortisol results that exceeded the upper normal limit of 95 micrograms/day was also much higher during high fluid intake (23/30 vs. 6/30, P < 0.005). By contrast, urine 17-hydroxycorticosteroids (high fluid vs. normal fluid: 5.3 +/- 1.5 vs. 5.0 +/- 1.7 mg/day, respectively, P = not significant) and urine creatinine (1.51 +/- 0.48 vs. 1.45 +/- 0.37 g/day, P = not significant) did not differ between the two study periods. We conclude that high fluid intake (5 liters/day) increases free cortisol excretion without an increase in urine 17-hydroxycorticosteroids. Thus, mild to moderate increases in urine cortisol excretion may not indicate hypercortisolism in individuals who have a high fluid intake and urine volume.

High Fluid Intake Increases Urine Free Cortisol Excretion in Normal Subjects

To test the hypothesis that increased fluid intake increases the urine free cortisol, we prescribed 5 liters of fluid intake per day or a normal fluid intake according to a randomized cross-over design in six normal volunteers. Each period lasted 5 days, with a 2-day washout period of normal fluid intake between the two periods. Urine free cortisol, 17-hydroxycorticosteroids, and creatinine were measured daily during each study period, and the average value over each 5-day period was calculated for each subject. High fluid intake caused a significant increase in the mean urine free cortisol [126 ± 33 (sd) vs. 77 ± 18 μg/day, P < 0.005]. The frequency of urine free cortisol results that exceeded the upper normal limit of 95 μg/day was also much higher during high fluid intake (23/30 vs. 6/30, P < 0.005). By contrast, urine 17-hydroxycorticosteroids (high fluid vs. normal fluid: 5.3 ± 1.5 vs. 5.0 ± 1.7 mg/day, respectively, P = not significant) and urine creatinine (1.51 ± 0.48 vs. 1.45 ± 0.37 g/day, P = not significant) did not differ between the two study periods. We conclude that high fluid intake (5 liters/day) increases free cortisol excretion without an increase in urine 17-hydroxycorticosteroids. Thus, mild to moderate increases in urine cortisol excretion may not indicate hypercortisolism in individuals who have a high fluid intake and urine volume.

Urinary free cortisol versus 17-hydroxycorticosteroids: a comparative study of their diagnostic value in Cushing's syndrome.

We evaluated the usefulness of the basal urinary 24-h excretion rates of free cortisol versus 17-hydroxycorticosteroids in the diagnosis of Cushing's syndrome. On an outpatient basis, both urinary free cortisol and 17-hydroxycorticosteroids levels were determined in 48 patients with Cushing's syndrome, as well as in 95 obese and 94 healthy control persons of normal weight. Determination of the urinary free cortisol content allowed a clear-cut distinction between the patients with hypercortisolism and the controls, resulting in a sensitivity of 100% and specificity of 98% for the diagnosis of Cushing's syndrome. The diagnostic accuracy of urinary free cortisol was distinctly superior to that of 17-hydroxycorticosteroids, which showed a wide overlap of values between the groups, with a sensitivity of 73% and a specificity of 94%. In conclusion, the measurement of basal urinary free cortisol provided an excellent diagnostic sensitivity and specificity in the assessment of adrenocortical function. This simple and accurate test thus seems to be particularly useful in the outpatient evaluation of patients with suspected Cushing's syndrome.

The Influence of Simvastatin on Adrenal Corticosteroid Production and Urinary Mevalonate during Adrenocorticotropin Stimulation in Patients with Heterozygous Familial Hypercholesterolemia*

The adrenal gland requires a continuous supply of cholesterol for the biosynthesis of adrenal corticosteroids, which can be supplied by low density lipoprotein receptor-mediated uptake or local synthesis. The present study examined whether hypolipidemic therapy with a potent HMG CoA reductase inhibitor, simvastatin, compromises the adrenal response to ACTH stimulation in adult patients with heterozygous familial hypercholesterolemia. The adrenal response to a 36-h continuous ACTH infusion was determined at baseline and after 2 months of simvastatin treatment (40 mg, twice daily) in eight patients. Simvastatin reduced total and low density lipoprotein cholesterol levels by 36% and 45%, respectively. The time course of the increase in serum cortisol concentrations with continuous ACTH infusion was the same before and during simvastatin therapy, as were the rates of urinary excretion of free cortisol, 17-hydroxycorticosteroids, and 17-ketosteroids. Urinary excretion of mevalonate, which correlates with rates of whole body cholesterol synthesis, decreased from 3.8 +/- 0.42 (+/- SEM) mu,ol/24 h at baseline to 2.75 +/- 0.56 on simvastatin; no significant changes were seen in the urinary mevalonate levels before and after simvastatin therapy during ACTH stimulation. We conclude that the hypolipidemic effects of simvastatin in patients with heterozygous familial hypercholesterolemia are paralleled by a decrease in urinary mevalonate, but that the drug does not adversely affect ACTH-stimulated adrenal corticosteroid production.

6β-Hydroxycortisol Excretion in Hypercortisolemic States*

Urinary 6beta-hydroxycortisol (6betaOHF) excretion was measured and compared with free cortisol and 17-hydroxycorticosteroid (17OH) excretion in normal children, patients with Cushing's syndrome or disease (CSD), and patients during cortisol therapy. Normal 6betaOHF excretion in children was 0.23 +/- 0.03 mg/m2/24 h (mean +/- SE). No sex difference was found. ACTH infusion (40 U/day for 5 days) and high dose cortisol altered the 6 betaOHF:17OH ratio so that it was indistinguishable from the ratio seen in CSD. The fact that both Cushing's disease and high dose cortisol therapy caused the same change in the 6 betaOHF:17OH ratio suggests that cortisol and not ACTH induced 6beta-hydroxylase in hypercortisolemic subjects. Since the 6betaOHF:17OH ratio in CSD patients was always well above the normal range, measurement of 6betaOHF excretion was a better and more rapid test for chronic hypercortisolemia than urinary 17OH or free cortisol. Thus, measurement of urinary 6betaOHF is suggested as a good diagnostic test for hypercortisolemic states.

Excretory Patterns of Urinary Free 11-Hydroxycortico-Steroids and Total Oestrogens Throughout the Normal Menstrual Cycle

The excretion of urinary free 11-hydroxycorticosteroids and total oestrogens was studied daily in morning urine specimens throughout the menstrual cycle of six normal women (age range 18-24 years). The follicular phase of the cycle was characterised by apparently random fluctuations in the excretion of urinary free corticosteroids. However, after the mid-cycle oestrogen peak there occurred a significant drop in corticosteroid excretion, which then rose to a peak eight to 10 days after ovulation and was synchronous with the second oestrogen peak during the luteal phase.

Changes in fractions of urinary free 17-hydroxycorticosteroids before and after operation.

Fractions (compound F, compound E, THE and THE) of urinary free and total 17-OHCS were determined in ten patients who underwent various operations, using thin layer chromatography and column chromatography, and the following results were obtained. The excretion pattern of fractions of urinary free 17-OHCS was quite different from fractions of total 17-OHCS which were measured after hydrolysis with B-glucuronidase. In addition, there was a limit to the excretion of free comp. E, THE and THE. The excretion amounts of free comp. F and comp. E were several tens percent of comp. F and comp. E which were determined as fractions of total 17-OHCS. However, the excretion amounts of free THE and THE were only several percent of THE and THE which were determined as fractions of total 17-OHCS. It was clear that comp. F and comp. E were also conjugated with glucuronic acid though the degrees of conjugation were much lower than their metabolites, THE and THE. After operation, the rates of conjugation of free comp. F and comp. E diminished as compared with those in control subjects, and the metabolism of cortisol shifted to THE pathway.

Plasma 17-Hydroxycorticosteroid Levels in Healthy and Diarrhoeic Calves

SUMMARY Plasma free 17-hydroxycorticosteroid (17-OHCS) levels of healthy calves of the red-spotted breed varied with age. The highest concentration was found in newborn calves during the first 16 hours of life. The 17-OHCS values observed in diarrhoeic calves also varied with age. However, the 17-OHCS concentration in diarrhoeic calves aged 1–14 days was significantly higher than that of healthy animals of the same age.

EFFECTS OF HALOTHANE ANAESTHESIA AND SURGERY ON ADRENOCORTICAL FUNCTION IN MAN

The present study was undertaken to investigate whether or not preanaesthetic medication and halothane anaesthesia affect adrenal cortical function in man as judged by their influence on the plasma levels of free 17-hydroxycorticosteroids. The blood samples were taken in the morning between 8:15 and 8.45 A.M. before induction of anaesthesia, during anaesthesia alone, during surgery, and in the recovery room. The determination of free 17-OHCS in plasma was made by Peterson’s modification of the Porter-Silber method. Twenty patients were given halothane anaesthesia and another eleven patients received premedication alone without anaesthesia.

Free 11-hydroxycorticosteroids in plasma and urine in pregnancy and in cases of stress

The free plasma 11-hydroxycorticosteroids and the free urinary 11-hydroxycorticosteroids were measured in normal nonpregnant women, in normal pregnant women, women with a suspected fetal death, and in women under a surgical stress. The highest values of free plasma 11-hydroxycorticosteroids were found in normal pregnancy. The highest values of free urinary 11-hydroxycorticosteroids were found in the stress cases.

Effects of halothane anaesthesia and surgery on adrenocortical function in man.

The present study was undertaken to investigate whether or not preanaesthetic medication and halothane anaesthesia affect adrenal cortical function in man as judged by their influence on the plasma levels of free 17-hydroxycorticosteroids. The blood samples were taken in the morning between 8:15 and 8.45 A.M. before induction of anaesthesia, during anaesthesia alone, during surgery, and in the recovery room. The determination of free 17-OHCS in plasma was made by Peterson’s modification of the Porter-Silber method. Twenty patients were given halothane anaesthesia and another eleven patients received premedication alone without anaesthesia.

THE LEVELS OF 17-HYDROXYCORTICOSTEROIDS IN THE PLASMA OF USERS OF ORAL CONTRACEPTIVES

The effect of Anovlar Lyndiol 2.5 Ortho-Novin Volidan and mestranol on the plasma levels of free and conjugated 17-hydroxycorticosteroids were studied in blood samples taken from 101 women about Day 15 of the menstrual cycle and on Day 217 of treatment. Mean free 17-hydroxycorticosteroids in the morning rose significantly (p .05) similar to the increase of free hydroxycorticosteroids at the end of the last trimester of pregnancy. Evening levels of free and conjugated hydroxycoricosteroids also rose signifcantly (ps < and .001 respectively) indicating that oral contraceptives do not seem to alter the diurnal rhythm of free 17-hydroxycorticosteroids although it leads to plasma retention of these substances. Prolongation of contraceptive intake resulted in higher free 17-hydroxycorticosteroid and lower conjugated 17-hydroxycorticosteroid levels in the samples taken in the evening (p < .01).

Free 11-hydroxycorticosteroids in plasma in normal pregnancies and in cases of fetal death and missed abortion

The plasma hydrocortisone levels of pregnant women were measured by the fluorimetric method of Mattingly. A gradual increase in values was noted in all normally developing pregnancies. In 60 cases of missed abortion or intrauterine death of the fetus values for plasma hydrocortisone were considerably lower than those found in normal pregnancies.With few precautions taken, a simple fast fluorimetric test is suggested to determine the presence of a missed abortion or fetal death.

Studies on Plasma Free 11-hydroxy corticosteroids in Infants and Children

Studies on Plasma Free 11-hydroxy corticosteroids in Infants and Children

Correlation between endocrine function and gastric secretion (I) influences of pepsinogen upon the pituitary-adrenocortical function

Various studies have been reported for secretory mechanism of gast r ic pepsinogen under influence of hypothalamo-pituitary-adreno cortical control, but the exact mechanism sti l l remains unknown. The present s tudies were under taken to invest igate physiological effect of pepsinogen in conjunction with the pi tu i tary adrenocortical function. Dogs were used under pentobarbi ta l anesthesia dividing into th ree groups. 1) Normal group: Exogenous pure crysta l l ine pepsinogen in saline (0"02 mg/kg) was adminis tered in t ravenously to normal subjects for 30 minutes to demonst ra te a marked increase in the serum pepsinogen level f rom the beginning, and increased excret ion of ur inary to ta l 17-hydroxyeorticosteroid and Uropepsin per hour were demonstra ted for the period of one hour af ter the intravenous infusion ended. Simultaneously, hourly out-put of acid and pepsin in the gast r ic juice was moderate ly increased. The resul ts were approximately three t imes than the average value. 2) In dogs with no blood supply to the s tomach devised by ligation, a s imi lar resul ts were obtained for the hourly excretion of uropepsin and ur inary total 17-hydroxycortieosteroids. In this group the plasma free 17-hydroxycorticosteroids levels f rom the adrenal blood increased markedly dur ing pepsinogen infusion. 3) In bi lateral ly adrenectomized dogs, hour ly excretion of uropepsin and output of acid and pepsin in the gastr ic juice were minimal. In s ta t is t ica l analysis significant correlat ion (p<0.05, 7=0.71) was noted between maximal excret ion of uropepsin per hour and ur inary total 17-hydroxycorticosteroids in normal subjects. In view of the above findings, it seems to assume f rom the homeosta t ic s tand point tha t there may be "nega t i ve feed back m e c h a n i s m " concerning to the re la t ion of the secret ion of pepsinogen and pi tui tary-adreno cortical function. Correlat ion between endocr ine funct ion and gast r ic secretion under effect of pepsinogen infusion on antrectomized-denervated-, or hypophysectomized dogs will be reported in future.

The relation between free cortisol in urine and 'urinary free 11-hydroxycorticosteroids' as measured by fluorescence.

SUMMARY The relation between free cortisol in urine and 'free Δ4·3 keto-11-hydroxycorticosteroids' has been explored by measurements in sixty urine samples. Free cortisol was measured by a method which involved paper chromatography and incorporated an internal isotopic standard to correct for losses. A rapid and simple technique, measuring fluorescence in ethanolic sulphuric acid, was used to determine the free 11-hydroxycorticosteroids. Although there was a good correlation between the two measurements (r = 0·96), the percentage contribution to the free 11-hydroxycorticosteroids made by true cortisol was small (mean 33 %) and varied from 8 to 73 % in different urine samples.

CORTICOSTEROIDS IN HUMAN BLOOD. II. FREE AND CONJUGATED 17-HYDROXYCORTICOSTEROIDS IN ESSENTIAL HYPERTENSION.

Metabolism of corticosteroids in hypertension was further investigated. Plasma levels of “free”, “polar free”, glucuronide, and solvolyzable conjugate fractions of 17-hydroxycorticosteroids (17-OH-CS) were estimated by methods recently developed in this laboratory in 17 patients with benign uncomplicated essential hypertension and 60 normotensive control subjects. The results obtained revealed the following statistically significant differences between the 2 groups: 1) levels of 17-OH-CS glucuronides were lower in hypertensives than in normotensives; 2) levels of solvolyzable (presumably sulfate) conjugated 17-OH-CS were higher in hypertensives; 3) the ratio of glucuronides to “sulfates,” which was therefore markedly lower in hypertensives, correlated fairly well (inversely) with the blood pressure of these patients; 4) the ratio of free to solvolyzable steroids in hypertensives was decreased; 5) the ratio of polar free to free 17-OH-CS in hypertensives was increased; 6) preliminary investigation into the...

The effect of ACTH on the plasma level of non-esterified fatty acids in healthy subjects and in diabetic patients

Following an intramuscular injection of ACTH to a group of healthy subjects a decrease was observed in the level of non-esterified fatty acids of the plasma, replaced by an increase in their concentration, whereas in a group of diabetes patients only a stage of decrease was observed under identical conditions. The most marked decrease in the concentration of non-esterified fatty acids in the plasma was noted in cases with a higher initial level. No quantitative relationship could be established between the decrease in the concentration of non-esterified fatty acids in the plasma was noted in cases with a higher initial level. No quantitative relationship could be established between the decrease in the concentration of non-esterified fatty acids and the increase in the amount of free 11-hydroxycorticosteroids of the plasma following an ACTH injection. The results of observations evidence the dependence of the ACTH administration effect on the initial condition of the organism, and agree with the hypothesis of a direct, extraadrenal mechanism of ACTH action on the level of non-esterified fatty acids of the plasma.