6-beta-hydroxycortisol and Cushing-Syndrome

Topics: ACTH Syndrome, Ectopic; Adrenal Cortex Diseases; Adrenocortical Hyperfunction; Biomarkers; Chromatography, High Pressure Liquid; Cushing Syndrome; Diagnosis, Differential; Humans; Hydrocortisone; Hyperaldosteronism; Immunoenzyme Techniques; Pituitary ACTH Hypersecretion; Radioimmunoassay; Reference Values; Specimen Handling

Topics: Adrenal Cortex; Adult; Amniotic Fluid; Animals; Arthritis, Rheumatoid; Cushing Syndrome; Depression; Female; Humans; Hydrocephalus; Hydrocortisone; Hydroxylation; Infant, Newborn; Infant, Premature; Liver Cirrhosis; Male; Meningomyelocele; Methods; Neoplasms; Pregnancy; Pregnancy Complications; Steroid Hydroxylases; Thyroid Diseases; Tissue Distribution

To evaluate the diagnostic value of serum levels of adrenal steroids for diagnosing and subtyping Cushing's syndrome. Patients diagnosed with endogenous Cushing's syndrome (34 and 19 patients with adrenal and pituitary Cushing's syndrome, respectively) and healthy controls (n = 34) were consecutively enrolled at Seoul National University from 2016 to 2020. Morning serum samples were collected before and 3 months after treatment. Serum steroids were profiled using liquid chromatography-mass spectrometry. The diagnostic value of each and the combination of steroids were assessed using the area under the curve of receiver operating characteristic (AUROC) and decision tree analysis. Tetrahydrocortisone and 6β-hydroxycortisol showed the highest AUROC (0.893 and 0.890, respectively) for the diagnosis of endogenous Cushing's syndrome. The decision tree composed of tetrahydrocortisone and 6β-hydroxycortisol correctly classified 79/87 (90.8 %) subjects. For subtyping into adrenal or pituitary Cushing's syndrome, dehydroepiandrosterone sulfate (DHEA-S) showed the highest AUROC (0.988), which was similar to that of plasma ACTH (0.994, P = 0.458). The decision tree composed of only DHEA-S correctly classified 51/53 (96.2 %) of the Cushing's syndrome subtype. DHEA-S showed a significant linear correlation with the plasma ACTH level, but not with the 24 -h urine free cortisol or dexamethasone suppression test results. All steroids, except allo-tetrahydrocortisol and tetrahydrocortisone, decreased significantly at 3 months post-treatment with similar patterns in both adrenal and pituitary Cushing's syndrome. Serum steroid profiling using a single morning serum sample provides valuable information for diagnosing and subtyping Cushing's syndrome.

Topics: Adrenal Glands; Adult; Case-Control Studies; Cushing Syndrome; Female; Humans; Hydrocortisone; Male; Middle Aged; Pituitary Gland; ROC Curve; Steroids; Tetrahydrocortisone

The aim of this study was to examine and compare the potential usefulness of plasma and salivary 6beta-hydroxycortisol measurements for assessing adrenocortical activity in patients with adrenocortical adenomas. Plasma and salivary cortisol as well as 6beta-hydroxycortisol determinations were performed by radioimmunoassay after extraction with ethyl acetate followed by chromatographic separation using a modified paper chromatographic system. Samples were obtained from 36 control subjects and 37 patients with non-hyperfunctioning adrenocortical adenomas in the morning at 8 a.m. after a low-dose of dexamethasone and after stimulation with synthetic depot ACTH. Basal and post-dexamethasone hormone levels were also measured in plasma and salivary samples of 4 patients with Cushing's syndrome from adrenal adenomas. In the baseline state, patients with non-hyperfunctioning adrenocortical adenomas had significantly higher plasma and salivary 6beta-hydroxycortisol levels (mean+/-SE, 79.0+/-7 and 17.1+/-2.2 ng/dl, respectively) compared to those measured in controls (62.0+/-4 and 7.7+/-0.6 ng/dl, respectively), whereas baseline plasma and salivary cortisol levels (9.6+/-0.5 microg/dl and 342+/-39 ng/dl, respectively) were similar to those measured in the control group (9.9+/-0.4 microg/dl and 366+/-24 ng/dl, respectively). In all groups, the changes in plasma and salivary 6beta-hydroxycortisol concentrations after dexamethasone suppression and ACTH stimulation were similar to the changes in plasma and salivary cortisol levels, although the differing ratios of 6betaOHF to cortisol indicated potentially important variations in the induction of 6beta-hydroxylase activity between the three groups. In patients with Cushing's syndrome, baseline plasma and salivary 6beta-hydroxycortisol concentrations (754+/-444 and 104+/-88 ng/dl, respectively) were more markedly increased than plasma and salivary cortisol levels (24.8+/-6.7 microg/dl and 1100+/-184 ng/dl, respectively), and all remained non-suppressible after dexamethasone administration. These results suggests that plasma and salivary 6beta-hydroxycortisol determinations may precisely detect not only overt increases of cortisol secretion in patients with Cushing's syndrome but also mild glucocorticoid overproduction presumably present in patients with non-hyperfunctioning adrenocortical tumors.

Topics: Adrenal Cortex Neoplasms; Adrenocortical Adenoma; Adrenocorticotropic Hormone; Adult; Cushing Syndrome; Dexamethasone; Female; Glucocorticoids; Humans; Hydrocortisone; Male; Middle Aged; Radioimmunoassay; Salivary Glands; Statistics, Nonparametric

To study the usefulness of 6beta-hydroxycortisol (6betaOHF) measurements for assessing hepatic drug metabolizing enzyme activity, plasma 6betaOHF and cortisol were measured in 22 patients with alcoholic liver disease after at least 2 weeks of alcohol abstinence, in 5 patients with severe Cushing's syndrome and in 12 healthy non-drinker subjects. Blood samples were drawn under resting conditions during midnight, in the morning at 0800 h, after a 1-mg overnight dexamethasone test and after ACTH administration. Plasma cortisol and 6betaOHF were determined with radioimmunoassay. In patients with alcoholic liver disease, the plasma cortisol levels at midnight and 0800 h, as well as after the administration of dexamethasone and ACTH were not different from corresponding values measured in non-drinker controls. In addition, these patients with alcoholic liver disease had similar plasma 6betaOHF levels at midnight, 0800 h and after dexamethasone administration as compared to corresponding values in controls. By contrast, ACTH administration in patients with alcoholic liver disease resulted in a significantly (p<0.05) larger increase of plasma 6betaOHF (from 106 +/- 22 to 1102 +/- 106 ng/dl, mean +/- SE) as compared to that found in controls (from 74 +/- 3 to 337 +/- 76 ng/dl). The markedly increased 6betaOHF response to ACTH administration in patients with alcoholic liver disease was similar to that measured in patients with severe Cushing's syndrome, in whom increased and non-suppressible plasma cortisol levels were accompanied by markedly elevated plasma 6betaOHF levels. These results indicate that alcohol abstinence in patients with alcoholic liver disease is associated with an exaggerated 6betaOHF response to ACTH and that this abnormality may prove to be a clinically useful parameter for a sensitive detection of altered drug metabolism present in these patients.

Topics: Adrenocortical Adenoma; Adrenocorticotropic Hormone; Adult; Cushing Syndrome; Dexamethasone; Humans; Hydrocortisone; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Liver Function Tests; Middle Aged; Radioimmunoassay; Time Factors

Topics: Adolescent; Adult; Age Factors; Aged; Child; Child, Preschool; Cushing Syndrome; Female; Humans; Hydrocortisone; Infant; Infant, Newborn; Male; Middle Aged; Phenobarbital; Reference Values; Sex Factors; Specimen Handling

A comparative study of urinary free cortisol and urinary 6 beta-hydroxycortisol levels as a diagnostic test for hypercortisolemic states was carried out by measuring the excretion in 24-h specimens from 289 apparently healthy subjects and 10 Cushing patients. The diurnal variations of both variables were examined in normal subjects and subjects with altered adrenal activities. Two of the 289 apparently normal subjects had high values of urinary free cortisol; one had a high, the other a normal 6 beta-hydroxycortisol level; they were later diagnosed as having Cushing's syndrome and infertility, respectively. Three other subjects had high values of the urinary variables, but during 5 years of follow-up did not show any clinical evidence of hypercortisolism. The two urinary variables gave no false-negative results in the Cushing patients. The diurnal variation revealed that levels of 6 beta-hydroxycortisol change in parallel with those of free cortisol in normal subjects and in subjects with altered adrenal activities. However, the ratio of 6 beta-hydroxycortisol to free cortisol during the diurnal variation varied from low values when free cortisol levels were high to high values when free cortisol levels were low. In normal subjects, 1 mg of dexamethasone taken orally at 23.00 h completely suppressed the levels of both variables on the following day. It is concluded that urinary 6 beta-hydroxycortisol is correlated to urinary free cortisol so that measurement of urinary 6 beta-hydroxycortisol levels can be used as a diagnostic test for hypercortisolism in a way comparable to the method using urinary free cortisol.

Topics: Adolescent; Adrenal Cortex; Adult; Circadian Rhythm; Cushing Syndrome; Follow-Up Studies; Humans; Hydrocortisone; Middle Aged

A method using high performance liquid chromatography for the quantitative determination of cortisol and 6 beta-hydroxycortisol in urine is described. Urine extracts were fractionated by high performance liquid chromatography. The peak of 6 beta-hydroxycortisol was identified on the basis of retention time (24.4 min) and its area was measured. The peak of cortisol could not be measured simultaneously with the same column because of interference by other absorbing materials. The cortisol and dexamethasone peaks were collected (Rt values 9.6 and 8.1 min, respectively), rechromatographed on another column system, and the concentration was determined. Precision and accuracy of the present method were within the range commonly achieved by other methods established for both steroids. Clinical utility of the present method was evaluated by measuring urinary cortisol and 6 beta-hydroxycortisol in normal subjects, in Cushing's syndrome and disease patients, and in patients receiving cortisol therapy.

Topics: 17-Hydroxycorticosteroids; Adult; Chromatography, High Pressure Liquid; Cushing Syndrome; Female; Humans; Hydrocortisone; Male; Middle Aged

A reliable radioimmunoassay for plasma and urine 6 beta-hydroxycortisol after chromatography on Sephadex LH 20 column has been described and evaluated. The antiserum used was raised in the rabbit injected with 6 beta-hydroxycortisol-3(O-carboxymethyl)oxime-bovine serum albumin. In control subjects, urine 6 beta-hydroxycortisol levels ranged from 137 to 348 micrograms/24 h (mean +/- SD: 238 +/- 66) in adult males (n = 14) and from 80 to 432 micrograms/24 h (mean +/- SD: 210 +/- 93) in adult females (n = 15). In plasma, 6 beta-hydroxycortisol levels ranged from 0.53 to 3.13 ng/ml (mean +/- SD: 1.14 +/- 0.57) in adult males (n = 17) and from 0.53 to 2.69 ng/ml (mean +/- SD: 1.22 +/- 0.53) in adult females (n = 19). In patients with Cushing's syndrome high levels were found in urine as well as in plasma. Finally the higher concentrations of 6 beta-hydroxycortisol found in adrenal effluent in comparison with those of the peripheral vein blood has clearly demonstrated that 6 beta-hydroxycortisol is secreted by the adrenal besides its extra-adrenal formation from cortisol.

Topics: Adolescent; Adult; Aged; Cushing Syndrome; Female; Humans; Hydrocortisone; Male; Middle Aged; Radioimmunoassay; Reference Values

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.

Topics: 17-Hydroxycorticosteroids; Adolescent; Adrenocorticotropic Hormone; Adult; Child; Cushing Syndrome; Female; Humans; Hydrocortisone; Male