[Evaluation of urinary excretion of free-cortisol in patients with abnormal pituitary-adrenal axes].

Since urine contains cross-reacting substances to anti-cortisol(F) antibody, we extracted F from urine samples with dichloromethane. Using this specific assay system for F with TDX dynapack(fluorescence polarization immunoassay system), clinical significance of urinary F was tested by measuring urine samples from normal subjects, patients with pituitary-adrenal diseases and liver cirrhosis, and pregnant. The circadian rhythm of urinary F was evaluated by using samples collected every 4-hour period. The amount of urinary excretions of F during 4 hours were the highest during between 4 a.m. to 8 a.m. and then gradually decreased reaching at the lowest level at between 0 a.m. and 4 a.m. in normal volunteers. Such circadian rhythm was not found in 6 patients with Cushing's syndrome. The urinary F levels in pregnant women started to increase in the samples from between 21 to 30 weeks of gestation, and the level was maintained even at the 5th day of post-partum. The urinary F levels in patients with liver cirrhosis were significantly lower than those for age-matched healthy volunteers. In patients with abnormal steroidogenesis, the urinary F levels in patients with Cushing's syndrome were extremely high, 165-3358 micrograms/day. On the other hands, those in patients with deficient steroid synthetase were 2.8-26.5 micrograms/day, and those in patients with neuroblastoma and pheochromocytoma were 7.8-43.8 micrograms/day. The urinary F levels were about 50-fold higher in the patients with Cushing's syndrome than those in the normal reference interval, whereas serum F and urinary 17-OHCS levels were only 2.5-fold and 7-fold higher than those healthy volunteers, respectively. These results indicate that the assay of urinary F extracted with dichloromethane is the most useful test for the diagnosis of Cushing's syndrome.

[Usefulness of measurement of hormones in urine samples collected during the night period for the diagnosis of endocrine disorders].

It has been known that the measurement of adreno-cortical and medullary hormones in urine for 24 hours is the best way for diagnosis of the disease state. This is not only because hormone release shows circadian rhythm but also because the urinary excretion of hormones reflects the total amount of hormones produced for 24 hours. However, the civilized people, particularly the workers, are too busy to correct urine for 24 hours. Therefore, we aimed to evaluate whether the urine sample corrected for only 12 hours during night is useful for the diagnosis of adrenal diseases. Circadian rhythm of hormones in urine was evaluated by correcting urine every 4 hours in 18 normal adults. Diurnal variations of 17-ketosteroids (17-KS) and 17-hydroxycorticosteroids (17-OHCS) were evident; they showed the peak value during 8 AM and 12 AM, and the trough during 0 AM and 4 AM. Similarly, urinary noradrenaline and adrenaline showed the peak value at day time and the trough at night time. However, the rhythm was not observed in urinary dopamine and vanilyl mandelic acid (VMA). Under consideration of outpatient's convenience, urine sampled during night (between 8 PM and 8 AM) was evaluated. The amount of hormones during night was 35.9-46.9% of the total amount of the day, and the deviation was relatively small. The reference intervals (mean +/- 2SD) were as follows; 17-KS (n = 56, 2.8 +/- 2.4 mg/12 hours), 17-OHCS (n = 56, 2.0 +/- 2.0 mg/12 hours), 17-KGS (n = 52, 3.6 +/- 3.6 mg/12 hours), free cortisol (n = 43, 14.3 +/- 14.2 micrograms/12 hours), noradrenaline (n = 50, 12.9 +/- 9.8 micrograms/12 hours), adrenaline (n = 50, 2.1 +/- 4.0 micrograms/12 hours), dopamine (n = 50, 134 +/- 194 micrograms/12 hours), VMA (2.3 +/- 2.4 mg/12 hours). When the night periods urine in patients with adrenal disorders such as virilizing adrenocortical carcinoma, pituitary adenoma, adrenocortical hydroxylase deficiency, Addison's disease, hypopituitarism, Cushing's syndrome, pheochromocytoma was analyzed, the respective urinary hormone level corresponding to each disease was significantly changed from the reference interval. These results indicate that analysis of urine sample during night has advantage over the daily urine because the difference between normal and disease state is augmented when measured urinary during night.