PubMed · 5658099
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J B Roberts, I E Bush, N B Gibree. 1968. Urinary steroid analysis: a comprehensive procedure.. https://doi.org/10.1016/0003-2697(68)90229-7
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The culture of human male foreskin fibroblasts was incubated with various concentrations of testosterone in the medium. It has been shown that the minimum formation of 17beta-oestradiol and 5alpha-dihydrotestosterone occurs at physiological testosterone concentrations. Any deviation of testosterone concentration, both up and down, was accompanied by an increase in the formation of 17beta-oestradiol and 5alpha-dihydrotestosterone.
We previously reported that the daily urinary unidentified ketosteroid glucuronide (US-G) level in patients with Cushing's syndrome was much higher than that in the healthy subjects. Furthermore, urine samples from patients with Cushing's syndrome, including those with pituitary adenoma and adrenal adenoma, yielded almost the same high excretion levels, despite the different sites of the adenomas. We extracted US obtained by hydrolysis of US-G in urine of patients with Cushing's syndrome, purified it, and analyzed its chemical structure. Molecular weight and molecular formula were analyzed by MS spectrometry, and the chemical structure was analyzed by NMR spectrometry, utilizing small quantities of refined US. The substance has a molecular weight of 304 Da, a molecular formula of C19H28O3, and its chemical structure is 3alpha,11beta-dihydroxyandrost-4-en-17-one.