Open-sandwich enzyme immunoassay for one-step noncompetitive detection of corticosteroid 11-deoxycortisol
Improved HPLC method for simultaneous analysis of cortisol, 11-deoxycortisol
, prednisolone, methylprednisolone, and dexamethasone in serum and urine.
Weiming Li, professor of fisheries and wildlife who helped lead the project, said: "By identifying 11-deoxycortisol
as a stress hormone in lamprey, it allows us to better understand how the endocrine system in vertebrates evolved into the complex systems we see in humans today.
Steroid concentrations in the medium and cell lysate were measured using liquid chromatography/mass spectrometry for 12 steroids [pregnenolone (PREG), 17[alpha]-hydroxypregnenolone (HPREG), dehy-droepiandrosterone (DHEA), progesterone (PROG), 17[alpha]-hydroxyprogesterone (HPROG), androstenedione (DIONE), testosterone (T), deoxycorticosterone (DCORTICO), corticos-terone (CORTICO), ALDO, 11-deoxycortisol
(DCORT), and CORT] and using enzyme-linked immunosorbent assay for two additional steroids [estrone ([E.
To eliminate endocrinological causes of hypertrichosis, serum levels of testosterone, DHEA-SO4, 17-OH progesterone, and 11-deoxycortisol
were measured and were found to be within normal limits.
Metyrapone inhibits the conversion of 11-deoxycortisol
to cortisol resulting in an increase in 11-deoxycortisol
(11DC; Reichstein s substance S),17-hydroxyprogesterone (17OHP), 17-hydroxypregnenolone (17OHPr), and pregnenolone (Pr), allows detection of these enzyme defects (6-9).
Recombinant CYP110 genes encode enzymes that can catalyze conversion of 11-deoxycortisol
to cortisol, 18-hydroxycortisol, and 18-oxocortisol.
This approach relies on measurement of 17-hydroxyprogesterone (17OHP), which is normally converted to 11-deoxycortisol
by CYP21 and is therefore increased when the enzyme is deficient.
Antibody specificity was evaluated by testing testosterone, cortisol, DHEA, cortisone, corticosterone, 11-deoxycortisol
, 21-deoxycortisol, prednisolone, prednisone, dexamethasone, triamcinolone, 17-[alpha]-hydroxyprogesterone, estriol, aldosterone, progesterone, and estradiol at 50 [micro]g/L or higher.
Apparent cortisol concentration at 10, 100, and 1000 nmol/L of the cross-reactant cortisone and 11-deoxycortisol
never exceeded, respectively, 0.
Aldosterone, cholesterol, cortisone, cortisol, danazol, dehydroepiandrosterone, dehydroepiandrosterone sulfate, estradiol, estradiol glucuronate, estriol, estrone, progesterone, corticosterone, testosterone, norethindrone, pregnenolone sulfate, 11-deoxycortisol
, 17[alpha]-hydroxypregnenolone, and 17[alpha]-hydroxyprogesterone at concentrations of [10.