C.J.P. Giroud

Insulin antagonism of cortisol action on lecithin synthesis by cultured fetal lung cells

In monolayer cel cultures from late gestation (28-day) rabbit fetal lungs, cortisol (5.5 X 10(-6)M) enhances lecithin synthesis and reduced cellular growth. The addition of insulin (25-100 muU/ml) abolishes the stimulatory effect of cortisol on lecithin synthesis but does not affect its growth-inhibiting activity.

The growth promoting effect of cortisol on human fetal lung cells

Abstract The addition of cortisol (5.5 × 10 −6 M) to the culture media of monolayer cultures of midgestation human fetal lung cells resulted in marked enhancement of growth as monitored by DNA accumulation. In contrast, the same molar concentration of cortisol led to growth inhibition of cultures of fetal larynx, trachea and esophagus (LTE) and of skin fibroblasts. Cortisone also promoted growth, but to a lesser extent than cortisol. The lung cells were capable of forming cortisol from cortisone, the magnitude of this conversion increasing with the length of time the cells were maintained in culture and being greater in cells which had previously been exposed to cortisol. These findings are interpreted as suggesting a role for cortisol and cortisone in human fetal lung growth.

Short term tissue culture of human midterm and term placenta: Parameters of hormonogenesis

Abstract Monolayer cultures of human midterm and term placentae have been established following trypsin dispersion of placental minces. Maintenance of endocrine function was monitored by the concentrations of specific hormones in the culture media. At either gestational age the cultures 1) secrete estradiol-17β(1) and estrone (in a ratio of about 1: 20) and aromatize 3H- or 14C-dehydroepiandrosterone sulfate and 14C-androstenedione, estrogen production being markedly enhanced by addition of dehydroepiandrosterone (10−6M) to the culture medium; 2) metabolize 3H-pregnenolone to progesterone and 14C-cortisol to cortisone; and 3) produce increasing amounts of chorionic gonadotropin and decreasing amounts of placental lactogen during the first week in culture. It is proposed that the model is highly suited to the study of factors affecting hormonogenesis by the human placenta whether they be of maternal or of fetal origin

Incorporation of 7α-3H-pregnenolone and 4-14c-progesterone into corticosteroids and some of their ester sulfates by human newborn adrenal slices.

Abstract Adrenal slices from a premature infant time of gestation: 22 weeks were incubated in the presence of equimolar amounts of 7α- 3 H-pregnenolone (7 α- 3 H-3β-hydroxypregn-5-en-20-one) and 4- 14 C-progesterone (4- 14 C-pregn-4-ene-3,20-dione). The incorporation of these two precursors into several steroids was studied as a function of the length of incubation 2.5 hrs. 17α-Hydroxyprogesterone (17α(-hydroxypregn-4-ene-3,20-dione), 11-deoxycortisol (17α(, 21-dihydroxypregn-4-ene-3, 20-dione) and cortisol (11β, 17α, 21-trihydroxypregn-4-ene-3, 20-dione) contained 14 C and 3 H in a ratio of 3.0–3.8, progesterone in a ratio of 31. 17-Deoxy-21-hydroxysteroids (11-deoxycorticosterone (21-hydroxy-pregn-4-ene-3, 20-dione) corticosterone (11β, 21-dihydroxypregn-4-ene-3, 20-dione) and their respective C-21 ester sulfates) contained low or undetectable amounts of 3 H. Pregnenolone sulfate (20-ketopregn-5-ene-3β -y1 sulfate) and dehydroisoandrosterone sulfate (17-ketoandrost-5-ene-3β-yl sulfate) were characterized as the main biotransformation products of 7α- 3 H-pregnenolone. The results are interpreted as illustrating pathways of steroid biogenesis leading to the formation of 17-hydroxy-corticosteroids from both progesterone and pregnenolone, the latter via 17α-hydroxypregnenolone (3β, 17α-dihydroxypregn-5-en-20-one), of 17-deoxycorticosteroids and some of their C-21 ester sulfates, mainly from progesterone and of steroids of the pregn-5-en and androst-5-ene-3β-y1 sulfate series from pregnenolone.

Simultaneous measurement of plasma cortisol, cortisone, corticosterone, corticosterone sulfate and 11-deoxycorticosterone sulfate by competitive protein binding assays during the perinatal period

Abstract A method has been developed for the simultaneous measurement of the main C-21 corticosteroids of the pregn-4-ene series characterized in umbilical cord plasma, namely: 11β,17 α,21-trihydroxypregn-4-ene-3, 20-dione (cortisol), 17α,21-dihydroxypregn-4-ene-3,11,20-trione (cortisone), 11β,21-dihydroxypregn-4-ene-3,20-dione (corticosterone), 11β-hydroxy-3,20-diketopregn-4-ene-21-yl sulfate (corticosterone sulfate) and 3,20-diketopregn-4-ene-21-yl sulfate (11-deoxycorticosterone sulfate). It involves extraction of plasma, before and following hydrolysis of steroid sulfates, and chromatography of the steroid alcohols thus obtained, prior to quantification by competitive protein binding assays, using dog plasma for cortisone and human male plasma in all other instances. 3 H-steroid alcohols and sulfates of high specific activity provide means of correcting for experimental losses. For each steroid the lower limit of sensitivity is in the order of 0.5 μg/100 ml of plasma. The results obtained compare favourably with double isotope derivative assays — being higher, however, for 11-deoxycorticosterone sulfate. The method is applicable to the assessment of adrenocortical function postnatally, and to the exploration of the factors affecting the plasma concentrations of the five steroids considered.

Characterization of the 21-yl sulfates of llβ,17α,21-trihydroxypregn-4-ene-3, 20-dione; 17α,21-dihydroxypregn-4-ene-3,11,20-trione; llβ,21-dihydroxypregn-4-ene-3, 20-dione; 21-hydroxypregn-4-ene-3,11,20-trione and 21-hydroxypregn-4-ene-3,20-dione in human cord plasma

Abstract The 21-yl sulfates of 113,17α,21-trihydroxypregn-4-ene-3,20-dione (cortisol); 17α,21-dihydroxypregn-4-ene-3,11,20-trione (cortisone); llβ, 21-dihydroxypregn-4-ene-3,20-dione (corticosterone); 21-hydroxypregn-4-ene-3, 11,20-trione (11-dehydrocorticosterone) and 21-hydroxypregn-4-ene-3,20-dione (11-deoxycorticosterone) were identified in pools of human mixed cord plasma by a method combining double isotope assay with crystallization. The data imply the presence of these conjugates in the fetal circulation, at term.

Production rate of corticosterone sulfate in normal human adults

Abstract Following a single intravenous injection of either 150 μg or of a tracer dose of 1,2-3H-corticosterone-21-sulfate (1,2-3H-11 β-hydroxy −3, 20-diketopregn-4-ene-21-yl-sulfate) to 4 normal adult males (age 20–40), its half life in plasma was calculated to be in the order of 451. Of the total radioactivity extractable from urine about 25% was released after mammalian β-glucuronidase hydrolysis, and 70–78% was subsequently extracted with ether-ethanol (sulfate fraction). The major steroid component of the latter fraction was corticosterone sulfate itself, from the specific activity of which production rates of 530 and 780 μg/day were calculated in two subjects. Following simultaneous injection of a tracer dose of 1,2-3H-corticosterone-21-sulfate and of 4-14 C-corticosterone in one subject, the calculated production rate of the latter steroid was 2.5 mg/day, that of the conjugate 270 μg. Purification of the glucuronide fraction led to the characterization of 14C-tetrahydrocorticosterone (3∝, 11β, 21-trihydroxy-5β-pregnan-20-one), 14C-a 11 o-tetrahydrocorticolsterone (3∝, 11β, 21-trihydroxy-5∝-pregnan-20-one) and 14C-tetrahydro-11-dehydrocorticosterone (3∝, 21-dihydroxy-5β-pregnan-11, 20-dione), which did not contain detectable amounts of tritium. Conversely the corticosterone sulfate fraction purified from the ether ethanol extract was devoid of 14C. The results are interpreted as indicating a significant secretion of corticosterone sulfate by the normal human adrenal.

Exploration of the human fetal pituitary adrenal axis: stimulation of cortisol and dehydroepiandrosterone sulfate biosynthesis by homologous pituitary in organ culture.

Adrenals obtained from human abortices at midpregnancy were kept under conditions of tissue culture and the production of cortisol and dehydroepiandrosterone sulfate (3beta-hydroxy-5-androsten-17-one, 3-sulfate; DHA-S) monitored by radioimmunoassays for up to 2 weeks. Basal production of dehydroepiandrosterone sulfate was considerably higher than that of cortisol. alpha1-24corticotrophin, alpha1-39corticotrophin, (a long acting porcine corticotrophin) at the concentration of 1 mU/ml of culture medium in both instances, and dibutyryl cyclic AMP (1mM) enhanced the production of both steroids some 3 to 30 times above control values. Medium harvested from homologous pituitary cultures had comparable corticotrophic activity. It is concluded that at midpregnancy the regulation of corticoidogenesis implies the existence of a corticotrophic factor either identical or closely related to ACTH.