Biomedical subjects

F Iohan

Publications and source records attributed to F Iohan.

7 recordsLinked to original sources

PubMed1991

High-performance liquid chromatographic determination of cortolic and cortolonic acids as pyrenyl ester derivatives.

A new procedure is described for the detection of the acidic metabolites of cortisol (cortoic acids) as the pyrenylmethyl-21-oic esters. The derivatizing reagent, diazomethylpyrene, was prepared by an improved procedure. The reagent was used at room temperature, required no catalyst, and was not restricted by stoichiometric requirements. The steroid esters were separated by reversed-phase high-performance liquid chromatography and analyzed simultaneously by their ultraviolet absorbance and fluorescence characteristics. Identities of the products were confirmed using the photodiode array detector to determine spectral profiles, absorbance maxima, and absorbance ratios. Further confirmation of identity of the cortoic acid esters used mass spectrometry under normal and collision-activated dissociation conditions. With the method described, a linear spectral response was obtained between 8 and 1680 fmol. Application of the technique to the analysis of steroid acids in human urine indicated the presence of cortoic acids.

Chromatography, High Pressure Liquid↗

PubMed1989

Synthesis of tritium labeled cortoic acids.

A procedure is described for the microsynthesis and purification of the high specific activity tritium labeled cortisol metabolites, 20 alpha- and 20 beta-cortolic acids and 20 alpha- and 20 beta-cortolonic acids.

Cortisone↗

PubMed1986

Estrogen and progesterone receptors in the organs of prenatal cynomolgus monkey and laboratory mouse.

The estrogen and progesterone receptors of several organs of the prenatal cynomolgus macaque and the fetal mouse were studied using a combination of the dextran-coated charcoal technique and high-performance liquid chromatography. This procedure permitted the concurrent measurement of both receptors in minute amounts of tissue. Estrogen receptors, but not progesterone receptors, were found in the fetal monkey and mouse uteri. No estrogen or progesterone receptors were detected in the lungs, liver, kidney, heart, brain, adrenal gland, or limbs of mouse or monkey fetuses. The nonspecific binding of radioactive ORG-2058 was not displaced by unlabeled progesterone, 17 alpha-hydroxyprogesterone caproate, or ORG-2058. Because the steroid receptors that are indispensable mediators of steroid hormone action were absent from the nonreproductive tissues, prenatal development of these organs and tissues cannot be adversely influenced by exposure to estradiol, progesterone, or their synthetic analogues.

17 alpha-Hydroxyprogesterone Caproate↗

PubMed1986

11 beta,20-dihydroxy-3-oxopregna-4,17(20)-dien-21-al: an intermediate in the biological 17-dehydroxylation of cortisol.

The role of 11 beta,20-dihydroxy-3-oxopregna-4,17(20)-dien-21-al (F enol aldehyde) as an intermediate in the biological conversion of cortisol to a 17-deoxy-21-oic acid was studied in vitro with mouse liver as the source of enzyme. The substrate, [1,2-3H]F enol aldehyde, was synthesized and found to contain cis and trans isomeric forms in a ratio of approximately 3:1. Tritium labeled F enol aldehyde was incubated with mouse liver homogenates. The metabolic products were analyzed by TLC and HPLC. 11 beta,20-Dihydroxy-3-oxopregna-4-en-21-oic acid, a 17-deoxy-21-oic acid, was identified as a quantitatively significant metabolite of the cis isomer. Metabolic conversion of the trans isomer to acid metabolites did not occur. The results are consistent with our hypothesis that the metabolic 17-dehydroxylation of cortisol requires an enol aldehyde intermediate and indicate that the conversion of the enol aldehyde to hydroxy acid is stereospecific.

Animals↗

PubMed1986

The syndrome of apparent mineralocorticoid excess: its association with 11 beta-dehydrogenase and 5 beta-reductase deficiency and some consequences for corticosteroid metabolism.

We describe the metabolism of cortisol (F) in three children, two of them siblings, with apparent mineralocorticoid excess (AME). As with prior patients with AME, oxidation of F to cortisone (E) was impaired, but reduction of E to F was not. We propose that this metabolic defect is caused by deficient 11-dehydrogenase associated with unimpaired 11-reductase. The following supporting observations were made: urinary C21 11-hydroxy metabolites exceeded C21 11-oxo metabolites: ratio of urinary cortols to cortolones, 6.6 +/- 2.8 (+/- SD; normal, 0.47); tetrahydrocortisol (THF) and alloTHF to tetrahydrocortisone, 14.6 +/- 5.6 (normal, approximately 1); normal subjects oxidized [11 alpha-3H]F with transfer of 3H to water; the patients did not; 11-hydroxy, but not 11-oxo, C19 steroids were excreted into the urine; and fibroblasts from patients had 5 times more 11-reductase activity than normal subjects, though fibroblasts from neither group had 11-dehydrogenase activity. Other defects of cortisol metabolism not directly associated with 11-dehydrogenase deficiency were found: impaired conversion of tetrahydro to hexahydro neutral steroids, indicating defective reductive metabolism of the side chain; depressed F production rate and increased half-life of circulating F, resulting in normal blood levels of F; increased excretion of unconjugated F metabolites; and decreased excretion of THF relative to alloTHF, consistent with a 5 beta-reductase defect. Excretion of acidic metabolites of F (cortoic acids) was within the normal range. However, little or no 20 beta-hydroxy acids were excreted, while the level of urinary 20 alpha-hydroxy acids was increased. The 11-hydroxy to 11-oxo ratio of acid metabolites was similar to values in normal subjects. The proportion of cortoic acids relative to neutral hexahydro metabolites was increased (0.37 to 1.27 in patients; 22 in normal subjects). We conclude that children with AME have multiple defects in the conversion of F to neutral metabolites, while metabolism to cortoic acids was less extensively affected. How the defects in cortisol metabolism and the symptoms of AME are related remains to be determined.

11-beta-Hydroxysteroid Dehydrogenases↗

PubMed1985

Measurement of ethynylestradiol and levonorgestrel incorporated into sustained release contraceptive formulations.

High performance liquid chromatography was used to measure the concentrations of ethynylestradiol (EE) and levonorgestrel (LNG) released from contraceptive devices into aqueous medium containing the cationic detergent, benzalkonium chloride. Most of the detergent was removed after solvent extraction of the steroid, although small amounts of it remained in the steroid phase. Over the course of many injections into octasilyl (C8) or octadecylsilyl (C18) reversed-phase columns, the chromatographic profile of EE was gradually altered with multiple peaks emerging. The profile of LNG was not changed. EE chromatographed as a double peak on a fully end-capped C18 column. Rechromatography of each peak yielded a mixture of the two. Mass spectral analysis showed that the peaks differed only in the gain or loss of an equivalent of water. Introduction of a cation exchange column before the analytical column removed residual benzalkonium ions, and by thus preventing deterioration of the column, permitted EE to consistently emerge as a single, symmetrical peak.

Chromatography, High Pressure Liquid↗

PubMedDate not supplied

Corticosteroid-calcium complexes.

Glucocorticoids and calcium ions are shown to interact to yield a complex with properties that are distinct from those of the reactants. Reaction of steroids with Ca2+ appears to require the dihydroxyacetone side chain, since other structures do not react. Evidence for complex formation are: increased aqueous solubility of cortisol when Ca2+ is added to an aqueous or a biphasic aqueous/chloroform (or ethyl acetate) system; increased rate of migration of cortisol during reversed-phase thin-layer chromatography and HPLC; chromatographic comigration of 45Ca2+ and 3H-labeled cortisol; coprecipitation of 45Ca2+-3H-cortisol complexes. After dissociation of the cortisol-calcium complex, the only steroid recovered was cortisol. By the above criteria, the properties of cortisol were not affected by Sr2+, Ba2+, or Mg2+. The cleavage patterns of cortisol in the mass spectrometer corresponded to that of 11 beta-hydroxyandrostenedione when Ca2+ was present, and to cortisol in its absence. We therefore postulate that the structure of the dihydroxyacetone side chain was transiently altered by Ca2+, resulting in a labile C17-C20 bond. These results support our earlier proposal that the chemical and physico-chemical properties of corticosteroids are modified by calcium ions.

Adrenal Cortex Hormones↗