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Query: UMLS:C0338671 (Steroids)
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Complete analysis of the proton nuclear magnetic resonance spectrum of desoxycorticosterone (DOC) has been made using selective double irradiation, two-dimensional experiments, relaxation rate, and nuclear Overhauser effect measurements in order to specify the structure and conformation of products encountered during the preparation of the specific antigen DOC-bovine serum albumin (BSA). It has been shown that DOC has the normal P conformation with ring A half-chair, and ring B chair. This confirms results previously obtained by circular dichroism measurements.
Steroids
PMID:Structural analysis of 21-hydroxypregn-4-ene-3,20-dione (deoxycorticosterone) derivative compounds. Part II: Proton NMR spectra. 321 60

A sensitive radioimmunoassay for dexamethasone 17,21-dipropionate and its four metabolites in human plasma and urine has been developed using single anti-dexamethasone antiserum. The antiserum was obtained by immunizing rabbits with dexamethasone-3-oxime-bovine serum albumin conjugate. All of the endogenous steroids tested cross-reacted less than 0.07%. Before radioimmunoassay, dexamethasone 17,21-dipropionate and dexamethasone 17-propionate were hydrolyzed to dexamethasone, and 6 beta-OH-dexamethasone 17-propionate was hydrolyzed to 6 beta-OH-dexamethasone in 3% ammonia/methanol at 5 C for 16 h. A standard curve was established with a useful range between 0.005 and 2 ng in the case of dexamethasone, between 0.05 and 5 ng in the case of 6 beta-OH-dexamethasone. Measurement of plasma concentrations and percent urinary excretion of the metabolites in healthy men was performed following occlusive dressing of dexamethasone 17,21-dipropionate cream and ointment. The main metabolites in plasma were dexamethasone 17-propionate and dexamethasone, which increased gradually and reached maximum levels (160-200 pg/mL) at 24-32 h after application. The major metabolites observed in urine were 6 beta-OH-dexamethasone 17-propionate and 6 beta-OH-dexamethasone. Total percentage of their urinary excretions within 72 h after application amounted to 0.28-0.50% of the dose administered.
Steroids
PMID:Radioimmunoassay for dexamethasone 17,21-dipropionate and its metabolites in plasma and urine after topical application. 324 67

5 alpha-[2,4-3H]Cholest-8(14)-en-3 beta-ol-15-one was administered to a series of male Sprague-Dawley rats by intragastric intubation in the form of an emulsion in a mixture of triolein, sodium taurocholate, bovine serum albumin, and glucose. [4-14C]Cholesterol was similarly administered to a second series of rats. The distribution of 3H and 14C was studied at 12 and 48 h after the administration of the sterols. The results demonstrated that the 15-ketosterol is absorbed and metabolized to material with the chromatographic properties of fatty acid esters of the 15-ketosterol, to cholesterol, and to fatty acid esters of cholesterol. The [3H]cholesterol formed from the 15-ketosterol was characterized by its behavior on silicic acid-Super Cel column chromatography, by the chromatographic behavior of its acetate derivative on alumina-AgNO3 column chromatography, and by purification by way of its dibromide derivative without significant change in specific activity. The general distribution of 3H was similar to that of 14C. No unusual concentration of 3H in any of the organs studied was observed.
Steroids
PMID:Inhibitors of sterol synthesis. Studies of the distribution and metabolism of 5 alpha-[2,4-3H]cholest-8(14)-en-3 beta-ol-15-one after intragastric administration to rats. 324 71

The preparation of high affinity and high specificity polyclonal and monoclonal antibodies to estradiol is described. Monoclonal antibodies were derived from BALB/c mice hyperimmunized with estradiol-3-O-carboxymethyl ether conjugated to bovine serum albumin. Spleen cells were hybridized with mouse myeloma cells. Quite a few monoclonal antibodies showed very good affinity for estradiol. Extended immunization and hyperimmunization were essential for producing a greater number of positive clones secreting high affinity antibodies. Binding constants of the antisera and their cross-reactivities with related steroids were calculated. Both polyclonal and monoclonal antibodies showed very high affinity for estradiol exhibiting little or no cross-reactivities with structurally related steroids indicating that this site of linkage is a good choice for discriminating between differences at the 16-17 position in the D-ring. This monoclonal antibody (44.28.6), having negligible cross-reactivity with estriol and estrone, can be used for diagnostic purposes.
Steroids
PMID:Production of highly specific polyclonal and monoclonal antibodies using estradiol-3-O-carboxymethyl ether as hapten. 325 32

Four strains of mice were immunized with 6 different conjugates of 3-O (carboxymethyl-oximino)-18-hydroxycortisol to bovine serum albumin (3 preparations), turkey serum albumin, porcine thyroglobulin, and keyhole limpet hemocyanin. Spleens from 7 of 48 mice immunized were fused with Fox/NY and/or HL-1 Friendly myeloma cell lines, yielding many positive clones for antibody formation. Short cross-reactivities were done in 293 culture supernatants and were found to have low cross-reactivity (less than 0.001%) to cortisol, but very high cross-reactivity to 18-hydroxy-11-deoxycortisol (70 to 140%). One clone showed over 100% cross-reactivity with all the 18-hydroxylated steroids studied. The major problem encountered in the generation of monoclonal antibodies was the low antibody response in the vast majority of mice injected. Half the mice developed no measurable titer, and the clones evaluated from those that did produce antibodies cross-reacted with other 18-hydroxylated steroids. Nevertheless, the antibody developed could serve in radioimmunoassay for 18-hydroxy-11-deoxycortisol separated chromatographically from other cross-reacting steroids. This is important as no synthetic 18-hydroxy-11-deoxycortisol is available.
Steroids
PMID:The production of a monoclonal antibody to 18-hydroxycortisol and other 18-hydroxylated steroids. 345 46

A specific radioimmunoassay (RIA) method is described for the determination of 21-deoxycorticosterone (21 DB) in human plasma. 21-Deoxycorticosterone-3-(O-carboxymethyl) oxime-bovine serum albumin conjugate was used to generate antisera in rabbits. Steroids which reacted significantly with the antisera were found to be progesterone, pregnenolone, corticosterone and 11-oxo progesterone. However, after extraction of plasma and column chromatography on Celite, all these steroids were separated from 21-deoxycorticosterone and consequently did not interfere with the radioimmunoassay. The intra- and interassays coefficients of variation were 8% and 11% respectively. Mean plasma 21-deoxycorticosterone level for healthy subjects was very low: 17.8 +/- 14.8 pmol/l (mean +/- SD) with no statistical difference between males and females. During the ACTH stimulation test, the 21-deoxycorticosterone levels of healthy subjects increased to 84.7 +/- 26.3 pmol/l (mean +/- SD) for males and 79.3 +/- 31.6 pmol/l (mean +/- SD) for females. Consequently high levels of plasma 21-deoxycorticosterone were found in treated patients suffering from congenital adrenal hyperplasia (CAH) with 21-hydroxylase deficiency, particularly in CAH salt-losers with high plasma renin activity (PRA), where the plasma level reached 40,545 pmol/l. Thus, 21-deoxycorticosterone may be a new marker for adrenal 21-hydroxylase deficiency.
...
PMID:The measurement of 11 beta-hydroxy-4-pregnene-3,20-dione (21-deoxycorticosterone) by radioimmunoassay in human plasma. 354 44

In order to develop a radioimmunoassay for the new progestagen dienogest (STS 557, 17 alpha-cyanomethyl-17 beta-hydroxy-estra-4,9-dien-3-one), bovine serum albumin (BSA) conjugates of STS 557-3-carboxymethyloxime and of STS 557-11-hemisuccinate were synthesized as antigens for the production of antisera. It was proved that an excess of isobutylchlorocarbonate in the coupling reaction using the "mixed anhydride method" results in an acylation of free NH2-groups in the BSA. By the immunization of rabbits with the STS 557-antigen-antisera of high specificity and affinity to STS 557 were produced. Endogenous steroids show no cross reaction with the STS 557-antisera. Steroids with a 17 alpha-CH2CN-group, being obtained by chemical synthesis or microbial transformation, compete with STS 557 for the binding positions of the antibodies to a different extent.
...
PMID:[Bovine serum albumin conjugates of the new progestagen dienogest, synthesis and immunogenic properties]. 379 51

In an extensive series of experiments, Balb/C mice and Lou rats were immunised with 3-O-(carboxymethyl)oximinocortisol conjugated to bovine serum albumin. The spleen cells from selected animals were fused with cells from mouse or rat plasmacytoma lines. Out of many hundreds of hybridomas screened, more than seventy produced antibody that bound 125I-labeled cortisol. These cultures were investigated further for stability of antibody production, affinity for cortisol and cross-reactivity with other steroids. An unexpected but consistent finding was that immunised rats produced antibody which cross-reacted with 11-deoxycortisol to a level greater than 100% and this characteristic was reproduced by rat-rat hybridomas. Strategies designed to improve the chances of generating non-cross-reactive anti-cortisol monoclonal antibodies did not appear to be successful. Nevertheless, several monoclonals were identified with properties that suggest they may be useful for the development of sensitive and specific cortisol assays.
Steroids 1985 Jun
PMID:The production and assessment of monoclonal antibodies to cortisol. 383 29

6-oxoestriol 6-carboxymethoxime, 6-oxoestradiol-17beta 6-carboxymethoxime, and 6-oxoestrone 6-carboxymethoxime were prepared and each was coupled to bovine serum albumin by means of the mixed anhydride technique. Standard methods were used to characterize the reaction products and intermediates. The results indicate that the estrone-, estradiol-17beta-, and estriol-6-albumin conjugates, as prepared in this study, can be used as antigens which induce specific antibodies to the corresponding haptens.
Steroids 1972 Apr
PMID:Determination of estrogens by radioimmunoassay with antibodies to estrogen-C6-conjugates. I. Synthesis of estrone-, estradiol-17 -, and estriol-6-albumin conjugates. 433 48

The development of a radioimmunoassay for medroxyprogesterone acetate (MPA) employing rabbit antibodies made against a bovine serum albumin (BSA) conjugate of the hemisuccinate ester of 11alpha-hydroxy MPA are described. The 11alpa-hydroxyl group was introduced into MPA by a series of chemical and microbiological transformations. The acid succinate MPA was coupled to BSA by reacting it with N,N'-carbonyldiimidazole. Cross-reactivities of several MPA analogs were compared suing rabbit antisera developed against the C-11 conjugate of MPA and goat antiserum developed against a C-3 conjugate of MPA. Antibodies formed against the C-11 conjugate showed increased specificity to steroid analogs which changes in the C-21 side chain. The profiles of serum MPA levels vs time, measured with both the C-3 and C-11 antisera, after oral drug administration to the monkey were similar. After intramuscular drug administration to dogs, serum MPA levels measured with the C-3 antisera were consistently greater as compared with those with C-11 antisera. Factors affecting precipitation of the antibody-antigen complex, assay precision, and assay sensitivity were evaluated.
Steroids 1974 May
PMID:Radioimmunoassay for medroxyprogesterone acetate (Provera) using the 11alpha-hydroxy succinyl conjugate. 436 69


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