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[Effect of gestagen therapy upon estradiol- and progesterone-receptor-level and 17beta-hydroxysteroid dehydrogenase in human endometrial adenocarcinoma (author's transl)].

Oestradiol was converted to oestrone about ten time more rapidly by subcellular fractions of normal human endometrium of the secretory phase than by tissue of the proliferative phase. In subcellular fractions of endometrial carcinoma the 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity decreased with decreasing differentiation of the tumour. Most of the 17beta-HSD activity was located in mitochondrial and microsomal fractions of both normal and neoplastic endometrium. After treatment of patients with gestagens only the well differentiated carcinomata significantly increased in 17beta-HSD activity demonstrating that the hormonal stimulus leads to similar effects on the 17beta-HSD activity as in normal endometrium. Furthermore quantitative aspects of the in vitro binding of 3H-oestradiol and 3H-progesterone to receptor components from normal endometrium and endometrial carcinoma cytoplasmic fractions have been studied. In normal tissue the number of cytoplasmic binding sites for both oestradiol and progesterone varied dramatically during the menstrual cycle: number of oestradiol binding sites were highest during the proliferative phase and fell during the secretory phase; for progesterone site the contrary was the case. In all endometrial carcinomata high oestradiol binding activity was observed. In contrast the number of progesterone sites in the tumours was related to the state of differentiation, which paralled the progestional sensitivity of these tumours.

Adenocarcinoma

A direct radioimmunoassay for estriol-16-glucuronide in urine for monitoring pregnancy and induction of ovulation.

Antibodies to estriol-16alpha-[beta-D-glucuronide] were raised in sheep with the use of keyhold limpet hemocyanin and bovine serum albumin conjugates of estriol-16alpha-[beta-D-glucuronide]. A simple, rapid method is presented for direct radioimmunoassay of estriol-16alpha-[beta-D-glucuronide] in urine with dextran-coated charcoal used for separation of free from bound and deionized water used for dilutions. The method is thrifty in its use of reagents. The assay has been evaluated in the pregnancy range, and the sensitivity has been extended into the range necessary for monitoring induction of ovulation with Pergonal.

Animals

Idiometric assay, the third way: a noncompetitive immunoassay for small molecules.

A novel, noncompetitive immunoassay applicable to the measurement of small molecules including ovarian steroids is described. Using monoclonal antibodies with the ability to recognize both beta-typic and alpha-typic binding sites, a new simplified, sensitive, and specific immunoassay system has been developed. The initial work of this development is presented, along with preliminary results for a novel immunoassay for estradiol.

Antibodies, Anti-Idiotypic

Adrenal steroids and the development of osteoporosis in oophorectomised women.

To explore the possibility that the wide variation in bone loss among oophorectomised women might be due to differences in adrenal androgens or their biosynthetic pathways, 18 women (10 with very fast and 8 with very slow bone loss) were selected. Serum levels of nine adrenal steroids, including the major androgens and cortisol, were measured under basal conditions and after overnight suppression followed by acute corticotropin stimulation. In addition, basal serum oestrone, oestradiol, dehydroepiandrosterone sulphate, sex-hormone-binding-globulin, corticosteroid binding globulin, and urinary free cortisol were measured. The only significant differences found were that women who lost bone rapidly had significantly higher urinary free-cortisol excretion (p less than 0.001) and a paradoxically diminished cortisol response to corticotropin. These data make it unlikely that endogenous adrenal androgens or oestrogens are a major factor in preventing bone loss after cessation of ovarian function; cortisol by its catabolic effect, however, may be a significant factor in causing osteoporosis.

17-Ketosteroids

Estrogen concentrations in bovine and porcine tissues.

Using an estrogen antiserum that measures biologically active estrogen (estradiol-17beta + estrone), a series of studies have been completed to describe the normal plasma levels of estrogen in cows and sows during various stages of reproduction. The levels are lower in the nonpregnant cow (less than 25 pg/ml) than in most species. Only during late pregnancy are the levels easily assayed. The levels in the nonpregnant sow are 2-3 times greater than in the cow. In both species there is a preovulatory peak of estrogen that is responsible for estrus. Plasma estrogen concentrations were also determined after induction of multiple ovulations by exogenous gonadotropin (PMSG) in the cow and sow and after treatment of cows with either a progestin (MGA) or a prostaglandin (PGF2alpha). (These two drugs have been used to synchronize estrus in the cow.) Estrogen levels rise greatly after treatment with PMSG but show little effect from MGA or PGF2alpha. Data are also presented on the validation of an assay for E2beta and E1 in beef tissue. Less than 20 pg of E2beta and E1 was measured per gram bovine muscle tissue. Considerable effort must be expended to apply radioimmunoassay to the assay of estrogens in tissues.

Animals

Estrogen profiles in young women: effect of maternal history of breast cancer.

To examine the hypothesis that familial breast cancer risk is related to estrogen metabolism, we analyzed urines of daughters of breast cancer patients and their matched controls for estrone (E1), estradiol (E2), and estriol (E3). From this, we computed estriol proportions (E3/E1 + E2 + E3). "Patient-daughters" and the matched controls showed no differences in estriol proportions. Our results failed to support the hypothesis that high-risk women (those with a family history of breast cancer) have relatively lower estriol proportions, and we concluded that whatever family history contributes to breast cancer risk, that risk is not likely to be transmitted by the estrogen profile.

Adolescent

Hormonal profiles in lactating and non-lactating women immediately after delivery and their relationship to breast engorgement.

Prolactin, human placental lactogen (HPL), oestrone, oestradiol and progesterone levels in plasma were measured before and during the first seven days after delivery in women who did not breast feed. The results confirmed the rapid clearance of placental steroids from the circulation after delivery. Plasma prolactin levels remained elevated during the early puerperium and the range of values were the same in non breast-feeding women and a group of breast feeding women. Of the 25 women studied, six developed breast engorgement. No difference in hormonal profiles were found leading to the conclusion that there is no endocrine basis for breast engorgement in non-breast feeding women.

Adult

Comparison of serum oestrogen concentrations in post-menopausal women taking oestrone sulphate and oestradiol.

Mean serum concentrations of oestradiol-17beta, oestrone, and oestrone sulphate in postmenopausal women were the same when measured up to six hours after treatment with either piperazine oestrone sulphate 1.5 mg or oestradiol valerate 2 mg. Maximum concentrations of oestradiol were less than those of oestrone, but oestrone sulphate reached concentrations about 30 times higher than those of oestrone. The rapid conversion of oestradiol valerate to oestrone and oestrone sulphate does not support the suggestion that in menopausal women oestradiol is less likely to be associated with a risk of endometrial carcinoma than oestrone sulphate, since the two preparations appear to become identical after ingestion.

Climacteric

Effect of a new LH-RH analogue (D-Ser(TBU)6-EA10-LH-RH) on gonadotrophin and gonadal steroid secretion in men.

The effect of a new analogue of the gonadotrophin-releasing hormone LH-RH, D-Ser(TBU)6-EA10-LH-RH, on the secretion of LH, FSH, as well as testosterone, oestradiol, HGH, prolactin, TSH, and cortisol was studied in normal men. The same subjects were injected intravenously in 4-day intervals with 1.0, 2.5, 5.0, and 10.0 mug of this substance. A significant LH but no FSH release was seen after doses of 1.0 and 2.5 mug LH-RH analogue, while after 5.0 and 10.0 mug dose-dependent increases of LH and imposed elevations of FSH were observed. Peak levels of LH were reached after 30 min, those of FSH after intravenous injection after 120 min. LH and FSH remained elevated for 8-10 h. LH peak levels after 5 mug of LH-RH analogue were comparable to those seen after injection of 100 mug of the decapeptide LH-RH. Following the release of LH and FSH after doses of 5.0 and 10.0 mug LH-RH analogue, there was a late stimulating effect in testosterone and oestradiol secretion. HGH, TSH, prolactin, and cortisol were not influenced by the LH-RH analogue.

Adult

15alpha-Hydroxyoestriol and other polar oestrogens in pregnancy monitoring.

Although oestriol measurements are well established for the assessment of 'at risk' pregnancies, there are a number of other oestrogens, excreted during pregnancy, which contain additional hydroxyl groups and might be more sensitive indicators of the condition of mother or fetus. Some of these result from the action of hydroxylases possibly present only in the fetus and others from maternal hydroxylations. We review the evidence for the biosynthesis of these polar oestrogens, summarise methods of measurement, and compare values obtained in normal and pathological pregnancies. There is as yet insufficient evidence to enable their potential value to be confirmed.

Catechols

[Preovulatory changes of steroidogenesis in isolated rabbit follicles (author's transl)].

In an attempt to investigate the effect of ovulating hormone on the steroidogenesis of mature follicles in the course of ovulation, transitory changes of steroidogenesis in isolated rabbit follicles have been studied at several intervals after injection of an ovulatory dose of human chorionic gonadotropin (hCG). Five to ten follicles of approximately 1-2 mm in diameter were isolated from ovaries of a mature rabbit (2.5-3.0 kg) under streomicroscope, before and at the 3rd, 6th, 9th and 12th hours after intravenous injection of of 100 IU/kg of hCG. Follicles were incubated with 100 muCi of acetate-1-14C in 2 ml of Krebs-Ringer bicarbonate buffer (pH 7.4) at 37 degrees C for 3 hours under 95% oxygen plus 5% carbon dioxide. Each incubation was terminated by quick freezing and stored forzen at -20 degrees C until eighty follicles had been collected for each time period before commencement of analysis. Incorporation of radioactive acetate into pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone., 20 alpha-dihydroxyprogesterone, dehydroepiandrosterone, androstenedione, testosterone, estrone and estradiol-17beta were analysed by the reverse dilution technique and identified in radiochemically pure form by recrystallization to constant specific activities. The steroidogenic activity of the follicles was evaluated by overall as well as fractionated incorporations. A peak in the overall incorporation of 14C- acetate into the ten steroids at the 3rd hour after hCG injection, followed by gradual decrease up to the 9th hour was observed. The incorporation decreased markedly to a minimum level at the 12th hour after hCG injection, which was below the level of preinjection control. Comparable quantitative fluctuations were found with the fractionated incorporation of 14C-acetate into the C21 and C18 steroids in the time sequence following hCG injection. However, the fractionated incorporation into C19 steroids reached to a maximum at the 6th hour after hCG injection. 5istribution patterns of incorporation among the individual steroids were varied at each interval of time. In the non-injected control, mature follicles synthesized predominantly estradiol-17beta, testosterone and androstenedione. Divergent steroids were formed from radioactive acetate at the 3rd hour after hCG injection. These included porgestogen, androgen and estrogen, but pregnenolone and 17hydroxyprogesterone were the two principal steroids produced. There was no essential difference in the steroidogenic patterns between the 6th and 9th hour, the major products being C21 and C19 steroids such as pregnenolone, 17hydroxyprogesterone, dehydroipiandrosterone and testosterone. The three androgens were the major steroids formed at the 12th hour after hCG injection. Thus the chages in the steroidogenic profile of the follicle was obvious in the course of ovulation. The basis of qualitative changes in follicular steroidogenesis during the process of ovulation have been discussed in connection with an accompanying effect of an ovulatory dose of hCG.

Acetates