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estradiol might be due, at the least in part, to prevention of β-cell apoptosis. Therefore we also evaluated the effect of oestriol, the major oestrogen produced during pregnancy, on progesterone-induced cell death. We report herein that progesterone, at
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Daily injections of oestriol, spanning a wide dose range, produced mitotic responses in the uterine luminal epithelium of mice. At the higher doses the response was comparable to that obtained with oestradiol. The maximum response occurred on day 1, after which mitosis was significantly reduced on days 2 and 3. This reduction in mitosis following an initial peak was seen at all doses and was not therefore consequent on a critical cell number being reached. Proliferation was also induced in the basal epithelium of the vagina, though to a lesser extent. Oestriol and progesterone together stimulated DNA synthesis and mitosis in the epithelium of the mammary gland.
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ABSTRACT
The concentration of oestriol and the proportion of this hormone not bound to plasma protein were measured using radioimmunoassay and centrifugal ultrafiltration respectively, in 55 samples of plasma obtained from 12 women in the last 2 to 7 weeks of uncomplicated pregnancy. Among individuals, the mean plasma concentration of oestriol varied from 25·8 to 94·8 nmol/l; in nine subjects, there was a tendency for oestriol concentrations to increase as delivery approached. The mean proportion of oestriol not bound to plasma protein in the different subjects varied from 13·1 to 18·9%, but values from any individual subject remained essentially constant during the periods of study. These measured values were used to calculate, for each sample, the apparent concentration of oestriol not bound to plasma protein. The results were combined with analogous values for oestradiol and progesterone obtained from the same plasma samples and described in a previous study. It was found that (i) the mean ratio of the concentration of oestriol and oestradiol was 0·75, (ii) the mean concentration of non-protein-bound oestriol was 8·7 times that of non-protein-bound oestradiol, and (iii) in individual subjects, there was no consistent trend as delivery approached in the ratio of the concentration of progesterone to that of oestriol in either the total or non-protein-bound form.
J. Endocr. (1986) 108, 75–80
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The assay of urinary oestriol as a measure of foeto—placental function is now a well-established procedure. The test is delayed for at least 24 h by the time taken for urine collections. In addition, urine collections mostly involve expensive hospitalization and are subject to many errors. Analysis of maternal blood or of amniotic fluid would obviate these disadvantages and may approach the endocrine state of the foetus more closely. The clinical usefulness of assays on amniotic fluid depends upon the concentration of oestriol in the fluid having a meaningful relationship to oestriol production in the foeto—placental unit. This possibility has been tested before by a comparison between the concentration in liquor and the urinary excretion (Berman, Kalchman, Chattoraj & Scommegna, 1968). This showed a positive correlation coefficient(r = 0·69) in a series of 24 patients whose stages of gestation ranged from 32–40 weeks and which included eight severely ill women.
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SUMMARY
There was no diurnal variation in plasma oestriol and diet, exercise and rest did not affect plasma oestriol levels in pregnant women. The 30-min, 3-h and day-to-day coefficients of variation compared favourably with that of 48-h urinary excretion.
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Sub-Department ofEndocrine Pathology, University of Liverpool, The Liverpool Clinic, Liverpool, L7 7DE
(Received 17 October 1974)
Although mild methods of hydrolysis are available, hot acid hydrolysis is frequently used to hydrolyse oestriol conjugates before estimation of the free steroid. Assessment of losses arising from hydrolysis, extraction and other manipulations is possible, using radioactive oestriol. Tritiated oestriol is preferable to 14C-labelled oestriol because of the high specific activity of tritium-labelled compounds. For radioimmunoassay, compounds of very high specific activity have been prepared, labelled in the 2,4 positions, with or without additional radioactivity at positions 6,7 or 6,9.
We have found that oestriol labelled in the 2,4 positions is unsuitable for following procedural losses if hot acid hydrolysis is used.
Tritiated steroids (Radiochemical Centre, Amersham) were checked for radiochemical purity by thin-layer chromatography, followed by scanning in a radiochromatogram scanner. No evidence of impurity was found. The steroid (either 20 000
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SUMMARY
Single injections of oestriol were partially effective in sensitizing the uterus to a decidual stimulus and inducing ovum-implantation in progesterone-treated spayed mice, whereas two injections (6 h apart) were fully effective. It seems that in the progestational uterus, as in the non-progestational organ, oestriol can induce a full oestrogenic response provided that its level in the target organ is maintained. It is also concluded that ovum-implantation is not triggered by some transient early effect of oestrogen, but requires about 12 h of sustained oestrogen action for its successful completion.
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SUMMARY
A method is described for estimating oestriol in amniotic fluid. The main steps of the method are acid hydrolysis, chemical purification and methylation, chromatography on alumina columns, acetylation and estimation by gas chromatography. A radioactive internal standard is employed to correct for losses during assay. Data on the reliability of the method are given.
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During normal human pregnancy alterations in liver function similar to those seen in cholestasis occur (Tindall & Beazley, 1965; Haemmerli & Wyss, 1967). Adlercreutz & Tenhunen (1970) concluded that oestrogens were primarily responsible for this effect. In a previous report from this laboratory (Tikkanen, 1972) progressive changes, including decreasing proportions of OE3-3Gl† and increasing proportions of OE3-3S,16Gl†, in the urinary excretion of oestriol conjugates during pregnancy were noted, which were regarded as reflections of changes in the excretory function of the liver. Since a clear consistent change in urinary oestriol conjugate pattern was not observed in all the pregnancies studied, it appears that some women are more susceptible than others to this action of oestrogens on liver function. In multiple pregnancies Beazley & Tindall (1966) noted changes in the excretory function of the liver that were similar to but greater than those previously observed in singleton pregnancies. Accordingly, it might
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SUMMARY
A method is described for estimating oestriol in 2–10 ml samples of human pregnancy peripheral plasma. It incorporates acid hydrolysis, chemical purification, methylation, chromatography on alumina columns, formation of a derivative and quantitative determination by gas chromatography. A radioactive internal standard was added to correct for procedural losses. Plasma oestriol determinations in five normal patients throughout pregnancy and delivery are reported.
