1. Evaluation of cytological cervico-vaginal smears by the Bethesda system enlisted cytodiagnostics among important laboratory methods which can be used also in risk pregnancies. 2. Vaginal cytology makes it possible to test at the same time the hormonal situation during pregnancy, which reflects the placental function, and to evaluate also the vaginal biocenosis. 3. The authors provided evidence that the large number of superficial cells on the cytological smear (more than 10%) is associated with low oestriol and pregnandiol levels which are warning signs of the approaching termination of pregnancy. 4. By the action of microorganisms on the vaginal epithelium typical morphological changes develop in the cell nucleus and in the cytoplasm. By polychromatic staining also the causal agents of inflammations and infections threatening the mother and foetus are apparent. 5. The authors assume that cytological examination and evaluation according to the Bethesda system should be included in the complex of antenatal examinations also in women without clinical symptoms of premature delivery or without signs of vaginal infection.

1. [16-(14)C]16-Epioestriol and [6,7-(3)H(2)]oestriol were incubated both simultaneously and separately at 40 degrees with hen liver homogenates obtained from birds in the laying stage. 2. Purification and identification of products was made by ether extraction, Girard separation, Celite partition and paper chromatography and crystallization to constant specific activity both with and without derivative formation. Percentage conversions of the radioactive 16-epioestriol and oestriol were calculated from these specific activities. 3. Oestriol yielded 16-epioestriol as early as 5min. after exposure of steroid to the tissue. No production of oestriol from 16-epioestriol was detectable in this time. 4. Considerably more 16-epioestriol was formed from oestriol than oestriol by the reverse reaction after 90min. of incubation. 5. 16-Oxo-oestradiol-17beta appeared in all cases to be the major identified intermediate in the interconversion of 16-epioestriol and oestriol.

The gonadotrophin-suppressing effect of 4 peroral oestrogen regimens (A: 2.5 mg piperazine oestrone sulphate; B: 1.25 mg piperazine oestrone sulphate + 5.0 mg oestriol; C: 2.0 mg oestradiol valerate; D: 10.0 mg oestriol; all administered daily) was studied in 7 post-menopausal women, in the absence, and then in the presence of daily oral doses of 250 micrograms of levonorgestrel. A randomized, complete cross-over design was used, and from each volunteer 80 blood samples were drawn during a period of 252 days for the assay of immunoreactive follicle stimulating hormone (FSH), luteinizing hormone (LH) and bioactive LH levels. All 4 formulations significantly diminished pretreatment gonadotrophin levels. Combination with levonorgestrel enhanced the suppressive effects. Subsequent placebo administration for a week restored gonadotrophin levels to those found previously during oestrogen administration, but not to pretreatment levels. Regimen D exhibited the relatively weakest and regimens A and B the strongest suppression, both in the absence and in the presence of levonorgestrel. Treatment with all oestrogen formulations decreased, and the addition of levonorgestrel increased the ratio of bioactive (B) to immunoreactive (I) LH. Seven days of placebo administration abolished the effect of levonorgestrel, but not that of the various oestrogens on the B/I ratios.

The metabolism of [3H]oestradiol by interstitial cells in culture prepared from 18-day old rat testes was investigated. Interstitial cells were able to convert [3H]oestradiol to [3H]oestriol as confirmed by recrystallization of oestriol to constant specific activity from samples containing cells and not from controls. This demonstrated for the first time the presence of 16 alpha-hydroxylase in rat testicular interstitial cells. The effect of in vitro FSH treatment on the cells in culture was also investigated. FSH failed to affect 16 alpha-hydroxylase activity since we could not demonstrate a significant difference between treated and untreated preparations. The 16 alpha-hydroxylation of phenolic steroids is widely regarded as the major pathway of oestrogen metabolism in mammals. This metabolic step significantly reduces the biological activity of oestradiol. The presence of 16 alpha-hydroxylase in interstitial cells suggests that it may play a role in inactivating the oestradiol that is produced in the Leydig cells and thus prevent its intracellular accumulation. Such activity may conceivably play a role in the overall local "fine tuning" of androgen biosynthesis.

A simple and sensitive high-performance liquid chromatographic method is described for the determination of three oestrogens (oestriol, oestrone and oestradiol) in pregnancy urine. Free oestrogens are extracted with chloroform from the urine sample. The phenolic group of each oestrogen in chloroform is formylated in the presence of an alkaline aqueous solution, and the resulting aldehyde is converted into a fluorescent derivative by reaction with 1,2-diamino-4,5-dimethoxybenzene. In order to determine free and conjugated oestrogens, conjugated oestrogens are hydrolysed by heating in hydrochloric acid before the extraction and then treated in the same way as free oestrogens. The fluorescent derivatives are separated on a reversed-phase column, TSKgel ODS-120T, with stepwise gradient elution using an aqueous methanol-containing phosphate buffer (pH 2.2). The lower limit of detection for each oestrogen is ca. 200 fmol per 100-microliters injection volume.

We described previously the in vivo immunoneutralization effects of a high affinity anti-oestradiol antibody clone 15 in blocking ovulation and synaptic remodeling in cycling female rats. In the present study we report the enhancing effects of this antibody. Treatment of ovariectomized female rats or female derived skeletal cell cultures in vitro with anti-E2 15 plus oestrogen (E2) potentiated the specific activity of the brain type creatine kinase (CK) response to E2 in the rat tissues or skeletal cells. The enhancing CK response of anti E2 15 plus E2 was time- and dose-dependent in the uterus, thymus, epiphysis and diaphysis of ovariectomized female rats. In the pituitary, on the other hand, anti-E2 15 blocked the stimulatory CK response to E2. Two other high affinity anti-E2 antibodies, clones 8D9 and 11B6, had no effect in augmenting the response of CK to E2 in rat tissues. Moreover, the enhancing CK response in rat tissues was specific to anti-E2 15 plus E2 since the intact anti-E2 in the presence of other oestrogen mimetics, such as oestriol or stilbestrol or tamoxifen did not potentiate the CK response in rat tissues. In this model system the Fab' monomer of anti-E2 15 abolished the CK response to E2 in rat tissues and not to anti-E2 15 plus E2 whereas tamoxifen completely blocked the CK response to anti E2 plus E2. Anti E2 15 may therefore serve as a specific carrier in delivering E2 to oestrogen sensitive rat tissues or cells containing functional oestrogen receptors and thereby increasing the magnitude of E2 effects in vivo and in vitro.

Ovariectomized adult rats with closed uteri were treated for 7 days with different oral and s..c. doses of oestradiol, oestrone, oestriol and ethinyl oestradiol. All treatments elicited the production of uterine fluid and the potencies of oestrogens were related to the amount of fluid secreted. Ethinyl oestradiol and oestradiol displayed similar activity when given s.c. A daily dose of 0-003 mg oestradiol/kg resulted in about 700 mg fluid. Oestrone was 3-10 times and oestriol about 100 times less active. Orally, ethinyl oestradiol was the most potent substance and 700 mg secretion was obtained with a dose of 0-03 mg/kg daily. Oestradiol was about 30 times, oestrone about 100 times and oestriol 50 times less active than ethinyl oestradiol by this route. The viscosity of the secretion was unaffected, remaining between 1-6 and 2-4 cP. The pH of the fluid did not change, but that of the uterine lumen diminished slightly. These effects of oestrogens were associated with an increase in the weight of the empty uterus and a decrease in body weight.

The uptake of [4-14C]oestriol by the isolated perfused rat liver is 3.8 times faster as compared to that of simultaneously perfused [6,9-3H2]oestriol 16 alpha-monoglucuronide. During perfusion the concentration of both radioactive oestrogens decreased exponentially in perfusion medium (apparent kel: 0.061 min-1 and 0.016 min-1, respectively). [6,9-3H2]Oestriol 16 alpha-monoglucuronide was metabolized only to a small extent; more than 92% was secreted unchanged into the bile where it was highly concentrated (1800 nmol/g). In contrast [4-14C]oestriol was extensively metabolized; it was mainly hydroxylated at C-atom 2, leading to a rapid increase in the concentration of 2-hydroxyoestriol in the perfused medium. This metabolite was quickly taken up by the liver during recirculation and subsequently either methylated or sulphated. 2-Hydroxyoestriol monosulphate was glucuronated to 2-hydroxyoestriol 16 alpha-monoglucuronide 3-sulphate, which was rapidly excreted into the bile. No double conjugate was formed when [6,9-3H2]oestriol 16 alpha-monoglucuronide was perfused; this is additional evidence for the correctness of the assumption that monoglucuronides cannot serve as precursors of sulphoglucuronides.

Serum oestradiol/oestrone ratios were measured during various oestrogen treatments in castrated women. Oral oestriol succinate therapy (8 mg/day) caused little change in the pre-treatment oestradiol/oestrone ratio. During oestradiol valerianate therapy (2 mg/day) serum total oestrogens and the E2/I1 ratio were considerably increased. One day after the injection of 10 mg of oestradiol valerianate and 2.5 mg of oestradiol benzoate + 10 mg of oestradiol phenylpropionate the E2/E1 ratio was similar to the ratio in middle of the normal ovulatory cycle. The change in serum oestriol was rather small after oral doses of 8 mg og oestriol succinate 15, 30 and 120 min after the application.

It is well known, that magnesium can positively influence preterm labour and that the magnesium level in the plasma is significantly lowered whereby magnesium excretion increases during pregnancy. These results lead to the question, whether the magnesium content in the myometrium is also reduced during pregnancy and whether blood parameters correlate with the concentration of magnesium in the myometrium. Myometrial tissues and blood samples from 127 patients, who underwent a Caesarean section, were analysed for magnesium, calcium, sodium and potassium. Erythrocytes, haemoglobin, haematocrit and the hormones oestradiol, oestriol and progesterone were analysed in the serum. We found a decrease in magnesium in the myometrium (p < 0.01) and in the plasma (p < 0.05) during pregnancy. Magnesium in the plasma correlates with magnesium in the myometrium (p < 0.001). These results indicate, that hypomagnesinaemia during pregnancy decreases the magnesium level in the myometrium.

In the years 1977 to 1981, 14 quality-control surveys for the determination of steroid hormones were performed in cooperation with the Deutsche Gesellschaft für Klinische Chemie. Hereby the laboratories participating could in each case analyze the following steroids: aldosterone, cortisol, oestradiol-17 beta, oestriol, progesterone, and testosterone. In the light of the results an investigation was made as to whether, in the course of time, an improvement in the accuracy or in the precision of the determinations had been attained, and to what extent the determinations depend on the qualities of the test material. A clear improvement in the accuracy of the results of the analyses could only be ascertained for oestradiol-17 beta. For aldosterone and cortisol, values were found in pool-plasma whose medians were significantly above the definitive values. An improvement in precision could be noted especially with oestradiol-17 beta and to lesser degrees with cortisol and oestriol. The kind of test material--plasma which contained only the hormones to be analyzed on the one hand, and, on the other hand, pool-plasma, which also contained all endogenous hormones--had no influence on the precision of the results from various laboratories. Low concentrations of the individual steroids led--on the basis of the methodological principle of radioimmunoassays--in almost all cases to a reduced interlaboratory precision in regard to values. The accuracy of the analysis values was considerably impaired only with aldosterone and oestradiol-17 beta by low concentrations: the medians here were in part twice as high as the definitive values.

Effects of prolonged administration (7 days) of Tamoxifen (Tx: 100 micrograms/day), oestradiol-17 beta (E2; 1 microgram/day) and oestriol (E3; silastic capsules) on the uterine weight, protein, DNA and progesterone receptor (RP) content of prepubertal rats have been studied. It has been observed when compared with E2 that Tx induces a dissociated action after two days of administration; the RP content is still increasing when the other parameters are at a constant level. E3 has a similar action to E2.

The biological effect of seven different oestrogen sulphates: oestrone-3-sulphate (E1-3-S), oestradiol-3-sulphate (E2-3-S), oestradiol-17-S), oestradiol-3,17-disulphate (E2 3,17-DS), oestriol-3-sulphate E3-3-S), oestriol-17-sulphate (E3-17-S), oestriol-3,16,17-trisulphate (E3-3,16,17-TS), was studied in the foetal uterus of guinea pig (55--65 days of gestation) after sc administration for 3 consecutive days of each sulphate to the mother. On day 4, two response parameters were investigated, the uterotrophic effect and the action on the progesterone receptor. The monosulphates at the C3 position of the oestrogens (E1-3-S, E2-3-S and E3-3-S) provoked an increase in uterine weight of 1.9--2.4 times in relation to the non-treated animals. These oestrogen sulphates also very significantly stimulated the number of progesterone specific binding sites, 7--10 times in relation to the non-treated animals. On the other hand, when the sulphate was at C17 of the oestrogens (E2-17-S, E3-17-S, E2-3-17-DS, E3-3,16,17-TS), very little or no effect on the foetal uterus was observed on the two parameters studied. It is concluded that oestrogen (oestrone, oestradiol, oestriol) sulphates in position C3 can be involved in the biological response to the hormone and it is suggested that the effect is carried out after the hydrolysis of the sulphate. On the other hand, sulphates in C17 are not hydrolysed and no significant biological effects were observed in the uterine growth or in the progesterone receptor.

The acute effects of salbutamol on the serum levels of hPL, hCG, oestriol and progesterone in diabetic and non-diabetic pregnant women in the last trimester were studied. I.V. administration of salbutamol in an increasing dose from 0 to 22.5 ug/min caused no change in hPL or hCG levels in any group. Progesterone showed a significant fall after 40 min in the diabetic group and a less pronounced, although still significant fall after 80 min in the non-diabetic group. The oestriol levels increased significantly after 40 min in the diabetic group while no significant change was noted in the non-diabetic group. It is speculated that the effects in the diabetic women might be explained by a redistribution of the available placental 5-pregnenolone due to an increased fetal conversion into oestriol. This might be due to an increased sensitivity of the adrenal cortex of these foetuses to a supposed stimulatory effect of salbutamol.

Human chorionic somatomammotrophin (HCS), progesterone, and unconjugated oestradiol and oestriol were measured in the plasma of 13 patients with intact hydatidiform moles from week 9 to 19 of pregnancy and in 89 normal women from week 5 to 20 of pregnancy. Plasma alpha-foetoprotein (AFP) was also measured in 9 out of 13 patients and in 23 of the normal women from week 13 to 20 of pregnancy. All the compounds were measured by radioimmunoassay. The plasma HCS concentration in 35 samples from 13 patients with hydatidiform moles ranged from 10 to 910 ng/ml; this was lower than that in normal pregnancy of corresponding duration in eight patients; within the normal range in four patients and high in one patient. The plasma progesterone concentration ranged from 17-5 to 79-2 ng/ml; it was higher than that in normal pregnancy in eight patients and within the normal range in five patients. The plasma unconjugated oestradiol concentration ranged from 1-82 to 8-10 ng/ml; it was higher than normal in six patients and within the normal range in seven patients. The plasma unconjugated oestriol concentration ranged from 0-168 to 1-37 ng/ml, the levels at 15-19 weeks of gestation being significantly lower than those in normal pregnancy at this time (P less than 0-005). Plasma AFP was not detectable in the nine patients (less than 10 ng/ml) whereas it ranged from 10 to 80 ng/ml in 18 out of 23 women in week 13-20 of normal pregnancy. The present results suggest that both plasma oestriol and AFP could be helpful in the diagnosis of hydatidiform mole at about 12-14 weeks though diagnosis could not be made with absolute certainty.

Progesterone receptors were measured in both cytosolic and nuclear fractions in vaginal tissue samples obtained from a total of 17 post-menopausal women. Ten of the women were given oestriol (E3) intravaginally 6-24 h before surgery, while the remaining 7 received no treatment. Cytosolic receptors were not detected in any of the 7 tissue samples from the untreated women, but nuclear receptors were present in two cases. In the 10 women treated with E3, it was possible to measure cytosolic receptors in 5 tissue samples and nuclear receptors in 7 samples. The mean receptor concentrations in the cytosolic and nuclear fractions were similar (45-65 fmol/mg protein) and the apparent dissociation constant of the receptors in all the samples was 1-2 nmol/1. These data would seem to indicate that oestrogen treatment results in the synthesis of new receptors, although not in all tissues, which compounds the scepticism regarding the validity of the classical two-step model for steroid hormone action.

The correlations between serum prolactin and progesterone, oestradiol-17-beta and oestriol were determined in 125 pregnant females between the sixth and fourteenth week of pregnancy. No significant correlations were found between prolactin and progesterone. Correlations between prolactin and oestradiol-17-beta and oestriol were mostly positive, but only significant during the twelfth week of pregnancy.

The enzymatic inactivation of noradrenaline was investigated in 25 fresh human placentae in vitro. Oxidative deamination by monoamine oxidase (MAO) was greater than enzymic O-methylation by catechol-O-methyltransferase (COMT). The addition of oestriol (E3) or progesterone to the organ bath significantly decreased (P less than 0.001) the activity of placental MAO. Oestrone (E1) and oestradiol (E2) showed no inhibitory effect. In addition, E3 significantly inhibited the activity of COMT (P less than 0.001), whereas oestrone and oestradiol had no effect on COMT activity. COMT was also inhibited by progesterone (P less than 0.05). The decrease in enzymic inactivation of noradrenaline caused by oestriol and progesterone suggests an activated adrenoceptor function.

Amniotic fluid concentrations of immunoreactive oestrogens and progesterone were measured at the time of caesarean section in 32 twin pregnancies; 25 women had an elective section and seven were in labour at the time of operation. No significant differences between concentrations in the amniotic fluid of the first and second twin were found in respect of conjugated and unconjugated oestrone, oestradiol, oestriol, oestetrol and unconjugated progesterone either before or during labour. It is unlikely that changes in oestrogens or progesterone in the amniotic fluid are responsible for the selective changes seen in prostaglandins and fetal adrenal steroid during labour in the first twin.

Previously we reported that administration of lipopolysaccharide (LPS) to mice increased the hepatic levels of putrescine (PUT) and N1-acetylspermidine (N1-acetyl-SPD). In the current study, we examined the in vivo effects of some steroid hormones on the LPS-induced increase in PUT and N1-acetyl-SPD. Corticosterone, hydrocortisone and dexamethasone suppressed the LPS-induced increase in PUT and N1-acetyl-SPD in mouse liver in a dose-dependent manner, dexamethasone being the most effective among them. On the other hand, oestrone and oestradiol-17 beta enhanced the LPS-induced increase in PUT and N1-acetyl-SPD in a dose-dependent manner. Oestradiol-17 alpha and 16 beta-ethyl-oestradiol, as an inactive oestradiol isomer and an antioestrogen, respectively, likewise enhanced the increase in PUT and N1-acetyl-SPD concentrations induced by LPS. 16 alpha-hydroxy-oestradiol (oestriol), 16 alpha-hydroxyestrone, 2-hydroxyoestradiol, 2-hydroxyoesterone, progesterone, testosterone, diethylstilboestrol and nonsteroidal antioestrogens such as tamoxifen and nafoxidine had no effect on the increase. Oestradiol-17 beta enhanced and corticosterone had little effect on the carbon tetrachloride-induced increase in PUT and N1-acetyl-SPD. These results suggest that glucocorticoids suppress the increase by preventing the immunological injury by Kupffer cells on hepatocytes and that the stimulatory effect of oestrogens may not be associated with their oestrogenic activities mediated by the oestrogen receptor system.

During the years 1971-1982 urinary oestriol excretion was tested in 38,536 patients (group 1). One or more low oestriol value was found in 11.0% of patients; in this group the stillbirth rate was 8 times higher, the neonatal death rate 4 times higher, and fetal growth retardation rate 4 times higher than in patients with normal oestriol values (all p less than 0.001). During the years 1982-1984 a further 12,887 patients were tested (group 2) and 9.5% had one or more low oestriol value. The perinatal mortality rate in patients with normal oestriol excretion fell from 0.8% in group 1 to 0.5% in group 2 (p less than 0.05), and in patients with low oestriol excretion from 4.7% in group 1 to 2.4% in group 2 (p less than 0.01). However, patients in group 2 with low oestriol values still had significantly unfavourable results, compared to those with normal oestriol values--stillbirth rate 4 times higher, neonatal death rate 5 times higher, and fetal growth retardation rate 4 times higher (all p less than 0.001). Although perinatal results have improved, fetal growth retardation and the risk of perinatal death are still identified by urinary oestriol assay.