[(3)H]Triamcinolone acetonide (15nm) was incubated with cytosol (150000g fraction) prepared from oviducts of egg-laying hens. The extent of steroid binding, as determined by charcoal assays, was greatest between 2-4h at 4 degrees C. A similar time curve was obtained when cytosol preparations were first fractionated with (NH(4))(2)SO(4) before labelling. The addition of 10mm-Na(2)MoO(4) or 10mm-ATP during the incubation of hen oviduct cytosol with [(3)H]triamcinolone acetonide lowered the extent of steroid binding. The presence of glycerol (20%), however, increased the extent of [(3)H]triamcinolone acetonide binding in cytosol fractions from chick (330%) and hen (160%) oviducts. The [(3)H]triamcinolone acetonide-receptor complex was stable for over 4h at 4 degrees C, but dissociated rapidly at 37 degrees C, exhibiting a half-life of about 10min. The presence of 10mm-Na(2)MoO(4) and 10mm-ATP or both had a small protective effect on the dissociation of [(3)H]triamcinolone acetonide-receptor complex. The receptor from hen oviduct showed significant affinity for unlabelled triamcinolone acetonide, cortisol, compound R5020 and dihydrotestosterone and, to a lesser extent, for oestradiol, oestrone and progesterone. Diethylstilboestrol treatment of immature chicks appeared to induce a more specific binder, which showed affinity for unlabelled triamcinolone acetonide, cortisol and compound R5020 only. Scatchard analysis of [(3)H]triamcinolone acetonide binding in hen oviduct cytosol revealed a K(d) value of 6.4nm. The steroid-receptor complex sedimented as a 7-8S and a 4S entity on low-salt (0.01m-KCl)- and high-salt (0.3m-KCl)-containing sucrose gradients respectively. The cytosol [(3)H]triamcinolone acetonide-receptor complex showed no affinity for ATP-Sepharose or DNA-cellulose, but acquired this ability on heat activation (23 degrees C, 40min). The data indicate the avian oviduct possesses a high-affinity binding molecule that fulfils the criteria of a glucocorticoid receptor.

An initial group of term (36-41 6/7 weeks), preterm (less than 36 weeks), and post-term (42 or more weeks) placentae were collected from women at delivery to determine the placental levels of important steroids and steroidogenic enzymes involved in the oestrogen synthesis pathway as a function of gestational age. A second group of placentae were obtained from women delivering at term before and after the onset of labour. Placentae were evaluated individually for cytosolic steroid hormone levels and microsomal steroidogenic enzyme activities. Oestradiol (E2), oestrone (E1), progesterone (P), and delta-4-androstenedione (A) were measured by radioimmunoassay in placental cytosols. Aromatase (AR), sulphatase (S), and 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta HSD) activities were assayed in placental microsomes. Cytosolic concentrations of E1, E2, P, and A did not differ with respect to gestational age. Correspondingly, the microsomal enzyme activities of 3 beta HSD, S, and AR did not vary as a function of gestational age. However, when patients at term who were in labour prior to delivery were compared to those who were not, the placental cytosolic level of E1 was found to be threefold higher in the non-labouring group (4572 versus 1427 pg/mg cytosolic protein, P < 0.025). Additionally, microsomal aromatase activity was also significantly higher in the non-labouring patients (46 versus 19 pM/min/mg protein, P < 0.025), while the E2 to P ratio in the labouring patients was twice that of the non-labouring group, a difference which was significant at the P < 0.025 level (Wilcoxon rank sum test). These data suggest that at term, prior to labour, the placental production of E1 by AR is high, and that AR activity and E1 levels fall significantly after the onset of labour. Also, the placental cytosolic concentration of the more active oestrogen, E2, demonstrates stable to rising levels with a significant increase in E2/P after the onset of labour. We theorize that in the term pregnancy prior to labour, E1 may represent a large but relatively inactive intracellular oestrogen pool which is maintained by high AR activity, and may function to protect the pregnant local uterine environment from the more oxytocic effects of E2.

Oestrone, oestradiol-17 beta and progesterone were measured by radioimmunoassay in daily urine samples after pairing and during subsequent pregnancy in a pied bare-face tamarin. On the basis of excretion profiles an ovarian cycle length of about 3 weeks and a gestation length of about 160 days are suggested. Oestrone was the predominant urinary oestrogen excreted by the non-pregnant and pregnant pied bare-face tamarin, the oestrone/oestradiol ratio being greater than 100:1. The results suggest that steroid monitoring can provide useful information about reproductive physiology in this species of tamarin.

Ten women were given dexamethasone intramuscularly in order to accelerate fetal lung maturation. Amniotic fluid and maternal serum were assayed for oestrone, oestradiol-17beta, oestriol, progesterone and chorionic gonadotrophin by specific radioimmunoassays before and after the treatment. No hormonal changes were observed in amniotic fluid. The serum levels of oestrogens 2 to 5 days after treatment were not significantly different from those before treatment, indicating that the previously reported suppression of oestrogen production by glucocorticoids disappears rapidly after treatment stops.

Progesterone, oestrone and oestradiol-17 beta levels in the muscle, fat, liver, kidney and plasma of steers treated with Synovex S, untreated steers, and cows during the normal oestrous cycle, were examined. Progesterone levels in female tissues reached a maximum in the luteal phase and fell to a minimum in the follicular phase. Oestrogen levels (oestrone and oestradiol-17 beta) did not change during the cycle. Progesterone and oestrogen levels in the tissues of steers treated with Synovex S and untreated steers were not statistically different. Tissue progesterone levels in steers were much lower than those in cows during the luteal phase. The highest oestrogen levels were found in the fat of female animals. These results indicate that residue levels of progesterone and oestrogen in the muscle and fat of steers treated with Synovex S were within the physiological range, and lower than those of cows.

In the mouse uterus the development of sensitivity to a decidualizing stimulus requires large amounts of progesterone and small amounts of oestradiol-17 beta. During the process progesterone induces changes in the sensitivity of the tissues of the uterus to oestrogen. From observations of human endometrium it has been suggested that progesterone modulates the biological activity of oestradiol-17 beta by stimulating the metabolic conversion of oestradiol-17 beta to oestrone. Accordingly [6,7-3H]oestradiol-17 beta was injected subcutaneously into ovariectomized mice at various stages of the development of sensitivity to a decidualizing injection of oil into the uterine lumen. Radioactivity was extracted 4 h later, fractionated and identified. There was no alteration in the amounts of oestradiol, oestrone or water-soluble metabolites in the uteri whether the mice were treated with progesterone or with progesterone plus oestrogen or whether the uterine horns were decidualized for 24 or 48 h. The results suggest that in vivo the metabolism of oestradiol-17 beta by the uterus is not stimulated by progesterone.

The presence of a protein that binds specifically and with high affinity to progesterone is found in the plasma of fetal guinea Pig. Progesterone and 5 alpha-dihydroxyprogesterone strongly compete for the 3H-progesterone-protein complex. 5 beta-dihydroxy progesterone, 20 beta-dihydroxyprogesterone, testosterone and synthetic progestagen, R-5020 (17,21-dimethyl-19-norpregna-4, 9-diene-3,20 dione), also compete but less intensely. 17-hydroxyprogesterone, aldosterone, cortisol, oestradiol and oestrone have no effect. The 3H-progesterone-protein, complex has an affinity of Kd = 8.8 +/- 3.5 x 10(-10) M and in sucrose density gradient the sedimentation coefficient is 4.6 S.

Whereas maternal venous levels of progesterone and oestrogens have been studied intensively in relation to parturition, little attention has been directed to fetal levels. In this study progesterone, oestrone (E1), oestriol (E2) and oestriol (E3) were measured in umbilical artery (UA) and umbilical vein (UV) serum at vaginal delivery (after induced or spontaneous onset of labour) and at elective Caesarean section. At least 14 samples were included in each group. All UV serum steroid levels were consistently higher than UA serum steroid levels. Cord serum levels of progesterone and arterio-venous differences were higher at vaginal delivery than at Caesarean section although maternal levels changed little. E1 and E2 levels were similar after spontaneous labour and at Caesarean section but were higher (P less than 0-05) after induced labour although E3 levels did not alter. E1 and E2 were higher and progesterone lower in primiparae than in multiparae. Fetal sex made no differences. These results suggest that induction of labour with oxytocin results in altered fetal hormone levels, that maternal hormone levels provide a poor reflection of fetal changes,and that current views about the mechanism of the onset of labour in animals may not be appropriate for man.

Ovaries from 19- and 20-day old rat embryos were cultured in the presence of [3H]testosterone and [3H]progesterone respectively, and the conversion of these precursors into [3H]oestrone and [3H]oestradiol was studied. By adding tracer amounts of [14C]oestrone and [14C]oestradiol to the culture media at the beginning of the analysis and crystallizing oestrone and oestradiol to constant specific activity, a conversion percentage could be calculated. There was no oestradiol formation either from progesterone or from testosterone. Oestrone was formed in measurable amounts from both precursors. It is concluded that in the ovary of the 19-20-day old rat embryo 17 beta-hydroxysteroid dehydrogenase favors the formation of oestrone.

The effects of oestradiol-17 beta, progesterone and oestrone-3-sulphate were studied in primary cultures of guinea-pig endometrial glandular epithelial cells. Comparative ultrastructural studies were performed by means of transmission electron microscopy on cells grown either without hormones or with oestradiol-17 beta (2 nmol/l), oestradiol-17 beta (2 nmol/l) plus progesterone (50 nmol/l), or oestrone sulphate (0.1 mumol/l). In the control medium, without steroid hormones, the majority of epithelial cells were poorly differentiated, although numerous small mitochondria were present and abundant lipid droplets could be observed. Oestradiol-17 beta stimulated metabolic activity in the cells. Progesterone added to oestradiol-17 beta-primed cells stimulated the development of the endoplasmic reticulum-Golgi system. Oestrone sulphate induced a higher level of differentiation characterized by large clear mitochondria, well-developed Golgi complexes, and active nuclei, suggesting secretory activity. In all cases, the cultured cells displayed deep invaginations of the nuclear membrane associated with nuclear pores, known as nucleolar channels. After treatment with oestrone sulphate these channels were associated with a characteristic reticular nucleolus. We conclude that cultured endometrial epithelial cells display secretory activity in response to treatment with oestradiol-17 beta plus progesterone, or with oestrone sulphate.

The plasma levels of progesterone and oestrogens were measured during treatment with an oral contraceptive containing 37.5 mug of ethinyl oestradiol and 1 mg of lynestrenol (= 3-desoxy-17alpha-ethinyl-19-nortestosterone) (Ovostat 1375, Organon, Oss, Holland). Blood samples were taken every day during 9 complete cycles in 5 healthy women. The plasma levels of progesterone were in the range of those found during the early follicular phase of the normal women. No ovulation seemed to have occurred. The mean plasma levels of oestrogens (oestradiol and oestrone) during treatment were below those found during early follicular phase of 34 normal cycles although overlapping of values occurred. The ratio between oestradiol and oestrone during treatment was 1:2 vs. 1:1 during the early follicular phase of the normal cycles. In two cycles treatment was started during the midcyclic rise of oestrogens. No rise of progesterone indicating corpus luteum formation was found. Despite the low amount of oestrogen in the drug the ovaries appeared to secrete minimal amounts of sex steroids. The lynestrenol content is likely to be partly responsible for the inhibition of ovulation and the additional suppression of steroidogenesis.

The high incidence of stillbirth in Swedish Holstein heifers has increased continuously during the last 15 years to an average of 11% today. The pathological reasons behind the increased incidence of stillbirth are unknown. The present experiment was undertaken to investigate possible causes of stillbirth and to study possible physiological markers for predicting stillbirth. Twenty Swedish Holstein dairy heifers sired by bulls with breeding values for a high risk of stillbirth (n = 12) (experimental group) and a low risk of stillbirth (n = 8) (control group, group B) were selected based on information in the Swedish AI-data base. The experimental group consisted of 2 subgroups of heifers (groups A1 and A2) inseminated with 2 different bulls with 3.5% and 9% higher stillbirth rates than the average, and the control group consisted of heifers pregnant with 5 different bulls with 0%-6% lower stillbirth rates than the average. The bull used for group A1 had also calving difficulties due to large calves as compared to the bull in group A2 showing no calving difficulties. The heifers were supervised from 6-7 months of pregnancy up to birth, and the pregnancies and parturitions were compared between groups regarding hormonal levels, haematology, placental characteristics and calf viability. In group A1, 1 stillborn, 1 weak and 4 normal calves were recorded. In group A2, 2 stillborn and 4 normal calves were registered. All animals in the control group gave birth to a normal living calf without any assistance. The weak calf showed deviating profiles of body temperature, saturated oxygen and heart rates, compared with the normal living calves. No differences of the placentome thickness, measured in vivo by ultrasonography were seen between the groups. The number of leukocytes and differential cell counts in groups A1 and A2 followed the profiles found in the control group. In group A1, a slight decrease of oestrone sulphate (E1SO4) levels was found in the animal delivering a stillborn calf from the first 24-h blood sampling at 6 weeks to the second at 3 weeks prior to delivery, while the levels of E1SO4 at both periods in the animal delivering a weak calf followed the profile in animals delivering a normal living calf. During late pregnancy and at the time of parturition, the levels of E1SO4 and PAGs in animals delivering a stillborn or weak calf (from group A1) followed the normal profiles found in animals delivering a normal living calf. In group A2, low levels of E1SO4 and pregnancy associated glycoproteins (PAGs) over 24 h at both 3 and 6 weeks prior to parturition (< 1.5 nmol/L) were recorded in animals delivering a stillborn calf. During late pregnancy and parturition, the levels of E1SO4 and PAGs were slightly lower during 30-50 days prior to delivery and increased with a lower magnitude at the time of parturition. In conclusion, our results indicate that the aetiology behind stillbirth varies depending on the AI-bulls used and is associated with dystocia or low viability of the calves. Deviating profiles of oestrone sulphate (E1SO4) and pregnancy associated glycoproteins (PAGs) in animals delivering a stillborn calf not caused by dystocia were observed, suggesting placental dysfunction as a possible factor. The finding suggests that the analyses of E1SO4 and PAGs could be used for monitoring foetal well-being in animals with a high risk of stillbirth at term.

Aminoglutethimide is an effective treatment for advanced postmenopausal breast cancer, acting in a novel way. It inhibits peripheral and tumour aromatase, which converts androgens to oestrogens. Marked suppression of oestrone and oestradiol is produced. Aminoglutethimide was used as an inhibitor of adrenal steroid biosynthesis, but the concentrations required to inhibit aromatase in vitro are 10 times lower. This has led to its clinical evaluation in much lower doses. Doses of 125 mg twice daily produce oestrogen suppression equivalent to conventional doses of 1 g daily, and also similar tumour responses. Toxicity is much reduced by the lower dose.

The influence of endogenously produced oestrogen on the growth of endometrial carcinoma was studied in postmenopausal 128 patients with endometrial adenocarcinoma. Plasma concentration of oestrone (E1) and oestradiol (E2) showed wide variations. Hormone levels were analysed in relation to growth rate, expressed as S-phase rate measured by flow cytometry, and to ploidy level. When the whole unclassified group was studied, no statistical relationship between E1 and E2 levels and S-phase rates were found. However, when peridiploid tumors (1.8-2.2 c) were divided according to histopathological grades, well differentiated tumors with low oestradiol concentrations (less than 60 pmol/I) had significantly lower S-phase rates than those with higher oestradiol levels (p less than 0.01). Aneuploid tumors showed high S-phase rates regardless of plasma oestradiol concentrations.

Steroid sulphatase (STS) activity was measured with tritiated dehydroepiandrosterone sulphate (DHEAS) and oestrone sulphate (OES) in leucocytes as well as in skin fibroblasts from 31 women who were presumed to be carriers of STS deficiency and recessive X-linked ichthyosis. Overall, 30 of the 31 women (96.8%) could be identified as heterozygotes in at least one of the four assay systems used, i.e. on the basis of having an STS activity below the 2.5 percentile calculated for normal control females. In the individual assay systems, the highest carrier detection rate was achieved with OES in leucocytes (96.2%), followed by DHEAS in leucocytes (80.8%), whereas a more pronounced overlap was present in the fibroblast systems. In leucocytes as well as in fibroblasts, the STS activity determined with DHEAS was positively correlated with the STS activity determined with OES (p less than 0.001) suggesting that a single sulphatase is responsible for the hydrolysis of both steroid sulphates.

Inhibition of oestrone sulphatase followed by oestrogen removal from tumour cells may be a new form of endocrine therapy of breast cancer in women. We investigated the inhibitory effect of the subchronic administration of oestrone-3-O-sulphamate (EMATE), a steroid sulphatase inhibitor, to ovariectomized rats, to evaluate this method for testing new nonsteroidal inhibitors. EMATE in DMSO was administered both orally and subcutaneously (s.c.) for 7 days at doses of 0.5 and 2.5 mg/kg. In addition the rats were injected s.c. with 0.5 mg oestrone sulphate/kg 26 and 2 h before decapitation under ether anaesthesia. Oestrone sulphatase activity (ESA) was measured radiometrically using [3H]oestrone sulphate as substrate for desulphuration in white blood cells, liver homogenate, microsomes and spleen homogenate. ESA in liver microsomes was found to be nearly 40 times higher than in white blood cells while in spleen ESA was nearly half of that found in liver homogenates and white blood cells. ESA can be inhibited by EMATE down to 50-1.5% of control activity depending on the dose and administration route. The inhibition was in the order, liver homogenate < spleen < liver microsomes < white blood cells, and was more pronounced after s.c. administration of the inhibitor than after oral administration. Ovariectomy was found to be not necessary for oestrone sulphatase-inhibiting studies. Two sequential s.c. injections of oestrone sulphate enhanced the enzyme activities significantly in liver and white blood cells, but not in spleen. In conclusion, white blood cells and liver microsomes of intact female rats can be used for ESA-inhibiting studies. Sulphate-conjugated oestrone can induce oestrone sulphatase in vivo in liver and white blood cells thereby enhancing oestrogen supply in the peripheral organs.

Conversion of oestrone sulphate to oestrone has been suggested to make a major contribution to the level of oestrone found in breast tissues. In order to examine the ability of breast tissues to take up oestrone sulphate (E1S), 3H E1S or E1-35S was infused into postmenopausal women with advanced breast cancer. For 3 subjects infusion of 3H E1S was repeated after treatment with Danazol, a potential inhibitor of oestrone sulphatase activity. After infusion of 3H E1S significant levels of 3H E1S were detected in normal and malignant breast tissues (tissue: plasma ratios 0.14 +/- 0.13 and 0.24 +/- 0.12 respectively, mean +/- S.D., n = 5). Similar 3H E1S tissue: plasma ratios were detected after infusions of 3H E1 indicating that the 3H E1S detected in breast tissues after infusion of 3H E1S may have originated from the hydrolysis of 3H E1S in tissues other than the breast, with subsequent uptake and sulphation in breast tissues. After infusion of E1-35S no significant levels of radioactivity were detectable in normal or malignant breast tissues. Treatment with Danazol had no significant effect on tissue levels of 3H E1S or on the CRE1S E1 or MCR-E1S. It is concluded that oestrone sulphate, as such, is not taken up by breast tissues and that any contribution that oestrone sulphate makes to the oestrogen content of breast tissues will depend upon prior hydrolysis.

The uptake and metabolism of 3H-oestradiol by breast tumour tissue was studied in three postmenopausal women before, after 1 week of treatment with ethynyl-oestradiol (EE, 10 micrograms/day) and again two weeks later after treatment with medroxyprogesterone acetate (MPA, 500mg/day, i.m.). Treatment with EE reduced the tissue: plasma gradients for both 3H-oestradiol and 3H-oestrone and little further change in these ratios occurred after MPA treatment. The proportion of oestrogen present as 3H-oestrone increased in tumours from 2/3 women after treatment with MPA but not EE, although no increase in the activity of oestradiol 17 beta-hydroxysteroid dehydrogenase (E2DH) was detected. Plasma and tissue concentrations of 5-androstenediol, dehydroepiandrosterone and dehydroepiandrosterone sulphate were all reduced after MPA treatment. The results from this study show that treatment with EE and MPA can reduce the uptake of oestradiol by breast tumours. No increase in E2DH activity was detected but it is possible that MPA influences other enzymes involved in oestradiol metabolism.

The authors hypothesized that IL-6 production by breast tumour tissues would correlate with OR-positivity, as only those tumours that contain oestrogen receptors (OR) and use oestrogen as a mitogen would benefit from locally increased oestrogen. IL-6 increases the activity of the 17beta-oxidoreductase, which converts oestrone to oestradiol, a process that may contribute to the increased concentration of oestrogen around breast tumours. IL-1alpha upregulates IL-6 production; therefore, the correlation between IL-1alpha and IL-6 immunoreactivity and OR-positivity in paraffin-embedded human breast tumours was further investigated.The results indicate IL-6 immunoreactivity in 40 of 66 paraffin embedded breast tumour specimens, a finding which did not correlate with the clinical evaluation of oestrogen receptor positivity (P=0.32 by Fisher's exact test). However, there was a correlation between IL-6 and IL-1alpha immunoreactivity (P<0.05). To study an in vitro model for this phenomenon, the IL-6 immunoreactivity in available cell lines was tested. Surprisingly, no production of IL-6 protein or mRNA could be detected in any of the cell lines, and this did not change with IL-1alpha stimulation. Therefore, none of the cell lines apparently reflected the immunological potential observed in the majority of surgical specimens.

BACKGROUND

Aminoglutethimide was the first aromatase inhibitor to be used successfully in breast cancer patients. However, this drug also inhibits mineralcorticoid and glucocorticoid synthesis, making co-medication with corticosteroids necessary, and it is often poorly tolerated. The primary objective of this trial was to evaluate the clinical efficacy and tolerability of vorozole, a new non-steroidal oral aromatase inhibitor, in postmenopausal breast cancer patients. The secondary objective was to evaluate the pharmacodynamic activity of the drug.

METHODS

Thirty-four postmenopausal patients previously treated with tamoxifen in the adjuvant setting and/ or for advanced disease were treated with vorozole, 2.5 mg once daily. Patients were monitored with respect to treatment efficacy and safety. Hormonal evaluations were performed at baseline and during the course of treatment in order to evaluate the pharmacodynamic efficacy and safety of vorozole.

RESULTS

According to UICC criteria, there were seven responders, one complete and six partial, for an overall response rate of 21% (95% confidence interval (CI) 9%-38%). The median duration of response was 9.6 months (95% CI 4.6-0), the median time to progression for the entire group was 4.7 months (95% CI 2.9-6.6) and the median survival time was 29.7 months (95% CI 19.1-0). Tolerability was excellent to good in 97% of the patients. Oestradiol and oestrone levels were suppressed to the limit of detection of the assays used. No effect was observed on the other endocrine parameters.

CONCLUSIONS

Our results suggest that vorozole is an effective and safe drug for the treatment of advanced postmenopausal breast cancer following tamoxifen failure.

Human peripheral blood adherent cells (PBAC) incubated with oestrone in high concentration (10(-5) M) release, on exposure to bacterial endotoxin, an amount of tumour necrosis factor (TNF) greater than do cells incubated without the steroid. The finding may imply a non-endocrine hormonal process leading to heightened local TNF responses. This may be the basis of endotoxin hypersensitivity in pregnancy.

The effect of oestrone acetate (in total doses of 5 and 10 mg) on systemic and renal haemodynamics and the renin-angiotensin system has been studied in adult female rats. The administration of 10 mg of oestrogen resulted in a significant fall in renal blood flow associated with significant rises in both renal vascular resistance and mean arterial pressure. No changes were noted in cardiac output or total peripheral resistance at either dose. Whilst the higher dose of oestrogen induced a significant increase in plasma renin activity, no change was noted in animals receiving 5 mg of oestrogen. Both regimens caused significant reductions in plasma and intrarenal renin concentrations. Although renal blood flow correlated with plasma renin activity in animals with a normal renal blood flow, no such correlation was noted in animals with oestrogen-induced reductions in renal blood flow. The present study demonstrates that oestrogen-induced reductions in renal blood flow result from a rise in intrarenal vascular resistance which cannot be accounted for by simultaneous changes in either plasma renin activity or renal renin concentration.

A large proportion of the beneficial effects that oestrogens demonstrate on the vasculature are believed to be mediated via direct effects on the vascular wall. In this study we compared a number of oestrogenic compounds isolated from pregnant mare's urine including 17beta-oestradiol and oestrone, in terms of their abilities to inhibit stimulated endothelin-1 release from normal human coronary artery endothelial cells (CAEC). We also examined their ability to stimulate expression of constitutive endothelial nitric oxide synthase (eNOS) and explored their effects on cellular angiotensin converting enzyme (ACE). All the oestrogens tested were able to inhibit serum-stimulated ET-1 release. Oestrone and 17alpha-dihydroequilenin failed to significantly affect cellular eNOS levels. 17Beta-oestradiol and oestrone significantly increased cellular ACE levels while 17beta,delta(8,9)-dehydroestradiol decreased cellular ACE. We discuss these observations in terms of their potential clinical relevance and use as a means of screening novel oestrogen-like compounds.