F. Franceschetti

Estrone sulfate, estrone and estradiol concentrations in normal and cirrhotic postmenopausal women

Circulating levels (mean +/- SD) of estrone sulfate (E1S), estrone (E1) and estradiol-17 beta (E2) were measured in normal and cirrhotic postmenopausal women matched for body weight and age. In cirrhotic postmenopausal women, the E1S concentrations (201 +/- 46 pg/ml), while both E1 and E2 levels showed an increase (46 +/- 7 and 30 +/- 8 pg/ml) compared to control subjects (32 +/- 6 and 18 +/- 7 pg/ml). These data suggest that the liver plays an important role on the control of estrogen sulfation.

Estrone sulphate plasma levels in postmenopausal women with and without endometrial cancer

Plasma estrone sulphate (E1S) and estrone (E1) concentrations were determined in healthy postmenopausal women and in postmenopausal women with endometrial cancer, matched for body weight, age, and years since menopause. E1S levels (mean ± SD) were significantly higher (P < 0.05) in cancer patients with normal weight (511 ± 200 pg/ml) than in control subjects (303 ± 99 pg/ml). E1S levels were also higher in obese cancer patients (691 ± 328 pg/ml) than in obese control subjects (610 ± 139 pg/ml). Both cancer groups showed similar plasma E1 levels as compared with their respective controls. The E1S/E1 ratio was higher in both groups of cancer patients than in control subjects. These data suggest that estrogen conjugates should be taken into account during studies on estrogen balance and endometrial cancer.

Uterine Contractility: Vaginal Administration of the β‐Adrenergic Agonist, Terbutaline

Spontaneous uterine contractility during the menstrual cycle is required for menstruation, gamete transport, and, most likely, embryo nidation. Abnormal uterine contractility has been linked to dysmenorrhea, a condition associated with painful uterine cramping. Based on previous studies with progesterone, we have postulated the existence of a portal system that is responsible for some degree of direct vagina‐to‐uterus transport of administered compounds (i.e., the “first uterine pass effect”). It is possible that treatment with uterorelaxing substances, particularly β‐adrenergic agonists, may alleviate the uterine discomfort that accompanies dysmenorrhea. However, side effects encountered with oral administration of β‐agonists limit their utility. Alternatively, vaginal delivery of β‐agonists could solve this dilemma by enhancing their efficacy and reducing side effects. Therefore, in the current study we used hysterectomy specimens and an in vitro uterine perfusion system to test the vagina‐to‐uterus transport of [3H]terbutaline, a well‐known β‐agonist. With the use of autoradiographic and scintillation counting techniques, our results clearly show progressive diffusion of labeled terbutaline from the rim of vaginal tissue through the uterus during the first 12 hours of perfusion. This indicates that uterine targeting of terbutaline can be accomplished through vaginal administration, suggesting a new therapeutic modality in womens health care.

Metabolic clearance rate of oestrone sulphate in post-menopausal women

The feasibility of using constant infusions of unlabelled oestrone sulphate (E1S) for the purposes of calculating its metabolic clearance rate (MCRE1S) and its conversion ratios to oestrone (E1) and oestradiol (E2) in post-menopausal women was exploited in this study. The results obtained by the infusion of unlabelled E1S were similar to those obtained by the infusion of labelled steroid. The MCRE1S values seen in our group of post-menopausal women fell within the range previously reported for fertile women. The contribution of E1S to circulating E1 averaged 18% (range 14-24%), indicating that the E1S-E1 equilibrium should be taken into account during studies on oestrogen balance in post-menopausal women.

Rapid and specific ria of serum estrone sulfate with selective solid phase extraction

The methods commonly used to evaluate conjugated steroids require hydrolysis and chromatographic purification. To avoid these steps, a simple method involving selective solid phase extraction and RIA using a highly specific antiserum for estrone sulfate (E1S) has been evolved. A Bond-Elut C2 cartridge was used for solid phase extraction of estrone (E1) and E1S; recoveries were 80 and 90% respectively. The intra- and inter assay precision of the assay at 3 serum levels, were 6.5, 10.4 and 4.4 and 12.7, 13.9 and 7.4% respectively. Accuracy, tested by linearity and recovery tests, was acceptable. A good correlation exists between a conventional enzymatic method and the proposed method. The latter is less time consuming and more reliable, thus providing a rapid assay to evaluate E1 and E1S in the same serum sample.

Treatment of Endometrial Hyperplasia with Cyproterone Acetate Histological and Hormonal Aspects

A regime of cyproterone acetate (CPA) (300 mg/‐day by the oral route for 30 days) has been used in 10 postmenopausal women with endometrial hyperplasia (8 atypical and 2 adenomatous). Androstenedione (A), estrone (E1), testosterone (T) and estradiol (E2) plasma levels were determined before and at the end of treatment. The regression of endometrial hyperplasia was ascertained histologically in all patients after 30 days of therapy. All steroids showed a significant decrease (p<0.05) as compared with their corresponding basal values. Moreover, the E1A ratio was significantly lowered (p<0.01) following CPA administration (5.9+2.9% to 2.5+0.6%). From these data it is evident that CPA can not only act as a progestin, but may also reduce the endogenous estrogen production, lowering either the adrenal production of A (the most important estrogen precursor in the post‐menopause) or the A to E1 peripheral conversion.

Reduced conversion of dehydroepiandrosterone into estrogens in women having hypogonadotropic hypogonadism associated with weight loss.

The purpose of this study was to evaluate, without using radioisotopes, the peripheral contribution of dehydroepiandrosterone (D) to estrogens and to androstenedione (A) in patients with hypogonadotropic hypogonadism associated with weight loss (HH) and in normal menstruating women (N). Unlabelled D was infused for 48 h in 12 normal women and in 12 women affected by HH. Plasma levels of D, dehydroepiandrosterone sulfate (DS), A, estrone (E1), estrone sulfate (E1s) and estradiol (E2) were measured before and after 48 h of infusion. Metabolic clearance rates of D (MCRD), production rates of D (PRD), and increases in plasma concentration of DS, A, E1, E1s and E2, relative to the corresponding increase in plasma concentration of D, were determined. The baseline plasma levels of all steroids studied were found to be significantly lower in the patient group than in the control. The MCRD in the normal and the HH groups were similar (1420 +/- 340 l/day versus 1670 +/- 569 l/day, P greater than 0.05). No significant difference was found in PRD between the 2 groups (mean +/- SD 10.3 +/- 5 versus 13.3 +/- 5.5 mg/day, P greater than 0.05). Administration of D increased the levels of estrogen in the normal group but not in the HH group. The relative increase in plasma levels of DS resulting from infusion of D (delta cDS/delta cD) was found to be larger in the HH group than in the normal group (40.4 +/- 17 versus 26.3 +/- 11.8, P less than 0.05). Furthermore, relative increases in plasma levels of A derived from infusion of D were larger in the HH group than in the normal group (0.0495 +/- 0.0021 versus 0.192 +/- 0.0071, P less than 0.001). We conclude from these results that in the HH patients there is a blockage of the peripheral conversion of D to E1 and E1s and an enhancement of the peripheral conversions of D to DS and to A. These metabolic changes may account for the androgenization of the patients under study.

Two new fluorescent derivatives of progesterone to use in fluoro immunoassay

Synthesis, fluorometric and immunological properties of two new fluorescent derivatives of progesterone are reported. Both compounds were obtained from 11 alpha-hydroxyprogesterone 11-hemisuccinate; the fluorescent molecules were joined to the steroid by bifunctional arms. The first of these is cysteamine whose thiol group was reacted with N-(3-fluoranthenyl) maleimide, and the second is tyramine whose phenolic group was reacted with 1-nitroso-2 naphthol.