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Abstract. Serum sex hormone binding globulin (SHBG) concentrations were measured by an immunoradiometric assay, as part of a longitudinal study of puberty in girls, and were related to age. pubertal stage, age at menarche, weight, nature of the menstrual cycle and serum concentrations of sex steroids. A slow but very significant decrease was seen in SHBG from 77 nmol/l at 8–10 years of age to about 50 nmol/l after 15 years of age. Serum SHBG concentrations showed weak negative correlations with those of androstenedione and testosterone during puberty. The closest associations found between SHBG and the parameters measured were negative correlations with weight and body fat percentage in both pre-menarcheal and post-menarcheal girls, even after the effect of age was accounted for by calculating partial correlation coefficients. Girls who experienced early menarche (before 13.0 years) had lower SHBG but higher oestradiol serum concentrations at 10.0–15.9 years of age compared to girls with later menarche. In ovulatory menstrual cycles, a significant increase in SHBG was found from the early to the late part of the cycle, whereas no changes took place in anovulatory cycles. Serum concentrations of SHBG showed positive correlations with those of oestradiol and progesterone in specimens taken in the late part of the cycle.In view of the weak relationships between serum SHBG and sex steroid concentrations, and the strong relationships between SHBG, weight and body fat percentage, factors other than steroids have to be considered in the regulation of SHBG levels during puberty.

Prostate tissues removed from patients with benign prostatic hypertrophy were separated into epithelial and stromal components and the concentrations of testosterone, 5α-dihydrotestosterone, 5α-androstane-3α,17β-diol, 4-androstene-3,17-dione, 5α-androstanedione stanedione and androsterone in these two fractions were determined by radioimmunoassays after the purification of solvent steroid extracts by Lipidex-5000 column chromatography. On a 'per cell' basis (i.e. relative to DNA), testosterone was equally distributed between the two components, while the other androgens measured were more abundant in the stroma. The observation that 5α-reduced androgens (especially 5α-dihydrotestosterone) were more concentrated in the stroma, and that significant correlations between concentrations of metabolically related androgens were more common in the stroma than in the epithelium, indicate that the stroma is an important site of androgen metabolism in benign prostatic hypertrophic tissues. The present data also support the suggestion that 5α-dihydrotestosterone produced in the prostatic stroma may be transferred to the epithelium by way of sex hormone binding globulin in the extracellular spaces of the prostate.

Prostate tissues removed from patients with benign prostatic hypertrophy were separated into epithelial and stromal components and nuclei purified from these. The concentrations of testosterone, 5α-dihydrotestosterone, 5α-androstane-3α,17β-diol, 4-androstene-3,17-dione, 5α-androstanedione and androsterone in pooled preparations of the purified nuclei were determined by radioimmunoassays after the purification of solvent steroid extracts by Lipidex-5000 column chromatography. The most abundant androgen measured was 5α-dihydrotestosterone and it was significantly (P< 0·05) more concentrated in the stromal nuclei than in the epithelial nuclei. The mean concentration of androsterone was also significantly (P<0·05) greater in the stromal nuclei whereas that of testosterone was equal in the two nuclear types. The concentrations of 5α-androstane-3α,17β-diol, 4-androstene-3,17-dione and 5α-androstanedione were below the sensitivity limits of the assays in the majority of cases, but the results indicated that when detectable the first two were more concentrated in the stromal than the epithelial nuclei. The results emphasize the importance of the prostatic stroma in androgen metabolism, and the relative concentrations of 5α-dihydrotestosterone and testosterone indicated identical 5α-reductase activities in the nuclei in comparison with respective whole cell epithelial and stromal preparations. 5α-Dihydrotestosterone concentrations were clearly higher than the nuclear androgen receptor levels previously reported from this laboratory.

The administration of danazol, 200 mg three times daily, from the 3rd to the 23rd day of the cycle to normally menstruating women exhibited the following actions on the human endometrium: significantly reduced cytosol oestrogen and progestin receptor concentrations, and declined 17β-hydroxysteroid dehydrogenase activity. Very similar results were obtained during medroxyprogesterone acetate (100 mg daily) treatment for the same period of time. Danazol administration did not decrease circulating gonadotrophin levels but clearly suppressed luteal serum oestradiol and progesterone concentrations. Danazol was found to bind in vitro to endometrial progestin receptor with an affinity approximately 3% of that of progesterone. These findings are compatible with the notion that a local progestin-like rather than a systemic action of danazol is the way by which its therapeutic effect is exerted. This may be potentiated by the suppression of circulating oestradiol levels.

Measurements of serum ferritin were correlated with bone marrow iron content and other laboratory hematological indices in 44 anemic (B-Hb < 100 g/l) pregnant women. Seventy-two% of patients classified as iron deficient by the absence of bone marrow iron had diagnostically low serum ferritin concentrations, which percentage is about double compared with those revealed by measurements of serum transferrin, TIBC, transferrin saturation, serum iron, blood hemoglobin and red cell indices. It is also important that serum ferritin was the only one of these measurements which was able to differentiate between iron deficiency, and anemia due to infection. Seven out of 8 patients with anemia due to infection had elevated serum ferritin levels. In 17% of patients no reason for the fall of blood hemoglobin could be shown. A plausible explanation for the low blood hemoglobin is extensive hemodilution.

Serum ferritin concentration was measured in 47 paired maternal and newborn (umbilical cord sample) blood specimens during delivery. The serum ferritin concentration was also measured in 31 of the above babies at the age of six months. Serum ferritin concentration of the newborn babies (mean 222 μg/l) born to the mothers with a low serum ferritin content at term (mean 28 μg/l) was significantly lower (p < 0.05) than that for those (mean 324 μg/l) born to the mothers with a normal ferritin concentration (mean 102 μg/l). The difference between the two groups of infants persisted for at least the whole study period of 6 months (mean at 6 months of age 99 μg/l and 150 μg/l, respectively).

The iron stores of 32 healthy pregnant women were evaluated longitudinally during pregnancy and 6 months post partum by serum ferritin assay and by bone marrow iron content. Half of the women were receiving oral iron while the others were not given iron supplementation.Women receiving iron could maintain their iron stores throughout the pregnancy. By contrast, women without iron therapy had low serum ferritin values, pointing to the absence of iron stores during the last trimester, and 6 of these 16 women developed anemia. This was confirmed by estimation of the quantity of stainable iron in the bone marrow. In addition serum iron, transferrin and red cell MCV values indicated iron deficient erythropoesis.During a 6-month period after pregnancy the women receiving supplemental iron during pregnancy had a significant increase in their serum ferritin concentrations, indicating restoration of iron stores. Women not receiving iron during pregnancy had exhausted iron stores at term and serum ferritin values stayed low even at 6 months after delivery. If iron therapy was instituted after parturition, serum ferritin assays indicated restoration of iron stores within the ensuing 6 month. To prevent iron deficiency anemia during pregnancy supplemental iron is advisable for all pregnant women in our country.

The faecal and urinary excretion of intravenously injected labelled dehydroepiandrosterone sulphate was studied in three healthy subjects on a standardized diet before and during oral cholestyramine administration. Faecal excretion was found to be an important pathway for the elimination of administered radioactivity. Thus, 28–45 per cent of the activity was recovered in the faeces during the control period. The urinary isotope excretion was slightly lower during cholestyramine administration. The plasma concentrations of »sulphates« of androsterone, dehydroepiandrosterone, epiandrosterone, androst-5-ene-3β,17β-diol and pregn-5-ene-3β,20α-diol and the urinary excretion of »sulphates« of the same steroids with the exception of epiandrosterone, were determined by gas chromatographic procedures. The individual and daily variations of the values were large and no change could be detected when cholestyramine was administered. The low renal clearance of steroid sulphates has been confirmed.

Steroid disulphate fractions were obtained by chromatography on Sephadex LH-20 from samples of amniotic fluid from three pregnancies complicated by anencephaly. Several of the steroids with a 3β-hydroxy-5-ene structure found in normal amniotic fluid could not be detected. 5-Androstene-3β,17α-diol, the main steroid disulphate in normal amniotic fluid was present in concentrations of <5% of the values at normal term. The disulphates of neutral steroids found in highest concentrations in the amniotic fluid of anencephalic pregnancies were those of isomeric pregnanediols; 5α-pregnane-3α,20α-diol, 5α-pregnane-3β,20α-diol and 5β-pregnane-3α,20α-diol. The quantities of these metabolites of progesterone in the amniotic fluid of anencephalic pregnancies were similar to those in normal pregnancies. A saturated 18-hydroxylated C19steroid, most probably 3α,18-dihydroxy-5α-androstan-17-one, was identified by gas—liquid chromatography and gas chromatography—mass spectrometry. Although this compound has been previously identified in human bile and in normal pregnancy urine, this appears to be the first identification of a saturated C19steroid in amniotic fluid.

Steroid monosulphate and disulphate fractions were obtained by chromatography on Sephadex LH-20 from human amniotic fluid collected at term. Only 16α-hydroxydehydroepiandrosterone could be identified in the monosulphate fraction, the detection limit being 5–8 μg./1000 ml. amniotic fluid. In the disulphate fraction 5-androstene-3β,17α-diol, 5-androstene-3β,17β-diol, 5-pregnene-3β,20α-diol, 5α-pregnane-3β,20α-diol, 16α-hydroxydehy-droepiandrosterone, 16β-hydroxydehydroepiandrosterone, 3β, 17β-dihy-droxy-5-androsten-16-one and 5-pregnene-3β,17α,20α-triol were identified by gas—liquid chromatography and gas chromatography—mass spectrometry. The concentrations of these steroids were determined in a pool of human amniotic fluid taken at term. The main compound was 5-androstene-3β,17α-diol.

Solvolyzable steroids in human peripheral plasma were separated by chromatography on Sephadex LH-20 into two fractions having the elution volumes of mono- and disulphates, respectively. The steroid conjugates in the latter fraction had the mobility of steroid disulphates on TLC. After solvolysis in ethyl acetate acidified with sulphuric acid the steroids were analyzed by gas chromatography and combined gas chromatography - mass spectrometry. Cholesterol, dehydroepiandrosterone, androsterone, epiandrosterone, aetiocholanolone, pregnenolone, pregn-5-ene-3β,20α-diol, androst-5-ene-3β,17β-diol, androst-5-ene-3β,17α-diol and an androstanediol, most probably 5α-androstane-3α,17β-diol were found in the monosulphate fraction. Androst-5-ene-3β,17β-diol, androst-5-ene-3β,17α-diol and pregn-5-ene-3β,20α-diol were found in the disulphate fraction. The ratio of C19to C21diols was higher in the disulphate than in the monosulphate fraction.

A disulphate fraction of the steroids in the urine of pregnant females was separated by chromatography on Sephadex LH-20. The steroids in this fraction showed the mobility of steroid disulphates on thin-layer chromatography, paper chromatography and high-voltage paper electrophoresis. After solvolysis, the free steroids were fractionated on silicic acid and by thin-layer chromatography. Fractions containing dihydroxylated C19and C21steroids were obtained. Using gas-liquid chromatography and gas chromatography-mass spectrometry, the following 18 steroids were identified in the disulphate fraction of neutral steroids in late-pregnancy urine: 5α-androstane-3α,17α-diol, androst-5-ene-3β,17α-diol, androst-5-ene-3β,17β-diol, 18-hydroxyandrosterone, 17α-hydroxydehydroepiandrosterone, 16β - hydroxydehydroepiandrosterone, 3β,17β-dihydroxyandrost-5-en-16-one, 5α-pregnane-3α,20α-diol, 5α-pregnane-3β,20α-diol, 5β-pregnane-3α,20α-diol, 5β-pregnane-3β,20α-diol, pregn-5-ene-3β,20α-diol, 3β,16α-dihydroxy-5α-pregnan-20-one, 16α-hydroxypregnenolone, 3α,21-dihydroxy-5α-pregnan-20-one, 3β,21-dihydroxy-5α-pregnan-20-one, 3α,21-dihydroxy-5β-pregnan-20-one and 21-hydroxypregnenolone. 5α-Pregnane-3β,20α-diol was the most abundant steroid in the fraction analysed, and preliminary quantitative data suggest that sulphate-conjugated pregnanediols and 21-hydroxypregnanolones are excreted mainly as disulphates in late pregnancy urine. The origin of these maternal urinary steroids is discussed.

Sulphate-conjugated neutral steroids in the intestinal contents of early and mid-term human foetuses (14–20 weeks of gestation) were investigated by gas—liquid chromatography and gas chromatography—mass spectrometry. Twenty-four neutral steroids were found in the monosulphate fraction and 21 in the disulphate fraction. The total concentration of these steroids varied between 9·6 and 15·3 mg/100 g meconium wet weight, about half being monosulphates and half disulphates. Steroids with a 3β-hydroxy-5-ene structure were found and also saturated steroids which carried hydroxyl groups at carbons 3, 11, 15, 16, 17, 18, 19, 20 or 21. Among them were steroids not previously detected in human foetal compartments. Thus, the foetal meconium is both qualitatively and quantitatively the richest foetal source of neutral steroid sulphates so far investigated.The present results together with those reported in the literature show that in the course of pregnancy there is a many-fold increase in the steroid concentration of the foetal intestinal contents. This is most obvious in the case of saturated C19steroids and polar pregnane derivatives. Reduction of the 20-ones to 20β-ols seems to be more extensive towards the end of pregnancy. 5, 16-Pregnadienes and 3β-hydroxy-5β-pregnanes are present in higher concentrations in early pregnancy.

The endogenous neutral steroids in early and mid-term (gestational age 10-14 weeks) human foetal kidneys were investigated. Fractions of unconjugated and mono- and disulphated neutral steroids were studied separately. No unconjugated compounds were detected, but the following steroid monosulphates were found: dehydroepiandrosterone, 5-androstene3β,17α-diol, 5-androstene-3β,17β-diol, 16α-hydroxydehydroepiandrosterone, 16β-hydroxydehydroepiandrosterone, 3β,17β-dihydroxy-5-androsten-16-one, pregnenolone, 5-pregnene-3β,20α-diol, 16α-hydroxypregnenolone and 17α-hydroxypregnenolone. In addition, the following compounds were present, as disulphates: 5-androstene-3β,17α-diol, 5-androstene-3β,17β-diol and 5-pregnene-3β,20α-diol.Quantitatively, dehydroepiandrosterone, 16α-hydroxydehydroepiandrosterone and pregnenolone in the monosulphate fraction and 5-androstene-3β,17α-diol in the disulphate fraction were the main steroids found. Furthermore, in vitroincubations with minced or homogenized foetal kidney tissue demonstrated the following enzyme activities: 16α-hydroxylase, 17β-reductase, 20α-reductase and sulphokinase. In the incubations, dehydroepiandrosterone, pregnenolone and their sulphate conjugates were used as substrates. The role of the foetal kidneys in the metabolism of neutral steroids during early and mid-pregnancy is discussed.

The endogenous neutral steroids in adrenal and liver tissue obtained from foetuses of 12–17 weeks gestation were investigated. After extractions, the steroids were fractionated into a monosulphate and a disulphate fraction on Sephadex LH-20. The conjugates were cleaved by solvolysis, and purified on silicic acid and their trimethyl silyl ethers were analysed by gas-liquid chromatography and gas chromatography-mass spectrometry. Only neutral steroid monosulphates were found. The following compounds were identified in the monosulphate fraction of adrenal tissue: dehydroepiandrosterone, 16α-hydroxydehydroepiandrosterone, pregnenolone and 17α-hydroxypregnenolone, and of liver tissue; dehydroepiandrosterone, 16α-hydroxydehydroepiandrosterone, androst-5-ene-3β,16α,17β-triol, pregnenolone, 17α-hydroxypregnenolone and 16α-hydroxypregnenolone. The identifications and the results of the quantitative determinations of endogenous neutral steroid sulphates suggest that pregnenolone is synthesised in vivoin the foetal adrenal and converted to 17α-hydroxypregnenolone and dehydroepiandrosterone sulphates. The latter compound is further transformed to 16α-hydroxydehydroepiandrosterone sulphate. No 16α-hydroxylation of pregnenolone was observed in adrenal tissue, whereas liver tissue seems to form 16α-hydroxypregnenolone. Reduction of a 17-keto group to 17β-hydroxyl and a 20-keto group to 20α-hydroxyl seems to take place in liver but not in adrenal tissue.