Twelve men, 19-40 years old, and 12 men, 60-86 years old, were given ACTH and in a second test series, HCG. Oestrone, oestradiol-17beta and cortisol were measured in the plasma before and after stimulation. Under basal conditions oestrone and oestradiol as well as the percentage binding of testosterone increased significantly with age, whereas cortisol remained unchanged. The administration of HCG resulted in higher levels of both plasma oestrone and oestradiol in older subjects than in younger ones, although the percentage increase was smaller. ACTH stimulation leads to higher levels of oestrone in the older group, whereas oestradiol remained unchanged. Cortisol rose 3.5 fold in both groups. Regarding absolute concentrations of oestrone and oestradiol the ability of the adrenals and of the testes to respond to exogenous gonadotrophin and adrenocorticotrophin appeared to be preserved despite ageing, though the relative responses of oestrone and oestradiol after HCG and of oestrone after ACTH were decreased in old as compared to young men.

The hormonal mechanisms involved in the development of gynaecomastia accompanying the treatment of multiple myeloma in adult men have been investigated by studying levels of circulating testosterone (T), oestrone (EI), oestradiol (E2), sex-hormone binding globulin (SHBG), prolactin (PRL) and the gonadotrophins LH and FSH, before, during and after development of gynaecomastia in 4 men. These have been compared with 5 closely matched men who did not develop gynaecomastia during similar treatment for myeloma. Levels of circulating T fell, and levels of E1 and E2 rose during treatment periods in all subjects, and the changes were statistically significant in subjects developing gynaecomastia, which resolved as levels of sex steroid returned towards normal following cessation of treatment. We conclude that treatment of adult men for myeloma results in testicular dysfunction with a reduction in circulating T and a rise in circulating oestrogens. These changes are most marked in subjects developing gynaecomastia in whom the normal breast tissue is stimulated by a subtle, transient oestrogen:androgen imbalance in favour of oestrogens.

In 3 testes of 2 adult Pony stallions under halothane anaesthesia, catheters were inserted into a vein and a lymphatic vessel in the spermatic cord and into a vein on the surface of the testis. Lymph and venous blood were collected from the catheters in the cord and p-aminohippurate (2% w/v, 0 . 1 ml/min) was infused into the vein on the testis to determine blood flow by dilution. After 1 h, 6000 i.u. hCG was injected i.v. and collections continued for 45 min. The testes weighed 126-176 g. Lymph flow was 20-150 microliter/min before hCG and 100-270 microliter/min after hCG; the range of blood flow, with a haematocrit of about 22%, was unchanged (27-47 ml/min) after hCG. The concentration of testosterone in spermatic venous blood rose from 6-20 ng/ml to 160-270 ng/ml within about 30 min after hCG. Peripheral levels rose from about 2 to 6 ng/ml and lymph levels increased from 9-20 to 34-150 ng/ml after hCG. In contrast, the concentrations of oestrone sulphate in spermatic venous blood were about 400 ng/ml, compared with about 250 ng/ml in peripheral blood, while lymph values ranged from 600 to 1500 ng/ml; hCG had no consistent effect. DHA and its sulphate were present in spermatic venous blood at concentrations (about 3 and 5 ng/ml respectively) slightly higher than in peripheral blood (1 . 7 and 2 . 5 ng/ml), but the sulphate was at a very much higher concentration in lymph (about 5 and 200 ng/ml respectively). These results indicate that testicular lymph is an important route for the secretion of conjugated steroids by the horse testis.

Peripheral serum samples were collected from 8 pregnant dromedary camels and hormone secretion patterns were examined at specific time intervals. Mean serum progesterone concentrations began to rise 3-4 days after ovulation and remained reasonably constant at 3-5 ng mL-1 for the first 90-100 days of gestation. Concentrations then showed a definite fall, but thereafter remained constant again at 2-4 ng mL-1 throughout the rest of pregnancy. In contrast, serum oestrogen concentrations showed pronounced fluctuations during the first 100 days of gestation. Mean oestradiol-17 beta concentrations increased at around Day 50 to about 100 pg mL-1 and then remained relatively constant from Day 90 to Day 300. Mean oestrone sulfate concentrations, however, showed two definite peaks in early gestation, each reaching about 10 ng mL-1, with the first peak occurring around Day 25 and the second peak around Day 75. Oestrogen production then remained fairly constant until around Day 300, after which concentrations of both oestrone sulfate and free oestradiol-17 beta rose steeply over the next 80 days to reach mean peak values of 46 ng ML-1 and 518.7 pg mL-1, respectively, at the time of parturition. Concentrations of 13,14 dihydro-15-keto prostaglandin F2 alpha (PGFM) remained low and reasonably steady at 100-200 pg mL-1 during the first 320 days of pregnancy; thereafter, PGFM concentrations rose steeply over the next 50 days, before an explosive further increase to a peak of 1900 +/- 141 pg mL-1 mean +/- sem on the day of calving. These results suggest that, as in the cow, a major change in steroid synthetic capability and/or enzyme content of the placenta may occur at around 80% (Day 300) of gestation in the pregnant camel.

Oestrone sulphate was extracted from pig serum with tetrahydrofuran and then hydrolysed with sulphatase. The radioimmunoassay of liberated oestrone was carried out directly on the reaction mixture. Oestrone sulphate concentrations were less than 60 pg/ml in non-pregnant sows. Detectable concentrations of oestrone sulphate were found in serum as early as the 18th day of gestation, and these rose to maximum levels of 2.1-3.5 ng/ml by Days 26-29 then fell to about 0.1 ng/ml by Day 37. Serum oestrone sulphate concentrations were not related to litter size, but were good indicators of early pregnancy.

Concentrations of oestradiol-17 beta, oestradiol-17 alpha, and oestrone in systemic plasma of pregnant pygmy goats (Capra hircus) remained low until about Day 60 and then rose to maximum values at Days 120-140 (parturition Day 140-145). Oestradiol-17 alpha was the predominant oestrogen. All 3 oestrogens at Days 100-130 were higher in females carrying 3 fetuses than in those carrying only one, but at Days 70-90 only oestrone values were higher. It is suggested that the feto-placental unit is the source of oestrogens during gestation in goats.

To examine the direct effect of STS 557 (17 alpha-cyanomethyl-17 beta-hydroxy-estra-4,9-dien-3-one) on the ovarian biosynthesis of steroids, tissue slices obtained from a cyclically functional human ovary were incubated with [4-14C]-pregnenolone as precursor. The steroid production was determined by measuring the concentration of eleven 14C-labelled steroids in the medium at the end of a 3 h-incubation. The addition of STS 557 to the incubation medium inhibited the biosynthesis of androstenedione, progesterone, 17 alpha-hydroxyprogesterone and oestrone, whereas dehydroepiandrosterone, 17 alpha-hydroxypregnenolone and delta 5-androstenediol increased. The principal manifestation of the effect of STS 557 is the apparent inhibition (87%) of the 3 beta-hydroxysteroid dehydrogenase-delta 5-4-isomerase activity in this experimental model. It is noteworthy, that the effects of STS 557 are similar to those effects previously published for other clinical used progestagens and the "pure antiandrogen" cyproterone. The results indicate, that STS 557 acts directly on the steroid biosynthesis in the human ovary in vitro.

During 3 consecutive calving seasons, calving performance, placental characteristics and endocrine profiles of total 98 pregnancies of late pregnant Swedish Red and White (SRB) and Swedish Holstein (SLB) dairy heifers and cows, were investigated. Ninety-four singleton pregnancies and 4 sets of twins were recorded. In animals with singleton pregnancy, 8 stillbirths, 7 weak calves, 3 premature parturitions and 1 abortion were registered. In the SLB heifers, 19% of stillbirth (5/26) were observed, while 5% (2/42) were noted for the SRB heifers. One stillborn calf derived from the SRB cows and none was found from the SLB cows. In the heifers and cows delivering a normal living calf with unassisted parturition, the placentome thickness monitored by ultrasonography was constant towards the end of pregnancy. The numbers of foetal cotyledons varied individually between animals but in total, fewer cotyledons were found in the foetal membranes of the SRB animals than in the SLB animals (69 +/- 19) vs. (88 +/- 29) (p < 0.05). No morphological and numerical differences of the placentome thickness in animals delivering a stillborn or weak calf, compared to animals delivering a normal living calf, could be observed. In animals with unassisted parturition and without birth complications, the levels of progesterone (P4), PGF2alpha metabolite (PG-metabolite), cortisol, oestrone sulphate (E1SO4) and pregnancy associated glycoproteins (PAGs) were not different by breeds and parities. In animals carrying stillbirth, higher levels of E1SO4 were found in 3 SRB animals and 1 SLB heifer, whereas lower levels of E1SO4 were recorded in 3 SLB heifers during the last week of pregnancy, compared to the profiles found in animals with unassisted parturition. Additionally, the levels of PAGs remained low and constant in 1 SRB cow (delivering a stillborn calf), 1 SRB heifer (giving birth prematurely), 4 animals (carrying twins) and 1 aborting SRB cow. Our results show a very high rate of stillbirth in especially SLB heifers and deviating profiles of E1SO4 and PAGs in animals with impaired parturition were recorded.

Estrus synchronization is important for optimal management of gilt reproduction in pig farms. Hormonal treatments, such as synthetic progestogens, are used on a routine basis, but there is a growing demand for non-hormonal alternative breeding tools. Before puberty, gilts exhibit a 'waiting period,' related to the ovarian development and gonadotrophin secretions, during which external stimulations, such as boar exposure, could induce and synchronize first ovulation. Practical non-invasive tools for identification of this period in farms are lacking. During this period, urinary oestrone levels are high, but urine sampling is difficult in group-housed females. The aim of this work was to search for specific biomarkers of the 'waiting period' in saliva and urine. In total, nine 144- to 147-day-old Large White gilts were subjected to trans-abdominal ultrasonography three times a week for 5 weeks until puberty detection (week -5 to week -1 before puberty). Urine and saliva samples were collected for oestrone assay to detect the 'waiting period' and for metabolome analysis using 1H-nuclear magnetic resonance spectroscopy to detect potential biomarkers of the 'waiting period.' Gilts were slaughtered 7 days after puberty detection for puberty confirmation. Results were consistent with ultrasonography data for six gilts. Urine and saliva samples from these six gilts were analyzed. Urinary estrone concentration significantly increased 2 weeks before puberty detection. Metabolome analysis of urine samples allowed the identification of 78 spectral bins, among them, 42 low-molecular-weight metabolites were identified. Metabolome analysis of salivary samples allowed the identification of 59 spectral bins, among them, 23 low-molecular-weight metabolites were detected and 17 were identified. No potential biomarker was identified in urinary samples. In saliva, butyrate and 2HOvalerate, 5.79 ppm (putatively uridine), formate, malonate and propionate could be biomarker candidates to ascertain the pre-puberty period in gilt reproduction. These results confirm that non-invasive salivary samples could allow the identification of the physiological status of the gilts and presumably the optimal time for application of the boar effect. This could contribute to synchronize puberty onset and hence to develop non-hormonal breeding tools.

Hyperprolactinaemic patients occasionally demonstrate hirsutism and elevated levels of DHA-S, a weak androgen of adrenal origin. Abnormal adrenal function is frequently observed in hirsute patients. These observations prompted speculation that prolactin may modulate normal adrenal secretion and that derangements of adrenal androgen secretion may be due to abnormalities in prolactin. In this study we examined the possibility that elevated prolactin levels may be involved in the pathogenesis of hyperandrogenaemia in hirsute patients. However, basal prolactin levels in hirsute women, with or without menstrual disturbances, 201 +/- 24.3 mU/l (mean +/- SE) and 192 +/- 24.3 mU/l respectively, were significantly suppressed below levels in normal women, 289 +/- 12.2 mU/l. The prolactin responses to stimulation with TRH and to suppression with L-dopa were also studied in hirsute patients. The prolactin response to TRH (maximum increment or integrated response) was exaggerated significantly in hirsute women with menstrual disturbances when compared to normal women, to hirsute women with normal menses or to normal men. This abnormal response may have been due to elevated oestrone levels present in patients with oligomenorrhoea (318 +/- 49.5 pmol/l compared to 191 +/- 12.1 pmol/l in normal women and 161 +/- 15.5 pmol/l in hirsute women with normal menses, P less than 0.05). There were no abnormalities detected in the suppression of prolactin in response to L-dopa in any of these groups. These findings do not support a role for prolactin in the pathogenesis of hyperandrogenaemia in hirsute patients. However, elevated androgen levels in women may bring about suppression of basal prolactin levels to values seen in normal men. (ABSTRACT TRUNCATED AT 250 WORDS)

An immunoquantification protocol based on an enzyme-linked immunosorbent assay was developed to measure the abundance of the microsomal enzyme steroid sulphatase (STS). The two-step sandwich immunoassay is sufficiently sensitive to detect 100-200 pg purified steroid sulphatase in a 50-microliters sample. The steroid sulphatase content in fibroblast, leukocyte and placental extracts correlates with the steroid sulphatase activity in these extracts. No steroid sulphatase protein was found in approximately 350 micrograms plasma proteins from a normal person. In three of four X-linked ichthyosis patients a complete gene deletion was found by Southern hybridization with the full-length STS cDNA as probe. Neither steroid sulphatase protein nor enzymatic activity was found in fibroblast extracts of these three patients. In a fibroblast extract of another X-linked ichthyosis patient, which had a normal Southern blotting pattern, no immunoreactive protein was detected. Residual activity of steroid sulphatase was also not found after prolonged incubation of this fibroblast extract with the natural substrate oestrone sulphate.

Many investigations of the regulation of prolactin synthesis and release are based on single plasma prolactin determinations. The purpose of the present experiment was to ascertain whether groups of rats (i.e. young or adult, male or female animals, being either intact, gonadectomized or gonadectomized and treated with oestrone), differing in age and/or endocrine status, will react to a single dose of perphenazine by an acute release of pituitary prolactin in proportion to their initial plasma prolactin levels. No consistent relation existed between the classification of the twelve groups of rats into three categories of basal plasma prolactin levels (i.e. less than 20, 25-50, greater than 125 ng/ml) and their response to perphenazine. Even though all groups showed a highly significant increase of plasma prolactin levels the magnitude of the maximum prolactin response at 30 min varied greatly within the groups of one category and thus was not related to the initial prolactin levels. The effect of 14 days of oestrone treatment in increasing plasma prolactin levels in gonadectomized animals was greatest in young and adult male rats, less in young females and not significant in adult females. The results obtained after perphenazine treatment in the latter group made it clear that the effect of oestrogen treatment on prolactin release can be completely blocked by increasing synthesis and/or release of the prolactin-release inhibiting factor (PIF). Since perphenazine induces decrease of pituitary prolactin and a concomitant increase of plasma prolactin levels through lowered PIF-action, the positive effect of oestrogens on prolactin release (as observed in gonadectomized male and young female rats) apparently is caused by a different mode of action. The implications of these findings for the regulation of prolactin release, as affected by the endocrine status of the rat, is discussed. Moreover, comparison of prolactin lost from the pituitary and gained in the circulation of the experimental animals, with amounts of prolactin that were observed to disappear from plasma during the experiment, provided suggestive evidence that the capacity to synthesize and/or eliminate prolactin, after a sudden provoked release of the hormone, differed among the groups. The rates of synthesis by the pituitary, of release from the pituitary into the circulation as well as of elimination of the hormone from the circulation (equally involved in determing actual plasma levels) are thought, therefore, to be far more important for the elucidation of prolactin regulation than single plasma prolactin determinations.

A bioavailability study was performed on ten oophorectomized women in a randomized cross-over design. The absorption of tablets containing 2 mg of micronized oestradiol and 1 mg of micronized oestriol (Estrofem) was compared to the absorption of the same micronized hormones administered in an aqueous suspension. Serum concentration values of oestradiol, oestriol and oestrone were measured by radioimmunoassay. The data obtained were analyzed both by univariate and multivariate analyses, and neither the serum concentrations at the various sample times, the maximum concentrations, the times for the maximum concentrations, nor the areas under the serum concentration curves disclosed any significant differences between the tablet and suspension administrations at the 5% level. The serum concentration values achieved by giving 2 mg of oestradiol and 1 mg of oestriol were of the same magnitude or higher than those of the normal menstrual cycle.