Search

Endocrine profiles were investigated in wild and captive mongoose lemurs (Eulemur mongoz; Lemuridae) by analysing faecal progestagens and oestrogens. Oestrous cycle characterization was not possible, as most females appeared to conceive during the first oestrus of the breeding season. Conception was preceded by a pseudo-oestrus with no discernible luteal phase. Pseudo-oestrus and oestrus identification was possible by investigating the oestrogen:progestagen ratio. Pregnancy was reliably determined approximately 47 days after conception, when progestagen and oestrogen excretion increased above breeding season concentrations. Gestation was further characterized by high progestagen concentrations and a decline in oestrogen excretion 70-80 days after conception. Post partum, progestagens declined, but oestrogen excretion increased to exceed breeding season concentrations. In the wild group, a nulliparous daughter conceived while still a member of her natal group and aborted after 70-80 days of pregnancy at which time progestagens had declined but oestrogens remained high. Comparisons with other strepsirhine primates suggest that pseudo-oestrus followed by conception at first oestrus may be prevalent in lemurs. Gestational progesterone profiles vary between species, but a delayed increase in oestrogens during pregnancy could be common to all strepsirhines, although oestrogen levels during the final trimester of pregnancy differ between lemurs and lorises. Methodological investigations showed that prolonged storage of faeces in ethanol is viable and that the presence of undigested vegetable matter in the faecal pellets had no effect on the interpretation of hormone profiles.

Techniques for manipulating the oestrous cycle of sable antelope, Hippotragus niger, were evaluated in a captive population of 24 females maintained at the Smithsonian Institution's Conservation and Research Center in Front Royal, VA, USA. A secondary objective was to demonstrate the effectiveness of fecal steroid monitoring techniques as a non-invasive method of tracking experimental manipulations. Controlled Internal Drug Releasing (CIDR) devices designed for cattle (type B, reduced in length by 5 cm to fit the sable antelope's smaller reproductive tract) were more effective than CIDR devices designed for goats (type G) at delivering progesterone into circulation, and maintained serum progesterone at levels up to 86.1+/-7.8% of normal luteal concentrations in females whose spontaneous ovarian activity had been inhibited with melengestrol acetate. Serum progesterone and fecal progestagen measurements were highly correlated (P<0.05). Synchronization treatments of prostaglandin (PG) F2alpha alone and in combination with modified CIDR-B devices (12-day insertion interval) were both effective in inducing synchronized ovulation, however the PGF2alpha/modified CIDR-B treatment resulted in more precise synchrony and a shorter latency to ovulation than did PGF2alpha alone. In a separate experiment to characterize the temporal relationship between synchronization treatment, behavioral oestrus and ovulation, onset of behavioral oestrus occurred 34.1+/-5.7 h following PGF2alpha/modified CIDR-B treatment. Mean duration of the induced oestrus was 24.9+/-4.3 h. The first detectable rise in fecal progestagens occurred 5.1+/-1.0 and 4.1+/-1.0 days following PGF2alpha/modified CIDR-B treatment in groups of females housed with and without an adult male, respectively, indicating that the presence of a male did not accelerate the onset of the induced cycle.

The objectives of this study were to determine whether ovarian function in Nkone cows could be monitored by measuring progestin concentrations in faeces and to assess the effect of duration of storage at room temperature on faecal progestin concentrations. Faecal and blood samples were obtained once a day for 26 days from 21 Nkone cows whose oestrous cycles had been synchronized. Faecal samples from each cow were divided into five portions that were kept at room temperature for 0, 6, 12, 24 and 48 h, respectively, and then frozen. After centrifuging the blood to recover plasma and extracting steroids from the faeces, analysis of progesterone (P4) was carried out using solid-phase radioimmunoassay. The faecal progestin and plasma progesterone profiles corresponded well and were positively correlated (r = 0.70, p < 0.01). Faecal progestin concentrations decreased with increasing duration of storage at room temperature during both the follicular and luteal phases (p < 0.01). In both cases, the decline in faecal progestin concentrations followed an exponential pattern. The progestin concentrations in faeces after 48 h of storage at room temperature were higher (p < 0.05) during the peak luteal phase than in the follicular phase.

In many primates species, female sexual attractivity is influenced by behavioral cues as well as by nonbehavioral cues (i.e., visual-morphological or chemical signals). Both kinds of cues are usually related to the ovarian cycle and female hormonal state. Female tufted capuchins (Cebus apella) lack any external morphological change in relation to the ovarian cycle and evidence of scent-marking behavior has never been reported. In addition, tufted capuchin males do not routinely investigate the female's body or urine. Instead, capuchin females are extremely active in sexually soliciting the male(s) and their courtship toward them involves a rich behavioral repertoire. In the present study we defined female tufted capuchin proceptivity and investigated its relationship with female reproductive state. Ovarian hormones were measured in urine and fecal samples from four captive females in order to (a) assess their reliability for monitoring female ovarian function and (b) provide information on the timing of the component cycle phases and in particular the periovulatory phase. Measurements of urinary and fecal progestin metabolites provided the best indicator of ovarian cyclicity and for timing of the periovulatory phase. Through a multivariate analysis of the behavioral data set we distinguished four behaviors (eyebrow raising with vocalization, touching-and-running, nuzzling and head cocking) which showed a marked cyclicity (21.3 days) that matched that of urinary progesterone (21.9 days). Data showed that each period of proceptive behaviors was 2.7 +/- 0.8 days long and the day of a defined luteal phase rise in urinary progesterone levels was markedly shifted toward the end of this period. Furthermore, the ejaculations observed always occurred within proceptive periods. The results clearly indicate that female behavior is a good indicator of the periovulatory phase and can enhance female attractivity., (Copyright 1999 Academic Press.)

Fecal progestagen analysis in okapis (Okapia johnstoni) was used for diagnosis of pregnancy and reproductive disorders, including a comparison of urinary and fecal progestagen analysis and endocrine data on the postpartum period. Data were generated on reliability of fecal progestagen analysis in early pregnancy diagnosis, and case reports were compiled involving single animals with missing luteal activity, abortion after twin pregnancy, and abortions due to deficient placental progestagen production. There was approximately 100-200-fold higher progestagen concentration in feces than in urine, thus explaining the high reliability of fecal progestagen evaluations in diagnosing luteal function and pregnancy. The postpartum period was characterized by lactational anestrus of several months duration, and a postpartum estrous cycle about 2-3 wk after parturition was observed in two of eight animals. An animal with five abortions due to deficient placental progestagen production was treated with altrenogest in a subsequent pregnancy and carried the fetus to term.

Fecal progesterone metabolite monitoring techniques were validated for the sable antelope and used to characterize ovarian cycle dynamics and reproductive seasonality in a captive population at the National Zoological Park's Conservation and Research Center. Hormone was homogeneously distributed within fecal samples. Longitudinal fluctuations in fecal progesterone metabolites were consistent with typical luteal phase patterns and corresponded closely with changes in serum progesterone. The lag time from an im injection of progesterone to peak excretion in feces was 16 h. The pattern of births showed a slight peak in the summer (May-July), but year-round endocrine monitoring of six nonpregnant females showed no evidence of seasonality in ovarian activity. Females exhibited 11-14 estrous cycles per year, averaging 24.2 +/- 0.9 days in length. Luteal and interluteal phases were 18.4 +/- 0.9 and 5.8 +/- 0.4 days in length, respectively. Although only a small number of hippotragine antelope species have been studied, data indicate that they exhibit considerable interspecific variability in estrous cycle length and reproductive seasonality and thus may be a useful group for further investigation of factors regulating fertility., (Copyright 1998 Academic Press.)

Oestrogen and progestagen metabolites were measured in the faeces of five female giant anteaters (Myrmecophaga tridactyla), to characterise the oestrous cycle and pregnancy. Faecal samples were collected twice weekly for a minimum of 6 months, and immunoreactive progestagens and oestrogens were analysed using enzyme immunoassays (EIA). For progestagens, two antibodies that cross-reacted with 20alpha-hydroxy- or 20-oxo-progestagens were used. Both assays effectively monitored ovarian cyclicity; however, the concentrations obtained using the antibody for 20alpha-hydroxy-progestagens were higher, and the hormonal changes were more pronounced. Regular ovarian cycles were identified in three of the five females. Average (+/-SEM) length of the oestrous cycle (n=10) was 51.4+/-5.6 days. Peak concentrations of 20alpha-hydroxy-progestagens ranged from 80-660 ng/g of faeces and those of oestrogens from 20-100 ng/g. Hormone concentrations were measured during parts of two pregnancies and during four post-partum periods. The length of one gestation (from oestrous oestrogen peak until parturition) was 184 days. In the second half of gestation, progestagen concentration started to increase above luteal phase values; in the week before parturition it was approximately 20 times higher than those during the luteal phase. Concentrations of excreted oestrogens began to increase after two thirds of gestation and exceeded that of the follicular phase by approximately 2.5-fold in the week before parturition. Onset of ovarian cyclicity after parturition varied from 4-11 weeks. In conclusion, the measurement of faecal immunoreactive progestagens and oestrogens in the giant anteater indicated an ovarian cycle of approximately 7 weeks in length and provided potentially useful data for successful breeding management.

Ultrasonography and radioimmunoassay (RIA) of serum oestradiol-17beta, luteinizing hormone (LH) and progesterone, and faecal oestrogen and progestin was used to assess ovarian activity in the scimitar-horned oryx (Oryx dammah). Ovarian examination using ultrasonography revealed maximal follicle and corpus luteum (CL) diameters of 15 and 32 mm, respectively. Steroid hormone metabolite distribution among individual faecal pellets within the same defaecation was relatively homogeneous with coefficients of variation averaging 10.2+/-1.8% and 16.2+/-4.6% for oestrogens and progestins, respectively. Elevated faecal oestrogen concentrations were associated with large (> 10 mm) antral follicles detected by ultrasonography. Periovulatory peaks in faecal oestrogen excretion, coincident with nadirs in progestin excretion, were detected in three females. Faecal progestin excretion exhibited a similar temporal pattern to serum progesterone concentrations, with a time lag of approximately 16 h. Faecal progestin concentrations corresponded with the presence of functional CL and proved useful for monitoring luteal function, spontaneous and prostaglandin-F2alpha analogue-induced luteolysis and anovulation. In summary, faecal steroid monitoring is a practical, noninvasive method for characterising ovarian steroid excretion and has potential for facilitating the application of assisted reproductive technologies in scimitar-horned oryx.

In this study we report on the content and bioactivity of plant (phyto) estrogens and progestins in various foods, herbs, and spices, before and after human consumption. Over 150 herbs traditionally used by herbalists for treating a variety of health problems were extracted and tested for their relative capacity to compete with estradiol and progesterone binding to intracellular receptors for progesterone (PR) and estradiol (ER) in intact human breast cancer cells. The six highest ER-binding herbs that are commonly consumed were soy, licorice, red clover, thyme, tumeric, hops, and verbena. The six highest PR-binding herbs and spices commonly consumed were oregano, verbena, tumeric, thyme, red clover and damiana. Some of the herbs and spices found to contain high phytoestrogens and phytoprogestins were further tested for bioactivity based on their ability to regulate cell growth rate in ER (+) and ER (-) breast cancer cell lines and to induce or inhibit the synthesis of alkaline phosphatase, an end product of progesterone action, in PR (+) cells. In general, we found that ER-binding herbal extracts were agonists, much like estradiol, whereas PR-binding extracts, were neutral or antagonists. The bioavailability of phytoestrogens and phytoprogestins in vivo were studied by quantitating the ER-binding and PR-binding capacity of saliva following consumption of soy milk, exogenous progesterone, medroxyprogesterone acetate, or wild mexican yam products containing diosgenin. Soy milk caused a dramatic increase in saliva ER-binding components without a concomitant rise in estradiol. Consumption of PR-binding herbs increased the progestin activity of saliva, but there were marked differences in bioactivity. In summary, we have demonstrated that many of the commonly consumed foods, herbs, and spices contain phytoestrogens and phytoprogestins that act as agonists and antagonists in vivo.

Recent research has demonstrated the potential of pregnancy diagnosis in elk (Cervus elaphus nelsoni) using immunoassays of fecal steroid concentration. However, multiple samples are required to insure accurate results, limiting its utility for free-ranging animals. We attempted to develop an accurate one-sample pregnancy diagnosis using 153 fecal samples that were collected from free-ranging, radio-collared, adult female elk in Yellowstone National Park (Wyoming, USA) and from captive elk maintained at the Starkey Research Facility (La Grande, Oregon, USA) February through April 1992 and 1997. The pregnancy status of each animal was diagnosed using serum pregnancy-specific protein B (PSPB) assays providing fecal samples from 38 nonpregnant and 115 pregnant animals. Fecal radioimmunoassay (RIA) indicated that mean (+/- SD) progestagens (P4) were elevated significantly in pregnant (2.96 +/- 1.49 micrograms/gm) compared to nonpregnant (0.43 +/- 0.26 microgram/gm) individuals. Confidence intervals (1.96 +/- SE) for the two groups were widely separated (nonpregnant 0.34-0.51, pregnant 2.69-3.24) with little overlap in the range of concentrations measured for each group (nonpregnant 0.09-0.98, pregnant 0.90-8.29). These results indicate that fecal progestagens RIA provides a reliable method of noninvasive pregnancy diagnosis using single fecal samples collected from elk during late gestation. However, independent validation of the suggested discrimination criteria should be performed before routine application.

A rapid and economical method for the determination in meat of androgens, estrogens, progestogens and corticoids, including some precursors and metabolites, has been developed. The extracted steroids are separated in a polar, a neutral, and a phenolic fraction by C8-SPE followed by a liquid-liquid extraction of the phenolates. Each fraction is separately purified by normal-phase SPE. The different steroid fractions can be analysed either together to obtain a comprehensive hormone pattern in one step or separately to enhance detection selectivity and sensitivity. Using a universally applicable silylation of the hydroxyl and keto groups, detection limits of 0.02-0.1 microg/kg are reached by GC-MS (EI) in the selected ion monitoring mode.

From 12 bitches of various breeds with fertile oestrus cycles faecal samples were collected daily from the onset of pro-oestrus till 20 days after the start of vulval bleeding, then once per week till about 1 week before term. Immunoreactive progesterone metabolites were extracted from the samples using methanol and measured using immunoassays. In a first experiment four different assays were compared in regard to the amounts of immunoreactive substances measured: the enzyme immunoassay against 20-oxo-3-hydroxypregnanes showed twice to four times higher values of immunoreactive material than another using an antibody against 6 beta-hydroxyprogesterone. An enzyme immunoassay for pregnanediol measured only low concentrations of immunoreactive material. Also a radio immunoassay using an antibody against 11 beta-hydroxyprogesterone detected only small amounts of reacting material. High performance liquid chromatography showed that in faeces of bitches the immunoreactive progesterone metabolites were present in unconjugated form, mainly as 3 alpha/beta hydroxylated progestagens with a 20-oxo group. In the second experiment the samples were measured with the assay system using the 20-oxo-3-hydroxypregnane antibody. A few days before mating the concentration of progesterone metabolites increased, reaching 5.77 mumol/kg faeces (median) at the day of mating. High levels (10.45 mumol/kg faeces) were measured till the end of the first month after mating. Thereafter, the concentrations decreased, reaching 2.68 mumol/kg (median) at the end of the second month.

Faecal samples were collected up to once daily from three female Nepalese red panda (Ailurus fulgens fulgens) prior to, during, and after the expected 1995 breeding season. Radioimmunoassay of faecal progestins and oestrogens showed hormone profiles that suggest that this species is a seasonally polyoestrous, induced ovulator. Examination of faecal extracts following high pressure liquid chromatography and using antisera of high specificity, demonstrated at least five faecal progestins but only one major oestrogen, probably oestradiol. Samples from males were collected up to once weekly during the breeding season in 1996, and androgens measured by radioimmunoassay. Faecal androgens fluctuated widely for all males.

Mammary secretions were collected from nine bitches of various breeds during the periparturient period. Initial mean progestogen concentrations were high and started to decline from 5 days before parturition, in a similar way to changes in plasma progesterone concentration. Values were basal by 3-4 days after parturition. Marked individual and day variations prevented this method from reliably indicating the time of impending parturition. Calcium concentrations in milk increased from 3-5 days after parturition, whereas a decline in magnesium concentration was noted from day 1 postpartum. There was an increase in the Ca:Mg ratio from 1 day after parturition, although this was not significant until day 3, and was therefore not useful for predicting impending parturition. There were no significant changes in sodium concentration in milk.

Measurements of reproductive hormones (progestins and estrogens) excreted in feces were used to discriminate between 25 conceptive and 76 nonconceptive (including undetected early abortion) cycles of free-ranging yellow baboons at Mikumi National Park, Tanzania. Conceptive cycles had significantly higher luteal-phase progestin and estrogen concentrations than nonconceptive cycles as early as Day 4 postovulation. However, mean early luteal-phase progestin concentrations in conceptive cycles were lower when conception occurred during ecologically optimal vs. suboptimal times, and among females of high compared to low dominance rank. Mean estrogen concentrations in conceptive cycles showed the opposite dominance rank pattern: mean luteal-phase estrogen concentrations were higher in conceptive cycles of high-compared to low-ranking females. None of these relations existed for nonconceptive cycles. These data suggest that successful implantation is facilitated by relatively high early luteal-phase progestin and estrogen concentrations. However, long-term environmental cues predicting the probability of offspring survival appear to influence the amount of progesterone required for successful implantation; progesterone concentrations necessary to facilitate successful implantation are higher during suboptimal seasons or among females of low dominance rank-cues that also suggest that offspring survival conditions are relatively poor. This may act as a reproductive filter, restricting conception to females whose immediate condition (e.g., low social stress and good physical health) enables them to compensate physiologically and behaviorally for effects associated with these relatively harsh offspring survival conditions.

The content of 10 naturally occurring steroidal sex hormones (androgens and progestogens), their biosynthetic precursors and metabolites was determined in 62 samples of beef (bulls, steers, heifers). After enzymatic hydrolysis of their conjugates, the steroids were extracted from the tissue by liquid-liquid extraction and purified by solid phase extraction. The identification and quantification was carried out by GC-MS of the trimethylsilyl ethers. The progestogens progesterone and pregnenolone were quantitatively dominant (43.7 and 6.5 micrograms/kg respectively). The highest steroid concentrations were determined in female cattle.

New generation-oral contraceptives containing desogestrel or gestodene, and possibly also norgestimate, are more or less similar with respect to contraceptive efficacy, cycle control and acceptability. They also show a more favourable metabolic profile in comparison with older preparations. The desogestrel-containing preparations Gracial and Marvelon, and possibly also the gestodene-containing preparation Gynera, have demonstrated a good efficacy in well-controlled studies in the treatment of mild to moderate acne and/or hirsutism. There may be differences between new generation oral contraceptives with respect to their effects on metabolic variables like high-density lipoprotein cholesterol and sex hormone-binding globulin. These differences are most probably modulated by variations in both the pharmacokinetics and selectivity of the progestogenic components. Of particular relevance here may be the recent finding that approximately 20% of administered norgestimate is metabolized into levonorgestrel. For use in clinical practice, it is of considerable help to have different preparations containing a range of oestrogen doses with the same progestogen. They allow the clinician to 'tailor make' the choice of oral contraceptives for those starting pill use or those switching to another combination due to symptomatology or changed circumstances, e.g. advancing age, smoking, etc. In this respect, desogestrel-containing oral contraceptives allow the most flexible approach.

Our objective was to determine the effect of progestin-only contraceptives on metabolic parameters, bleeding patterns, and weight changes during the first year of use. Seventy-one women (> 95% Caucasian), who were advised regarding contraception alternatives, self-selected levonorgestrel implants (n = 44), depo-medroxyprogesterone acetate (n = 22), or oral norethindrone (n = 5). One year later, 11 levonorgestrel implant and five depomedroxyprogesterone acetate patients were randomly selected to compare (pre- and post-progestin use) levels of cholesterol, triglycerides, low density lipoprotein (LDL), high density lipoprotein (HDL), very low density lipoprotein (VLDL), apolipoproteins A-1 and B-100, bilirubin, and sex hormone binding globulin. Monthly bleeding and spotting records were kept in each group. Body weights were also monitored in each group. No statistically significant differences in metabolic parameters were found between pre- and post-progestin use in the levonorgestrel implant and depo-medroxyprogesterone acetate groups. Continued bleeding patterns were more prominent in the levonorgestrel implant and oral norethindrone groups than in patients receiving depo-medroxyprogesterone acetate. No significant weight gain was detected in any group. No changes in metabolic parameters or weight were noted over the one year of use of levonorgestrel implants or depo-medroxyprogesterone acetate. Depo-medroxyprogesterone acetate had the highest incidence of amenorrhea.

The objectives of the study were to (a) develop a simple fecal progestin extraction and radioimmunoassay method to measure immunoreactive progestin in porcine feces and (b) to characterize fecal progestin profiles during the estrous cycle and postpartum. A simple extraction method was developed in trial 1 and the mean (+/- SD) progestin recovery of the method was 84.3 +/- 3.5%. Progesterone levels measured at five different spiked concentrations (50, 100, 200, 400, and 500 ng/0.5 g feces) showed no systematic error. The sensitivity of the assay was 0.16 nmol/L of the extract. Trial 2 involved collecting fecal samples from six cycling sows every second or third day, beginning on the day of estrus (day 0) and continuing until day 22. The mean (+/- SD) fecal progestin concentrations of these sows determined by the above assay during days 0-5, days 6-10, days 11-15, and days 16-21 were 87.1 +/- 17.5, 262.6 +/- 102.1, 1188.2 +/- 454.1, and 897.3 +/- 274.1 x 10(-3) nmol/g feces, respectively. In trial 3, fecal samples from six postpartum sows were collected at weekly intervals beginning from day 7 after farrowing until day 50. The mean (+/- SD) fecal progestin concentrations were 111.0 +/- 61.1, 74.1 +/- 21.3, 66.5 +/- 26.1, 122.7 +/- 58.8 and 533.5 +/- 244.2 x 10(-3) nmol/g feces, during days 7-10, days 11-20, days 21-30, days 31-40, and days 41-50 postpartum, respectively. The results indicate that simple fecal progestin extraction and assay are feasible alternatives to the standard blood progesterone assays for monitoring reproductive function in swine.

Women who have used combined oral contraceptives (COC) have a reduced risk of endometrial cancer relative to that of women who have never used oral contraceptives, but it is unclear whether the size of the reduction is influenced by the progestin content of the preparation. We analyzed data from two population-based case-control studies of endometrial cancer to investigate this question. Among women aged 40 to 59 years who were residents of King or Pierce Counties, Washington (United States), incident cases who were diagnosed during 1975-77 or 1985-87 were identified. Personal interviews were conducted with 316 such women and their responses compared with those of 501 controls who were selected by household surveys or random-digit dialing. A reduced risk of endometrial cancer associated with COC use was present only among users of five or more years' duration, and even then only in women who were not long-term users of unopposed postmenopausal estrogens. Among these women, the relative risk (RR) of endometrial cancer did not differ according to the progestin potency of the COC used: it was equally low for women who had used a COC with low progestin content (RR = 0.2, 95 percent confidence interval [CI] = 0.1-0.8) as for women who had used a COC with high progestin content (RR = 0.3, CI = 0.1-0.9). Our results argue that, if the reduced risk of endometrial cancer in long-term users of COCs is due to the progestins contained in these preparations, that amount of progestin in most COCs exceeds the threshold amount needed to produce this beneficial effect.

Measurement of immunoreactive progesterone, pregnanediol and oestradiol in faeces collected throughout ovarian cycles in three species of callitrichid primates is reported. Faecal hormone concentrations were compared with plasma progesterone profiles during PGF2 alpha-controlled (n = 7) and natural (n = 8) cycles in Callithrix jacchus and Saguinus fuscicollis, respectively, and with urinary oestrone conjugates during five cycles in Saguinus oedipus. Unconjugated steroids, which predominated over enzyme hydrolysable conjugates in samples from all species, were used to generate cycle profiles. According to results from HPLC, oestrone and oestradiol accounted for virtually all oestrogen immunoreactivity, and oestradiol most often predominated, whereas large amounts of nonspecific immunoreactivity were detected by both progesterone and pregnanediol assays. Faecal progestins were excreted in a cyclic manner in all species; luteal phase values were on average five- to tenfold higher than corresponding follicular phase values. Significant increases in mean amounts of faecal progestins were seen within 48 h of the post-ovulatory rise in plasma progesterone. Although a similar trend was also seen for faecal oestradiol, a clear and consistent luteal phase increase was seen only in Callithrix jacchus and this generally occurred later than that of progestins. The results indicate that faecal progestin analysis provides a useful method for noninvasive reproductive assessment in callitrichid primates. In particular, measurement of immunoreactive pregnanediol enables a multispecies application of a single assay methodology for comparative studies on callitrichid reproductive function.

A study was conducted in captive baboons to determine (i) the impact of cereal dietary fibre on faecal progestogen excretion, and (ii) whether means of controlling dietary effects could be identified. Blood was collected on 3 days per week and faeces on 5 days per week from four unanesthetized cyclic female baboons, consecutively fed three diets of 5, 10 and 20% fibre for 90 days per diet. A 2 day lag time was detected before progesterone in the blood appeared in the faeces, regardless of diet (mean correlation was 0.62, P = 0.002). Increased dietary fibre had a negative effect on progestogen excretion (P < 0.004). Correspondence between blood and faecal progestogens was consistently greatest and the effect of dietary fibre least when faecal progestogens were expressed g-1 dry faeces. Several means of indexing faecal steroid excretion rates were examined including dehydroepiandrosterone (DHEA) and a number of byproducts of cholesterol metabolism. The cholesterol metabolite, cholestanone, was positively correlated with dietary fibre (r = 0.27; P < 0.04). Multiplying faecal progestogen concentration by the cholestanone g-1 dry faeces concentration increased the correlation between serum and cholestanone-indexed faecal progestogens (r = 0.78, P = 0.0001) compared with nonindexed progestogens (r = 0.71, P = 0.0001). We conclude that expressing faecal progestogens g-1 dry faeces may be sufficient and the most cost-effective method for controlling for most dietary effects when the objective is monitoring longitudinal endocrine status in baboons. However, it may be appropriate to express faecal progestogens by cholestanone concentrations when increased precision is needed to overcome the effects of profound variations in dietary fibre.

The interrelations of steroid hormone levels in plasma and amniotic fluid from mothers and their undisturbed fetuses at early midgestation of human pregnancy have not been defined previously. We, therefore, studied 12 healthy mothers and their fetuses undergoing termination of pregnancy for social reasons at 16-20 weeks gestation. Fetal arterial and venous blood was obtained by direct vessel puncture through a fetoscope in the conscious sedated mothers immediately before termination of pregnancy. Simultaneously, maternal peripheral venous blood and amniotic fluid were collected. Aldosterone (Aldo), corticosterone (B), 11-deoxycorticosterone, progesterone (P), 17-hydroxyprogesterone (17OHP), 11-deoxycortisol, cortisol (F), and cortisone were simultaneously determined by specific RIA after extraction and chromatography. Positive fetal arterio-venous differences were found for F, B, and Aldo, whereas arteriovenous differences were negative for P and 17OHP. In amniotic fluid, six of the eight corticosteroids showed significantly lower levels during fetoscopy than during routine amniocentesis, as reported previously using the same analytical methods. The present study demonstrates that the undisturbed human fetus at 16-20 weeks gestation actively secretes the most important gluco- and mineralocorticoids, F, B, and Aldo, independent of the mother. P and 17OHP were shown to be primarily derived from placental production and supplied to the fetus as a source of F and Aldo biosynthesis. The fetoscopy procedure with premedication seemed to give rise to less stress to the fetus than routine amniocentesis without sedation. Fetoscopy is, therefore, an ideal method to study feto-maternal steroid interrelations in human pregnancy.

Desogestrel is the most selective progestogen used in oral contraceptives (OCs). The clinical characteristics of the monophasic combined OC containing 150 micrograms desogestrel and 30 micrograms EE per tablet (Marvelon) are in accordance with the strong progestogenic and minimal androgenic effects of desogestrel: a very high contraceptive efficacy is combined with minimal and, in the case of lipid metabolism, even potentially positive effects on metabolic parameters. Through increasing the plasma levels of sex hormone binding globulin, and thereby decreasing the plasma levels of free testosterone, the desogestrel-containing OC also has substantial beneficial effects on acne.

Faecal samples were collected at weekly intervals from pregnant Lipizzan mares during Weeks 7-16 following mating and from Lipizzan, Trotter and Thoroughbred mares during the last 3 months of gestation. After parturition, samples were taken daily from the Thoroughbred mares for another 6 days. Non-pregnant mares served as controls. The concentrations of unconjugated oestrogens (Eg), 20 alpha-OH-progestagens (20 alpha-G) and 20 beta-OH-progestagens (20 beta-G) were measured by enzyme immunoassay. In the faeces of Lipizzan mares, immunoreactive progestagens were significantly (P less than 0.01) elevated above the levels in non-pregnant mares by Week 11, and Eg by Week 13 of pregnancy onwards. During the last 3 months of gestation, concentrations of Eg were significantly higher in Trotter mares than in Lipizzan and Thoroughbred mares. Concentrations of 20 alpha-G and 20 beta-G increased to maximal values in the last month of gestation. There was no significant difference among the 3 breeds with respect to 20 alpha-G but, during the 10 weeks before parturition, concentrations of 20 beta-G in the Lipizzan mares were significantly lower (P less than 0.05) than those in the Thoroughbred mares. They were also significantly lower than those of the Trotter mares during the last 4 weeks of gestation. After parturition, the concentrations of Eg and progestagens had declined to baseline values by Days 3 and 4 respectively. From these results we conclude that high concentrations of progestagens with 20 alpha- and 20 beta-hydroxyl groups are present in the faeces of pregnant mares, especially during the last month of gestation.

Current views favor the notion that resident ovarian macrophages play an in situ role in the regulation of ovarian function through the local secretion of regulatory molecule(s) (i.e., cytokines). Herein we report on the potential ovarian relevance of one such macrophage product, tumor necrosis factor (TNF) alpha, a polypeptide capable of oncolytic as well as pleiotropic noncytotoxic biologic activities. Our findings suggest that the ability of TNF alpha to diminish the gonadotropin-supported accumulation of progesterone by granulosa/luteal cells is largely due to attenuation of key biosynthetic steps leading to progesterone production. These findings are in keeping with the notion that TNF alpha, possibly of intraovarian (e.g., macrophage or granulosa cell) origin, may comprise the centerpiece of a regulatory loop designed to attenuate gonadotropin hormonal action. Acting at or adjacent to its site of synthesis, TNF alpha may thus partake in the modulation of ovarian progestin economy, possibly in connection with the death of the corpus luteum.

The synthesis and use of stable isotope-labeled analogs of various steroids have made it possible to undertake a study of follicular fluid (FF) aspirated from mature and preovulatory follicles. Our previous results have been brought together here in order to review quantitative work done by gas chromatography-mass spectrometry. A positive gradient between peripheral plasma and FF concentrations of a steroid suggests the possibility of ovarian biosynthesis. This is particularly relevant to the catecholestrogens, 19-norsteroids, and some corticosteroids.

The present study investigates the levels of a large series of 18 non-conjugated or conjugated steroids in 71 samples of human breast cyst fluid (BCF) as divided into three groups corresponding to different electrolyte composition. In the type 1 group, the K+/Na+ ratio was higher than 1.5, while in type 2 it was lower than 0.66 and finally type 3 had an intermediate ratio. Pregnenolone (PREG) and progesterone (PROG) levels were approximately 2-fold higher (P less than 0.05) in the type 2 than in the type 1 group while both 17-OH-pregnenolone (17-OH-PREG) and 17-OH-progesterone (17-OH-PROG) concentrations were similar in these two groups. Most of the C-19 steroids analyzed, namely dehydroepiandrosterone sulfate (DHEAS), androst-5-ene-3 beta, 17 beta-diol (5-ene-DIOL), testosterone (TESTO), dihydrotestosterone (DHT), androstane-3 alpha, 17 beta-diol (3 alpha-DIOL), androsterone (ADT), androstane-3 alpha, 17 beta-diol glucuronide (3 alpha-DIOL-G) and androsterone glucuronide (ADT-G) were 180-360% (P less than 0.05) higher in type 1 than in type 2 cysts while no difference in C-18 steroid and C-18 steroid glucuronide levels was observed. A small or no difference was seen in steroid levels between types 2 and 3. We conclude that the arbitrary division according to the electrolyte composition of BCF permits identification of different patterns of steroid concentrations in BCF. However, the mechanism responsible for both sets of parameters remain unclear.

The production of song in passerine birds is under the control of steroid hormones, and brain regions involved in song production have been shown to contain androgen and/or estrogen receptors. Studies to date, however, have not considered the possible role of progestins in this behavior. As one approach to this question, the autoradiographic method was used to investigate the distribution of progestin-concentrating cells in the brain of the adult male zebra finch (Poephila guttata) after injection of the radiolabeled synthetic progestin [17 alpha-methyl-3H]-promegestone. In the telencephalon, identifiable groups of progestin-accumulating cells were found in the hyperstriatum dorsale, at the medial edge of the lobus parolfactorius, and in the medial septum. In the diencephalon, labeled groups of cells were found in the preoptic area, through much of the medial hypothalamus--including nucleus periventricularis magnocellularis, nucleus medialis hypothalami posterialis, and area infundibularis--and in the medial spiriform nucleus and dorsomedial thalamus. In the myelencephalon, labeled cells are described at the dorsal edge of the medulla and scattered lateral to nXII. These findings offer no support for the hypothesis that progestin acts on any of the known song regions, but do suggest areas of progestin action in the avian central nervous system outside of the known song system. Not surprisingly, these include many areas of the medial hypothalamus and other midline structures.

A method is presented based on capillary GLC using both a thermionic and a flame ionization detector to simultaneously analyse all major unconjugated steroids in ovarian follicular fluids (FF). Although specificity can not always be guaranteed for the smaller concentrations of androstenedione and cortisol, accuracy and reproducibility are excellent for the major progestagens and estrogens (progesterone, 17- and 16 alpha-hydroxyprogesterone, pregnenolone, 20 alpha-dihydroprogesterone, estradiol and estrone). Above all the analysis is performed with relatively cheap instrumentation and products. Apart from the "profiles" of unconjugated steroids, a semi-quantitative analysis of steroid conjugates is possible if a preliminary group separation with disposable anion exchanger columns is included.

The present report deals with the concentrations of C-21, C-19 and C-18 steroids as well as steroid glucuronides, namely androstane-3 alpha,17 beta-diol glucuronide, androsterone glucuronide, estradiol glucuronide, and estrone glucuronide (E1-G) in breast cyst fluid (BCF). The concentration of the gross breast cystic disease protein 15 (GCDFP-15) was also measured and its value was correlated with concentrations of unconjugated steroids as well as steroid conjugates. The present data indicate that for a large number of unconjugated steroids (namely pregnenolone, progesterone, dehydroepiandrosterone (DHEA), androsterone, androstane-3 alpha,17 beta-diol, estrone and estradiol) there is an important accumulation in BCF. Our data permit us to demonstrate a statistical relationship between the concentrations of DHEA and its metabolites (namely, androstenedione [4-ene-Dione]), thus suggesting an activity of the enzyme 3 beta-hydroxysteroid dehydrogenase-4-ene-5-ene-isomerase in the breast tissue. Further examination of the relationship between the concentrations of 4-ene-Dione and its metabolites, namely testosterone and 5 alpha-reduced steroid metabolites, as well as their glucuronide derivatives, strongly suggests that the metabolism of androgens in the breast tissue occurs mainly via 5 alpha-reductase and glucuronyl transferase activities. The concentration of GCDFP-15 in BCF found in the present investigation was 2745 +/- 234 micrograms/ml and we have shown that a negative relationship exists in the BCF between E1-G and GCDFP-15 levels.

Ovarian or uterine lymph was collected continuously for periods of up to 25 days from 16 cows cannulated at stages of pregnancy ranging from 96 to 278 days post coitum. Blood samples were taken acutely from the ovarian and uterine veins during surgery and periodically from the jugular vein during the course of lymph collection. The flow rate and cell content of lymph was measured and blood and lymph plasma samples were analysed for progesterone, pregnenolone, pregnenolone sulphate, androstenedione, testosterone, oestrone, oestrone sulphate, oestradiol-17 beta, prostaglandin (PG) F-2 alpha, total protein and albumin. There was a high flow rate of protein-rich lymph from luteal ovaries with rates up to 101.7 ml/h occurring in individual lymphatics over short periods. Peripheral ovarian and uterine lymph contained a low concentration of cells (mean less than 10(5) cells/ml) comprising about 82-87% lymphocytes, 11-14% macrophages and monocytes and 2-4% other cells. At all stages of pregnancy, the concentration of progestagens and androgens was higher in ovarian lymph than in uterine lymph or blood plasma. The differences were greatest for progesterone and androstenedione which occurred at 200-fold and 60-fold greater concentration respectively in ovarian lymph than in jugular plasma. When serial 10 min samples were collected over a 12-h period, the concentration and output of progesterone in ovarian lymph varied in a phasic manner, ranging from 3.5 to 7.6 microM and from 31.7 to 293.1 nmol/h respectively. There was a positive correlation between the output of progesterone in lymph and the progesterone concentration in jugular blood samples taken every 20 min. During most of pregnancy there was little difference between the concentration of oestrogens in ovarian lymph, ovarian venous plasma and jugular plasma but, during the 3-5 days before calving, these hormones occurred at slightly higher concentration in ovarian lymph. Apart from pregnenolone and androstenedione, all steroids occurred at lower concentrations in uterine lymph than in jugular plasma. Shortly before parturition there was an abrupt increase in the concentration of PGF-2 alpha in uterine lymph. Lymph reflects more accurately the milieu of tissue cells than efferent blood and further analysis of differences in the concentration of substances in lymph relative to the output in the ovarian and uterine arterial and venous blood may lead to the identification of factors important in local regulatory mechanisms in the reproductive tract.

In a 16-year-old girl with left upper abdominal pain of two months' duration fine-needle biopsy (guided by computed tomography) revealed a solid-cystic acinar-cell tumour of the pancreas. The tail of the pancreas was resected and the spleen removed. On inspection, the tumour--about 9 cm in diameter--was encapsulated and consisted of soft, brownish, partly haemorrhagic tissue. Histologically and immunohistochemically the diagnosis of an acinar-cell tumour was confirmed. Analysis of steroid hormone receptors revealed a high progestogen content. Since the operation the patient has remained symptom-free without any sign of tumour recurrence.

A routine procedure is described for the dependable detection of various anabolic residues in tissues or urine contaminated at levels as low as 0.5-10 ppb (10 parts per 10(9)). A suitable extraction and clean-up procedure was developed, permitting adequate recovery (60-80%) of various anabolics from tissue samples (50 g) or urine (50 ml). Following two-dimensional thin-layer chromatography, the presence of the anabolic residues are detected by sulphuric acid-induced fluorescence. The detection limit of most anabolics is of the order of 1-10 ng.

The influence of androgens on the FSH modulation of progestin biosynthetic enzymes was studied in vitro. Granulosa cells obtained from immature, hypophysectomized, estrogen-treated rats were cultured for 3 days in a serum-free medium containing FSH (20 ng/ml) with or without increasing concentrations (10(-9)-10(-6)M) or 17 beta-hydroxy-5 alpha-androstan-3-one (dihydrotestosterone; DHT), 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-diol), or the synthetic androgen 17 beta-hydroxy-17-methyl-4,9,11-estratrien-3-one (methyltrienolone; R1881). FSH treatment increased progesterone and 20 alpha-hydroxy-4-pregnen-3-one (20 alpha-OH-P) production by 10.2- and 11-fold, respectively. Concurrent androgen treatment augmented FSH-stimulated progesterone and 20 alpha-OH-P production in a dose-related manner (R1881 greater than 3 alpha-diol greater than DHT). In the presence of an inhibitor of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), the FSH-stimulated pregnenolone (3 beta-hydroxy-5-pregnen-20-one) production (a 20-fold increase) was further enhanced by co-treatment with R1881, 3 alpha-diol or DHT. Furthermore, FSH treatment increased 4.4-fold the activity of 3 beta-HSD, which converts pregnenolone to progesterone. This stimulatory action of FSH was further augmented by concurrent androgen treatment. In contrast, androgen treatment did not affect FSH-stimulated activity of a progesterone breakdown enzyme, 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD). These results demonstrate that the augmenting effect of androgens upon FSH-stimulated progesterone biosynthesis is not due to changes in the conversion of progesterone to 20 alpha-OH-P, but involves an enhancing action upon 3 beta-HSD/delta 5,delta 4-isomerase complexes and additional enzymes prior to pregnenolone biosynthesis.

Four readily identifiable changes in appearance of corpora lutea (Stages I to IV) occur during a bovine estrous cycle. Accuracy of estimating the stage of an estrous cycle by appearance of corpora lutea was determined in a double-blind study. One investigator observed estrus in a group of heifers while another with no prior knowledge of reproductive histories of the heifers estimated stage of the estrous cycle by visual inspection of their corpora lutea. Stage of the estrous cycle (Stage I, days 1 to 4; Stage II, days 5 to 10; Stage III, days 11 to 17; Stage IV, days 18 to 20) was estimated correctly in 41 of 48 heifers. The correlation between estimated and actual days of the estrous cycle was .81. In addition, concentrations of progestins and weights of corpora lutea during estimated stages of the estrous cycle were similar to many other investigations. Stages of the estrous cycle in heifers can be estimated from appearance of corpora lutea.

The topographic distribution of progestin concentrating cells in the female hamster brain and pituitary was studied by thaw-mount autoradiography. Fifteen minutes after injection of [3H]R5020, a synthetic progestin, nuclear uptake and concentration of radioactivity was found in certain cells of the forebrain and midbrain, as well as in the anterior pituitary. Competition studies with unlabeled R5020 abolished and with progesterone reduced the nuclear uptake of radioactivity. In the forebrain, radioactively labeled cells are observed in n. septi lateralis, n. interstitialis striae terminalis, n. preopticus medialis and lateralis, n. periventricularis hypothalami, organum subfornicale, n. arcuatus hypothalami, n. ventromedialis hypothalami, n. dorsomedialis hypothalami and n. premammillaris ventralis. In the midbrain a few labeled cells are found in the griseum centrale. In the anterior pituitary, labeled cells are identified by immunostaining as luteinizing hormone-producing cells. The results of the autoradiographic study demonstrate nuclear progestin binding sites and suggest genomic action of progestin on the labeled structures, probably related to the modulation of gonadotropin secretion and sexual behavior in the brain and pituitary.

The effect of the antifungal agent, ketoconazole, on rat ovarian concentration of progestins and androgens was investigated. A single oral dose of 20 mg/kg on the morning of proestrus resulted 3h later in a significant decrease. Control values at this time for progesterone, 17 alpha-hydroxyprogesterone, androstenedione and testosterone were 1173, 53, 40 and 2.5 ng/100 mg tissue respectively. In ketoconazole treated rats, the corresponding concentrations were 837, 34, 23 and 0.7 ng/100 mg tissue respectively. At a dose of 5 mg/kg ketoconazole, the concentrations of these steroids tended to be lower but the differences from control were not statistically significant.