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Biomedical subjects

E Möstl

Publications and source records attributed to E Möstl.

At least 37 records · Page 2Linked to original sources

Fecal glucocorticoids document stress in female Barbary macaques (Macaca sylvanus).

Patterns of received aggression and the endocrine response were related to an increase in fecal glucocorticoid metabolites in an intact semifree-ranging group of Barbary macaque females in order to quantify the social stress incurred over a 20-week observation period. The novel result showed that immunoreactive cortisol and 11-oxoetiocholanolone found in the feces can indeed determine the endocrine response of the adrenal gland after a social stressor. After HPLC separation of pooled fecal samples, EIA analyses using three different assays (corticosterone, cortisol, and 11-oxoetiocholanolone) to quantify immunoreactive steroids showed that the corticosterone EIA had no distinctive immunoreactive peaks. Cortisol and 11-oxoetiocholanolone immunoassays showed respectively four and two immunoreactive substances. Time series analyses revealed a behaviorally initiated increase in concentrations of cortisol and 11-oxoetiocholanolone equivalents. Furthermore, both hormone curves exhibit comparable time functions. Either antibody is very suitable for determining glucocorticoid secretion after periods of stress.

Animals↗

Seasonal patterns of sex steroids determined from feces in different social categories of Greylag geese (Anser anser).

Seasonal patterns of fecal 17beta-OH-androgen, estrogen, and progesterone equivalents of male and female greylag geese (Anser anser) were analyzed in a flock of free-living geese. These were compared among social categories determined by pairbond status and breeding success. The annual cycle was divided into 13 phases. Phasewise intra-sexual comparisons were made between social categories. The seasonal variation obtained from feces was in general agreement with the literature on plasma patterns in geese and other temperate-zone birds. However, there were distinct differences in seasonal hormone patterns among the social categories. In unpaired males, androgen was elevated for a longer period of time during sexually active phases compared with paired males. In male geese, high levels of androgen did not interfere with parenting but were related to pairbond status, whereas in females, androgen and progesterone were positively related to parental behavior. In the Fall, androgen, progesterone, and estrogen peaked only in unpaired males. In unsuccessful females, estrogen started to increase earlier in the Winter and was higher in amplitude and duration than that in females guarding offspring. In general, fecal steroids showed a clear-cut difference only between sexually active and parental phases of the year in the successfully breeding pairs, whereas unpaired males retained a hormonal state closer to sexually active phases throughout the year.

Androgens↗

Noninvasive fecal monitoring of glucocorticoids in spotted hyenas, Crocuta crocuta.

The aim of this study was to validate a method for measuring glucocorticoids noninvasively in feces of spotted hyenas (Crocuta crocuta). Three established enzyme immunoassays (EIA) for cortisol, corticosterone, and 11-oxoetiocholanolone were tested, but proved unsatisfactory. A new EIA using another corticosterone antibody was established and was used for all subsequent analyses; this EIA was validated by demonstrating parallelism between serial dilutions of spotted hyena fecal extracts and dilutions of standard corticosterone and by the recovery of corticosterone added to fecal extracts. High-performance liquid chromatography (HPLC) fractions analyzed by EIA showed various immunoreactive substances with polarities of unconjugated steroids. The physiological relevance of fecal glucocorticoid metabolites was further validated by demonstrating that (1) injection of exogenous ACTH to four males and two females led to a significant increase in fecal glucocorticoid metabolites within 24-50 h, (2) the translocation of a male spotted hyena to a new enclosure resulted in a fivefold increase compared to baseline concentrations, and (3) agonistic social interactions and physical conflict resulted in large increases of fecal glucocorticoid metabolites in both protagonists. Fecal steroid assessment is therefore of use in monitoring adrenal activity in spotted hyenas.

Adrenocorticotropic Hormone↗

Measurement of glucocorticoid metabolite concentrations in faeces of domestic livestock.

After 14C-labelled cortisol infusion in ponies and pigs, faecal samples were collected. Extraction of 0.5 g faeces with 5 ml 80-90% methanol yielded the highest radioactivity in the supernatant. Most of the metabolites were ether soluble. After high performance liquid chromatography (HPLC), the presence of immunoreactive metabolites was demonstrated by measuring each HPLC fraction using enzyme immunoassays for cortisol, corticosterone and 11-oxoaetiocholanolone. Only the assay for 11-oxoaetiocholanolone revealed peaks with co-eluting radioactivity. For biological validation of the test system, adrenocorticotrophic hormone (ACTH) and dexamethasone were injected intravenously successively in both species (n = 6). Cortisol concentration in blood and the 11-oxoaetiocholanolone immunoreactive substances in faeces were determined. In horse faeces, basal values of 2.3-35.2 nmol/kg were measured. After ACTH administration, an increase (more than 200% above basal values) of these metabolites was seen about 1 day after ACTH administration. After dexamethasone injection the levels decreased, reaching minimum concentrations 2 days after administration. In pigs, an increase in these metabolites was measured in only three animals after ACTH; dexamethasone did not cause a decrease. The stability of the samples after defecation was tested by storing samples from cows, horses and pigs at room temperature. It was shown that there was a significant increase in the concentration of measured cortisol metabolites in bovine, equine and porcine faeces after storage for 1 h, 4 h and 24 h, respectively. In frozen samples this effect was diminished after thawing samples at 40 degrees C; thawing the samples at 95 degrees C prevented an increase in immunoreactive substances.

Androstanes↗

Perineal swellings: a social and endocrine advantage for Barbary macaque females (Macaca sylvanus).

This study focuses the relationship between social rank, intersexual behavior, age, fecal cortisol equivalent excretion rates and perineal swelling size in contraceptive-treated Macaca sylvanus females. Behavioral data and fecal samples were collected during a twenty-week period from 24 females. Females were categorized with regard to perineal swelling (enlarged or reduced), and to their dominance rank (high or low); we found that swelling size was not associated with it. However, females with an enlarged perineum received more male grooming interactions, had closer spatial relations to males, and received significantly more interventions from males than did the others. Endocrinologically, females with enlarged swelling showed decreased fecal cortisol equivalent excretion rates. Multiple regression models showed a positive relationship between the extent of the swelling size and grooming, inspection and spatial relation. There was a negative relationship between the extent of swelling size and fecal cortisol equivalents. These results imply that enlarged perineal swellings among implanted Barbary macaque females have a greater impact on intersexual contact and adrenocortical activity than does social rank or age.

Animals↗

Uterine androgen receptor mRNA expression in metestrous and anestrous bitches being healthy or suffering from Pyometra.

The importance of androgens for the female reproductive system has been investigated for decades and a number of androgen sensitive processes has now been identified in female reproductive organs. For carnivore species no data were available so far about uterine androgen sensitivity and its regulation. The present study therefore aimed to investigate whether androgen receptors (AR) are present in the dog uterus, whether they are regulated throughout the ovarian cycle and whether pyometra affects their expression rate. Uterine tissue samples were collected from 28 bitches of different ages and various breeds. The samples were grouped according to the stage of estrous cycle (metestrus ME or anestrus AE) and the pathological status of the uterus (i.e. suffering from pyometra or not). Androgen receptor mRNA (AR mRNA) was quantified from 500 ng of total RNA isolated from the tissue samples using an internally standardized reverse transcription polymerase chain reaction (RT-PCR) described previously. The amount of total RNA extractable per g tissue was elevated during pyometra. The successful amplification of the expected 172 bp fragment from canine uterine RNA together with the confirmation of the identity of this fragment by sequence analysis, demonstrates that AR is expressed in this particular tissue. Comparing the expression rates in uteri from bitches during ME or AE being healthy (H) or suffering from pyometra (P), the only significant (p < 0.01) difference was found between H and P uteri during ME with 3.5-fold lower expression rates in P. Although the same seems true for AE bitches, a significant difference could not be demonstrated due to the low number (n = 2) of diseased animals in the AE group. There was no evident effect of the stage of ovarian cycle on uterine AR mRNA levels.

Anestrus↗

Comparison of different progestagen assays for measuring progesterone metabolites in faeces of the bitch.

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.

Animals↗

A competitive enzyme immunoassay for the pyrrolizidine alkaloids of the senecionine type.

We developed an immunoassay with antibodies against retrorsine to detect the closely related senecionine, the main alkaloid in various Asteraceae. Sensitivity is about 23 pg (68 fmol) for senecionine. Cross reactivity of monocrotalin, retrorsine N-oxide, senkirkine (< 0.1%), and seneciphylline (3.6-34.5%) was resolved. The alkaloid content in leaves of Petasites hybridus (Asteraceae) was determined to be 3.86 ppm, calculated as senecionine, which was far less than in rhizomes (104.8 ppm). Additionally, we proved specificity by taking immunograms from different parts of Petasites hybridus. The test is a suitable tool for assessing the toxic potential of medicinal plants containing senecionine.

Animals↗

The horse feto-placental unit.

In the pregnant mare two different groups of oestrogens are produced by the placenta. The precursor of "classical" oestrogens (oestrone, oestradiol-17 beta and oestradiol-17 alpha) is dehydroepiandrosterone which originates from the fetal gonads. The ring B unsaturated oestrogens (equilin and equilenin and their derivatives) derive from farnesyl pyrophosphate by a pathway not involving cholesterol.

Animals↗

Measurement of urinary oestrogens and 20 alpha-dihydroprogesterone during ovarian cycles of black (Diceros bicornis) and white (Ceratotherium simum) rhinoceroses.

The development of a sensitive enzyme-immunoassay for 20 alpha-dihydroprogesterone (20 alpha-DHP) and its use in determining reproductive status in black and white rhinoceroses is reported. 20 alpha-DHP in hydrolysed urine diluted in parallel to standards, and high-performance liquid chromatography (HPLC) confirmed the presence of 20 alpha-DHP and the absence of pregnanediol-3 alpha-glucuronide (PdG) in urine collected from rhinoceroses after oestrus. Conjugated oestrone was identified by HPLC as the major urinary oestrogen in the black rhineroceros and conjugated oestradiol-17 beta was the most abundant in the white rhinoceros. In African species, the black (Diceros bicornis), and northern (Ceratotherium simum cottoni) and southern (Ceratotherium simum simum) white rhinoceroses, excretion of 20 alpha-DHP and oestrogen followed a cyclic pattern. Excretion of 20 alpha-DHP was low before mating, at the time of peak oestrogen excretion, but high after oestrus. In the black rhinoceros, the follicular phase was 3-4 days and the luteal phase was 18 days, suggesting a cycle of 21-22 days. The interoestrus interval in the northern subspecies of white rhinoceros was 25 days, which correlated well with the interval between peaks of oestradiol-17 beta excretion. The interval between urinary oestrogen peaks in the southern subspecies of white rhinoceros suggested a cycle length of 32 days. This paper provides the first description of the pattern of excretion of urinary oestrogens and progesterone metabolites in African rhinoceroses.

20-alpha-Dihydroprogesterone↗

Concentrations of progestagens and oestrogens in the faeces of pregnant Lipizzan, trotter and thoroughbred mares.

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.

Animals↗

Conversion of steroids in bovine blood in vitro.

Blood samples collected from eight Braunvieh cows between the sixth and eighth month of gestation were allowed to stand with and without anticoagulant at 20 degrees C and 0 degrees C for different time periods. In these samples the degree of in vitro conversion of gestagens, androgens and estrogens was investigated. The concentrations were measured by radioimmunoassay. After 24 h at 20 degrees C, the levels of pregnenolone, progesterone, 17alpha-hydroxyprogesterone, androstenedione, dehydroepiandrosterone and estrone decreased to 62, 29, 25, 10, 34 and 44%, respectively, of the initial value and those of 17alpha, 20beta-dihydroxyprogesterone, epitestosterone and estradiol-17alpha increased to 385, 800 and 852%, respectively. The conversion was slower in clotted blood. The concentrations of testosterone and estradiol-17beta were consistent over the 24 h period. There was no marked decrease of the steroid concentration after 24 h of incubation of whole blood at 0 degrees C and of plasma at 20 degrees C. After the addition of (3)H-steroids, conversion could be demonstrated by thin-layer chromatography and autoradiography. These results demonstrate that all investigated hormones except testosterone and estradiol-17beta were metabolized by bovine blood cells.

Journal Article↗

Dehydroepiandrosterone and epitestosterone in the blood of cows at term.

From Day 268 of gestation till two days after parturition blood samples were collected from the jugular vein of 5 cows with normal and 4 cows with flumethasone induced parturition and centrifuged immediately after sampling. In another experiment blood samples were taken from the jugular and uterine vein of 3 cows during the last week before normal parturition. The dehydroepiandrosterone (DHA) and epitestosterone (ET) concentrations in plasma were measured using radioimmunoassay. In the control group the DHA concentration was in the range of 2 to 5 nmol/l, ET levels varied from 4 to 8 nmol/l before term. In both groups, there was an increase in DHA concentrations during the last periparturient period whereas ET levels increased only after flumethasone induced parturition. The concentrations of both androgens declined after the expulsion of the placenta and they were higher in the uterine vein than in the jugular vein. It is concluded, that both androgens are secreted by the bovine placenta and that the delta 5-pathway of steroidogenesis is active in vivo.

Animals↗

Low sensitivity to oestrogens in bovine placenta at term.

To elucidate the importance of elevated oestrogen levels at term for bovine placental maturation, the sensitivity of placental tissue to oestrogens was investigated by estimating oestrogen receptor levels. Unlabelled oestrogen receptors were bound to heparin sepharose in a small column and the gel was washed to remove coenzymes, cosubstrates and non-binding proteins. After subsequent incubation with 3H-oestradiol and removal of unbound steroids, a valid receptor quantitation was possible. In the maternal compartment of the mature bovine placenta only low oestrogen receptor levels (5-14 fmol/mg protein) were detectable, whereas the fetal compartment presumably contains no receptors (less than 1 fmol/mg). Due to the low oestrogen sensitivity of the bovine placenta we conclude that unconjugated oestrogens are of little or no importance for placental maturation. The final oestrogen peak prior to parturition may be a marker for maturation with a questionable physiological importance for the placentome.

Animals↗

[Catecholamine and cortisol concentrations in plasma from cattle at parturition].

Cows with spontaneous parturition (n = 8) and cows with flumethasone (n = 3) or prostaglandin (n = 3) induced parturition were used in this study. Catecholamine concentrations from cows in the periparturient period were measured in order to evaluate the stress of birth on the dam and the calf. The plasma content of both norepinephrine and epinephrine in late pregnancy was not different from control values in the 3rd to 6th month of pregnancy (norepinephrine 3.27 +/- 0.99 nmol/l plasma and epinephrine 0.98 +/- 0.45 nmol/l plasma), though both parameters appear elevated in the pregnant group. Norepinephrine concentrations rose to a value of 4.50 nmol/l immediately after birth, whereas epinephrine levels did not show an increase post partum. In two neonates norepinephrine and epinephrine values were 2-3 times higher than in the dams and decreased to the values of the dams within 48 hours. Synthetic glucocorticoids can mimic the increase of cortisol, which leads to birth. Cortisol concentrations in the cows remained unaffected by induction of parturition by prostaglandins. Newborn calves showed values of about 330 nmol/l, which also declined within 48 hours to values similar to those measured in the dam (9-28 nmol/l). As stress and exogenous glucocorticoids may affect antibody production, the IgG-content of the colostrum was examined. Prostaglandin induction of birth did not alter the IgG-content of colostrum, but flumethasone produced a decrease of about 14.5%. It is therefore concluded that pre-term birth induction does not produce negative effects on the supply of IgG.

Animals↗