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Steroid metabolism in racing greyhoundsBiddle, Simon January 2014 (has links)
The metabolism of androgenic anabolic steroids has been studied in the racing greyhound. Various drug preparations have been investigated utilising different derivatisation techniques, coupled with gas chromatographic analysis, to enable the identification of key metabolites in canine post administration samples. This has led to an increased understanding of some of the generic routes of steroid metabolism that take place in the greyhound. This valuable information can help to support metabolism studies in the future. The identification of specific metabolites for each compound investigated, has provided a means for controlling the misuse of these compounds, and contributed valuable enhancements to screening protocols utilised in the canine sports drug testing industry. Utilisation of the techniques described, resulted in the identification of specific major metabolites of the anabolic steroid methyltestosterone, namely 17??-methyl-5??- androstan-3??-17??-diol and 17??-methyl-5??-androstan-3??,16??,17??-triol. 16??- hydroxylation was shown to be a major phase I metabolic pathway in the canine along with phase II conjugation with glucuronic acid. Similar results were obtained during the metabolism study of the progestatgenic steroid norethisterone. Several di- and trihydroxy metabolites were detected in the glucuronic acid fraction of the post administration urines from this study. The norethisterone metabolism study also provided some insight, into the area of trace contaminants of pharmaceutical preparations. Low levels of nandrolone metabolites were also detected in the norethisterone post administration urine samples, leading to the discovery that the administered pharmaceutical tablets contained small quantities of nandrolone and 19- norandrostenedione, albeit below FDA approved contaminant levels. Modern methods of drug screening employ such highly sensitive techniques, that they allow for the detection of metabolites of such trace contaminants, following administration of the drug preparation to the greyhound. It is therefore important to have a broad understanding of the metabolism of various drug preparations, both banned and permitted substances alike; as detection of a trace amount of a banned substance metabolite, arising from the administration of a permitted medication, whose iii metabolite profile is unknown, and therefore potentially not detected, could present an interesting case. In conjunction with research into controlling the use of banned substances for the purposes of suppressing oestrus in the greyhound bitch, an investigation into normal/reference levels of endogenous hormones has been carried out. The endogenous steroid levels in a population of 212 greyhound bitches have been studied with a view to establishing a method for the detection of the exogenous administration of the endogenous anabolic steroid testosterone. The major urinary metabolites investigated were epiandrosterone, 5??-androstane-3??,17??-diol and 5??-androstane-3??,17??-diol. Statistical evaluations have been carried out to support the implementation of a suitable threshold for the key testosterone metabolites, namely 5??-androstane-3??,17??-diol and epiandrosterone. The detection of 5??-androstane-3??,17??-diol was found to be a very good indicator of the exogenous administration of testosterone to the greyhound bitch, when compared with the reference population data for this metabolite. However, further statistical/analytical data evaluation was deemed necessary before an absolute threshold could be implemented for this analyte, for the purposes of controlling the misuse of testosterone in the racing greyhound bitch. To support the understanding of endogenous steroid levels in the female greyhound, yet further, the endogenous reproductive steroid profiles were measured throughout the entire oestrus cycle of a cohort of 33 racing bitches. The results of the study clearly indicate a surge in androgen metabolites during the first 7-10 days of the oestrus cycle, in particular epiandrosterone and 5??-androstane-3??,17??-diol. This unique set of data has provided detailed information regarding the fluctuating concentrations of androgen and progesterone metabolites (following ovulation), at key stages of the canine oestrus cycle. The information obtained from this research can be used to support regulatory decisions regarding the misuse of testosterone in the racing greyhound bitch.
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Effect of the oestrous cycle, pregnancy and uterine region on the responsiveness of the isolated mouse uterus to prostaglandin F(2alpha) and the thromboxane mimetic U46619.Griffiths, A.L., Marshall, Kay M., Senior, J., Fleming, C., Woodward, D.F. 03 November 2009 (has links)
No / Previous studies in this laboratory have suggested that the isolated uterus from non-pregnant mice has a prostaglandin F and a thromboxane receptor population similar to that found in human myometrium. The aim of this study was to investigate any regional variation in myogenic activity ) and the and responsiveness to prostaglandin F(2alpha) (PGF(2alpha) thromboxane mimetic U46619 in the mouse uterus taken during different stages of the oestrous cycle and during pregnancy. Uterine samples from BKW mice were taken from different anatomical segments along the length of each uterine horn and set up for superfusion at 2 ml/min with Krebs solution (containing 1 microM indometacin) at 37 degrees C, and gassed with 95%O(2)/5%CO(2). Responses (area under the curve) are expressed as a percentage of the final contraction induced by hypotonic shock. Data are expressed as the means +/- s.e.m. of n=5-12 and were analysed using Student's paired t-test or two-way ANOVA with a Bonferroni post hoc test. Regional variation in myogenic activity was observed in all tissues studied except those taken during labour. These tissues displayed significantly greater myogenic activity than tissues taken at late gestation and at all stages of the oestrous cycle. Tissues from pregnant animals were generally more responsive to U46619 and PGF(2alpha) than tissues taken from non-pregnant animals. Tissues taken from the upper segment of the uterine horn were more responsive to both agonists during the oestrous cycle. The findings demonstrated that the hormonal milieu and site of excision are important for myogenic activity and responsiveness.
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The relationships between ovarian antral follicle dynamics, luteal function and endocrine variables in ewesBartlewski, Pawel Mieczyshaw 01 January 2001 (has links)
Transrectal ovarian ultrasonography and hormone measurements were used to study ovarian antral follicular dynamics and development of luteal structures during the middle portion of the breeding season in non-prolific cross-bred Western white-faced ewes and prolific Finn sheep. Studies were also done on ovarian activity in Western white-faced ewes during the transition to seasonal anoestrus and at the onset of the breeding season. Lastly, two experiments were carried out to examine ovulatory responses and subsequent luteal function in Western white-faced ewes treated with luteolysin (PgF 2á) and progestogen (medroxyprogesterone acetate-MAP) during the luteal phase of the oestrous cycle and after ovulation induction with gonadotrophin-releasing hormone (GnRH) in mid-anoestrus. The results of the present experiments showed that the growth of ovine antral follicles reaching ovulatory sizes of >=5 mm in diameter occurred in a wave-like pattern throughout the oestrous cycle in both breeds of sheep under study. There were typically 3 or 4 waves of follicle production throughout the 17-day interovulatory period. Ovarian follicular emergence, or beginning of growth from the pool of 3-mm follicles, appeared to be primarily controlled by changes in circulating concentrations of follicle-stimulating hormone (FSH). In cyclic ewes, the largest ovarian follicles acquired the ability to secrete oestradiol from the day of emergence and a peak of oestradiol secretion occurred about the time they reached their maximum diameter. The high ovulation rate in prolific Finn sheep appeared to be achieved mainly by the ovulation of follicles emerging in the last two waves of the interovulatory interval. Interestingly, prolific Finn ewes produced more but smaller corpora lutea (CL) and had lower serum concentrations of progesterone during the luteal phase of the oestrous cycle as compared to non-prolific Western white-faced ewes. During the transition into seasonal anoestrus in Western white-faced ewes, FSH secretion resembled that during the breeding season but the pattern of emergence of sequential follicular waves was dissociated from FSH and oestradiol secretion. Prior to the first ovulation of the breeding season, there was a distinct elevation in circulating concentrations of progesterone produced by luteinized unovulated follicles and/or interstitial tissue of unknown origin. This increase in serum levels of progesterone, heralding the resumption of ovulatory cycles, did not alter the rhythmic pattern FSH secretion or follicular wave emergence. Treatment of non-prolific Western white-faced ewes with PgF2á and MAP applied late in the oestrous cycle changed follicular dynamics and increased ovulation rate to resemble that in prolific Finn sheep. Effects of MAP on the recruitment and growth of ovulatory follicles in Western white-faced ewes did not have a clear gonadotrophic dependancy, suggesting a possible local regulation of ovarian activity by progestins in ewes. Following the induction of ovulation with GnRH in anoestrous Western white-faced ewes, an array of ovarian responses were detected with ultrasonography, including failure of ovulation of large antral follicles, normal (fall-lifespan) and short-lived CL post-ovulation, and luteinized cystic-like follicles. The normal luteinization of ovulated follicles appeared to be related to the amplitude of episodic elevations in daily serum FSH concentrations before induction of ovulation and characteristics of the preovulatory LH surge.
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