The absorption, distribution and excretion of anabolic agents

The metabolic fate of the anabolic agents, diethylstilbestrol, hexestrol, trenbolone acetate, zeranol and the endogenous steroids are discussed under the headings absorption, distribution and excretion. There is an optimum concentration of anabolic agent in the systemic circulation that results in a maximum increase in growth rate of farm animals. This optimum blood concentration should be maintained over a long period. However, there is rapid metabolism and excretion of anabolic agents with short half-lives in blood, and metabolic clearance rate equals entry rate. The rate of absorption of the agent, which is determined by formulation and site of administration, is most important and is best achieved by the use of slow release implants. The pattern of exponential absorption from compressed pellets of single anabolic agents is not ideal, and a more constant payout of drug, in particular estradiol-17 beta, is best achieved in combined preparations of agents or from silicone rubber implants impregnated with the agent. The high metabolic clearance rate and rapid excretion of anabolic substances influences the distribution of residues. Outside the site of administration, less than 1% of the administered dose is present in the animal. The lowest concentrations of residues are found in muscle and fat, higher concentrations are present in liver and kidney and the highest concentrations are in the bile, urine and feces.     

Excretion patterns of fecal progestagens, androgen and estrogens during pregnancy, parturition and postpartum in okapi (Okapia johnstoni)

The aim of the present study was to establish a simple method to monitor ovarian activity and non-invasively diagnose pregnancy in okapi (Okapia johnstoni). The feces of a female okapi were collected daily or every 3 days for 28 months. Steroids in lyophilized feces were extracted with 80% methanol, and the fecal levels of immunoreactive progestagens (progesterone and pregnanediol-glucuronide), androgen (testosterone), and estrogens (estradiol-17beta and estrone) were determined by enzyme immunoassays with commercially available antisera. Using the progesterone profiles, the durations of the luteal phase, follicular phase, and estrous cycle were determined to be 11.1 +/- 0.4, 5.3 +/- 0.6, and 16.5 +/- 0.7 days (n=22), respectively. Fecal levels of immunoreactive progesterone, pregnanediol glucuronide, and testosterone gradually increased from early pregnancy and peaked several months before parturition. More pregnanediol glucuronide was excreted in feces than progesterone during late pregnancy, but not during the estrous cycle. Although the fecal concentrations of immunoreactive estradiol-17beta and estrone change a little throughout pregnancy and non-pregnancy, they rose sharply and temporarily on the day following parturition. The present study indicates that fecal assays with commercial antisera for progesterone and pregnanediol glucuronide are useful for evaluating luteal activity and diagnosing pregnancy and indicates that estrogens might have some role as a trigger of parturition.     

Plasma concentrations of immunoreactive (ir)-inhibin, gonadotropins and steroid hormones during the ovulatory cycle of the duck

The present study was undertaken to determine changes in circulating levels of immunoreactive (ir)-inhibin, FSH, LH, estradiol-17beta, progesterone, and testosterone during the ovulatory cycle of Shao ducks. Serial blood samples were taken from two groups of laying ducks for measurement of ir-inhibin, gonadotropins, and steroid hormones at 2 h intervals for 24 h. Plasma concentrations of ir-inhibin did not change significantly during the ovulatory cycle. The highest level of plasma ir-inhibin was observed 6 h prior to ovulation, which coincided with a decreased level of plasma FSH. One FSH surge was found 12 h after ovulation. Estradiol-17beta, progesterone, and testosterone were also determined during the ovulatory cycle. Two peak values were detected for estradiol-17beta 8 h before ovulation and 4 h after ovulation, while progesterone started to increase 4 h before ovulation and reached a peak at ovulation. The highest level of plasma testosterone was detected around the time of ovulation. These results suggest that inhibin may be involved in the control of FSH secretion during the ovulatory cycle. In addition, both LH and progesterone are of importance in the ovulation process of Shao ducks.     

Characterization of urinary metabolites of testosterone, methyltestosterone, mibolerone and boldebone in greyhound dogs

Androgenic steroids are used in female greyhound dogs to prevent the onset of estrus; moreover, these steroids also have potent anabolic activity. As anabolic steroids increase muscle mass and aggression in animals, the excessive use of these agents in racing greyhounds gives an unfair performance advantage to treated dogs. The biotransformation of most anabolic steroids has not been determined in greyhound dogs. The objective of the present study was to identify the urinary metabolites of testosterone, methyltestosterone, mibolerone, and boldenone in greyhound dogs. These steroids were administered orally (1 mg/kg) to either male or female greyhound dogs and urine samples were collected pre-administration and at 2, 4, 8, 12, 24, 72, and 96 h post-administration. Urine extracts were analyzed by high-performance liquid chromatography/mass spectrometry (HPLC/MS) to identify major metabolites and to determine their urinary excretion profiles. Major urinary metabolites, primarily glucuronide, conjugated and free, were detected for the selected steroids. Sulfate conjugation did not appear to be a major pathway for steroid metabolism and excretion in the greyhound dog. Phase I biotransformation was also evaluated using greyhound dog liver microsomes from untreated dogs. The identification of several in vivo steroid metabolites generated in this study will be useful in detecting these steroids in urine samples submitted for drug screening.     

Age-related changes of reproductive hormones in young Meishan boars

Meishan pigs are known for their early sexual maturity. On the other hand, they grow slowly. There is no information currently available about the combination of these two characteristics in Meishan pigs. To study the developmental characteristics of Meishan pigs, the plasma concentrations of LH, FSH, inhibin, testosterone, estradiol-17beta, progesterone and insulin-like growth factor-I (IGF-I) in young Meishan boars were determined using RIA and ELISA. Inhibin decreased with age in weeks, while testosterone and estradiol-17 beta increased. Testosterone increased gradually, and an increase in estradiol-17beta occurred after sexual maturity. IGF-I increased before puberty and subsequently decreased just after puberty like a pubertal IGF-1 surge. FSH, LH and progesterone did not change with age. There was no significant correlation among the hormones. During the experimental period, the Meishan boars showed large individual differences. These differences may depend on the fact that Meishan boar reach maturity at 12 weeks of age and continue to grow thereafter.     

Drug metabolism: in vitro biotransformation of anabolic steroids in canines

Forensic drug testing of anabolic steroids in racing animals is required because of the potential for steroid abuse. Often when the metabolic products of an administered compound have not been identified, the analysis and verification of the administered compound is difficult. The objective of this study was to qualitatively identify the in vitro phase I biotransformation products of anabolic steroids that have a high potential for abuse in canines. The investigated steroids included testosterone, methyltestosterone, mibolerone and boldenone. Steroid biotransformation products were generated using beagle liver microsomes and analysed by high performance liquid chromatography (HPLC)/mass spectrometry (MS) with an electrospray ionization source. Characterization of steroid metabolites was based on HPLC retention, UV and mass spectra. The major testosterone metabolites were identified as androstenedione and 6beta- and 16alpha-hydroxytestosterone. 6beta-Hydroxymethyltestosterone was identified as a major metabolite in the methyltestosterone microsomal incubations. Several mibolerone metabolites were identified as monohydroxylated mibolerones as well as an oxidized mibolerone metabolite. Boldenone metabolites were identified as monohydroxylated boldenones, oxidized boldenone, and testosterone. This information should assist in the determination of anabolic steroid use in canines through the correlation of the urinary metabolites to the administered drug.     

Efficiency of fecal steroid hormone measurement for assessing reproductive function in the Hokkaido brown bear (Ursus arctos yesoensis)

The present study aimed to establish simple systems for measuring fecal steroid hormones in order to monitor the reproductive profiles of captive Hokkaido brown bears. The efficiency of fecal sample processing at the steps of dehydration and extraction and the correlation between steroid concentrations in matched fecal and blood samples were studied. Then, monthly changes in fecal estradiol-17 beta and progesterone in female bears, and testosterone in male bears were examined. The procedure was finalized as follows. Fecal samples were dried at 100 degrees C for 3 hr and extracted with diethyl ether. The diethyl ether in the extracts was evaporated and residues were reconstituted in ethanol for the assays. Hormone concentrations were quantified using enzyme immunoassays. Concentrations of progesterone and testosterone in fecal and plasma samples were correlated in the systems. The changes in fecal progesterone and testosterone concentrations were similar to those in serum concentrations of bears as reported previously. In contrast, fecal estradiol concentrations did not correlate with plasma levels probably because of the time lag in excretion. However, the changes in estradiol-17 beta concentrations in feces in the present study were similar to those reported in serum. In conclusion, fecal progesterone and testosterone assay systems appear practical for monitoring ovarian and testicular activities without immobilization, though methodological improvements and further validation may be required. For the fecal estradiol-17 beta assay, there is a need to solve the problem of excretion time lag before the system can be used in the study of reproductive physiology.     

Intramuscular collagen and serum hydroxyproline as related to implanted testosterone, dihydrotestosterone and estradiol-17 beta in growing wethers

Relationships of implanted testosterone, dihydrotestosterone and estradiol-17 beta to collagen degradation and intramuscular collagen concentration and stability were determined. Intramuscular collagen content, solubility and shrinkage temperature and serum hydroxyproline were analyzed in groups of six rams, wethers, and wethers implanted with various levels of testosterone or dihydrotestosterone and groups of 10 rams, wethers and wethers implanted with estradiol-17 beta, dihydrotestosterone or a combination of these two steroids. Intramuscular collagen content in both experiments was higher (P less than .05) in muscles of rams than in muscles of wethers. Administration of the highest level of testosterone to wethers raised (P less than .05) total and insoluble intramuscular collagen to concentrations noted in rams. Administration of the testosterone metabolite, dihydrotestosterone, to wethers had no effect on intramuscular collagen. Administration of estradiol-17 beta to wethers tended to raise concentrations of intramuscular collagen so that they were no longer lower (P less than .05) than those in rams. Collagen stability as measured by solubility and thermal shrinkage temperature did not differ among rams, wethers or implanted wethers (P greater than .05). Increases in collagen accretion due to hormone administration were observed to be the result of increases in the insoluble portion of the intramuscular collagen (P less than .05).     

In vitro metabolism of ovarian steroids by bovine whole blood and plasma

Experiments were conducted to evaluate the effects of time and temperature on the potential of bovine whole blood (WB) or plasma (PL) to metabolize the ovarian steroids progesterone, estradiol-17β and testosterone. During a radioimmunoassay study (Experiment 1), we observed a temperature and time-dependent reduction (P<0.001) of plasma progesterone concentrations in samples incubated as WB at 5, 15, 25, or 35C for up to 48 hr. Most notable was the observation that 27% of progesterone present in controls was lost when WB was incubated at 5C for 48 hr and a 17% reduction was observed when PL samples were incubated at 35C for 48 hr. Immunoreactive estradiol-17β concentrations (Experiment 2) in PL and WB incubates were not affected by time or temperature. However, immunoreactive testosterone concentrations increased more than 3-fold by 48 hr in WB incubates held at 35C. To examine the latter observation further, ³H-progesteone was incubated with WB at 35C, followed by extraction and thin-layer chromatography (Experiment 3). Results generally supported RIA findings and revealed the presence of significant 17α-hydroxylase, 17–20 lyase and aromatase activity. Heretofore this has not been considered to occur outside major steroid metabolizing organs.     

Oxytocin-induced secretion of prostaglandin F2alpha in postpartum beef cows: effects of progesterone and estradiol-17beta treatment

The purpose of the present study was to determine the effect of progesterone or progesterone + estradiol-17beta on oxytocin-induced prostaglandin F2alpha (PGF2alpha) secretion in postpartum beef cows. Thirty-four anestrous postpartum beef cows were ovariectomized (d 32 [Groups 1 to 3] or d 23 [Groups 4 to 6] postpartum [d 0 = parturition]) and allotted to six treatments (Group 1; negative control) to simulate short (Groups 2 through 5) or normal (Group 6) length estrous cycles. Steroid treatments for the respective groups were as follows: Group 1) no estradiol-17beta or progesterone treatment (n = 8; negative control); Group 2) progesterone (d 34 to 40; n = 6); Group 3) estradiol-17beta (d 32 to 33) and progesterone (d 34 to 40; n = 6); Group 4) progesterone (d 23 to 29), no estradiol-17beta (d 32 to 33), and progesterone (d 34 to 40; n = 5); Group 5) progesterone (d 23 to 29), estradiol-17beta (d 32 to 33), and progesterone (d 34 to 40; n = 5); and Group 6) progesterone (d 23 to 29), estradiol-17beta (d 32 to 33), and progesterone (d 34 to 50; n = 4; positive control). Oxytocin (100 IU) was injected (i.v.) at the end of each treatment to test the ability of the postpartum uterus to secrete PGF2alpha as measured by a stable metabolite of PGF2alpha, 15keto-13,14 dihydro-PGF2alpha (PGFM). Peak concentrations ofPGFM (P < 0.08) and total PGFM secreted (area under the curve; P < 0.05) were increased on d 6 following first (Group 2) or second (Group 4) exposure to progesterone and were similar to peak concentrations and total PGFM secreted 16 d following a simulated normal estrous cycle (Group 6). Administration of estradiol-17beta before first progesterone exposure (Group 3) did not reduce peak concentrations of PGFM or total PGFM secreted relative to the preceding groups. Peak concentrations of PGFM (P < 0.08) and total PGFM secreted (P < 0.05) were reduced following a second progesterone exposure, provided that cows were pretreated with estradiol-17beta (Group 5). In summary, oxytocin-induced release of PGFM was inhibited on d 6 following second exposure to progesterone only when cows were pretreated with estradiol-17beta. Therefore, estradiol-17beta and progesterone were both associated with the timing of PGF2, secretion in postpartum cows.     

Relationship between endogenous estradiol-17 beta and estrous behavior in heifers

Thirty-five Holstein heifers were used to examine the relationship between endogenous estradiol-17 beta and estrous traits. During a non-superovulation period (NSP), estrous cycles were synchronized and during the periovulatory stage blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. In addition, continuous observation for estrous behavior was performed for 98 h. A gonadotropin-induced superovulation period (SP) was begun 12 d after estrus was detected during NSP. Heifers were injected with FSH twice daily for 4 d and single injections of prostaglandin were given on d 14 and 15. Beginning at d 14, blood samples were collected every 6 h for 120 h for subsequent determination of estradiol-17 beta and progesterone. Continuous observation for estrous behavior was performed for 98 h. Peak estradiol-17 beta was greater during SP than during NSP (49.0 +/- 3.1 vs 12.9 +/- 3.0 pg/ml serum). Thirty-three and 31 of the 35 heifers were in estrus during NSP and SP, respectively; duration of estrus was 2.3 h longer during SP than during NSP. However, number of behavioral interactions during estrus did not differ between NSP and SP. In conclusion, estrous traits were similar, whereas peak estradiol-17 beta concentrations were markedly different between NSP and SP.     

Estrogen receptor in bovine skeletal muscle

In connection with investigations of the anabolic action of estrogens, we examined skeletal muscle of veal calves for estradiol receptors. The high speed supernatant of muscle homogenate was incubated with .5 nM 3H-estradiol and for the determination of nonspecific binding with .5 nM 3H-estradiol plus 13 nM estradiol at 0 C overnight. After treatment with charcoal two times, the supernatant was analyzed by agar gel electrophoresis. Specific binding was found in the typical position of cytosolic estradiol receptor. Ninety percent of 3H-estradiol binding was suppressed by estradiol-17 beta, zeranol, estrone or diethylstilbestrol, but was not affected by testosterone, dihydrotestosterone, trenbolone or progesterone. The specific binding activity varied between .3 and 2.0 fmol/mg protein and the dissociation constant of the receptor was Kd = 60 pM. After an enrichment up to 42 fmol/mg cytosolic protein using heparin sepharose, the receptor remained unchanged as determined by agar gel electrophoresis. Although uterine tissue generally contains 1,000 times more estradiol receptors, these results clearly demonstrate that skeletal muscle also contains estradiol receptors with identical properties. This indicates that one possible component of the anabolic action of estrogens may be the direct stimulation of the muscle via the estradiol receptor.     

Residue levels of endogenous estrogens in beef tissues

Endogenous estrogen concentrations in edible tissues of cows, bulls and steers were compared with those of steers with estrogen implants. Concentrations are expressed as pg estrogen/g of tissue, wet weight. In addition, depletion rates of estradiol-17 beta (E2 beta) estrone (E1) and estradiol-17 alpha (E2 alpha) from these tissues and blood plasma were determined. In muscle, the main estrogen was E2 beta, regardless of sexual status (nonpregnant cow, bull or steer); however, concentrations approached the lower limits of analytical sensitivity. In liver and kidney, E2 beta and E1 were equimolar (25 to 40 pg/g for each) in heifers and steers, whereas in kidney fat, concentrations of E1 exceeded those of E2 beta. Concentrations of E1 in fat were slightly higher in bulls than in cows or steers. The predominant estrogen in fat during pregnancy was E1, with concentrations 150 times greater than those of nonpregnant heifers or nonimplanted steers and 75 times the concentration found in steers with E2 beta implants. In kidney of pregnant cows, E1 rose 40-fold and in liver 10-fold over that of implanted steers. Concentrations of E2 alpha were low and depleted rapidly after withdrawal of an E2 beta implant. Tissue depletion studies of the three estrogens demonstrated that E1 disappeared from plasma, fat, liver and kidney more slowly than E2 beta or E2 alpha. Depletion of E2 beta from the tissue can be manipulated as shown by the faster rate of depletion in implanted steers than in nonimplanted steers. Because E1 is cleared from fat more slowly than E2, and E1 concentrations are higher, this estrogen-tissue combination should be used to monitor estrogen implants as anabolic agents.     

Pfaffia paniculata-induced changes in plasma estradiol-17beta, progesterone and testosterone levels in mice

The present study undertook chemical analysis of components of Pfaffia paniculata roots. In addition, an animal experiment was conducted in which mice had ad libitum access to water enriched with powdered P. paniculata root for 30 days. Changes in plasma concentrations of estradiol-17beta and progesterone in female mice and of testosterone in male mice were ascertained. The results revealed that P. paniculata roots contain two types of phytosteroids, beta-sitosterol and stigmasterol, in addition to other compounds such as pfaffic acid, allantoin, saponins, beta-sitosteryl-beta-D-glucoside, and stigmasteryl-beta-D-glucoside. Regarding changes in plasma concentrations of hormones, levels of the sex hormones estradiol-17beta, progesterone and testosterone were clearly higher for mice that drank P. paniculata root-enriched water than for mice that drank plain water. Powdered P. paniculata root is easily dissolved in feed or water, and as no adverse reactions were seen in mice within 30 days of oral intake, consumption of P. paniculata for long periods of time appears safe.     

The effect of intramuscular injections of boar pheromone 5alpha-androstenol on the hormonal regulation of the estrous cycle in hypoosmatic gilts

Until 1999 it was accepted that pheromones act exclusively by stimulating the dendritic receptors present in olfactory epithelium. Cycling gilts with an experimentally-disrupted neural olfactory pathway were used to test the hypothesis that boar pheromone 5alpha-androstenol may affect the secretion of hormones involved in the regulation of the estrous cycle by the humoral pathway. On day 12 of the estrous cycle the nasal cavity of gilts (n=15) was irrigated with zink sulfate solution. From day 16 to 20, the experimental group (n=10) was injected intramuscularly with 5alpha-androstenol (20 microg) twice a day. Blood samples were collected from the jugular vein at 4 h intervals on days 17-21 to estimate plasma concentration of LH, oxytocin, estradiol-17beta, testosterone and progesterone. The experimental group displayed a significantly lower mean concentration of LH than the control animals (P<0.0001). The decrease in concentration of LH was accompanied by the reduction of oxytocin (P<0.001), estradiol-17beta (P<0.001) and testosterone (P<0.01) secretion. These results demonstrated that 5alpha-androstenol influenced hormonal regulation by humoral pathway and might be considered to be the priming pheromone in gilts.     

Serum concentrations of IGF-I, estradiol-17beta, testosterone, and relative amounts of IGF binding proteins (IGFBP) in growing boars, barrows, and gilts

The purpose of this research was to determine whether serum concentrations of steroids, IGF-I, and relative amounts of serum IGF-binding proteins (IGFBP) differ in growing boars (n = 11), barrows (n = 11), and gilts (n = 12) from 70 to 140 d of age. Pigs of similar age and weight were housed in pens of three or four and given ad libitum access to a 17% CP corn-soy diet and water. Pigs were weighed and blood samples were collected every 14 d beginning at 70 d of age. Serum concentrations of IGF-I and steroids were determined by RIA and relative amounts of IGFBP were determined by ligand blot analysis. By 84 d of age and continuing through 140 d of age, mean serum concentrations of IGF-I were greater (P < .05) in boars than in barrows or gilts. Relative amounts of 46-kDa IGFBP3 and 28-kDa IGFBP-4 were similar (P > .05) among pigs at 70 d of age; however, boars and barrows had greater (P < .05) relative amounts of 24-kDa IGFBP-4 and 41-kDa IGFBP-3 than gilts. Relative amounts of IGFBP-2 were greater (P < .01) in barrows than in gilts or boars at 70 d of age. From 84 d of age through 140 d of age, relative amounts of both forms of IGFBP-3 and the 28-kDa IGFBP-4 were greater (P < .05) in boars than in gilts or barrows. Relative amounts of IGFBP2 were greater (P < .05) in barrows than in gilts or boars at 98 d of age, but by 140 d of age relative amounts were greater (P < .05) in boars and barrows than in gilts. Mean serum concentrations of estradiol-17beta were similar (P > .05) between gilts and boars at 70 d of age, but by 98 d of age, and continuing through 140 d of age, mean serum concentrations of estradiol-17beta were greater (P < .05) in boars than in gilts. Mean serum concentrations of testosterone in boars increased (P < .05) with increasing age and were greatest at 128 and 140 d of age. Serum concentrations of testosterone were negatively correlated (P < .01) with relative amounts of serum IGFBP-2 but positively correlated (P < .01) with serum concentrations of IGF-I and estradiol-17beta. Serum concentrations of estradiol-17beta were positively correlated (P < .01) with serum concentrations of IGF-I in boars. Changes in serum concentrations of IGF-I and relative amounts of IGFBP resulting from changes in serum concentrations of estradiol-17beta and testosterone may contribute to growth differences observed among sexes.     

Behavioral, ovarian and endocrine relationships in the pubertal bitch

Behavioral, ovarian and endocrine correlates were determined in seven bitches experiencing first estrus, and the results compared with previous data obtained on multiparous females. One or more atypical reproductive patterns were observed in certain bitches at the pubertal estrus. The ability of pubertal dogs to display normal reproductive relationships appeared to be related to age, because animals that exhibited normal sexual behavior and endocrine profiles tended to be older than females that produced aberrant patterns. For the latter group, atypical observations included lack of sexual receptivity and reduced or inconsistent patterns of circulating concentrations of estradiol-17 beta, luteinizing hormone (LH) or progesterone. Ovulation was confirmed in all four pubertal dogs subjected to laparoscopy, including two bitches that failed to demonstrate estrous behavior. In individual animals, it appeared that inadequate or sporadic elevations in titers of estradiol-17 beta during the preovulatory phase of the cycle were associated with reduced or abbreviated LH surge concentrations, and the latter were subsequently related to an absent or only minor preovulatory rise in serum progesterone. Reduced preovulatory estradiol-17 beta concentrations and a delayed increase in circulating progesterone were also associated with a lack of sexual receptivity. These results (1) suggest that hormonal insufficiencies or irregularities exist in certain bitches at the pubertal estrus and that such atypical activity appears more characteristic of the first than later adult estrous periods, and (2) are consistent with the theory that estrogen preconditioning and a decrease in the endogenous estrogen to progesterone ratio are important in the initiation of sexual receptivity in this species.     

Non-genomic actions of progesterone and estrogens in regulating reproductive events in domestic animals

It has been established that nuclear receptors mediate the action of estrogens and progestins in regulating gene expression in the hypothalamic-hypophyseal-gonadal axis of domestic animals during various reproductive states. Results of recent in vitro studies suggest that estradiol-17beta and progesterone can act non-genomically to affect signal transduction responses in target cells by binding to receptors in the plasma membrane. The genomic action of steroids is generally detectable in hours to days whereas non-genomic responses of cells occur in seconds to minutes. The nature of the plasma membrane receptors for estrogens and progesterone has been explored but has not been conclusively established for all cell types studied. In the ewe, estradiol-17beta or estradiol-bovine serum albumin conjugate has been shown by in vitro and in vivo approaches to act non-genomically to suppress luteinizing hormone secretion by gonadotropes and stimulate production of nitric oxide by uterine arterial endothelial cells. Progesterone has been shown to inhibit oxytocin (OT) binding to its receptor in isolated ovine endometrial plasma membranes. This non-genomic action of progesterone blocks OT activation of the phosphoinositide cascade and production of prostaglandin F(2alpha) by ovine and bovine endometrium. The acrosome reaction of caprine and porcine spermatozoa is activated by the non-genomic action of progesterone. Further research is required to define the biological significances of the non-genomic actions of estrogens and progestins.     

Pituitary-testicular responses of estradiol-17 beta-implanted bull calves to continuous versus pulsatile infusion of luteinizing hormone releasing hormone

The luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone response of bull calves implanted with estradiol-17 beta to continuous and pulsatile infusion of luteinizing hormone releasing hormone (LHRH) has been examined. Estradiol-17 beta reduced serum LH and FSH concentrations and suppressed testosterone secretion and testicular growth when compared with sham-implanted bulls. Pulsatile iv infusion of LHRH [500 ng every 2 h (6 micrograms/d)] for a 4-wk period to estradiol-17 beta-implanted bulls resulted in elevated mean serum LH and testosterone concentrations that were characterized by discrete secretory episodes. Mean serum FSH was also increased by LHRH pulse infusion, but LHRH-coupled secretory episodes were not apparent. Continuous infusion of LHRH (6 micrograms/d) did not increase the low serum gonadotropin levels observed in estradiol-17 beta-implanted calves. Testicular growth was normal in LHRH pulse-infused calves, but was markedly curtailed in continuously infused calves. These results suggest that estradiol-17 beta inhibits testicular development by blocking gonadotropin release at the level of the hypothalamus because pulsatile administration of LHRH can override the inhibitory effect by increasing LH and FSH secretion.     

The effect of intrauterine infusion of inflammation-provoking factors on proinflammatory cytokines and hormones in rat peripheral blood

The aim of the study was to determine, using the rat model, whether uterine infections cause an increase in cytokine concentrations in peripheral blood, and whether this increase is accompanied by changes in the pituitary-ovarian axis function. The levels of tumor necrosis factor-alpha, interleukin-1beta, luteinizing hormone, follicle-stimulating hormone, prolactin, progesterone, testosterone and estradiol-17beta in blood plasma as well as the weight of the uterus were determined after intrauterine infusion of lipopolysaccharide (15 microg), peptidoglycan (1 mg) and Escherichia coli (10(6) cfu) suspension on the day of metaestrus. On days 3, 7 and 10 after treatment the rats were sacrificed to collect the blood samples. Inflammation of uterus and vaginal discharge developed in all rats after treatment. The administration of lipopolysaccharide, peptidoglycan and Escherichia coli induced considerable changes in ovarian cyclic activity, mainly diestrus was observed. Application of all these factors resulted in an increase (P<0.05, P<0.01) of plasma levels of tumor necrosis factor-alpha, interleukin-1beta, mainly on day 3 and 7. In the rats receiving pathological factors, the plasma levels of luteinizing hormone, follicle-stimulating hormone, prolactin and estradiol-17beta decreased (P<0.05, P<0.01) whereas progesterone and testosterone increased (P<0.05). These results indicate that in rats, the developing inflammatory process of the uterus following lipopolysaccharide, peptidoglycan and Escherichia coli infusions is connected with an increase of tumor necrosis factor-alpha, interleukin-1beta concentrations in peripheral blood, and is accompanied by changes in the pituitary-ovarian axis function.