Copyright (c) 2012 All rights reserved. http://works.bepress.com/hugo_eiler Recent documents in Hugo Eiler DVM, MS, PhD en-us Sat, 24 Nov 2012 11:21:11 PST 3600 http://works.bepress.com/hugo_eiler/16 http://works.bepress.com/hugo_eiler/16 Thu, 26 Jan 2012 10:44:37 PST Objective - To characterize the physiologic response to IV bolus injection of glucose and insulin for development of a combined glucose-insulin test (CGIT) in horses. Animals - 6 healthy mares and 1 mare each with pituitary adenoma and urolithiasis. Procedure - Horses were given a CGIT (glucose, 150 mg/kg; insulin, 0.1 U/kg); results were compared with a singular IV glucose tolerance test (GTT; 150 mg/kg) and a singular IV insulin sensitivity test (IST; 0.1 U/kg). Healthy horses were also given a CGIT after receiving xylazine and undergoing stress. Results - Physiologically, the CGIT resulted in a 2-phase curve with positive (hyperglycemic) and negative (hypoglycemic) portions; the positive phase came first (250% of baseline at 1 minute). The descending segment declined linearly to baseline by approximately 30 minutes and to a nadir at 58% of baseline by 75 minutes. After a 35-minute valley, a linear ascent to baseline began. Addition of insulin in the CGIT increased glucose utilization by approximately 4.5 times during the positive phase but not during the negative phase. The diseases' effects and experimental inhibition of insulin secretion with xylazine and stress were detectable by use of the 2 phases of the CGIT. Only a single positive phase resulted from the GTT and a single negative phase from the IST. Conclusions and Clinical Relevance - The CGIT resulted in a consistent, well-defined glycemia profile, which can be disrupted experimentally or by a disease process. The CGIT has clinical potential because it provides integrated information and more information than either the singular GTT or IST.

]]> Hugo Eiler et al. Selected Articles http://works.bepress.com/hugo_eiler/15 http://works.bepress.com/hugo_eiler/15 Thu, 26 Jan 2012 10:44:34 PST Hugo Eiler et al. Contributions to Books http://works.bepress.com/hugo_eiler/14 http://works.bepress.com/hugo_eiler/14 Thu, 26 Jan 2012 10:44:33 PST Hugo Eiler Contributions to Books http://works.bepress.com/hugo_eiler/13 http://works.bepress.com/hugo_eiler/13 Thu, 26 Jan 2012 10:44:31 PST Hugo Eiler Contributions to Books http://works.bepress.com/hugo_eiler/12 http://works.bepress.com/hugo_eiler/12 Thu, 26 Jan 2012 10:44:28 PST Antonio Horvath et al. Books http://works.bepress.com/hugo_eiler/11 http://works.bepress.com/hugo_eiler/11 Thu, 26 Jan 2012 10:44:27 PST To investigate the in vitro effect of the combination of lignan enterolactone (ENL) or lignan enterodiol (END) with melatonin on steroid hormone secretion and cellular aromatase content in human adrenal carcinoma cells. SAMPLE: Human adrenocortical carcinoma cells. PROCEDURES: Melatonin plus ENL or END was added to cell culture medium along with cAMP (100μM); control cells received cAMP alone. Medium and cell lysates were collected after 24 and 48 hours of cultivation. Samples of medium were analyzed for progesterone, 17-hydroxyprogesterone, androstenedione, aldosterone, estradiol, and cortisol concentration by use of radioimmunoassays. Cell lysates were used for western blot analysis of aromatase content. RESULTS: The addition of ENL or END with melatonin to cAMP-stimulated cells (treated cells) resulted in significant decreases in estradiol, androstenedione, and cortisol concentrations at 24 and 48 hours, compared with concentrations in cells stimulated with cAMP alone (cAMP control cells). The addition of these compounds to cAMP-stimulated cells also resulted in higher progesterone and 17-hydroxyprogesterone concentrations than in cAMP control cells; aldosterone concentration was not affected by treatments. Compared with the content in cAMP control cells, aromatase content in treated cells was significantly lower. CONCLUSIONS AND CLINICAL RELEVANCE: The combination of lignan and melatonin affected steroid hormone secretion by acting directly on adrenal tumor cells. Results supported the concept that this combination may yield similar effects on steroid hormone secretion by the adrenal glands in dogs with typical and atypical hyperadrenocorticism.

]]> Kellie Fecteau et al. Selected Articles http://works.bepress.com/hugo_eiler/10 http://works.bepress.com/hugo_eiler/10 Thu, 26 Jan 2012 10:44:25 PST OBJECTIVE: To determine whether tetanus antitoxin, equine serum, and acetylcysteine, which are currently used in the treatment of equine corneal ulcer, inhibit the digestion of equine corneal collagen when exposed to collagenase in vitro. ANIMALS STUDIED: Corneas from 40 adult horses. PROCEDURES: Sections of equine corneas were incubated with saline, a solution of bacterial collagenase in saline, bacterial collagenase in saline plus equine tetanus antitoxin, bacterial collagenase in saline plus equine serum, or bacterial collagenase in saline plus acetylcysteine. Each one of the collagenase inhibitors was tested at different concentrations. The degree of corneal collagen digestion was determined by concentrations of hydroxyproline released into the incubation media and/or by weight loss of the cornea. RESULTS: Corneas exposed to collagenase released a significant (0.05 level) large amount of hydroxyproline (43.1 +/- 2.3 microg/mL/100 mg cornea/5 h) and decreased cornea weight by up to 89%. Blood serum (200 microL/mL), purified albumin or globulin fractions of serum, tetanus antitoxin (120 units/mL), and acetylcysteine (20 mg/mL) when used at the highest concentrations blocked collagenase digestive activity by approximately 50%. Dilution of inhibitors decreased corneal protection and linearly increased corneal weight loss. Purified equine serum albumin and globulin fractions were equally effective in protecting corneas. CONCLUSIONS: This experiment indicates that tetanus antitoxin, serum and acetylcysteine equally protected corneas from collagenase digestion, in vitro. However, a clinical trial is needed to establish relative therapeutic value.

]]> Kellie Fecteau et al. Selected Articles http://works.bepress.com/hugo_eiler/9 http://works.bepress.com/hugo_eiler/9 Thu, 26 Jan 2012 10:44:22 PST The purpose of this study was twofold: (1) to determine whether there is a profile of 5-hydroxytryptamine (5-HT, serotonin) concentrations in fetal bovine blood and tissues during pregnancy, parturition and the early neonatal period and (2) to determine whether 5-HT has a 'mitogenic' effect on cultured placentome cells in bovine. Results revealed a 5-HT concentration profile in fetal blood. Overall concentrations of 5-HT in fetal blood were 6.6 times (P< 0.001) that of adult cows and 2.8 times (P< 0.001) that of blood collected during caesarean section (from umbilical veins) and from 18-72 h old calves. Mid-term and full-term pregnancy fetuses were not statistically different from each other. Overall concentrations of 5-HT in the intestinal wall of the fetus were 4.4 times higher (P< 0.05) than in the 24 h calf. Concentration of 5-HT in full-term muscle was 3.5 times higher than in mid-term muscle and 2.8 times higher than in 24 h old calf muscle (P< 0.05). Concentrations of 5-HT in mid-term and full-term cotyledon were 4.4 times higher (P< 0.05) than in post partum cotyledon. Characterized trophoblast cells and a heterogeneous population of bovine cotyledon cells treated with 5-HT (2.5, 5.0 and 10.0 microm) incorporated between 2.0 and 3.0 times more(3)[H]-thymidine than untreated controls, indicating a dose-dependent (r=0.94) positive mitogenic effect of 5-HT. Both groups of cultured cells responded equally. Five-HT treatment did not affect either cell number or cell size. It was concluded that a 5-HT concentration profile exists in fetal bovine blood and tissues and that 5-HT has the ability to act as a mitogen in bovine placental cells.

]]> Kellie Fecteau et al. Selected Articles http://works.bepress.com/hugo_eiler/8 http://works.bepress.com/hugo_eiler/8 Thu, 26 Jan 2012 10:44:20 PST The purpose of this study was to determine to what degree bacterial collagenase may digest human placentae compared to equine and bovine placentae. Placenta samples from human, equine and bovine were incubated with bacterial collagenase solution at various concentrations. The degree of hydrolysis and collagen breakdown was measured by the release of total proteins and hydroxyproline into the incubation media. Also, whole placentae were injected via umbilical cord arteries with collagenase solution (200 U/ml, 200 ml total volume in human and 1000 ml in equine) and hydrolysis determined chemically and subjectively. Human and equine placental collagens were the most sensitive to collagenase digestion. Overall mean collagenase activity determined by the release of hydroxyproline from human placenta was 1.6 times and in equine placenta three times greater than in bovine placenta, while the breakdown of non-collagenous proteins remained negligible. When injected into whole placenta, the collagenase digested placentae evenly within 6-12 h. At 24 h, placentae were liquefied, although, umbilical blood vessels resisted collagenase digestion. Bacterial collagenase was highly effective in breaking down human placenta collagen. Intraplacental injections of collagenase via umbilical cord arteries may help to detach retained placenta in women as it does in mares and cows.

]]> Kellie Fecteau et al. Selected Articles http://works.bepress.com/hugo_eiler/7 http://works.bepress.com/hugo_eiler/7 Thu, 26 Jan 2012 10:44:17 PST In the cow, cesarean section delivery is often followed by retention of fetal membranes. Hypothetically, the retention of fetal membranes could be prevented by intraplacental injections of the enzyme collagenase. However, the infusion of this potent proteolytic enzyme into a uterus traumatized by surgery can lead to uterine damage, including perforation. Thus, the objective of this research was to evaluate tolerance of intraplacental treatment of bacterial collagenase. A cesarean section was performed on 10 experimental cows undergoing induced delivery or diagnosed with dystocia. During the surgical procedure, 200,000 units of bacterial collagenase in 1 L of saline were infused via the umbilical arteries. A cesarean section was also performed on control cows (n = 25) affected by dystocia, but these received no collagenase. The collagenase-treated cows showed no clinical or laboratory signs of abnormality over a 3- to 4-wk observation period post treatment. When membrane retention time was set at 36 h post surgery, 20% of the experimental cows and 60% of the control cows had retained the fetal membranes. It was concluded that intraplacental administration of collagenase during cesarean section is safe. However, treatment effectiveness and economic benefits for commercial application need further study.

]]> Hugo Eiler et al. Selected Articles http://works.bepress.com/hugo_eiler/6 http://works.bepress.com/hugo_eiler/6 Thu, 26 Jan 2012 10:44:15 PST A significant percentage of cows (11%) fail to release the placenta within 12 h postpartum. Failure of collagen breakdown seems to be related to the retention of placentas. Sections of placentomes incubated with bacterial collagenase caused an increase in placentome proteolysis (6.6-fold) and placentome collagenolysis (94-fold) within 4 h in a dose-related fashion (r = 0.94). Injections of collagenase (825 U/cc) into the placentomes, via umbilical vessels, decreased the cotyledon-caruncle binding force (determined by manometry) to 30 +/- 5 mm Hg from 97 +/- 2 mm Hg, and increased proteolysis by 42% within 8 h (r = -0.95). Hyaluronidase at various concentrations (400-8 250 U/cc) and at various incubation times (up to 8 h) was not effective. Hyaluronidase (825 U/cc) and collagenase (825 U/cc) were not synergistic in loosening cotyledon-caruncle attachment. A single 15-min collagenase pulse, given prior to perfusion with collagenase-free blood, was as effective in loosening cotyledon attachment as was a sustained 2-h perfusion of blood with collagenase added. It was concluded that collagenase caused collagenolysis and loosening of cotyledon from caruncle, but collagenolysis and cotyledon-caruncle separation were not facilitated by the presence of hyaluronidase.

]]> Hugo Eiler et al. Selected Articles http://works.bepress.com/hugo_eiler/5 http://works.bepress.com/hugo_eiler/5 Thu, 26 Jan 2012 10:44:12 PST Bovine placentome collagen was quantified (P<0.01) at four gestational stages (90, 150, 210 and 270 d, n = 8 d ), at 2 h post partum without (n = 4) and at 2 and 12 h post partum with (n = 8) experimentally-induced placental retention. Placentome sections were fixed and stained for collagen. Fetal cotyledonary (FC) collagen volume fraction (V(V)) increased over days of gestation studied (V(V)=0.03+/-0.01, 0.06+/-0.01, 0.13+/-0.01 and 0.19+/-0.01). Fetal cotyledonary hydroxyproline (3.15+/-0.41, 4.55+/-0.41 and 7.04+/-0.41 mg/g) and FC protein (432.0+/-17.1, 479.9+/-17.1, 585.4+/-17.1 mg/g) increased over Days 90, 150 and 210 and were similar on Days 210 and 270. Fetal cotyledonary collagen V(V) and hydroxyproline did not differ between Day 270, retained and nonretained cotyledons. Protein concentration was higher in 2 h (578.1+/-18.5 mg/g) and 12 h (526.0+/-18.5 mg/g) retained versus nonretained (400.4+/-36.2 mg/g) cotyledons. Maternal caruncular (MC) collagen V(V) and protein concentration were higher on Days 90 and 150 than on Days 210 and 270. Maternal caruncular hydroxyproline was similar from Day 90 to 210 and increased from Day 210 to 270. Maternal caruncular collagen V(V), hydroxyproline and protein concentrations were similar on Day 270 and in 2 h and 12 h retained membrane caruncles. Gestational increases in placentome collagen occurred from FC sources. No difference in FC or MC collagen V(V) existed between Day 270, retained and nonretained placentomes.

]]> K L. Sharpe et al. Selected Articles http://works.bepress.com/hugo_eiler/4 http://works.bepress.com/hugo_eiler/4 Thu, 26 Jan 2012 10:44:10 PST Artificial vagina (AV) and electroejaculation (EE) are the 2 methods used to obtain semen from bulls. The purpose in the present study was to evaluate these 2 methods of collection when 2 markers, cortisol and progesterone, were injected IV. During period 1 (control measurement), semen was obtained by EE at 0, 20, 60, 120, and 180 minutes. In period 2, bulls were injected (3 days later) with a mixture of cortisol (113 mg) and progesterone (100 mg), and then samples were obtained by EE. In period 3, cortisol and progesterone were injected (3 weeks later), and samples were obtained by AV. Seminal plasma concentrations of cortisol and progesterone were maximal at 20 minutes in EE and AV collections. Seminal plasma concentrations of progesterone and cortisol were roughly 50% less in EE than in AV collection. However, the total excretion of progesterone and cortisol per collection was similar in both techniques. Excretion of cortisol was 14 to 33 times greater than that of progesterone. It was concluded that concentrations of markers in the EE were significantly less than those in AV collection. For this reason, total excretion and concentration of marker in semen should be accounted for when conducting excretion studies.

]]> Hugo Eiler et al. Selected Articles http://works.bepress.com/hugo_eiler/3 http://works.bepress.com/hugo_eiler/3 Thu, 26 Jan 2012 10:44:07 PST A study was designed to evaluate the response of blood cortisol content in dogs tentatively diagnosed as having hyperadrenocorticism by using the combined dexamethasone suppression/ACTH stimulation test procedure. Four groups of abnormal responses were identified in 54 dogs. In group I (14.8% of the dogs with abnormal responses), the only abnormality was partial suppression with dexamethasone (clinically normal dogs suppressed to less than 10 ng/ml). In group II (29.6%), 2 abnormalities were found: partial suppression with dexamethasone and hyperreactivity to the ACTH stimulation test. In group III (typical pituitary-dependent hypercortisolism, 48.1%), 3 abnormalities were found: base-line hypercortisolemia, partial suppression with dexamethasone, and hyperreactivity to the ACTH stimulation test. In group IV (7.4%), 2 abnormalities were found: base-line hypercortisolemia and partial suppression with dexamethasone. Base-line blood cortisol content was normal in 44.4% of the adrenopathic dogs. A normal response to ACTH stimulation was seen in 25.9% of the dogs, and 74.1% of the dogs hyperreacted to the ACTH stimulation test. All of the adrenopathic dogs were found to suppress partially with dexamethasone. Failure to suppress the adrenal gland completely (less than 10 ng/ml) with dexamethasone was the most consistent finding in adrenopathic dogs when using the combined dexamethasone suppression/ACTH stimulation test procedure. It was concluded that the test procedure is feasible, flexible, and convenient for clinical situations. Also, these results suggested that there may be several stages in the negative feedback failure associated with hyperadrenocorticism in dogs.

]]> Hugo Eiler et al. Selected Articles http://works.bepress.com/hugo_eiler/2 http://works.bepress.com/hugo_eiler/2 Thu, 26 Jan 2012 10:44:06 PST Hugo Eiler United States Patent http://works.bepress.com/hugo_eiler/1 http://works.bepress.com/hugo_eiler/1 Thu, 26 Jan 2012 10:44:04 PST Background: A combined dexamethasone (DEX) suppression/thyrotropin-releasing hormone (TRH) test (DEX/TRH test) has been developed to evaluate horses for presence of a pars intermedia pituitary adenoma (PIPA), but to the authors' knowledge, the accuracy of this test has not been previously determined. Hypothesis: The sensitivity and specificity of the DEX/TRH test can be determined by comparing test results with histopathologic examination findings. Animals: Age of 42 horses of various breeds ranged from 2 to 33 years. Methods: Plasma cortisol concentration was measured before and 24 hours after IV administration of 40 g of DEX/kg of body weight, and before and 30 minutes after IV administration of 1 mg of TRH that had been given 3 hours after the injection of DEX. Results of the DEX/TRH test were considered positive if either the plasma cortisol concentration exceeded 10 ng/mL 24 hours after DEX administration, or if the change in plasma cortisol concentration 30 minutes after injection of TRH was 66% above the 3-hour baseline. Diagnosis of PIPA was determined by histologic examination of the pituitary gland. Results: PIPA was detected in 17 of 42 (40%) horses. The DEX/TRH test had sensitivity, specificity, positive predictive value, and negative (NPV) predictive value of 88, 76, 71, and 90%, respectively. Conclusions and Clinical Importance: The combined DEX/TRH test was more sensitive than either of its component tests and had a high NPV, but was not as specific as the TRH component alone (92%). The DEX/TRH test should be used to screen older horses for PIPA.

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