Objective—To apply an in vitro model for assessment of the solid-phase binding capacity of acetaminophen and thus assess the reliability of this marker for evaluation of solid-phase gastric emptying in vivo in animals.
Sample Population—4 test meals.
Procedures—A spectrophotometric method for detection of acetaminophen was validated and applied for assessment of the percentage retention of acetaminophen in the solid phase of 4 test meals. The gastric milieu was simulated by incubating each meal in artificial gastric juice for 2 hours in a shaking water bath maintained at 37°C. Solid-phase retention was then assessed 3 times by measuring the amount of acetaminophen that had leached into the liquid phase.
Results—Acetaminophen was poorly retained in the solid phase of all the test meals examined in the study. There was also a large degree of variability in the percentage retention for each meal when the experiment was repeated 3 times.
Conclusions and Clinical Relevance—Analysis of the results of this in vitro study confirmed that acetaminophen may not be an appropriate marker of solid-phase gastric emptying. The acetaminophen gastric emptying test should be applied only for the assessment of liquid-phase emptying in animals.
Objective—To assess the anti-inflammatory effects of an adenosine analogue on lipopolysaccharide (LPS)-stimulated equine neutrophils.
Sample Population—Neutrophils obtained from 10 healthy horses.
Procedures—An adenosine analogue (5′-N-ethylcarboxamidoadenosine [NECA]) was tested for its ability to inhibit production of reactive oxygen species (ROS) in LPS-stimulated equine neutrophils. Selective adenosine receptor antagonists were used to identify the receptor subtype responsible for effects. To assess the mechanism of action of NECA, cAMP concentrations were measured, and effects of dibutyryl cAMP (a stable analogue of cAMP) and rolipram (a type 4 phosphodiesterase inhibitor) were investigated.
Results—NECA elicited concentration-dependent inhibition of ROS production that was inhibited by ZM241385, a selective adenosine A2A receptor antagonist; this effect of NECA was not affected by the adenosine A2B receptor antagonist MRS1706. Also, ZM241385 blocked NECA-induced increases in cAMP concentrations, whereas MRS1706 did not alter this effect of NECA. Rolipram potentiated NECA-induced inhibition of ROS production, and dibutyryl cAMP also inhibited ROS production.
Conclusions and Clinical Relevance—Activation of adenosine A2A receptors inhibited ROS production by LPS-stimulated equine neutrophils in a cAMP-dependent manner. These results suggest that stable adenosine A2A receptor agonists may be developed as suitable anti-inflammatory drugs in horses.
Objective—To determine the effects of the protein kinase C (PKC) inhibitor, Ro-31-8220, on agonist-induced constriction of laminar arteries and veins obtained from horses.
Sample Population—Laminar arteries and veins obtained from 8 adult mixed-breed horses.
Procedures—Laminar arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Concentration-response curves were then obtained for the vasoconstrictor agonists phenylephrine, 5-hydroxytryptamine, prostaglandin F2α, and endothelin-1. All responses were measured with or without the addition of Ro-31-8220 (3μM).
Results—Laminar veins were more sensitive to vasoconstrictor agonists than laminar arteries, and incubation of laminar veins with Ro-31-8220 resulted in significantly smaller agonist-induced contractile responses for all agonists tested. In contrast, Ro-31-8220 had no effect on agonist-induced contractile responses of laminar arteries.
Conclusions and Clinical Relevance—Results of the study were consistent with activation of PKC being confined to agonist-induced contraction of laminar veins isolated from the laminar dermis of horses. Consequently, the possible involvement of PKC in the venoconstriction observed during the development of laminitis is worthy of further investigation.
Objective—To evaluate the clinical and immunologic response in healthy dogs to infusions of human serum albumin (HSA).
Animals—9 healthy purpose-bred mixed-breed dogs.
Procedures—Each dog was administered a 25% HSA solution once or twice. Various physical examination and laboratory variables were serially evaluated. Antibody against HSA was assayed before and after infusion by use of an ELISA. Intradermal testing was also conducted. A repeated-measures ANOVA or Friedman repeated-measures ANOVA on ranks was used to compare results for the variables.
Results—Adverse clinical reactions were observed after the first or second infusion in 3 dogs. Anaphylactoid reactions were observed in 1 of 9 dogs during the first infusion and in 2 of 2 dogs administered a second infusion. Two dogs developed severe edema and urticaria 6 or 7 days after an initial infusion. All dogs developed anti-HSA antibodies. Positive responses for ID tests were observed in 8 of 9 dogs. Short-term increases were detected in blood protein, total bilirubin, and calcium concentrations after HSA infusion. Serum cholesterol concentrations and platelet counts decreased after HSA infusion.
Conclusions and Clinical Relevance—Administration of HSA resulted in profound reactions in 2 of 9 dogs administered a single infusion and in 2 of 2 dogs administered a second infusion. This indicates that there is risk of life-threatening adverse reactions to HSA infusion in healthy dogs.
Objective—To determine whether prolonged exercise by conditioned sled dogs affects urine concentrations of homovanillic acid (a metabolite of dopamine), vanillylmandelic acid (a metabolite of norepinephrine and epinephrine), and cortisol.
Animals—24 conditioned Alaskan sled dogs (2 to 8.5 years old) that were in training for a multiday endurance race.
Procedures—Voided urine samples were collected from 4 groups of dogs (randomly selected from 54 dogs) after no exercise (control group; n = 6 dogs), completion of a 160km run (group A; 3), completion of a 420-km run (group B; 7), and completion of a 560-km run (group C; 6). Urine cortisol concentrations were determined by use of an immunoassay technique; urine vanillylmandelic acid and homovanillic acid concentrations were measured via high-performance liquid chromatography.
Results—Compared with the control group, urine cortisol concentration in groups A, B, and C was significantly different (5.33 × 10−4 ± 2.62 × 10−4 μg/dL vs 1.04 × 10−4 ± 2.31 × 10−5 μg/dL, 8.88 × 10−4 ± 5.49 × 10−4 μg/dL, and 6.31 × 10−4 ± 5.09 × 10−4 μg/dL, respectively). Urine homovanillic acid concentration did not differ among the 4 groups. Vanillylmandelic acid was not detected in any urine samples.
Conclusions and Clinical Relevance—Results indicated that prolonged exercise by sled dogs did not affect urine homovanillic acid concentration but did increase urinary cortisol secretion, which is indicative of adrenocortical stimulation. The apparent lack of vanillylmandelic acid in voided urine samples requires further investigation.
Objective—To determine whether there are increased concentrations of 25-hydroxyvitaminn D3 in red-eared slider turtles (Trachemys scripta elegans) after exposure to UV radiation.
Animals—12 yearling turtles recently removed from aestivation.
Procedures—Turtles were randomly allocated to 2 groups (6 turtles/group). An initial blood sample was collected from all turtles for measurement of 25-hydroxyvitamin D3 concentrations. Turtles of 1 group were then provided no supplemental lighting, whereas turtles of the other group were exposed to full-spectrum coil bulbs at a distance of 22.86 cm. The UV-A and UV-B radiation generated by the supplemental lighting was measured by use of a radiometer-photometer at weekly intervals. Measurements were collected 2.54 and 22.86 cm from the bulb surface. The study was continued for a 4-week period. At the end of the study, a second blood sample was collected from all turtles for measurement of 25-hydroxyvitamin D3.
Results—Mean ± SD 25-hydroxyvitamin D3 concentrations differed significantly between turtles provided supplemental UV radiation (71.7 ± 46.9 nmol/L) and those not provided UV radiation (31.4 ± 13.2 nmol/L).
Conclusions and Clinical Relevance—Appropriate husbandry recommendations for raising and maintaining red-eared slider turtles should include use of sunlight that is unobstructed by UV-B filtering material or provision of an artificial source of UV-B radiation.
Objective—To evaluate whether bronchoalveolar lavage (BAL) alters respiratory mechanics of horses with recurrent airway obstruction (ie, heaves) over a 48-hour period.
Animals—6 horses affected with heaves.
Procedures—Horses were subjected to a complete BAL procedure, which included sedation with xylazine and butorphanol, intratracheal administration of lidocaine, and instillation and aspiration of two 250-mL boluses of saline (0.9% NaCl) solution through an endoscope (study 1). To evaluate the effects of saline solution, horses were subjected to the same procedure without saline solution instillation and aspiration (study 2). Lastly, the endoscope was similarly introduced into the lower airways, without sedation or saline instillation and aspiration (study 3). Respiratory mechanics were performed at baseline (time 0) and at 3, 6, 12, 24, and 48 hours after each procedure.
Results—In study 1, BAL induced a significant decrease in pulmonary resistance lasting up to 6 hours. This may have resulted from clearance of mucus in large airways. We also observed a significant increase in lung elastance and transpulmonary pressure at 12 hours after BAL in all 3 studies, which may be attributed to a circadian effect.
Conclusions and Clinical Relevance—Our results indicate that the temporal effects of BAL procedures on lung mechanics should be taken into account when designing research protocols involving horses with heaves. Future studies should address the immediate effects of BAL on lung function.
Objective—To determine distribution of urokinase plasminogen activator-like protein and urokinase plasminogen activator receptor-like protein in urinary tract tissues of healthy dogs.
Animals—11 healthy dogs.
Procedures—Necropsy specimens from kidney, ureter, bladder, urethra, prostate, and testis were obtained from 4 sexually intact female dogs, 5 sexually intact males, and 2 castrated males; dogs ranged in age from juvenile to adult. Urokinase plasminogen activator-like protein and urokinase plasminogen activator receptor-like protein in tissue lysates from kidney, prostate, and testis were identified by use of SDS-PAGE, western blot analysis, and immunoprecipitation. Urokinase plasminogen activator-like protein and urokinase plasminogen activator receptor-like protein in kidney, ureter, urinary bladder, urethra, prostate, and testis were identified by use of immunohistochemical staining of tissue sections.
Results—Urokinase plasminogen activator-like protein and urokinase plasminogen activator receptor-like protein in the molecular-weight range published for urokinase and urokinase receptor (53 and 33 kd for urokinase and 60 to 65 kd for urokinase receptor) were identified. Distribution of the proteins identified by use of immunohistochemical staining was comparable with published information for humans and mice for the urinary tract. Staining of these proteins was detected in more tissue types than reported in healthy humans.
Conclusions and Clinical Relevance—Urokinase plasminogen activator-like protein and urokinase plasminogen activator receptor-like protein were detected in the urinary tract of healthy dogs. This information is important for further evaluation of the functions of urokinase and urokinase receptor in the canine urinary tract and the pathophysiologic features of urinary tract disease.
Objective—To determine whether administration of isoflupredone acetate (ISO) to healthy cows increases the frequency of severe hypokalemia and whether dexamethasone (DEX) has detectable mineralocorticoid properties.
Animals—33 cows at 20 to 25 days of lactation.
Procedures—Cows were randomly allocated to 5 treatment groups and received 2 IM injections (on days 0 and 2) of sterile saline (0.9% NaCl) solution (10 mL each), an injection of ISO (20 mg) or DEX (20 mg) followed by 10 mL of saline solution, or 2 injections of ISO or DEX. Milk production was measured, physical examinations were performed, and blood and urine samples were collected daily on days 0 through 7.
Results—Physical examination parameters did not differ among groups; however, 1 cow developed atrial fibrillation on day 4. Both corticosteroids significantly increased plasma glucose concentrations, and ISO significantly decreased plasma potassium concentrations and increased total carbon dioxide concentrations with time. One dose of ISO decreased mean plasma potassium concentration by 25% on day 2, compared with day 0, and severe hypokalemia (serum potassium concentration < 2.3 mEq/L) developed in 1 of 6 cows. Mean plasma potassium concentration was 46% lower on day 3 than on day 0 in cows receiving 2 doses of ISO, and 5 of 7 cows became severely hypokalemic. Mean urinary fractional excretion of potassium significantly increased from that on day 0 in cows receiving 2 doses of ISO.
Conclusions and Clinical Relevance—Both corticosteroids had glucocorticoid activity; however, only ISO had mineralocorticoid activity. Compared with saline solution, administration of 2 doses of ISO significantly increased the frequency of severe hypokalemia.
Objective—To investigate the effects of preventive angiotensin converting enzyme inhibitor treatment with ramipril in dogs with progressively severe experimentally induced heart failure.
Procedures—Dogs were randomly allocated to receive no treatment (control) or ramipril (0.125 mg/kg, PO, daily) for 7 weeks. Physical examination, repetitive catheterization of the right side of the heart, and echocardiography were performed before the study (day 0) and weekly for 7 weeks. Renal plasma flow (RPF) as determined by para-aminohippuric acid clearance and glomerular filtration rate (GFR) as determined by creatinine and iohexol clearances were measured on day 0 and at weeks 4 and 7.
Results—Overpacing induced a progressive increase in right atrial pressure (RAP) and pulmonary artery pressure, occluded (PAPO), with a decrease in systemic arterial pressure. There were progressive alterations of echocardiographic indices of diastolic and systolic ventricular function. The RPF and GFR decreased before cardiac output decreased, and filtration fraction increased. The logarithm of the urinary sodium–to–potassium concentration ratio (log10[Na+/K+]) decreased. Significant effects of ramipril included a delay in clinical signs of heart failure, a late decrease in RAP and PAPO, and increases in the sodium excretion fraction and log10(Na+/K+). There was a satisfactory agreement between the creatinine and iohexol clearance measurements.
Conclusions and Clinical Relevance—Results suggest that, in this rapid-evolving, dilated cardiomyopathy, activation of the renin-angiotensin system contributes to the pathophysiology of heart failure late in the disease and essentially by an activation of renal salt and water retention.