Objective—To evaluate expression of matrix metalloproteinase (MMP)-2 and -9 and membrane-type 1 MMP (MT1-MMP) in melanocytomas and malignant melanomas of dogs, analyze in vitro production of MMPs by canine melanoma cell lines and primary dermal fibroblasts, and investigate mutual communication between tumor cells and fibroblasts and the influence of collagen on MMP regulation. Sample—35 biopsy specimens from melanocytic tumors and primary dermal fibroblasts of dogs and 3 canine melanoma cell lines (CML-1, CML-10c2, and CML-6M). Procedures—MMP-2, MMP-9, and MT1-MMP were detected in tumor samples by use of unohistochemical analysis. In vitro production was analyzed via reverse transcriptase-PCR assay, immunocytochemical analysis, zymography, and immunoblotting. Results—MMP-9 was overexpressed in malignant melanomas, compared with expression in melanocytomas, whereas no significant differences in MMP-2 and MT1-MMP immunostaining were detected. Stromal cells also often had positive staining results. In vitro, all 3 melanoma cell lines and dermal fibroblasts had evidence of MMP-2 and MT1-MMP, but only melanoma cells had evidence of MMP-9. Coculture of CML-1 or CML-10c2 cells and dermal fibroblasts induced an increase in expression of the active form of MMP-2. Culture of melanoma cells on type I collagen increased the activation state of MT1-MMP. Conclusions and Clinical Relevance—MMP-9 expression was increased in malignant melanomas of dogs. Stromal cells were a source for MMPs. Stromal cells, in combination with matrix components such as type I collagen, can interact with tumor cells to regulate MMP production. Information about MMP production and regulation could help in the development of new treatments.

This project evaluated the use of an ingestible temperature sensor to measure body core temperature (Tc) in exercising dogs. Twenty-five healthy, unconditioned Labrador retrievers participated in an outdoor 3.5-km run, completed in 20 min on a level, 400-m grass track. Core temperature was measured continuously with a telemetric monitoring system before, during, and after the run. Data were successfully collected with no missing data points during the exercise. Core temperature elevated in the dogs from 38.7 +/- 0.3 degrees C at pre-exercise to 40.4 +/- 0.6 degrees C post-exercise. While rectal temperatures are still the standard of measurement, telemetric core temperature monitors may offer an easier and more comfortable means of sampling core temperature with minimal human and mechanical interference with the exercising dog.

Objective—To compare responses of equine digital arteries (EDAs) and veins (EDVs) to human-acalcitonin gene-related peptide (hαCGRP), evaluate effect of the endothelium, and characterize receptors and sources of endogenous CGRP. Sample—Palmar digital vessels (5 to 9/experiment) from healthy adult horses killed at an abattoir. Procedures—Vessel rings were mounted under tension in organ baths containing Krebs-Henseleit solution at 30°C, with relaxation responses examined in vessels preconstricted with a thromboxane-mimetic (3 × 10(−8)M). Responses of endothelium-intact (+e) and -denuded (−e) EDAs and EDVs to hαCGRP C10−10 to 3 × 10(−7)M) were compared. Following incubation with an hαCGRP receptor antagonist (hαCGRP8–37; 1μM), responses of EDA(−e) and EDV(−e) to hαCGRP (10(−7)M) were obtained. Responses of endothelium-intact and -denuded arteries and veins to hαCGRP (3 × 10(−7)M) or capsaicin (10(−5)M) were evaluated as well as responses of endothelium-intact and -denuded EDA and EDV to hαCGRP (10(−10) to 10(−6)M) after incubation with endothelin-1 (ET-1; 10(−12)M). Results—hαCGRP resulted in nonendothelium, concentration-dependent relaxation in EDAs and EDVs, with greater responses in EDAs. Treatment with hαCGRP8–37 had minimal effect on responses to hαCGRP in either vessel type. Capsaicin induced relaxation in both vessel types. There were no differences between responses to hαCGRP for vessels pretreated with ET-1 or vehicle. Conclusions and Clinical Relevance—Both hαCGRP and capsaicin induced digital vasodilation unaffected by a functional endothelium. This suggested that endogenous CGRP likely emanates from sensory-motor nerves and may contribute to digital vasodilation.

Objective—To provide a detailed computed tomography (CT) reference of the anatomically normal equine stifle joint. stifle joints. Procedures—CT of the stifle joint was performed on 8 hind limbs. In all limbs, CT was also performed after intra-articular injection of 60 mL of contrast material (150 mg of iodine/mL) in the lateral and medial compartments of the femorotibial joint and 80 mL of contrast material in the femoropatellar joint (CT arthrography). Reformatted CT images in the transverse, parasagittal, and dorsal plane were matched with corresponding anatomic slices of the 8 remaining limbs. Results—The femur, tibia, and patella were clearly visible. The patellar ligaments, common origin of the tendinous portions of the long digital extensor muscle and peroneus tertius muscle, collateral ligaments, tendinous portion of the popliteus muscle, and cranial and caudal cruciate ligaments could also be consistently evaluated. The cruciate ligaments and the meniscotibial ligaments could be completely assessed in the arthrogram sequences. Margins of the meniscofemoral ligament and the lateral and medial femoropatellar ligaments were difficult to visualize on the precontrast and postcontrast images. Conclusions and Clinical Relevance—CT and CT arthrography were used to accurately identify and characterize osseous and soft tissue structures of the equine stifle joint. This technique may be of value when results from other diagnostic imaging techniques are inconclusive. The images provided will serve as a CT reference for the equine stifle joint

Objective—To directly assess microcirculatory changes associated with induced hemorrhagic shock by use of sidestream dark field microscopy (SDM) and correlate those values with concurrently measured macrovascular and blood gas variables in healthy anesthetized dogs. Animals—12 adult dogs. Procedures—Dogs were anesthetized and splenectomized. Instrumentation and catheterization were performed for determination of macrohemodynamic and blood gas variables. Hemorrhagic shock was induced via controlled hemorrhage to a mean arterial blood pressure (MAP) of 40 mm Hg. Dogs were maintained in the shock state (MAP, 35 to 45 mm Hg) for 60 minutes. An SDM device was used to image microcirculation of buccal mucosa, and vascular analysis software was used to determine microcirculatory variables. These values were compared with other cardiovascular and blood gas variables to determine correlations. Results—Following hemorrhage, there was a significant decrease in microvascular variables (mean ± SD), including proportion of perfused vessels (82.77 ± 8.32% vs 57.21 ± 28.83%), perfused vessel density (14.86 ± 2.64 mm/m2 vs 6.66 ± 4.75 mm/m2), and microvascular flow index (2.54 ± 0.52 vs 1.59 ± 0.85). Perfused vessel density individually correlated well with macrovascular variables, with heart rate (zero order, partial correlation, and part correlation coefficients = −0.762, −0.884, and −0.793, respectively) and oxygen extraction ratio (−0.734, −0.832, and −0.746, respectively) being the most important predictors. Conclusions and Clinical Relevance—SDM allowed real-time imaging of the microvasculature and has potential as an effective tool in experimental and clinical applications for monitoring microcirculatory changes associated with hemorrhagic shock and resuscitation in dogs.

Objective—To quantify changes in hoof wall strain distribution associated with exercise and time in Standardbreds. Animals—18 young adult Standardbreds. Procedures—9 horses were exercised 4 d/wk for 30 to 45 minutes at a medium trot for 4 months; 9 nonexercised horses served as the control group. Rosette strain gauges were used to measure the principal surface strains at the toe, lateral quarter of the hoof wall (LQ), and medial quarter of the hoof wall (MQ) of the right forefoot at the beginning and end of the experiment. Midstance maximal (msepsilon1) and minimal (msepsilon2) principal and peak minimal principal (pkepsilon2) surface strains were measured; SDs of each of those variables were also calculated. Results were compared through ANOVA of time and exercise effects between and within the groups. Results—Both the exercised and nonexercised groups had changes in strain distribution in their hooves over time. The msepsilon1 did not change significantly with exercise; however, it changed significantly in both groups at both hoof quarters over time. At the beginning of the study, mean msepsilon2 and pkepsilon2 values were significantly higher in the exercised group than in the control group at the MQ and LQ but not at the toe. At the end of the study, these values were significantly higher in the control group than in the exercised group at the toe but not at the MQ or LQ. Conclusions and Clinical Relevance—Detected changes in hoof wall surface strain may indicate the ability of hoof capsule material to respond to exercise. A better understanding of hoof adaptation to applied forces may allow implementation of proper trimming and shoeing techniques to promote adaptation to exercise loads in horses.

Objective—To compare the efficacy and adverse effects of sustained-release (SR) buprenorphine following SC administration and buprenorphine following oral transmucosal (OTM) administration in cats undergoing ovariohysterectomy. Animals—21 young healthy female cats. Procedures—As part of anesthetic premedication (0 hours), 10 cats received buprenorphine (0.02 mg/kg) via OTM administration with additional doses at 12, 24, 36, 48, and 60 hours and 11 cats received an equivalent total dose as a single SC injection of SR buprenorphine (0.12 mg/kg). The SR product contained buprenorphine hydrochloride in a proprietary SR matrix. All other anesthetic drugs and a single postoperative dose of meloxicam were administered similarly to all cats. Behavioral and physiologic variables were recorded, and signs of pain were assessed by use of 2 pain assessment scales and von anesthesia (RFA), and at 12, 24, 36, 48, 60, and 72 hours. Results—Heart rate increased and temperature (determined via microchip transponder thermometry) decreased from baseline values during RFA in both groups. Compared with baseline values, pain scores were increased during RFA and at the 12- and 24-hour time points in both groups; von Frey scores were higher during RFA. Behavioral and physiologic variables did not differ significantly between groups at any time point. Conclusions and Clinical Relevance—In cats undergoing ovariohysterectomy, SC administration of a preoperative dose of SR buprenorphine appeared to have comparable efficacy and adverse effect profile as that of twice-daily OTM administration of buprenorphine before and after surgery.

Objective--To establish a computed tomography (CT)-angiography protocol and measure the diameters of major arteries in parrots. Animals--13 Hispaniolan Amazon parrots (Amazona ventralis). Procedures--16-slice CT scanning was used to measure the apparent diameter of the ascending aorta, abdominal aorta, pulmonary arteries, and brachiocephalic trunk. Before scanning, all birds underwent ECG and echocardiographic assessment and were considered free of detectable cardiovascular diseases. Each bird was anesthetized, and a precontrast helical CT scan was performed. Peak aortic enhancement was established with a test bolus technique via dynamic axial CT scan over a predetermined single slice. An additional bolus of contrast medium was then injected, and a helical CT-angiography scan was performed immediately afterward. Arterial diameter measurements were obtained by 2 observers via various windows before and after injection, and intra- and interobserver agreement was assessed. Results--Reference limits were determined for arterial diameter measurements before and after contrast medium administration in pulmonary, mediastinal, and manual angiography windows. Ratios of vertebral body diameter to keel length were also calculated. Intraobserver agreement was high (concordance correlation coefficients ≥ 0.95); interobserver agreement was medium to high (intraclass correlation coefficients ≥ 0.65). Conclusions and Clinical Relevance--CT-angiography was safe and is of potential diagnostic value in parrots. We recommend performing the angiography immediately after IV injection of 3 mL of iohexol/kg. Arterial diameter measurements at the described locations were reliable.

Objective: To evaluate the in vitro susceptibility of various field isolates of Mycobacterium avium subsp paratuberculosis (MAP) to gallium nitrate. Sample: 10 isolates of MAP, including 4 isolated from cattle, 2 isolated from bison, 1 isolated from an alpaca, and 3 isolated from humans. Procedures: The in vitro susceptibility to gallium nitrate was tested by use of broth culture with detection of MAP growth by means of a nonradiometric automated detection method. For each MAP isolate, a series of 7 dilutions of gallium nitrate (concentrations ranging from 200 to 1,000μM) were tested. Gallium nitrate was considered to have caused 90% and 99% inhibition of the MAP growth when the time to detection for culture of the MAP stock solution and a specific concentration of gallium nitrate was delayed and was similar to that obtained for culture of the MAP stock solution (without the addition of gallium nitrate) diluted 1:10 and 1:100, respectively. Results: Gallium nitrate inhibited MAP growth in all 10 isolates. The susceptibility to gallium nitrate was variable among isolates, and all isolates of MAP were inhibited in a dose-dependent manner. Overall, the concentration that resulted in 90% inhibition ranged from < 200μM for the most susceptible isolates to 743μM for the least susceptible isolates. Conclusions and Clinical Relevance: Gallium nitrate had activity against all 10 isolates of MAP tested in vitro and could potentially be used as a prophylactic agent to aid in the control of MAP infections during the neonatal period.

Objective—To examine the changes in monocarboxylate transporter (MCT) 1 and MCT4 content and in indicators of energy metabolism in the gluteus medius muscle (GMM) of Thoroughbreds during growth. Animals—6 Thoroughbreds (3 males and 3 females). Procedures—Samples of GMM were obtained when horses were 2, 6, 12, and 24 months old. Muscle proteins were separated via SDS-PAGE; amounts of MCT1 and MCT4 and peroxisome proliferator-activated receptor-γ coactivator-1α content were determined by use of western blotting. Muscle activities of phosphofructokinase and citrate synthase were measured biochemically; lactate dehydrogenase isoenzymes were separated by agarose gel electrophoresis and quantified. Results—Compared with findings when horses were 2 months old, MCT1 protein content in GMM samples obtained when the horses were 24 months old was significantly higher; however, MCT4 protein content remained unchanged throughout the study period. Peroxisome proliferator-activated receptor-γ coactivator-1α content was significantly increased at 24 months of age and citrate synthase activity was increased at 6 and 24 months of age, compared with findings at 2 months. Phosphofructokinase activity remained unaltered during growth. The percentage contributions of lactate dehydrogenase 1 and 2 isoenzymes to the total amount of all 5 isoenzymes at 12 and 24 months of age were significantly higher than those at 2 months of age. Conclusions and Clinical Relevance—Changes in protein contents of MCTs and the lactate dehydrogenase isoenzyme profile in GMM samples suggested that lactate usage capacity increases with growth and is accompanied by an increase in the oxidative capacity in Thoroughbreds.