Objective-To determine localization of tartrate-resistant acid phosphatase (TRAP) and cathepsin K in ruptured and healthy cranial cruciate ligaments (CCL) in dogs. Animals-30 dogs with ruptured CCL, 8 aged dogs without ruptured CCL, and 9 young dogs without ruptured CCL. Procedure-The CCL was examined histologically and cells containing TRAP and cathepsin K were identified histochemically and immunohistochemically, respectively. Results-Cathepsin K and TRAP were detected within the same cells, principally within the epiligamentous region and to a lesser extent in the core region of ruptured CCL. Numbers of cells containing TRAP and cathepsin K were significantly greater in ruptured CCL, compared with CCL from young or aged dogs, and numbers of such cells were greater in CCL from aged dogs, compared with those of young dogs. In aged dogs, small numbers of cells containing TRAP and cathepsin K were seen in intact CCL associated with ligament fascicles in which there was chondroid transformation of ligament fibroblasts and disruption of the extracellular matrix. Conclusion and Clinical Relevance-Ruptured CCL contain greater numbers of cells with the proteinases TRAP and cathepsin K than CCL from healthy, young, or aged dogs. Results suggest that cell-signaling pathways that regulate expression of these proteinases may form part of the mechanism that leads to upregulation of collagenolytic ligament remodeling and progressive structural failure of the CCL over time.

Objective-To determine the effect of experimental intraluminal distention on microvascular perfusion of the small colon in horses. Animals-6 mixed-breed healthy horses (mean age [+/- SD], 9.1 +/- 2 years). Procedure-Under general anesthesia, the small colon was exposed by celiotomy and 3 segments were demarcated. In 1 of these segments, intraluminal obstruction was created by placement of a latex balloon inflated to a pressure of 40 mm Hg (obstructed segment). The other segments were the sham-operated segment and the control segment. Microvascular perfusion was evaluated in the mucosal, submucosal, muscular, and serosal layers by injection of 15--µm-diameter colored microspheres into branches of the caudal mesenteric artery. Recovery of microspheres was performed by tissue digestion, washing, and centrifugation. Distribution of microspheres in the intestinal layers was evaluated by direct observation of stained frozen sections by light microscopy. Results-A significant reduction was observed in total microvascular perfusion of obstructed segments, which was 26.4% of that of control segments. This reduction was not evident in the mucosal layer. Conclusion and Clinical Relevance-Intraluminal distention of the equine small colon wall can promote ischemia by a reduction in microvascular perfusion in the intestinal wall. Intestinal layers do not seem to be affected to the same extent, because the absolute value for mucosal perfusion did not decrease in the obstructed segment.

Objective-To evaluate effects of age and body size of dogs on intestinal permeability (unmediated diffusion) as measured by the ratio of urinary lactulose to L-rhamnose (L:R) and absorption (carrier-mediated transport) as measured by the ratio of urinary D-xylose to 3-O-methyl-D-glucose (X:MG) and to determine whether these variables correlated with fecal quality. Animals-6 Miniature Poodles, 6 Standard Schnauzers, 6 Giant Schnauzers, and 6 Great Danes. Procedure-A solution that contained lactulose and rhamnose or xylose and 3-O-methyl-D-glucose was administered orally to dogs that were 12, 22, 36, and 60 weeks old. Urine was collected 6 hours later, and urinary L:R and X:MG were calculated. Fecal moisture and scoring were recorded during the same periods. Results-Age and breed did not affect intestinal absorption, and we did not detect a relationship between X:MG and fecal variables. In contrast, we detected significant effects of age and body size on intestinal permeability. Puppies (12 weeks old) and large dogs had higher intestinal permeability than adult (60 weeks old) and small dogs. The increased intestinal permeability in large dogs was associated with lower fecal quality as indicated by the significant positive correlations between L:R and fecal moisture (r, 0.61) and L:R and fecal scores (r, 0.86) in adult dogs. Conclusion and Clinical Relevance-These results indicate that age and body size should be considered when assessing intestinal permeability by use of the L:R urinary excretion test in dogs. High intestinal permeability could be a possible cause of poor fecal quality in large dogs.

Objective-To evaluate splenic mast cell tumors (MCT) of cats for activating mutations in the protooncogene c-kit. Sample Population-10 formalin-fixed, paraffinembedded splenic MCT from cats in the pathology database of the Veterinary Medical Teaching Hospital at the University of California, Davis. Procedure-Genomic DNA was isolated from tumor specimens, and the polymerase chain reaction (PCR) procedure was performed for exons 11, 12, and 17. The PCR products were analyzed by use of agarose gel electrophoresis and then directly sequenced. Results-We did not identify mutations in the juxtamembrane domain (encoded by exons 11 and 12) or catalytic domain (encoded by exon 17) of c-kit in any of the splenic MCT specimens. Conclusions and Clinical Relevance-Although mutations in the proto-oncogene c-kitoccur frequently in naturally developing MCT in dogs and aggressive mastocytosis in humans, the data reported here documented that dysregulation of Kit function through activating mutations is unlikely in splenic MCT of cats. Therapeutic strategies aimed at inhibiting Kit signaling (ie, kinase inhibitors such as imatinib [STI571]) may not be of benefit for the treatment of this disease in cats.

Objective-To provide an epidemiologic investigation of the seroprevalence of antibodies to Toxoplasma gondii in populations of cats and wild rodents in Rhode Island and to address the possible epidemiologic role of wild rodents in the spread of toxoplasmosis. Animals-200 cats and 756 small wild rodents. Procedure-Serum samples were obtained from 84 cats in animal shelters and 116 cats in veterinary hospitals. Serum samples were also obtained from 756 small wild rodents from multiple sites in Rhode Island. Sera from rodents and cats were assayed for antibodies to T gondii by use of the modified agglutination test Results-Overall, 42% (84/200) of cats had serum antibodies to T gondii. Seroprevalence was not significantly different between stray (50%; 42 /84) versus client-owned (36%; 42/116) cats, between male (43%; 40/94) versus female (42%; 39/93) cats, or between indoor (26%; 7/27) versus outdoor (39%; 35/89) cats. Seroprevalence rate of trapped rodents was 0.8% (6/756). Six rodents captured in Washington County accounted for of the seropositive rodents. Four of 6 of the seropositive rodents were trapped at a single site in Washington County (an abandoned barn). Five stray cats, known to have resided at the same site in Washington County as 4 of the seropositive rodents, were also found to be seropositive for antibodies to T gondii. Conclusions and Clinical Relevance-Seroprevalence rate in rodents was not correlated with the seroprevalence rate in cats. Stray cats, especially those known to be feral, may be more likely to perpetuate the cat-mouse cycle of T gondii than clientowned cats.

Objective-To determine whether alterations in myoplasmic calcium regulation can be identified in muscle cell cultures (myotubes) and intact muscle fiber bundles derived from Thoroughbreds affected with recurrent exertional rhabdomyolysis (RER). Animals-6 related Thoroughbreds with RER and 8 clinically normal (control) Thoroughbred or crossbred horses. Procedures-Myotube cell cultures were grown from satellite cells obtained from muscle biopsy specimens of RER-affected and control horses. Fura-2 fluorescence was used to measure resting myoplasmic calcium concentration as well as caffeine- and 4-chloro-m-cresol (4-CMC)-induced increases in myoplasmic calcium. In addition, intact intercostal muscle fiber bundles were prepared from both types of horses, and their sensitivities to caffeine- and 4-CMC-induced contractures were determined. Results-Myotubes of RER-affected and control horses had identical resting myoplasmic calcium concentrations. Myotubes from RER-affected horses had significantly higher myoplasmic calcium concentrations than myotubes from control horses following the addition of ≥ 2mM caffeine; however, there was no difference in their response to 4-CMC (greater than 1mM). Caffeine contracture thresholds for RER and control intact muscle cell bundles (2 vs 10mM, respectively) were significantly different, but 4-CMC contracture thresholds of muscle bundles from RER-affected and control horses (500µM) did not differ. Conclusions and Clinical Relevance-An increase in caffeine sensitivity of muscle cells derived from a family of related RER-affected horses was detected in vitro by use of cell culture with calcium imaging and by use of fiber bundle contractility techniques. An alteration in muscle cell calcium regulation is a primary factor in the cause of this heritable myopathy.

Detection of the scrapie-associated protease-resistant prion protein (PrPres) in sheep brains in the early phase after intracerebral inoculation of the scrapie agent has not been documented. Fourteen 4-mo-old, genetically susceptible lambs (QQ homozygous at codon 171 of the PrP gene) were obtained for this study. Twelve lambs were inoculated intracerebrally with a brain suspension from sheep naturally affected with scrapie, and 2 served as uninoculated controls. Two inoculated animals were euthanized at each of 6 times postinoculation (1 h to 6 wk), and their brains were collected for histopathological study, for detection of PrPres by the Western blot technique and an immunohistochemical (IHC) method, and for the detection of scrapie-associated fibrils (SAF) by negatively stained electron microscopy (EM). Microscopic lesions associated with introduction of the inoculum were seen in the brains of inoculated animals at all 6 times. However, both the Western blot and IHC techniques did not detect PrPres after the initial 3 d postinoculation, nor did EM detect SAF in any of the samples. From these findings, it is presumed that until host amplification has occurred, the concentration of PrPres in inoculum is insufficient for detection by currently available techniques.

Objective-To clone segments of the canine liver alkaline phosphatase (LALP) and corticosteroidinduced alkaline phosphatase (CIALP) genes and use those clones to determine the tissue source of CIALP, the kinetics of LALP and CIALP mRNA expression for glucocorticoid-treated dogs, and the correlation between LALP and CIALP transcript concentrations and isoenzyme activities. Sample Population-Tissues obtained from 7 dogs treated with prednisone (1 mg/kg, SC, q 24 h) for up to 32 days and 1 untreated (control) dog. Procedure-Gene segments of LALP and CIALP were obtained by reverse transcription-polymerase chain reaction (RT-PCR) assay. The tissue source of CIALP and IALP mRNA was determined by northern blot analysis of tissues from 1 of the glucocorticoidtreated dogs. Hepatic tissues and serum samples were obtained from the 6 remaining glucocorticoidtreated dogs on days 0, 2, 5, 10, and 32 of prednisone treatment, and relative expression of LALP and CIALP mRNA was correlated with LALP and CIALP activity. Results-A 2,246-base pair (bp) segment of canine LALP and a 1,338-bp segment of CIALP were cloned. Northern blot analysis revealed CIALP mRNA expression in hepatic tissues only after glucocorticoid treatment. Kinetics of LALP and CIALP mRNA expression in the liver of glucocorticoid-treated dogs paralleled liver and serum activities of LALP and CIALP. Conclusions and Clinical Relevance-The liver is the most likely source for CIALP in dogs. Analysis of kinetics of serum and hepatic LALP and CIALP mRNA suggests that after glucocorticoid treatment, both are regulated by modification of mRNA transcript concentrations, possibly through differing mechanisms.

Objective-To determine whether changes in amplitude of the reflex-evoked muscle action potential (REMP) elicited by noninvasive dental dolorimetry (electrical stimulation of the tooth pulp) in anesthetized dogs may be used to objectively evaluate the effectiveness of IV and intrathecal (IT) administration of morphine. Animals-6 male Beagles that were 2 to 6 years old. Procedure-Dogs were used in a crossover design with at least a 5-day washout period between treatments. Each dog received morphine, saline (0.9% NaCl) solution, and oxytocin via the IV and IT routes of administration; however, only results for morphine and saline treatments were reported here. Dogs were anesthetized and prepared for noninvasive dental dolorimetry. After IV or IT administration, electrical stimulation was applied to a tooth, and REMPs of the digastricus muscle were recorded at 5-minute intervals for 60 minutes. To determine differences in REMP amplitude between treatments, a linear regression line was fitted for each dog-treatment combination. Results-The IV administration of morphine significantly inhibited REMP amplitude, compared with IV administration of saline solution. Intrathecal administration of morphine significantly inhibited REMP amplitude, compared with IT administration of saline solution. Conclusions and Clinical Relevance-Noninvasive dental dolorimetry in anesthetized dogs has promise as a technique for use in evaluating the analgesic potential of drugs administered IV and IT through evaluation of their effect on REMP amplitude recorded for the digastricus muscle.

Objective-To correlate tissue distribution with development of lesions after experimental infection with a virulent strain of noncytopathic bovine viral diarrhea virus (BVDV) type 2 in calves. Animals-Ten 14-day-old and two 2-month-old colostrum-deprived calves. Procedure-Calves were intranasally inoculated with BVDV type-2 strain 1373 from an outbreak of clinically severe bovine viral diarrhea (BVD). Two 14-day-old calves served as noninfected controls. Two calves each were euthanatized on postinoculation days 3, 6, and 12, and 1 each on days 8, 9, 13, and 14. Tissues were collected for immunohistologic and histologic examination. Results-Inoculated calves developed nonspecific clinical signs characterized by high fever and decreased numbers of leukocytes and thrombocytes. Viral antigen was detected focally in lymphoid tissues on day 3. On days 6, 8, 9, 12, and 14, viral antigen became increasingly widespread throughout organs and tissues. Viral antigen in lymphoid tissues was associated with severe depletion of all compartments. Lesions in other tissues were not well correlated with distribution of viral antigen. Depletion of lymphoid tissues was observed in a calf on day 13, but viral antigen had been cleared from most tissues and was detected in vascular walls only. Conclusions and Clinical Relevance-Infection with a virulent BVDV strain resulted in wide dissemination of viral antigen in host tissues. Severe lymphoid depletion developed in lymphoid tissues, whereas viral antigen was generally not associated with lesions in other tissues. Findings suggest that development of lesions in acute BVD is not solely a function of viral replication and is also attributable to host reaction to infection.