Objective—To evaluate effects of transplantation of bone marrow stromal cells (BMSCs) into the CSF for the treatment of chronic spinal cord injury in dogs that had not responded by 1 month after decompressive surgery. Animals—23 dogs. Procedures—Dogs with paraplegia and loss of nociception in the pelvic limbs for at least 1 month after decompressive surgery were assigned to transplantation or control groups. Dogs in the transplantation group received BMSCs injected into the CSF 1 to 3 months after decompressive surgery. Dogs in the control group did not receive additional treatments. Improvements in gait, proprioceptive positioning, and nociception were evaluated by use of the Texas Spinal Cord Injury Scale for ≥ 6 months after BMSC transplantation. Results—6 of 10 dogs in the transplantation group regained the ability to walk, whereas only 2 of 13 dogs in the control group regained the ability to walk. Scores for the Texas Spinal Cord Injury Scale in the transplantation group were significantly higher than scores in the control group at the endpoint of the study (6 months after BMSC transplantation or after decompressive surgery for the transplantation and control groups, respectively). Only 1 dog (transplantation group) recovered nociception. All dogs from both groups had fecal and urinary incontinence. No complications were observed in relation to BMSC transplantation. Conclusions and Clinical Relevance—Injection of BMSCs into the CSF caused no complications and could have beneficial effects on pelvic limb locomotion in dogs with chronic spinal cord injuries.

Objective—To compare in vitro axial compression, abaxial compression, and torsional stiffnesses of intact and plated radii from small- and large-breed dogs. Sample—Radii from 18 small-breed and 9 large-breed skeletally mature dogs. Procedures—3 groups were tested: large-breed dog radii plated with 3.5-mm limited-contact dynamic compression plates (LCDCPs), small-breed dog radii plated with 2.0-mm dynamic compression plates (DCPs), and small-breed dog radii plated with 2.0/2.7-mm cut-to-length plates (CTLPs). The axial compression, abaxial compression, and torsional stiffnesses of each intact radius were determined under loading with a material testing machine. An osteotomy was performed, radii were plated, and testing was repeated. The stiffness values of the plated radii were expressed as absolute and normalized values; the latter was calculated as a percentage of the stiffness of the intact bone. Absolute and normalized stiffness values were compared among groups. Results—The absolute stiffnesses of plated radii in axial and abaxial compression were 52% to 83% of the intact stiffnesses in all fixation groups. No difference was found in torsion. There was no difference in normalized stiffnesses between small-breed radii stabilized with CTLPs and large-breed radii stabilized with LCDCPs; however, small-breed radii stabilized with DCPs were less stiff than were any other group. Conclusions and Clinical Relevance—Plated radii of small-breed dogs had normalized stiffnesses equal to or less than plated radii of large-breed dogs. The complications typically associated with plating of radial fractures in small-breed dogs cannot be ascribed to an overly stiff bone-plate construct.

Objective--To investigate the roles of transforming growth factor-β (TGF-β) isoforms and matrix metalloproteinases (MMPs) in development of chronic mitral valvular disease (CMVD) in dogs. Sample Population--12 mitral valve leaflets collected from cadavers of 5 clinically normal dogs and from 7 dogs with CMVD. Procedures--Expression of TGF-β isoforms 1, 2, and 3; MMPs 1, 2, 3, and 9; TGF-β receptor II (TβR-II); and α smooth muscle actin (αSMA) in mitral valves of dogs with CMVD was compared with that in mitral valves from clinically normal dogs. Additionally, responses of valvular interstitial cells (VICs) to TGF-β3, MMP-3, and angiotensin-converting enzyme inhibitor (ACEI) as a suppressor of TGF-β3 were examined in vitro. Results--Expression of TGF-β3, TβR-II, αSMA, and MMP-3 was only detected in mitral valves of dogs with CMVD. Concentrations of αSMA and proteoglycans in cultured VICs were significantly increased following incubation with TGF-β3; treatment with MMP-3 resulted in increased amounts of active and total TGF-β3, and total TGF-β3 in VICs was significantly decreased by incubation with ACEI. Conclusions and Clinical Relevance--Findings suggested that increased TGF-β3 and MMP-3 contribute to the pathogenesis of valvular degeneration associated with CMVD. In addition, it is possible that the use of ACEI could effectively block pathological alterations in VICs associated with CMVD in vitro. Impact on Human Medicine--CMVD is associated with primary mitral valve prolapse and Marfan syndrome in humans. Results of the study reported here will help to elucidate the molecular mechanisms of CMVD in dogs and humans.

Objective: To obtain quantitative variables of the abdominal aorta and both kidneys on the basis of time-attenuation curves (TACs) and to measure glomerular filtration rate (GFR) for each kidney and the global GFR in clinically normal cats by use of dynamic computed tomography (CT) and Patlak analysis. Animals: 9 healthy cats. Procedures: All the cats were anesthetized with propofol. Anesthesia was maintained by administration of isoflurane, and CT examination was performed in the anesthetized cats. The TACs and renal volume were measured by use of the baseline precontrast and single-slice dynamic scans. The CT-GFR of each kidney and the global CT-GFRs were calculated via Patlak plot analysis. Results: CT-GFR results from 7 cats were valid. Peak aortic enhancement was detected between 9.0 and 14.0 seconds after iohexol injection, and the initial peak time of renal parenchymal enhancement was 15 to 24 seconds after iohexol injection. Mean ± SD global GFR was 2.06 ± 0.62 mL/min/kg. Mean ± SD CT-GFR of the right and left kidneys was 0.97 ± 0.32 mL/min/kg and 1.05 ± 0.31 mL/min/kg, respectively. Conclusions and Clinical Relevance: The CT-GFR method can be rapidly and conveniently performed in clinically normal cats. This combined structural-functional approach provided physiologic and morphological information on the kidneys of cats.

Changes in luteinizing hormone (LH) secretion after 17beta-estradiol (E2) injection were evaluated during sexual maturation in 10 prepubertal Nelore heifers. Heifers were divided into 2 groups: intact (I) and ovariectomized (OVX). 17beta-estradiol (2 micrograms/kg) was administered to both groups at 10, 13, and 17 mo of age. Only at 10 mo of age was there a greater mean LH concentration in OVX heifers (1.33 +/- 0.29 ng/mL) compared with the I group (0.57 +/- 0.15 ng/mL). At 13 and 17 mo of age there was no significant difference between the 2 groups in any of the evaluated variables (number of peaks, total peak area, greatest peak area, and time to greatest peak occurrence). This suggests a decrease in negative E2 feedback associated with an increase in positive feedback to LH secretion during sexual maturation, and these were likely the key factors that determined the time of first ovulation in Nelore heifers.

Objective—To evaluate the ability of industrial polystyrene foam insulation pads to redistribute loads placed on clinically normal weight-bearing structures of the foot and shift the location of the center of pressure palmarly in horses. Animals—25 nonlame mature horses. Procedures—Both forefeet from each horse were evaluated. Center of pressure data and solar load distribution patterns were recorded during a 5-second trial by use of a commercial pressure measurement system prior to placement of foam sole support and at 0, 6, 12, 24, and 48 hours after placement. Total contact surface area, contact pressure, peak contact pressure, and center of pressure positions were compared by use of a linear mixed model with repeated measurements. Results—Total contact surface area was increased significantly at all time points, whereas contact pressure and peak contact pressure were significantly decreased at all time points following application of foam sole supports. Immediately following application of sole support, the position of the center of pressure was significantly moved cranially. However, by 48 hours, the center of pressure was significantly positioned more palmarly than prior to application of the foam supports. Conclusions and Clinical Relevance—Results indicated that the use of foam sole supports may be an effective, economical, and immediate treatment for acute laminitis.

Objective—To identify and characterize cytochrome P450 enzymes (CYPs) responsible for the metabolism of racemic ketamine in 3 mammalian species in vitro by use of chemical inhibitors and antibodies. Sample—Human, canine, and equine liver microsomes and human single CYP3A4 and CYP2C9 and their canine orthologs. Procedures—Chemical inhibitors selective for human CYP enzymes and anti-CYP antibodies were incubated with racemic ketamine and liver microsomes or specific CYPs. Ketamine N-demethylation to norketamine was determined via enantioselective capillary electrophoresis. Results—The general CYP inhibitor 1-aminobenzotriazole almost completely blocked ketamine metabolism in human and canine liver microsomes but not in equine microsomes. Chemical inhibition of norketamine formation was dependent on inhibitor concentration in most circumstances. For all 3 species, inhibitors of CYP3A4, CYP2A6, CYP2C19, CYP2B6, and CYP2C9 diminished N-demethylation of ketamine. Anti-CYP3A4, anti-CYP2C9, and anti-CYP2B6 antibodies also inhibited ketamine N-demethylation. Chemical inhibition was strongest with inhibitors of CYP2A6 and CYP2C19 in canine and equine microsomes and with the CYP3A4 inhibitor in human microsomes. No significant contribution of CYP2D6 to ketamine biotransformation was observed. Although the human CYP2C9 inhibitor blocked ketamine N-demethylation completely in the canine ortholog CYP2C21, a strong inhibition was also obtained by the chemical inhibitors of CYP2C19 and CYP2B6. Ketamine N-demethylation was stereoselective in single human CYP3A4 and canine CYP2C21 enzymes. Conclusions and Clinical Relevance—Human-specific inhibitors of CYP2A6, CYP2C19, CYP3A4, CYP2B6, and CYP2C9 diminished ketamine N-demethylation in dogs and horses. To address drug-drug interactions in these animal species, investigations with single CYPs are needed.

Objective—To evaluate the elimination pharmacokinetics of a single IM injection of a long-acting ceftiofur preparation (ceftiofur crystalline-free acid [CCFA]) in healthy adult helmeted guineafowl (Numida meleagris). Animals—14 healthy adult guineafowl. Procedures—1 dose of CCFA (10 mg/kg) was administered IM to each of the guineafowl. Blood samples were collected intermittently via jugular venipuncture over a 144-hour period. Concentrations of ceftiofur and all desfuroylceftiofur metabolites were measured in plasma via high-performance liquid chromatography. Results—No adverse effects of drug administration or blood collection were observed in any bird. The minimal inhibitory concentration (MIC) for many bacterial pathogens of poultry and domestic ducks (1 μg/mL) was achieved by 1 hour after administration in most birds and by 2 hours in all birds. A maximum plasma concentration of 5.26 μg/mL was reached 19.3 hours after administration. Plasma concentrations remained higher than the MIC for at least 56 hours in all birds and for at least 72 hours in all but 2 birds. The harmonic mean ± pseudo-SD terminal half-life of ceftiofur was 29.0 ± 4.93 hours. The mean area under the curve was 306 ± 69.3 μg•h/mL, with a mean residence time of 52.0 ± 8.43 hours. Conclusions and Clinical Relevance—A dosage of 10 mg of CCFA/kg, IM, every 72 hours in helmeted guineafowl should provide a sufficient plasma drug concentration to inhibit growth of bacteria with an MIC ≤ 1 μg/mL. Clinical use should ideally be based on bacterial culture and antimicrobial susceptibility data and awareness that use of CCFA in avian patients constitutes extralabel use of this product.

Objective—To compare the toxic effects of a Delphinium occidentale chemotype containing -(methylsuccinimido) anthranoyllycoctonine (MSAL)—type alkaloids and a D occidentale chemotype lacking MSAL-type alkaloids in mice and cattle. Animals—225 male Swiss Webster mice and 11 Black Angus steers. Procedures—4 collections of larkspur containing MSAL-type alkaloids and 4 collections of larkspur lacking MSAL-type alkaloids were used. From each collection, total alkaloid extracts (0.05 to 0.20 mL) were administered via tail-vein injection in 27 to 29 mice. Dried, finely ground plant material from 1 collection with and 1 collection without MSAL-type alkaloids (doses equivalent to 37.6 mg of total alkaloids/kg) were each administered to 8 cattle via oral gavage in a crossover experiment; 3 cattle received a single dose equivalent to 150.4 mg of total alkaloids/kg (no MSAL-type alkaloids). In mice, clinical effects were monitored; in cattle, heart rate was monitored before (baseline) and 24 hours after treatment. At the 24-hour time point, cattle were exercised as a measure of muscle weakness. Results—In mice, mean LD50 associated with alkaloid extracts prepared from plants that did or did not contain MSAL-type alkaloids was 2.3 and 54.2 mg/kg, respectively. In cattle at 24 hours after treatment, plant material containing MSAL-type alkaloids significantly increased heart rate from baseline and was associated with exercise-induced collapse; plant material lacking MSAL-type alkaloids had no similar effects. Conclusions and Clinical Relevance—Taxonomic classification of D occidentale alone was not a good indicator of the toxic risk to grazing cattle.

Objective: To determine accuracy of the use of triaxial accelerometry for measuring daily activity as a predictor of maintenance energy requirement (MER) in healthy adult Labrador Retrievers. Animals—10 healthy adult Labrador Retrievers. Procedures: Dogs wore an accelerometer for two 2-week periods, with data on daily activity successfully collected for 24 to 26 days. These data, along with body weight, were used as independent variables in a multiple linear regression model to predict the dependent variable of daily MER. The predictive accuracy of the model was compared with that of a model that excluded activity. Dietary energy intake at a stated amount of body weight stability was used as an equivalent measure of MER in these analyses. Results: The multiple linear regression model that included body weight and daily activity as independent variables could be used to predict observed MER with a mean absolute error of 63.5 kcal and an SE of estimation of 94.3 kcal. Removing activity from the model reduced the predictive accuracy to a mean absolute error of 129.8 kcal and an SE of estimation of 165.4 kcal. Conclusions and Clinical Relevance: Use of triaxial accelerometers to provide an independent variable of daily activity yielded a marked improvement in predictive accuracy of the regression model, compared with that for a model that used only body weight. Improved accuracy in estimations of MER could be made for each dog if an accelerometer was used to record its daily activity.