Objective-To evaluate subchondral bone density patterns in elbow joints of clinically normal dogs by use of computed tomographic (CT) osteoabsorptiometry. Sample Population-20 cadaver forelimbs from 10 clinically normal dogs. Procedure-Each elbow joint was imaged in parasagittal and transverse planes of 1.5-mm thickness. Slice data were converted to dipotassium phosphate equivalent density (PPED) values. Sagittal, parasagittal, and transverse medial coronoid process topographic maps were constructed. Defined zones were created for each of the 3 CT planes, and confluence and peak PPED values were determined. Results-The lowest PPED value was 340 mg/ml (articular and subchondral confluence), and the highest was 1780 mg/ml (peak subchondral density). Detectable effects of joint laterality were not found in the confluence or peak PPED measurements or in the peak-to-confluence PPED ratio for all 3 CT planes. Significant differences were found among zones in all 3 planes for confluence and peak PPED measurements and between sagittal and transverse planes for peak-to-confluence PPED ratios. Subjectively, the pattern of density distribution among dogs was fairly consistent for the sagittal and parasagittal slices. Three specific patterns of density distribution were apparent on the transverse topographic maps of the medial coronoid process that corresponded to conformational differences. Conclusions and Clinical Relevance-The use of CT osteoabsorptiometry provides a repeatable technique that can be used to noninvasively examine bone density and the effects of stress acting on joints in vivo. Variability in density values for any of the CT planes was not identified among clinically normal dogs.

Objectives-To evoke and measure the nociceptive withdrawal reflex (NWR) by use of electromyographic recordings and to score the behavioral nociceptive responses to electrical pulses in standing nonsedated horses. Animals-10 adult horses. Procedure-The lateral palmar digital nerve of the forelimb was transcutaneously stimulated, and surface electromyographic responses were recorded from the ulnaris lateralis, extensor carpi radialis, and common digital extensor muscles. Stimuli consisted of a 25-millisecond train of 5 constant-current pulses delivered by a computer-controlled stimulator. The 80- to 250-milliseconds poststimulation interval was analyzed to detect the NWR. The current intensity was increased in steps of 0.5 mA until the NWR threshold intensity (It) was reached. The stimulus at It was repeated twice. Latency and amplitude of the NWR, together with the behavioral reaction of horses, were analyzed. The latter was scored according to a scale from 0 (no reaction) to 5 (vigorous reaction). Finally, 3 suprathreshold stimuli at 1.2 × It were analyzed. Results-The median It to elicit NWR was 2.5 mA. Median onset latency of the NWR was 96.0 milliseconds at It and 89.6 milliseconds for suprathreshold stimuli. The amplitude of the reflexes was higher for suprathreshold stimulations, and behavioral reactions were slightly stronger when stimulus intensity increased. Conclusions and Clinical Relevance-Results of our study indicate that it is possible to record NWR in conscious standing horses, to define a reflex threshold, and to measure reflexes in response to increasing stimulus intensity.

Objective-To determine the ratio of ventral-to-dorsal transverse diameters between the wings of the sacrum on ventrodorsal radiographic views of the pelvis in large dogs and to validate the reliability of this morphometric analysis for functional interpretation. Sample Population-Pelvic specimens from 40 large-breed dogs and radiographs of 113 large-breed dogs. Procedure-In an anatomic and radiographic evaluation, the transverse dorsal diameter (TVDS) and transverse ventral diameter (TVV) between the wings of the sacrum were evaluated in sacrum specimens and on corresponding radiographs of the pelvis and sacrum. The ratio between TVV and TVDS (VD ratio) was calculated. Intraobserver reliability was determined by calculation of the coefficient of variation. In a retrospective radiographic evaluation, the VD ratio was determined in Rottweilers, Golden Retrievers, and German Shepherd Dogs. Correlations between VD ratio and breed, age, and sex were tested. Results-The VD ratio was significantly higher in Rottweilers than in Golden Retrievers and German Shepherd Dogs, denoting an oblique alignment of the sacral wings in Rottweilers (ie, the dorsal aspects of the sacral wings were located more medially than the ventral aspects) and an almost sagittal alignment in the other breeds. The VD ratio was significantly associated with age but not with sex. Conclusion and Clinical Relevance-Sagittal alignment of the wings of the sacrum is considered to be biomechanically less efficient. These results provide a basis for further studies to evaluate radiographic assessment of the sacroiliac joints similar to the evaluation for hip dysplasia.

Objective-To investigate whether upper airway sounds of horses exercising with laryngeal hemiplegia and alar fold paralysis have distinct sound characteristics, compared with unaffected horses. Animals-6 mature horses. Procedure-Upper airway sounds were recorded in horses exercising on a high-speed treadmill at maximum heart rate (HRMAX) under 3 treatment conditions (ie, normal upper airway function [control condition], and after induction of left laryngeal hemiplegia or bilateral alar fold paralysis) in a randomized crossover design. Fundamental frequency, spectrograms using Gabor transform, and intensity characteristics of acquired sounds (peak sound level [soundpeak] and highest frequency of at least –25 dB sound intensity [F25max]) were evaluated. Results-Evaluation of the fundamental frequency of the time domain signal was not useful. Sensitivity and specificity (83 and 75%, respectively) of spectrograms were greatest at maximal exercise, but the exact abnormal condition was identified in evaluation of only 12 of 18 spectrograms. Increased accuracy was obtained using soundpeak and F25max as discriminating variables. The use of soundpeak discriminated between control and laryngeal hemiplegia conditions and F25max between laryngeal hemiplegia and alar fold paralysis conditions. This increased the specificity of sound analysis to 92% (sensitivity 83%) and accurately classified the abnormal state in 92% of affected horses. Conclusions and Clinical Relevance-Sound analysis might be a useful adjunct to the diagnosis and evaluation of treatment of horses with upper airway obstruction, but would appear to require close attention to exercise intensity. Multiple measurements of recorded sounds might be needed to obtain sufficient accuracy for clinical use.

Objective-To determine the effects of orally administered glucosamine on concentrations of markers of bone and cartilage metabolism in Standardbred horses during race training. Animals-Twenty 16- to 20-month-old Standardbreds beginning race training. Procedure-Horses were randomly assigned to 2 groups. One group received glucosamine hydrochloride (4 g, PO, q 12 h), and the second (control) group received glucose (4 g, PO, q 12 h). Serum samples were obtained prior to onset of the study (baseline) and at regular intervals for 48 weeks for determination of concentrations of keratan sulfate (KS), osteocalcin (OC), and pyridinoline crosslinks (PYD). Results-Osteocalcin concentrations changed significantly with time; mean serum concentrations were significantly higher than baseline values for samples obtained at 24 to 48 weeks after onset of the study. Although a significant effect of time was observed for mean concentration of KS, concentrations did not differ significantly from baseline values at any time during the study when groups were analyzed separately. However, pooled analysis revealed significant increases of mean serum KS concentration at weeks 24 and 30. Significant changes in serum PYD concentrations were not detected. Oral administration of glucosamine did not significantly affect serum concentrations of any of the markers. Conclusions and Clinical Relevance-Increased serum OC in clinically normal Standardbreds during race training may reflect bone formation that accompanies adaptive remodeling of the appendicular skeleton. For these experimental conditions, glucosamine did not appear to exert a detectable influence on serum concentrations of these 3 markers of connective tissue metabolism.

Objective-To study the antiviral activity of genistein, a soya isoflavone, on in vitro replication of bovine herpesvirus type 1 (BHV-1). Sample Population-Madin-Darby bovine kidney (MDBK) cells. Procedure-Effects of genistein on the magnitude and kinetics of inhibition of BHV-1 phosphorylation of glycoprotein E (gE) and in vitro replication of BHV-1 in MDBK cells were evaluated. Antiviral activity of genistein was compared with 2 compounds, estradiol-17β (EST) and tamoxifen (TAM), that have estrogenic and antiestrogenic activity, respectively. High-performance liquid chromatography (HPLC) was used to determine the concentration of genistein in medium from infected and uninfected MDBK cultures. Results-Genistein reduced BHV-1, but not gE-deleted BHV-1 (BHV-1gEΔ3.1), replication by 90% at 18 hours after inoculation. This inhibition was not sustained through 24 hours after inoculation. The genistein concentration in media from MDBK cells was decreased by 40% during BHV-1 infection, compared with 16% for uninfected cells, at 24 hours after inoculation. Genistein inhibited gE phosphorylation and BHV- 1 replication in a dose-dependent manner. Dosing with 25 µMgenistein at 0 and 12 hours after inoculation of BHV-1 was optimal for decreasing BHV-1 replication. Estradiol-17β EST and TAM did not affect BHV-1 replication. Conclusions and Clinical Relevance-The decrease in genistein concentration was a viral infection-dependent event. Genistein is an inhibitor of BHV-1 replication because of its ability to inhibit tyrosine kinase activity. A possible application may be for the control of BHV-1 infection in cattle by feeding soya products rich in genistein prior to or during periods of stress.

Objective-To determine significant molecular and cellular factors responsible for differences in secondintention healing in thoracic and metacarpal wounds of horses. Animals-6 adult mixed-breed horses. Procedure-A full-thickness skin wound on the metacarpus and another such wound on the pectoral region were created, photographed, and measured, and tissue was harvested from these sites weekly for 4 weeks. Gene expression of type-I collagen, transforming growth factor (TGF)-β1, matrix metalloproteinase (MMP)-1, and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by quantitative in situ hybridization. Myofibroblasts were detected by immunohistochemical labeling with α-smooth muscle actin (α-SMA). Collagen accumulation was detected by use of picrosirius red staining. Tissue morphology was examined by use of H&E staining. Results-Unlike thoracic wounds, forelimb wounds enlarged during the first 2 weeks. Myofibroblasts, detected by week 1, remained abundant with superior organization in thoracic wounds. Type-I collagen mRNA accumulated progressively in both wounds. More type-I collagen and TGF-β1 mRNA were seen in forelimb wounds. Volume of MMP-1 mRNA decreased from day 0 in both wounds. By week 3, TIMP-1 mRNA concentration was greater in thoracic wounds. Conclusions and Clinical Relevance-Greater collagen synthesis in metacarpal than thoracic wounds was documented by increased concentrations of myofibroblasts, type-I collagen mRNA, TGF-β1 mRNA, and decreased collagen degradation (ie, MMP-1). Imbalanced collagen synthesis and degradation likely correlate with development of exuberant granulation tissue, delaying healing in wounds of the distal portions of the limbs. Factors that inhibit collagen synthesis or stimulate collagenase may provide treatment options for horses with exuberant granulation tissue.

Objective-To determine the effects of etodolac administration on results of thyroid function tests and concentrations of plasma proteins in clinically normal dogs. Animals-19 healthy random-source mixed-breed dogs. Procedure-Blood samples for measurement of serum thyroxine (T4), 3,5,3'-triiodothyronine (T3), free T4 (fT4), and endogenous canine thyroid stimulating hormone (cTSH) were measured twice before as well as on days 14 and 28 of etodolac administration (mean dosage, 13.7 mg/kg, PO, q 24 h). Plasma total protein, albumin, and globulin concentrations and serum osmolality were measured once before as well as on days 14 and 28 of etodolac administration. Results-Etodolac administration did not significantly affect serum T4, T3, fT4, or cTSH concentrations or serum osmolality. Significant decreases in plasma total protein, albumin, and globulin concentrations were detected on days 14 and 28 of administration. Conclusions and Clinical Relevance-Results of thyroid function tests are not altered when etodolac is administered for up to 4 weeks. Therefore, interpretation of results of these tests should accurately reflect thyroid function during etodolac treatment. Plasma total protein, albumin, or globulin concentrations that are less than the respective reference range in a dog administered etodolac for greater than 2 weeks may be an effect of treatment rather than an unrelated disease process. A decrease in plasma protein concentrations may reflect subclinical injury of the gastrointestinal tract.