Ouabain, a cardiac glycoside, binds to the Na+-K+i-adenosine triphosphatase (Na+ pump) and prevents active transport of Na+ and K+ across cell membranes. We used [3H]ouabain to quantify the number and affinity of Na+ pumps in skeletal muscle from Quarter Horses with the muscular disorder hyperkalemic periodic paralysis (HYPP). [3H]Ouabain-binding properties of gluteal muscle from clinically normal and affected horses were used to determine whether altered Na+ pump number or affinity could contribute to the pathologic features of muscle in affected horses. Foals and adult horses with HYPP were compared with age-matched clinically normal horses. The number of [3H]ouabain-binding sites in adult gluteal muscle was not different between the 2 types of horses (85.7 +/- 8.9 pmol of [3H]ouabain-binding sites/g [wet muscle weight] in horses with HYPP vs 100.2 +/- 8.8 pmol/g in clinically normal adult horses). Gluteal muscles in HYPP-affected and clinically normal foals also contained a similar number of [3H]ouabain-binding sites (222.3 +/- 21.0 pmol/g vs 225.3 +/- 24.2 pmol/g, respectively). The affinity of these binding sites for ouabain was not different, between adults or foals, in clinically normal or affected horses. Our results indicate that membrane events underlying the periodic episodes of paralysis in horses with HYPP are not attributable to quantitative changes in Na+ pump number or affinity. Our data cannot exclude the possibility that the specific activity of the Na+ pump is altered in muscle from HYPP-affected horses.

OBJECTIVE To evaluate the effect of bilateral ventriculocordectomy via ventral laryngotomy on laryngeal airway resistance (LAR) in canine cadaver larynges. SAMPLE 6 clinically normal canine cadaver larynges. PROCEDURES LAR was determined for each specimen before (baseline) and after bilateral ventriculocordectomy with the epiglottis open and closed. After ventral laryngotomy was performed, the vocal cords were sharply excised, and the incised mucosal edges were apposed with 4-0 glycomer 631 suture in a simple continuous pattern. The thyroid cartilage was apposed with 3-0 polypropylene suture in a simple continuous pattern. RESULTS With the epiglottis closed, baseline median LAR was 27.6 cm H2O/L/s (range, 21.2 to 30.6 cm H2O/L/s), which did not differ significantly from the median LAR after bilateral ventriculocordectomy (24.7 cm H2O/L/s [range, 20.6 to 27.7 cm H2O/L/s]). With the epiglottis open, baseline median LAR was 7.3 cm H2O/L/s (range, 5.4 to 7.8 cm H2O/L/s), which did not differ significantly from the median LAR after bilateral ventriculocordectomy (7.2 cm H2O/L/s [range, 6.6 to 7.6 cm H2O/L/s]). CONCLUSIONS AND CLINICAL RELEVANCE Bilateral ventriculocordectomy did not affect LAR with an open epiglottis in canine cadaver larynges. Therefore, it may not be an effective treatment for laryngeal paralysis. It also did not affect LAR with a closed epiglottis, which may indicate protection against aspiration pneumonia.

OBJECTIVE To evaluate gene expression and DNA copy number in adipose tissue-derived stromal cells (ADSCs) and in ADSC-derived neurosphere-like cell clusters (ADSC-NSCs) generated from tissues of chronically paraplegic dogs. ANIMALS 14 client-owned paraplegic dogs. PROCEDURES Dorsal subcutaneous adipose tissue (< 1 cm3) was collected under general anesthesia; ADSCs were isolated and cultured. Third-passage ADSCs were cultured in neural cell induction medium to generate ADSC-NSCs. Relative gene expression of mesenchymal cell surface marker CD90 and neural progenitor marker nestin was assessed in ADSCs and ADSC-NSCs from 3 dogs by quantitative real-time PCR assay; expression of these and various neural lineage genes was evaluated for the same dogs by reverse transcription PCR assay. Percentages of cells expressing CD90, nestin, glial fibrillary acidic protein (GFAP), and tubulin β 3 class III (TUJ1) proteins were determined by flow cytometry for all dogs. The DNA copy number stability (in samples from 6 dogs) and neural cell differentiation (14 dogs) were assessed with array-comparative genomic hybridization analysis and immunocytochemical evaluation, respectively. RESULTS ADSCs and ADSC-NSCs expressed neural cell progenitor and differentiation markers; GFAP and microtubule-associated protein 2 were expressed by ADSC-NSCs but not ADSCs. Relative gene expression of CD90 and nestin was subjectively higher in ADSC-NSCs than in ADSCs. Percentages of ADSC-NSCs expressing nestin, GFAP, and TUJ1 proteins were substantially higher than those of ADSCs. Cells expressing neuronal and glial markers were generated from ADSC-NSCs and had no DNA copy number instability detectable by the methods used. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested ADSCs can potentially be a safe and clinically relevant autologous source for canine neural progenitor cells. Further research is needed to verify these findings.

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.

Clinical, morphologic, and morphometric findings are reported in 14 young Dalmatians with laryngeal paralysis. Neurologic signs, including megaesophagus, were observed in 13 of 14 dogs. Electromyographic abnormalities included fibrillation potentials and positive sharp waves in laryngeal, esophageal, facial, and distal appendicular muscles. Neurogenic atrophy was detected in intrinsic laryngeal and appendicular skeletal muscles. A diffuse, generalized polyneuropathy, dominated by axonal degeneration, was observed in recurrent laryngeal and appendicular peripheral nerves. Results of quantitative studies, using single teased fiber and cross-sectional nerve preparations, indicated that changes were more severe in distal parts of peripheral nerves, with preferential loss of medium sized (5.5 to 8 micrometers) and large-caliber (8.5 to 12 micrometers) myelinated nerve fibers. Ultrastructural alterations were observed in myelinated and unmyelinated nerve fibers. The term laryngeal paralysis-polyneuropathy complex is proposed for this apparent dying-back disorder, which is clinically, electrophysiologically, and pathologically different from laryngeal paralysis in young Bouvier des Flandres and Siberian Huskies. Prognosis for Dalmatians with laryngeal paralysis-polyneuropathy complex is guarded to poor. The condition is believed to be inherited.

Ouabain, a cardiac glycoside, binds to the Na+-K+i-adenosine triphosphatase (Na+ pump) and prevents active transport of Na+ and K+ across cell membranes. We used [3H]ouabain to quantify the number and affinity of Na+ pumps in skeletal muscle from Quarter Horses with the muscular disorder hyperkalemic periodic paralysis (HYPP). [3H]Ouabain-binding properties of gluteal muscle from clinically normal and affected horses were used to determine whether altered Na+ pump number or affinity could contribute to the pathologic features of muscle in affected horses. Foals and adult horses with HYPP were compared with age-matched clinically normal horses. The number of [3H]ouabain-binding sites in adult gluteal muscle was not different between the 2 types of horses (85.7 +/- 8.9 pmol of [3H]ouabain-binding sites/g [wet muscle weight] in horses with HYPP vs 100.2 +/- 8.8 pmol/g in clinically normal adult horses). Gluteal muscles in HYPP-affected and clinically normal foals also contained a similar number of [3H]ouabain-binding sites (222.3 +/- 21.0 pmol/g vs 225.3 +/- 24.2 pmol/g, respectively). The affinity of these binding sites for ouabain was not different, between adults or foals, in clinically normal or affected horses. Our results indicate that membrane events underlying the periodic episodes of paralysis in horses with HYPP are not attributable to quantitative changes in Na+ pump number or affinity. Our data cannot exclude the possibility that the specific activity of the Na+ pump is altered in muscle from HYPP-affected horses.

A nerve muscle pedicle (NMP) graft was placed in the cricoarytenoideus dorsalis (CAD) muscle of 6 horses with induced left laryngeal hemiplegia. The NMP graft was created by use of the first cervical nerve and omohyoideus muscle. In 1 horse (control), the first cervical nerve was transected after placement of the NMP graft. One year after the surgical procedure, horses were examined endoscopically and then anesthetized. While the larynx was observed endoscopically, the first cervical nerve was stimulated. Horses were subsequently euthanatized, and the larynx was harvested. Prior to anesthesia, the endoscopic appearance of the larynx of all horses was typical of laryngeal hemiplegia. During anesthesia, stimulation of the first cervical nerve produced vigorous abduction of the left arytenoid in principal horses but not in the control horse. The right cricoarytenoideus lateralis and CAD muscles were grossly and histologically normal. Also, the left cricoarytenoideus lateralis was atrophic in all horses as was the left CAD muscle of the control horse. In contrast, the left CAD muscle harvested from principal horses had evidence of reinnervation with type 1 or type 2 fiber grouping. One year after the NMP graft procedure, horses with left laryngeal hemiplegia had reinnervation of the left CAD muscle. In another study, reinnervation was sufficient to allow normal laryngeal function during exercise. Combined, these data suggest that the NMP graft procedure is a viable technique for the treatment of left laryngeal hemiplegia in horses.