OBJECTIVE To assess effects of speed on kinematic variables measured by use of extremity-mounted inertial measurement units (IMUs) in nonlame horses performing controlled exercise on a treadmill. ANIMALS 10 nonlame horses. PROCEDURES 6 IMUs were attached at predetermined locations on 10 nonlame Franches Montagnes horses. Data were collected in triplicate during trotting at 3.33 and 3.88 m/s on a high-speed treadmill. Thirty-three selected kinematic variables were analyzed. Repeated-measures ANOVA was used to assess the effect of speed. RESULTS Significant differences between the 2 speeds were detected for most temporal (11/14) and spatial (12/19) variables. The observed spatial and temporal changes would translate into a gait for the higher speed characterized by increased stride length, protraction and retraction, flexion and extension, mediolateral movement of the tibia, and symmetry, but with similar temporal variables and a reduction in stride duration. However, even though the tibia coronal range of motion was significantly different between speeds, the high degree of variability raised concerns about whether these changes were clinically relevant. For some variables, the lower trotting speed apparently was associated with more variability than was the higher trotting speed. CONCLUSIONS AND CLINICAL RELEVANCE At a higher trotting speed, horses moved in the same manner (eg, the temporal events investigated occurred at the same relative time within the stride). However, from a spatial perspective, horses moved with greater action of the segments evaluated. The detected changes in kinematic variables indicated that trotting speed should be controlled or kept constant during gait evaluation.

OBJECTIVE To assess insulin, glucagon, and somatostatin expression within pancreatic islets of horses with and without insulin resistance. ANIMALS 10 insulin-resistant horses and 13 insulin-sensitive horses. PROCEDURES For each horse, food was withheld for at least 10 hours before a blood sample was collected for determination of serum insulin concentration. Horses with a serum insulin concentration < 20 μU/mL were assigned to the insulin-sensitive group, whereas horses with a serum insulin concentration > 20 μU/mL underwent a frequently sampled IV glucose tolerance test to determine sensitivity to insulin by minimal model analysis. Horses with a sensitivity to insulin < 1.0 × 10(−4) L•min−1•mU−1 were assigned to the insulin-resistant group. All horses were euthanized with a barbiturate overdose, and pancreatic specimens were harvested and immunohistochemically stained for determination of insulin, glucagon, and somatostatin expression in pancreatic islets. Islet hormone expression was compared between insulin-resistant and insulin-sensitive horses. RESULTS Cells expressing insulin, glucagon, and somatostatin made up approximately 62%, 12%, and 7%, respectively, of pancreatic islet cells in insulin-resistant horses and 64%, 18%, and 9%, respectively, of pancreatic islet cells in insulin-sensitive horses. Expression of insulin and somatostatin did not differ between insulin-resistant and insulin-sensitive horses, but the median percentage of glucagon-expressing cells in the islets of insulin-resistant horses was significantly less than that in insulin-sensitive horses. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that, in insulin-resistant horses, insulin secretion was not increased but glucagon production might be downregulated as a compensatory response to hyperinsulinemia.

OBJECTIVE To characterize expression profiles of circulating microRNAs via genome-wide sequencing for dogs with congestive heart failure (CHF) secondary to myxomatous mitral valve degeneration (MMVD). ANIMALS 9 healthy client-owned dogs and 8 age-matched client-owned dogs with CHF secondary to MMVD. PROCEDURES Blood samples were collected before administering cardiac medications for the management of CHF. Isolated microRNAs from plasma were classified into microRNA libraries and subjected to next-generation sequencing (NGS) for genome-wide sequencing analysis and quantification of circulating microRNAs. Quantitative reverse transcription PCR (qRT-PCR) assays were used to validate expression profiles of differentially expressed circulating microRNAs identified from NGS analysis of dogs with CHF. RESULTS 326 microRNAs were identified with NGS analysis. Hierarchical analysis revealed distinct expression patterns of circulating microRNAs between healthy dogs and dogs with CHF. Results of qRT-PCR assays confirmed upregulation of 4 microRNAs (miR-133, miR-1, miR-let-7e, and miR-125) and downregulation of 4 selected microRNAs (miR-30c, miR-128, miR-142, and miR-423). Results of qRT-PCR assays were highly correlated with NGS data and supported the specificity of circulating microRNA expression profiles in dogs with CHF secondary to MMVD. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested that circulating microRNA expression patterns were unique and could serve as molecular biomarkers of CHF in dogs with MMVD.

OBJECTIVE To determine the in vitro effect of calcitriol on indicators of immune system function in endotoxin-primed blood samples from healthy dogs. SAMPLE Blood samples from 6 healthy adult dogs. PROCEDURES Leukocytes were primed by incubation of blood samples with lipopolysaccharide (LPS; endotoxin) or PBS solution (unprimed control group) for 1 hour. Following priming, blood samples were incubated with calcitriol (2 × 10(−7)M) or ethanol (control substance) for 24 hours. After sample incubation, LPS-stimulated leukocyte production of tumor necrosis factor (TNF) and interleukin-10 (IL10) was measured with a canine-specific multiplex assay, and apoptosis and toll-like receptor 4 (TLR4) expression were evaluated via flow cytometry. RESULTS LPS stimulation of unprimed leukocytes but not endotoxin-primed leukocytes resulted in a significant increase in TNF and IL10 production, confirming the presence of endotoxin tolerance in dogs in vitro. Endotoxin priming significantly increased neutrophil viability with no effect on lymphocyte viability or TLR4 expression by neutrophils and monocytes. Calcitriol exposure significantly decreased LPS-stimulated production of TNF by unprimed and endotoxin-primed leukocytes. Conversely, calcitriol exposure had no effect on IL10 production by unprimed leukocytes but did significantly increase IL10 production by endotoxin-primed leukocytes. Calcitriol had no significant effect on the degree of neutrophil or lymphocyte apoptosis, nor was neutrophil and monocyte TLR4 expression affected in unprimed or endotoxin-primed leukocytes. CONCLUSIONS AND CLINICAL RELEVANCE These data indicated that calcitriol induced an anti-inflammatory shift in unprimed and endotoxin-primed canine leukocytes in vitro, without compromising neutrophil and monocyte TLR4 expression or altering the viability of neutrophils and lymphocytes in canine blood samples.

OBJECTIVE To quantify fatigue-induced electromyographic changes in hind limb muscles in horses. ANIMALS 8 Thoroughbreds. PROCEDURES The left and right hind limb longissimus dorsi, tensor fasciae latae, gluteus medius, and biceps femoris muscles were instrumented for surface electromyography. Hoof strain gauges were attached to confirm stride cycle. Each horse was galloped on a treadmill (grade, 3%) at a constant speed (12.6 to 14.7 m/s) to achieve fatigue after approximately 360 seconds. Before and after this exercise, the horses were trotted at 3.5 m/s. At 30-second intervals during galloping an integrated electromyography (iEMG) value for a stride and the median frequency of muscle discharge (MF) in each limb were measured. The mean of stride frequency (SF), iEMG value, and MF of 5 consecutive strides at the start and end of galloping for the lead and trailing limbs were compared. For trotting, these variables were compared at 60 seconds before and after galloping. RESULTS The mean ± SD value for SF decreased over time (2.14 ± 0.06 to 2.05 ± 0.07 stride/s). In both the lead and trailing limbs, fatigue decreased the iEMG values of the gluteus medius and biceps femoris muscles but not those of the longissimus dorsi and tensor fasciae latae muscles. The MF did not change for any muscle during galloping with fatigue. The SF, iEMG value, and MF did not change during trotting with fatigue. CONCLUSIONS AND CLINICAL RELEVANCE Fatigue induced by high-speed galloping decreased the gluteus medius and biceps femoris muscles' iEMG values in Thoroughbreds. Fatigue of these less fatigue-resistant hind limb muscles would affect a horse's speed.

OBJECTIVE To evaluate and compare surface and cross-sectional structure as well as localized electrochemical corrosion and ion release for cast stainless steel (SS) tibia plateau leveling osteotomy (TPLO) plates retrieved from dogs with and without osteosarcoma (OSA) and to compare these findings with similar variables for forged SS TPLO plates retrieved from dogs. SAMPLE 47 TPLO plates explanted from 45 client-owned dogs (22 cast plates from dogs with OSA, 22 cast plates from dogs without OSA, and 3 forged plates from dogs without OSA). PROCEDURES Histologic evaluations of tissue samples collected from implant sites at the time of plate retrieval were performed to confirm implant site tumor status of each dog. Surfaces and metallographic cross sections of retrieved plates were examined, and the microcell technique was used to obtain local electrochemical corrosion and ion release measurements. RESULTS Findings indicated that all cast SS plates demonstrated high spatial variability of their electrochemical surface properties and inhomogeneous superficial and cross-sectional composition, compared with forged plates. Greater metal ion release was observed in cast plates than in forged plates and in cast plates from dogs with OSA than in cast or forged from dogs without OSA. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that accumulation of metal ions from implants could be a trigger for neoplastic transformation in neighboring cells. Metal ion release caused by corrosion of implants that do not comply with recommended standards of the American Society for Testing and Materials International or the International Organization for Standardization could potentially place patients at increased risk of tumor development.

The purpose of this study was to determine the pharmacokinetics of cannabidiol (CBD) in healthy dogs. Thirty, healthy research dogs were assigned to receive 1 of 3 formulations (oral microencapsulated oil beads, oral CBD-infused oil, or CBD-infused transdermal cream), at a dose of 75 mg or 150 mg q12h for 6 wk. Serial cannabidiol plasma concentrations were measured over the first 12 h and repeated at 2, 4, and 6 wk. Higher systemic exposures were observed with the oral CBD-infused oil formulation and the half-life after a 75-mg and 150-mg dose was 199.7 ± 55.9 and 127.5 ± 32.2 min, respectively. Exposure is dose-proportional and the oral CBD-infused oil provides the most favorable pharmacokinetic profile.

OBJECTIVE To compare sedation in cockatiels (Nymphicus hollandicus) after intranasal administration of midazolam and midazolam-butorphanol. ANIMALS 9 healthy adult cockatiels. PROCEDURES A randomized, controlled, blinded, complete crossover study was conducted. Birds were assigned to 3 treatment groups. Midazolam (3 mg/kg), midazolam-butorphanol (3 mg/kg for each drug), or sterile saline (0.9% NaCl) solution (control treatment) was administered intranasally. Sedation quality was assessed at 3 time points by use of eye and body position; response to visual, auditory, and tactile stimulation; and response during manual restraint on the basis of eye position and struggling intensity. To evaluate attenuation of the manual restraint–induced stress response, heart rate, respiratory rate, and cloacal temperature were measured over a 15-minute period. Treatments were repeated after a minimum washout period of 7 days. RESULTS Median onset of first sedation effects was 85 seconds (range, 60 to 120 seconds) for midazolam and 90 seconds (range, 45 to 180 seconds) for midazolam-butorphanol. Midazolam-butorphanol resulted in significantly less vigorous struggling during restraint than did midazolam or the control treatment. Heart rate did not differ significantly among treatments. The stress-induced increase in respiratory rate was significantly attenuated by midazolam and midazolam-butorphanol, whereas the increase in cloacal temperature was not attenuated by midazolam or midazolam-butorphanol. CONCLUSIONS AND CLINICAL RELEVANCE Intranasal administration of midazolam and midazolam-butorphanol resulted in a rapid onset of sedation in cockatiels. Midazolam-butorphanol resulted in deeper sedation in both restrained and unrestrained birds than did midazolam alone. Midazolam and midazolam-butorphanol both provided safe and effective sedation in cockatiels.

OBJECTIVE To evaluate effects of various concentrations of collagenase and dimethyl sulfoxide (DMSO) on yield of equine adipose-derived multipotent stromal cells (ASCs) before and after cryopreservation. SAMPLE Supragluteal subcutaneous adipose tissue from 7 Thoroughbreds. PROCEDURES Tissues were incubated with digests containing 0.1%, 0.05%, or 0.025% type I collagenase. Part of each resulting stromal vascular fraction was cryopreserved in 80% fetal bovine serum (FBS), 10% DMSO, and 10% Dulbecco modified Eagle medium F-12 and in 95% FBS and 5% DMSO. Half of each fresh and cryopreserved heterogeneous cell population was not immunophenotyped (unsorted) or was immunophenotyped for CD44+, CD105+, and major histocompatability complex class II (MHCII; CD44+-CD105+-MHCII+ cells and CD44+-CD105+-MHCII− cells). Cell proliferation (cell viability assay), plasticity (CFU frequency), and lineage-specific target gene and oncogene expression (reverse transcriptase PCR assays) were determined in passage 1 cells before and after culture in induction media. RESULTS Digestion with 0.1% collagenase yielded the highest number of nucleated cells. Cell surface marker expression and proliferation rate were not affected by collagenase concentration. Cryopreservation reduced cell expansion rate and CD44+-CD105+-MHCII− CFUs; it also reduced osteogenic plasticity of unsorted cells. However, effects appeared to be unrelated to DMSO concentrations. There were also variable effects on primordial gene expression among cell isolates. CONCLUSIONS AND CLINICAL RELEVANCE Results supported the use of 0.1% collagenase in an adipose tissue digest and 5% DMSO in cryopreservation medium for isolation and cryopreservation, respectively, of equine ASCs. These results may be used as guidelines for standardization of isolation and cryopreservation procedures for equine ASCs.

OBJECTIVE To develop contact time (ConT) and withers height-normalized relative ConT (ConT*) for force platform gait analysis of dogs. ANIMALS 29 healthy client-owned dogs. PROCEDURES Height at the most dorsal aspect of the shoulders (withers) was measured with a framing square. Dogs were trotted across a force platform at their preferred velocity with controlled acceleration (± 0.5 m/s2). Ranges of ConT and ConT* centered on the population mean ConT were created. Variance effects on ground reaction forces (GRFs) for 4 thoracic limb and 4 pelvic limb ConT and associated ConT* ranges were examined. Efficiency of trial capture and effects of velocity ranges on GRF variance were determined. RESULTS Individual dogs had the greatest effect on GRF variance for thoracic and pelvic limbs. Narrow ConT and ConT* ranges had few significant effects on GRFs but were inefficient at capturing trials. The ConT ranges of 0.22 to 0.29 seconds and 0.19 to 0.25 seconds for thoracic and pelvic limbs, respectively, provided the most efficient rates of trial capture with the fewest significant effects on GRFs. Compared with ConT and ConT* ranges, relative velocity ranges had higher efficiency and smaller GRF variance effects. CONCLUSIONS AND CLINICAL RELEVANCE Dogs of various morphologies have differing limb velocities. Use of ConT as a surrogate for limb velocity may improve GRF data quality. We identified ConT and ConT* ranges associated with low GRF variance. However, relative velocity ranges captured data more efficiently. Efficient capture of data may help avoid worsening of lameness during gait analysis of dogs.