While heart failure is a primary cause of death for many in-transit-loss (ITL) pigs, the underlying cause of these deaths is not known. Cardiomyopathies are considered a common cause of heart failure in humans and often have a genetic component. The objective of this study was to determine if genes associated with cardiomyopathies could be identified in ITL pigs. Samples from the hearts of pigs that died during transport to an abattoir in Ontario, Canada were collected and genotyped along with samples from pigs that did not die during transport (ILT hearts: n = 149; non-ITL/control hearts: n = 387). Genome-wide analyses were carried out on each of the determined phenotypes (gross cardiac lesions) using a medium density single nucleotide polymorphism (SNP) chip and 500 kb windows/regions for analysis, with 250 kb regions of overlap. The distribution derived by a multidimensional scaling (MDS) analysis of all phenotypes demonstrated a lack of complete separation between phenotypes of affected and unaffected animals, which made diagnosis difficult. Although genetic differences were small, a few genes associated with dilated cardiomyopathy (DCM) and arrhythmogenic right ventricular cardiomyopathy (ARVM) were identified. In addition, multiple genes associated with cardiac arrhythmias and ventricular hypertrophy were identified that can possibly result in heart failure. The results of this preliminary study did not provide convincing evidence that a single, heritable cardiomyopathy is the cause of heart failure in ITL pigs.

OBJECTIVE To investigate the effects of recombinant equine IL-1β on function of equine endothelial colony-forming cells (ECFCs) in vitro. SAMPLE ECFCs derived from peripheral blood samples of 3 healthy adult geldings. PROCEDURES Function testing was performed to assess in vitro wound healing, tubule formation, cell adhesion, and uptake of 1,1′-dioctadecyl-3,3,3′,3′ tetramethylindocarbocyanine perchlorate–labeled acetylated low-density lipoprotein (DiI-Ac-LDL) by cultured ECFCs. Cell proliferation was determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. Effects on function test results of different concentrations and exposure times of recombinant equine IL-1β were assessed. RESULTS Challenge of cultured ECFCs with IL-1β for 48 hours inhibited tubule formation. Continuous challenge (54 hours) with IL-1β in the wound healing assay reduced gap closure. The IL-1β exposure did not significantly affect ECFC adhesion, DiI-Ac-LDL uptake, or ECFC proliferation. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested a role for IL-1β in the inhibition of ECFC function in vitro. Functional changes in ECFCs following challenge with IL-1β did not appear to be due to changes in cell proliferative capacity. These findings have implications for designing microenvironments for and optimizing therapeutic effects of ECFCs used to treat ischemic diseases in horses.

OBJECTIVE To develop a technique for isolation and culture of canine insulinoma cells and assess expression of cellular hexokinases (glucokinase and hexokinase I) and expression and secretion of insulin from these cells in vitro. SAMPLE Pancreatic insulinomas and normal pancreatic tissue from 4 and 3 dogs, respectively. PROCEDURES Tissues were collected by surgical excision or at necropsy. Insulinoma cells from 2 dogs were cultured for up to 10 weeks with standard techniques; insulin synthesis in vitro was confirmed by immunohistochemical analysis of freshly prepared slides of cultured cells, and insulin secretion was assessed by measurement of insulin concentrations in culture medium with an ultrasensitive mouse insulin ELISA. Expression of cellular hexokinases in insulinomas and adjacent normal (nontumor) pancreatic tissue from the same dog (n = 3) was examined by quantitative reverse transcriptase PCR assay. RESULTS Insulinoma cells survived for up to 10 weeks but did not proliferate in culture. Insulin was detected in isolated cells and secreted into culture medium for up to 10 weeks. Both cellular hexokinases were expressed; glucokinase appeared to be overexpressed in insulinomas, compared with normal pancreatic tissue from the same dogs. CONCLUSIONS AND CLINICAL RELEVANCE Canine insulinomas expressed hexokinases responsible for glucose responsiveness. Insulinoma cells were successfully maintained in short-term culture; cultured cells remained functional for 10 weeks as evidenced by cellular insulin content and had detectable secretion of insulin into the culture medium for ≥ 5 weeks. Apparent glucokinase overexpression by insulinomas suggested a possible mechanism underlying excessive insulin release by these tumors.

OBJECTIVE To determine the effect of epitendinous suture (ES) caliber on the tensile strength of flexor tendon repairs in cadaveric specimens from dogs. SAMPLE 60 cadaveric superficial digital flexor tendons (SDFTs) from 30 skeletally mature dogs. PROCEDURES Specimens were randomly assigned to 5 suture caliber groups (n = 12 SDFTs/group). After sharp transection, SDFTs were repaired by placement of a simple continuous circumferential ES created with size-0, 2–0, 3–0, 4–0, or 5–0 polypropylene suture. Constructs were preloaded to 2 N and load tested to failure. Loads at yield, peak, and failure and mode of failure were compared among groups by statistical methods. RESULTS Yield, peak, and failure loads for SDFT repair constructs were positively correlated with ES caliber and did not differ between the size-0 and 2–0 groups on pairwise comparisons. Yield load was significantly greater for size-0, 2–0, and 3–0 groups than for the 4–0 and 5–0 groups. Peak and failure loads were significantly greater for the size-0 and 2–0 groups than for the remaining groups. Most size-0 (12/12), 2–0 (12/12), and 3–0 (10/12) group constructs failed because of ES pull-through; several constructs in the 4–0 group (5/12) and most in the 5–0 group (11/12) failed because of ES breakage. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested size-0 and 2–0 sutures should be considered when placing an ES for flexor tendon repairs in dogs. However, in vivo studies are needed determine the effects of increasing ES caliber on clinical outcomes for dogs undergoing these procedures.

OBJECTIVE To evaluate synoviocentesis of the equine forelimb digital flexor tendon sheath (DFTS) via a basilar sesamoidean approach (BSA) or distal approach (DA). ANIMALS 21 healthy adult horses without DFTS-related lameness. PROCEDURES The forelimbs of each horse underwent the BSA or DA (21 limbs/approach) performed by 1 individual. The volume of synovial fluid (SF) aspirated, time from skin puncture to collection of SF, and number of attempts to place a needle in the DFTS were compared between approaches. RESULTS An SF sample was successfully aspirated from 16 of 21 (76%) limbs with the BSA and 20 of 21 (95%) limbs with the DA. For the BSA and DA, the number of attempts to obtain SF was 2 and 1, respectively; the median volume of SF obtained was 0.4 and 0.7 mL, respectively; and the median time to SF collection was 17.91 and 18.48 seconds, respectively. Between the approaches, the number of limbs with SF successfully aspirated and number of attempts to collect SF differed significantly, whereas the volume of SF aspirated and time to SF collection did not. CONCLUSIONS AND CLINICAL RELEVANCE Regarding SF collection from forelimb DFTSs in horses without DFTS-related disease, use of the DA had a greater success rate with fewer attempts, compared with findings for the BSA, which may reflect the relative ease of identifying anatomic landmarks for the DA. Results suggested that a DA for DFTS synoviocentesis in horses appears efficient and effective and may minimize limb trauma by requiring fewer attempts for SF sample collection, compared with a BSA.

OBJECTIVE To compare intraoperative and short-term postoperative variables pertaining to laparoscopic ovariectomy (LapOVE) and open ovariectomy (OVE) in rabbits (Oryctolagus cuniculus). ANIMALS Twelve 4− to 5-month-old female New Zealand White rabbits. PROCEDURES Rabbits were randomly assigned to undergo LapOVE (n = 6) or OVE (6), with a vessel-sealing device used to seal and transect the ovarian pedicles. Laparoscopic ovariectomy was performed with a 3-port approach. Variables were measured during surgery (surgery and anesthesia times and incision lengths) and for up to 7 days after surgery (food consumption, feces production, body weight, vital parameters, blood glucose and cortisol concentrations, abdominal palpation findings, facial grimace scale scores, and ethograms). RESULTS Mean surgery (43.2 vs 21.7 minutes) and anesthesia (76.2 vs 48.8 minutes) times were longer and mean incision length was shorter (24.0 vs 41.5 mm) for LapOVE versus OVE. No significant differences in postoperative variables were identified between groups. During LapOVE, small intestinal perforation occurred in 1 rabbit, which was then euthanized. Postoperative complications for the remaining rabbits included superficial incisional dehiscence (LapOVE, 1/5; OVE, 2/6), subcutaneous emphysema (LapOVE, 1/5; OVE, 0/6), and seroma formation (LapOVE, 1/5; OVE, 0/6). CONCLUSIONS AND CLINICAL RELEVANCE Surgery time for LapOVE was twice that of OVE, and LapOVE resulted in unique complications in rabbits. No evidence of a reduction in pain or faster return to baseline physiologic status was found for LapOVE. Further evaluation of LapOVE in rabbits is warranted, with modification to techniques used in this study or a larger sample size.

OBJECTIVE To evaluate stiffness of the liver parenchyma in healthy adult cats by means of point shear wave elastography (PSWE). ANIMALS 18 client-owned adult (1- to 6-year-old) healthy cats. PROCEDURES Echogenicity and echotexture of the liver parenchyma were assessed by means of conventional B-mode ultrasonography. The shear wave velocity (Vs) of the right and left portions of the liver were measured by means of PSWE. RESULTS B-mode ultrasonography revealed no abnormalities in echotexture or echogenicity of the liver parenchyma in any cat. Mean (95% CI) Vs in the liver parenchyma was 1.46 m/s (1.36 to 1.55 m/s) for the right portion, 1.36 m/s (1.26 to 1.47 m/s) for the left portion, and 1.43 m/s (1.35 to 1.51 m/s) overall. The difference in mean Vs between the 2 portions of the liver was significant. No significant correlation was found between Vs and body weight or between Vs and the depth at which this variable was measured. CONCLUSIONS AND CLINICAL RELEVANCE Quantitative PSWE of the liver was feasible in healthy adult cats. The obtained values for Vs may be useful for interpretation of and comparison with values measured in cats with liver disease. Additional research is needed to explore the potential usefulness of PSWE for diagnostic purposes.

The cross-pin technique for the treatment of distal femoral physis fractures (specifically, Salter-Harris Type I fractures) was investigated using femurs collected from beagle cadavers. The pin was inserted from the medial surface of the femur at an inclination of approximately 30 to 45° relative to the long axis of the femur in the anteroposterior direction; the pin exit was set proximal to the origin of the long digital extensor tendon. Digital and radiographic images of the femur in the anteroposterior and lateral directions were obtained. In both types of images, the insertion angle of the pin relative to the long axis was measured. Results suggest that when inserting a pin proximal to the fracture line, the ideal position can be achieved by inclining the pin approximately 20° cranially relative to the long axis of the lateral direction of the femur, in addition to the previously described criteria.

Bone repair in horses implies invasive surgeries and increased cost. Research on musculoskeletal disorders therapy in horses includes cell-based therapy with mesenchymal stromal cells (MSCs). Mesenchymal stromal cells can be obtained from bone marrow (BMMSCs). Unfortunately, BMMSCs have limited cell replication in vitro. The objective of this study was to develop a biologically immortalized equine stem cell line derived from bone marrow, with unlimited in-vitro proliferation and the ability to differentiate into bone cells. Equine BMMSCs were transfected and immortalized with human telomerase reverse transcriptase (hTERT) gene. Cell passages from equine immortal BMMSCs were characterized by the presence of stemness CD markers and expression of multi-potent differentiation genes (OCT-4, SOX2, and NANOG). Equine immortal BMMSCs were incubated in osteogenic medium and bone cell differentiation was determined by alkaline phosphatase and von Kossa staining, and osteogenic gene expression (osteocalcin, Runx2, and osterix). Telomerase activity was determined by telomeric repeat amplification technique. Results showed that equine immortal BMMSCs were able to replicate in-vitro up to passage 50 and maintain stem cell characteristics by the presence of CD90 and expression of multi-potent genes. Equine immortal BMMSCs were able to differentiate into bone cells, which was confirmed by the positive osteogenic staining and gene expression. Equine BMMSCs were successfully immortalized and maintained characteristics of stem cells and readily differentiated into osteogenic cells. Extending the life span of equine BMMSCs by transfection of the hTERT gene will revolutionize the clinical use of MSCs by making them available to orthopedic surgeons "off the shelf."

OBJECTIVE To investigate the effects of novel legwear designed to limit metacarpophalangeal joint (MCPJ) extension and redirect loading forces from the flexor apparatus through analyses of 2-D kinematic and kinetic data. ANIMALS 6 adult horses without musculoskeletal disease. PROCEDURES Horses were subjected to 4 treatments: control (no legwear), inactive legwear (unlimited legwear extension), and active legwear with mild (30°) and moderate (20°) legwear extension limitation. Two-dimensional kinematic data were collected for the right forelimb (FL) during walk and trot and from leading and trailing FLs during canter on a treadmill. Ground reaction force (GRF) data were collected from FLs during overground walk and trot. Peak MCPJ angle and angular velocity were calculated from kinematic data, and peak force and average loading rate were calculated from vertical GRF data during the stance phase of the gait. Interactions between gait and treatment were determined via ANOVA. RESULTS Interactions between gait and treatment for peak MCPJ angle were significant. Significant reductions in MCPJ angle were noted between the control treatment and legwear with moderate extension limitation for trot and canter (leading and trailing FL) and between inactive legwear and legwear with moderate extension limitation for trot and leading FL during canter. Interactions among peak MCPJ angular velocity, peak vertical GRF, and average loading rate of the vertical GRF showed nonsignificance. CONCLUSIONS AND CLINICAL RELEVANCE Significant reductions in MCPJ extension without significant alterations to peak vertical GRF suggested the legwear's ability to redistribute internal forces. Findings suggested that the legwear may be beneficial for horses rehabilitating from flexor apparatus injuries.