Marc A. Kinsley

Analgesic and gastrointestinal effects of epidural morphine in horses after laparoscopic cryptorchidectomy under general anesthesia

OBJECTIVE To evaluate the hypothesis that epidural morphine (0.1 mg kg(-1) ) decreases pain in horses after laparoscopic surgery without adversely affecting gastrointestinal (GI) motility. STUDY DESIGN Randomized clinical trial. ANIMALS Eighteen horses undergoing laparoscopic cryptorchidectomy under general anesthesia. METHODS Horses were randomly assigned to receive either epidural morphine (0.1 mg kg(-1) ) or no epidural before the start of surgery. Pain behaviors were assessed during the first two post-operative days using a numerical rating scale. Barium-filled spheres were administered through a nasogastric tube before anesthesia. GI motility was assessed by recording manure production, by quantitating the spheres in the manure, and by abdominal auscultation of intestinal sounds. Heart rates and cortisol concentrations were also measured during the post-operative period. RESULTS Pain scores increased for 12 hours after surgery in the control group and were significantly higher than in the morphine group for the first 6 hours. Pain scores remained unaltered in the morphine group throughout the observation period. Heart rate and plasma cortisol concentrations did not differ between groups or with time. No signs of colic were observed in any horse. CONCLUSION AND CLINICAL RELEVANCE Epidural morphine (0.1 mg kg(-1) ) did not adversely affect GI motility in horses after laparoscopic surgery under general anesthesia.

Use of Plain Radiography in the Diagnosis, Surgical Management, and Postoperative Treatment of Obstructive Urolithiasis in 25 Goats and 2 Sheep

Objective To describe use of plain radiography for diagnosis, surgical management, and postoperative treatment of obstructive urolithiasis in small ruminants. Study Design Retrospective case series. Animals Small ruminants (n = 27; 25 goats, 2 sheep). Methods Medical records (January 2002–November 2011) and radiographs for all small ruminants diagnosed with obstructive urolithiasis and having plain abdominal radiographs were reviewed. Signalment, surgical procedures, radiographic findings, ultrasonographic findings, position of calculi, and how plain radiography influenced surgical management and postoperative treatment were recorded. Results Radiopaque urinary calculi were detected in 23 (85%) plain radiographic studies. Location of uroliths determined by plain radiography included: cystic only (n = 5), distal to the sigmoid flexure and cystic (5), subischial (5), distal to the sigmoid flexure only (3), sigmoid flexure (3), and subischial and cystic (2). In 8 of these animals, postoperative radiographs revealed residual calculi in the urethra and were essential for their targeted removal by urethrotomy in 7 animals. Conclusions In regions where radiopaque calculi (calcium carbonate, calcium oxalate, silica) are commonly encountered in small ruminants, plain radiographs are recommended to determine the appropriate surgical approach(es) and to confirm resolution of the obstruction.OBJECTIVE To describe use of plain radiography for diagnosis, surgical management, and postoperative treatment of obstructive urolithiasis in small ruminants. STUDY DESIGN Retrospective case series. ANIMALS Small ruminants (n = 27; 25 goats, 2 sheep). METHODS Medical records (January 2002-November 2011) and radiographs for all small ruminants diagnosed with obstructive urolithiasis and having plain abdominal radiographs were reviewed. Signalment, surgical procedures, radiographic findings, ultrasonographic findings, position of calculi, and how plain radiography influenced surgical management and postoperative treatment were recorded. RESULTS Radiopaque urinary calculi were detected in 23 (85%) plain radiographic studies. Location of uroliths determined by plain radiography included: cystic only (n = 5), distal to the sigmoid flexure and cystic (5), subischial (5), distal to the sigmoid flexure only (3), sigmoid flexure (3), and subischial and cystic (2). In 8 of these animals, postoperative radiographs revealed residual calculi in the urethra and were essential for their targeted removal by urethrotomy in 7 animals. CONCLUSIONS In regions where radiopaque calculi (calcium carbonate, calcium oxalate, silica) are commonly encountered in small ruminants, plain radiographs are recommended to determine the appropriate surgical approach(es) and to confirm resolution of the obstruction.

Wnt/β‐catenin signaling of cartilage canal and osteochondral junction chondrocytes and full thickness cartilage in early equine osteochondrosis

The objective of this study was to elucidate gene and protein expression of Wnt signaling molecules in chondrocytes of foals having early osteochondrosis (OC) versus normal controls. The hypothesis was that increased expression of components of Wnt signaling pathway in osteochondral junction (OCJ) and cartilage canal (CC) chondrocytes would be found in early OC when compared to controls. Paraffin‐embedded osteochondral samples (7 OC, 8 normal) and cDNA from whole cartilage (7 OC, 10 normal) and chondrocytes surrounding cartilage canals and osteochondral junctions captured with laser capture microdissection (4 OC, 6 normal) were obtained from femoropatellar joints of 17 immature horses. Equine‐specific Wnt signaling molecule mRNA expression levels were evaluated by two‐step real‐time qPCR. Spatial tissue protein expression of β‐catenin, Wnt‐11, Wnt‐4, and Dkk‐1 was determined by immunohistochemistry. There was significantly decreased Wnt‐11 and increased β‐catenin, Wnt‐5b, Dkk‐1, Lrp6, Wif‐1, Axin1, and SC‐PEP gene expression in early OC cartilage canal chondrocytes compared to controls. There was also significantly increased β‐catenin gene expression in early OC osteochondral junction chondrocytes compared to controls. Based on this study, abundant gene expression differences in OC chondrocytes surrounding cartilage canals suggest pathways associated with catabolism and inhibition of chondrocyte maturation are targeted in early OC pathogenesis.

Age-related differential gene and protein expression in postnatal cartilage canal and osteochondral junction chondrocytes

Wnt/β-catenin, Indian hedgehog (Ihh)/Parathyroid-related peptide (PTHrP) and retinoid signaling pathways regulate cartilage differentiation, growth, and function during development and play a key role in endochondral ossification. The objective of this study was to elucidate the gene and protein expression of signaling molecules of these regulatory pathways in chondrocytes surrounding cartilage canals and the osteochondral junction during neonatal and pre-adolescent development. This study revealed cell-specific and age-related differences in gene and protein expression of signaling molecules of these regulatory pathways. A trend for higher gene expression of PTHrP along the cartilage canals and Ihh along the osteochondral junction suggests the presence of paracrine feedback in articular-epiphyseal cartilage. Differential expression of canonical (β-catenin, Wnt-4, Lrp4, Lrp6) and noncanonical Wnt signaling (Wnt-5b, Wnt-11) and their inhibitors (Dkk1, Axin1, sFRP3, sFRP5, Wif-1) surrounding the cartilage canals and osteochondral junction provides evidence of the complex interactions occurring during endochondral ossification.

Expression of pro-apoptotic markers is increased along the osteochondral junction in naturally occurring osteochondrosis

Osteochondrosis (OC) is a naturally occurring disease of the articular-epiphyseal cartilage and subchondral bone layers, leading to pain and decreased mobility. The objective of this study was to characterize gene and protein expression of apoptotic markers in chondrocytes surrounding cartilage canals and along the osteochondral junction of osteochondrosis (OC)-affected and normal cartilage, using naturally occurring disease in horses. Paraffin-embedded osteochondral samples (6 OC, 8 normal controls) and cDNA from chondrocytes captured with laser capture microdissection (4 OC, 6 normal controls) were obtained from the lateral trochlear ridge of femoropatellar joints in 14 immature horses (1–6 months of age). Equine-specific caspase-3, caspase-8, caspase-10, Fas, Bcl-2, BAG-1, TNFα, cytochrome C, thymosin-β10, and 18S mRNA expression levels were evaluated by two-step real-time quantitative PCR. Percentage of cell death was determined using the TUNEL method. Protein expression of caspase-10, Fas, cytochrome C, and thymosin-β10 was determined following immunohistochemistry. Statistical analysis was performed using the Wilcoxon rank sum test or two-sample t-test (p < 0.05). In OC samples, there was significantly increased gene expression of caspase-10, Fas, cytochrome C, and thymosin-β10 in chondrocytes along the osteochondral junction and increased Fas gene expression in chondrocytes adjacent to cartilage canals, compared to controls. In OC samples, higher matrix Fas and cytochrome C protein expression, lower mitochondrial cytochrome C protein expression, and a trend for higher cytoplasmic caspase-10 protein expression were found. Collectively, these results suggest that both extrinsic and intrinsic apoptotic pathways are activated in OC cartilage. Increased apoptosis of osteochondral junction chondrocytes may play a role in OC, based on increased gene expression of several pro-apoptotic markers in this location.

Correlation of dickkopf-1 concentrations in plasma and synovial fluid to the severity of radiographic signs of equine osteoarthritis

OBJECTIVE The purpose of this study was to determine whether there was a correlation between circulating and intra-synovial Dkk-1 and radiographic signs of equine osteoarthritis. METHODS Circulating and intra-synovial Dkk-1 levels were measured in clinical cases using a commercially available human Dkk-1 ELISA. Radiographs were performed of the joints from which fluid was collected and these were assessed and scored by a boarded radiologist for joint narrowing, subchondral bone sclerosis, subchondral bone lysis, and periarticular modelling. Comparisons were made between radiographic scores and the concentrations of Dkk-1 using a Kruskal-Wallis one-way ANOVA. Correlations were calculated using Kendalls statistic. RESULTS A total of 42 synovial fluid samples from 21 horses were collected and used in the analysis. No significant correlation was identified between Dkk-1 concentrations and radiographic signs of osteoarthritis. Intra-synovial Dkk-1 concentrations were significantly greater (p <0.001) in low motion joints (mean concentration, 232.68 pg/mL; range, 109.07-317.17) when compared to high-motion joints (28.78 pg/mL; 0.05-186.44 pg/mL) (p <0.001). CLINICAL SIGNIFICANCE Low motion joints have significantly higher concentrations of Dkk-1 compared to high motion joints. Further research is needed to establish the importance of this finding and whether potential diagnostic or therapeutic applications of Dkk-1 exist in the horse.