Canine hip dysplasia is predictable by genotyping.

OBJECTIVE: To establish a predictive method using whole genome genotyping for early intervention in canine hip dysplasia (CHD) risk management, for the prevention of the progression of secondary osteoarthritis (OA), and for selective breeding. DESIGN: Two sets of dogs (six breeds) were genotyped with dense SNPs covering the entire canine genome. The first set contained 359 dogs upon which a predictive formula for genomic breeding value (GBV) was derived by using their estimated breeding value (EBV) of the Norberg angle (a measure of CHD) and their genotypes. To investigate how well the formula would work for an individual dog with genotype only (without using EBV), a cross validation was performed by masking the EBV of one dog at a time. The genomic data and the EBV of the remaining dogs were used to predict the GBV for the single dog that was left out. The second set of dogs included 38 new Labrador retriever dogs, which had no pedigree relationship to the dogs in the first set. RESULTS: The cross validation showed a strong correlation (R>0.7) between the EBV and the GBV. The independent validation showed a moderate correlation (R=0.5) between GBV for the Norberg angle and the observed Norberg angle (no EBV was available for the new 38 dogs). Sensitivity, specificity, positive and negative predictive values of the genomic data were all above 70%. CONCLUSIONS: Prediction of CHD from genomic data is feasible, and can be applied for risk management of CHD and early selection for genetic improvement to reduce the prevalence of CHD in breeding programs. The prediction can be implemented before maturity, at which age current radiographic screening programs are traditionally applied, and as soon as DNA is available.

Single nucleotide polymorphisms refine QTL intervals for hip joint laxity in dogs.

Hip laxity is one characteristic of canine hip dysplasia (CHD), an inheritable disease that leads to hip osteoarthritis. Using a genome-wide screen with 250 microsatellites in a crossbreed pedigree of 159 dysplastic Labrador retrievers and unaffected greyhounds, we previously identified putative (P < 0.01) QTL on canine chromosomes 11 and 29 (CFA11 and CFA29). To refine these QTL locations, we have genotyped 257 dogs including 105 Labrador retrievers, seven greyhounds, four generations of their crossbreed offspring and three German shepherds for 111 and 171 SNPs on CFA11 and CFA29 respectively. The distraction index (DI, a measure of maximum hip laxity) was used as an intermediate phenotype that predicts whether a hip joint will or will not develop osteoarthritis. Using a multipoint linkage analysis, significant evidence (95% posterior probability) was found for QTL contributing to hip laxity in the 16.2-21 cM region on CFA11 that explained 15-18% of the total variance in DI. Evidence for an independent QTL on CFA29 was weaker than that on CFA11. Identification of the causative mutation(s) will lead to better understanding of biochemical pathways in both dogs and humans with hip laxity and dysplasia.

Noninvasive Assessment of Neuromuscular Disease in Dogs: Use of the 6-minute Walk Test to Assess Submaximal Exercise Tolerance in Dogs with Centronuclear Myopathy.

BACKGROUND: Noninvasive methods of quantitating exercise tolerance in dogs with neuromuscular disease are needed both for clinical and research use. The 6-minute walk test (6MWT) has been validated as a reliable test of exercise tolerance in dogs with pulmonary and cardiac disease, but not in dogs with neuromuscular disease. HYPOTHESIS/OBJECTIVES: Distance walked and number of steps taken during 6MWT will differ between Labrador retriever dogs with centronuclear myopathy (CNM) and control (ie, healthy) littermates. ANIMALS: Eight purebred Labrador retrievers were drawn from a purpose-bred research colony (status: 3 clear, 2 carrier, and 3 homozygous mutants for the protein tyrosine phosphatase-like A (PTPLA) gene mutation associated with CNM). METHODS: Pilot, prospective, Case-controlled study. Researchers were blinded to disease status. Each dog was leash-trained and acclimatized to the testing area (length, 12.8 m). At the start of testing, each animal was fitted with a pedometer, a timer was started, and dogs were allowed to walk at their own pace for 6 minutes. Distance walked and pedometer readings were recorded. RESULTS: Degree of paresis varied among affected dogs, and was reflected by significant differences in distance walked between CNM-affected dogs and those with clear and carrier genotypes (P = .048). Pedometer readings did not vary according to genotype (P = .86). CONCLUSIONS: The 6MWT appears to differentiate between the ambulatory capacity of normal and CNM-affected dogs. Additional studies are needed to confirm this relationship in a larger number of dogs, and to evaluate the ability of the 6MWT to differentiate between dogs with variable severity of neuromuscular disease-associated exercise intolerance.

In vitro biomechanical comparison of limited contat dynamic compression plate and locking compression plate.

The locking compression plate (LCP) supports biological osteosynthesis by functioning as an internal fixator, rather than as a full or limited contact bone plate which must be adequately contoured and affixed directly to the bone for stable internal fixation of the fracture. In order to help justify the use of the LCP in our veterinary patients, in vitro biomechanical testing was performed comparing the LCP to the conventional limited contact dynamic compression plate (LC-DCP) in canine femurs. We hypothesized that the LCP construct would be at least as stiff under bending and torsional loads as the LC-DCP. The LCP and LC-DCP were applied over a 20-mm osteotomy gap to contralateroal bones within each pair of 14 femora. Non-destructive four-point bending and torsion, and cyclical testing in torsion were performed. The constructs were then loaded to failure in torsion. In medial-lateral and lateral-medial structural bending, significant differences were not found between the LCP and LC-DCP, however, at the gap, the LCP construct was stiffer than the LC-DCP in lateral-medial bending. Significant differences in behaviour over time were not noted between the plate designs during cyclical testing. When loading the constructs to failure in internal rotation, the LC-DCP failed at a significantly lower twist angle (P = .0024) than the LCP. Based on the similar performance with loading, the locking compression plate is a good alternative implant for unstable diaphyseal femoral fracture repair in dogs.

Effects of exercise and polysulfated glycosaminoglycan on repair of articular cartilage defects in the equine carpus.

Our aim was to determine if mild to moderate postoperative exercise and intra-articular polysulfated glycosaminoglycan result in improved repair of large, experimentally induced osteochondral defects in a weight-bearing surface of equine joints. Arthroscopic debridement was used to produce full-thickness defects in a weight-bearing area of the radial carpal bones in 18 ponies. The ponies were randomly assigned to two groups balanced for age: nine animals in the exercise and nine in the no exercise group. Six ponies in each group were medicated weekly for 5 weeks with an intra-articular injection of polysulfated glycosaminoglycan in one middle carpal joint beginning at the time of operation. Walking (twice daily) was begun 6 days postoperatively, and by the twelfth week postoperatively the ponies were trotting for 25 min and walking for 15 min twice daily. At the time of the ponies' death, 17 weeks postoperatively, each defect had an average of 50-75% coverage with repair tissue. Exercised, medicated joints had a significantly smaller area of coverage with repair tissue than exercised, nonmedicated joints. Cartilaginous repair tissue from exercised ponies contained significantly more glycosaminoglycan and type-II collagen (r = 0.53, p < 0.05). The ratio of hydroxylysine to hydroxyproline was significantly lower and the ratio of collagen content to total protein was significantly higher in the repair tissue of medicated joints than in the repair tissue of nonmedicated joints; this is consistent with the presence of less type-II collagen in the repair tissue in medicated joints. We concluded that postoperative exercise was beneficial and that the immediate postoperative use of intra-articular polysulfated glycosaminoglycan was detrimental to the development of cartilaginous repair tissue in large osteochondral defects of equine joints.

Corticosteroids alter the differentiated phenotype of articular chondrocytes.

Experimental evidence suggests that recommended dosages of some corticosteroids used clinically as antiinflammatory agents for treating arthropathies damage articular cartilage, but low dosages may be chondroprotective. The purpose of this study was to evaluate how different concentrations of methylprednisolone affect chondrocyte function and viability. Articular cartilage and chondrocytes were obtained from young adult horses, 1.5-3.5 years of age. Corticosteroid-induced changes in collagen expression were studied at the transcriptional level by Northern blot analyses and at the translational level by measuring [3H]-proline incorporation into [3H]-hydroxyproline. Fibronectin mRNA splicing patterns were evaluated with ribonuclease protection assays. Cytotoxicity was studied using erythrosin B dye exclusion. Steady-state levels of type II procollagen mRNA decreased without concurrent changes in type I procollagen expression as the medium methylprednisolone concentrations were increased from 1 x 10(1) to 1 x 10(8) pg/ml, dropping below 10% of control values by 1 x 10(5) pg/ml. Cytotoxicity occurred as methylprednisolone levels were increased further from 1 x 10(8) to 1 x 10(9) pg/ml. Changes in total collagen (protein) synthesis were less pronounced, but also demonstrated significant suppression between 1 x 10(4) and 1 x 10(8) pg/ml. Corticosteroid-induced changes in fibronectin isoform levels were evaluated in articular cartilage samples without in vitro culture. The cartilage-specific (V + C)(-) isoform was suppressed in both normal and inflamed joints by a single intraarticular injection (0.1 mg/kg) of methylprednisolone. Combined, these data indicate that methylprednisolone suppresses matrix protein markers of chondrocytic differentiation. Decreased and altered chondrocyte expression of matrix proteins likely contributes to the pathogenesis of corticosteroid-induced cartilage degeneration.

Chondrocyte-fibrin matrix transplants for resurfacing extensive articular cartilage defects.

Cartilage resurfacing by chondrocyte implantation, with fibrin used as a vehicle, was examined in large (12 mm) full-thickness articular cartilage defects in horses. Articular chondrocytes, isolated from a 9-day-old foal, were mixed with fibrinogen and injected with thrombin, in a 1:1 mixture, into 12 mm circular defects on the lateral trochlea of the distal femur of eight normal horses. The contralateral femoropatellar (knee) joint served as a control in which the defect was left empty. Synovial fluid from the femoropatellar joints was sampled on days 0, 4, 7, 30, 120, and 240 postoperatively. Groups of four horses were killed at 4 or 8 months postoperatively, and the repair tissue was evaluated by gross and histologic examination with use of hematoxylin and eosin and safranin O staining and by autoradiography. Biochemical analyses included quantitation of proteoglycan, total collagen, and type-II collagen in the repair tissue. Grossly, grafted defects had improved filling of the cartilage lesions; histologically, these areas consisted of differentiated chondrocytes in the deep and middle zones. The cellular arrangement in these zones resembled that of hyaline cartilage. The control defects contained poorly attached fibrous tissue throughout. Grafted tissue at 8 months had increased proteoglycan synthesis evident by both safranin O staining and autoradiography. Glycosaminoglycan quantitation by dye-binding assay confirmed a significantly elevated glycosaminoglycan content in grafted defects (58.8 micrograms/mg of dry weight) compared with control defects (27.4 micrograms/mg; p < 0.05). Similarly, the levels of chondroitin sulfate/dermatan sulfate was significantly elevated in the grafted defects, and this was the predominant glycosaminoglycan epitope present. There was a statistically significant (p < 0.05) increase in type-II collagen in the grafted tissue at 8 months (61.2% grafted; 25.1% control). This resurfacing attempt with use of allograft chondrocytes, secured in large full-thickness articular defects with polymerized fibrin, resulted in an improved cartilage surface in comparison with the control defects, a significantly greater aggrecan level, and a significantly higher proportion of type-II collagen.

Could tourist boots act as vectors for disease transmission in Antarctica?

BACKGROUND: Over the last decade there has been a rapid increase in the number of visitors landing at wildlife sites on the Antarctic continent, and concern has been raised that tourists may transmit important pathogens to or between wildlife colonies. The aim of this study was to determine if tourist activities pose a potential threat to Antarctic wildlife, or possibly to human populations through carriage of pathogens on boots. METHODS: In two trips conducted to Antarctica in the summer season of 2000/2001, swabs were collected from tourist boots: prior to landing, to determine baseline level of bacterial flora on the boots (A isolates); immediately on return to the ship, to quantify the level of contamination (B isolates); and after the boots were washed in seawater to determine the recovery of the organisms after cleaning (C isolates). Swabs were cultured for coliforms, and isolates identified using the API system. RESULTS: Twenty organisms resembling coliforms were isolated from 15 of 72 pairs of boots. Two isolates were recovered from group A, 4 from group B, and 14 from group C. Of these 20 isolates, 11 could be identified using the API identification method. The remaining 9 isolates all produced an unknown but identical profile number. CONCLUSION: These results indicate that current practices for cleaning the boots of tourists visiting Antarctic wildlife colonies may not be sufficient to prevent the transmission of pathogens, and indicate that further studies are needed to define the best method of disinfection.

Cytology of polychrome-stained equine synovial fluid smears. Comparison with clinical findings, histologic specimens, Wright-Giemsa-stained smears and outcome.

Polychrome-stained equine synovial fluid specimens from 34 normal joints and 129 joints with clinical abnormalities were examined cytologically. The smears from joints with abnormalities were categorized as within normal limits (4.7%), slight abnormality (27.9%), proliferative synovitis (21.7%), neutrophilic pattern (20.2%), elongated cell pattern (10.1%), other moderate to marked abnormality (11.6%) and unsatisfactory (3.9%). Cytologic abnormalities that were not restricted to a single category included spindle cells, crystals, stellate cells and cartilage fragments. Multinucleate cells and mononucleate cells with dense cytoplasm and a delicate periphery were seen in smears from cases with clinical diagnoses of osteochondrosis or fracture; interpretation of these cells as osteoclasts and their mononucleate precursors was supported by positive staining with tartrate-resistant acid phosphatase. Smears within the same cytologic category were not found to correspond with a single clinical diagnosis. The identification of several cytologic patterns in cases with the same clinical diagnosis suggests that multiple stages of disease were sampled. Except in cases with the cytologic neutrophilic pattern, there was not a consistent relationship between the histologic features in synovial biopsy specimens and the cytologic findings; the morphologic variation within synovial membrane sections and between sections from different locations was sometimes marked. When compared with air-dried, Wright-Giemsa-stained smears, the polychrome-stained smears were more sensitive in the detection of cytologic abnormalities and were less often falsely negative or unsatisfactory. Following surgery, cases with clinical diagnoses of osteochondrosis (29 cases) and fracture (25 cases) were analyzed according to clinical outcome and cytologic category. While 80% of the horses with proliferative synovitis in cytologic specimens were sound, only 67% of those with the elongated cell pattern, 50% of those with slight abnormality and 33% of those with other moderate to marked abnormality were sound. A statistically significant relationship (P less than .02) was found in cases with a diagnosis of osteochondrosis: animals with a proliferative synovitis pattern were almost three times as likely to be sound as compared to those with slight abnormality. These findings indicate that polychrome-stained equine synovial fluid smears (1) provide information that is different from that found in corresponding histologic sections and (2) are superior to air-dried, Wright-Giemsa-stained smears for cytologic examination. The polychrome-stained equine synovial fluid smears were found to provide information supportive of clinical, radiographic and prognostic data.

Gastric rupture in horses: a review of 54 cases.

The historical, clinical, laboratory, surgical and necropsy findings in 54 cases of gastric rupture in horses are described. Eleven per cent of the deaths of horses undergoing exploratory coeliotomy for colic during the period of the study were a result of gastric rupture. Comparison with all horses which had exploratory coeliotomies for colic over an eight year period did not show that horses with gastric rupture were different from these reference horses regarding age, breed or season. There were fewer stallions than expected in the gastric rupture group. Horses with histories of both acute and chronic (more than 36 h) colic were susceptible to gastric rupture. Primary and idiopathic causes of gastric dilation and rupture accounted for about one-third of the horses. All but one of these cases resulting from secondary causes fell into three aetiologically-related groups: obstructive, peritoneal and enteric, with approximately equal numbers of horses in each group. Most of the ruptures occurred along the greater curvature of the stomach. At least six horses ruptured their stomachs postoperatively in the presence of an indwelling nasogastric tube. The presence or absence of gastric reflux following nasogastric intubation was not a reliable indicator, on its own, of gastric dilation. Horses that later died from gastric rupture had markedly elevated heart rate, hypochloraemia, peritoneal exudative effusion (particularly with evidence of sepsis), pre- and/or postoperative gastric reflux and small or large intestinal disease. However, no distinctive feature of these horses was shown to place them at risk of gastric rupture.

Preliminary studies on long distance, retrograde transport of horseradish peroxidase in equine peripheral nerves.

As a prelude to studies on retrograde axonal transport of neurotoxin (ie, so-called suicide transport) as a means to prevent post neurectomy neuroma formation, preliminary studies were conducted with an innocuous enzymatic marker, horseradish peroxidase (HRP). The proximal stumps of resected medial and lateral palmar digital nerves in six ponies were injected via a tuberculin syringe and needle with 50 micron 1 of a 30 per cent solution of HRP in order to assess long distance retrograde axonal transport. The dorsal root ganglion of the cervical spinal enlargement (ie, C6, C7, C8, T1, T2) were removed at post injection intervals of two, four, six, eight, 10 and 12 days. These were sectioned serially and reacted by the tetramethylbenzidine method to demonstrate transported enzyme in the ganglionic cell bodies which give rise to sensory fibres of the palmar digital nerves. Enzyme, retrogradely transported over axon lengths of 115 cm, was first demonstrated in spinal ganglia four days after injections of the palmar digital nerves. The calculated transport velocity of 287 mm/day, although almost certainly an underestimate, greatly exceeded rates of 72 to 120 mm/day recorded previously with HRP in the peripheral nerves of small laboratory animals. The intensity of the HRP reaction product in ganglionic neurons was strong at four days and it remained unabated in ganglia examined at six, eight, 10 and 12 days post injection. The major sources of the sensory fibres of the palmar digital nerves appeared to be the ganglia of the C8 and T1 spinal segments which contained more than 90 per cent of all labelled neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Attempts to prevent equine post neurectomy neuroma formation through retrograde transport of two neurotoxins, doxorubicin and ricin.

Digital neurectomies, performed to relieve pain and lameness, are often complicated postoperatively by formation of painful neuromas. In this study attempts were made to deliver lethal doses of neurotoxin to the cell bodies of the transected digital nerve fibres via long-distance retrograde axon transport and, thereby, prevent the regenerative changes that lead to neuroma formation. After applying doxorubicin in various ways to the digital nerve stumps of ponies, degenerating or necrotic neurones appeared only sporadically in the spinal ganglia. Although doxorubicin was largely ineffective in retrograde destruction of cell bodies, when absorbed in pledgets on the stumps it exerted a sustained action which prevented Schwann cell proliferation and axon sprouting. Ricin, in contrast to doxorubicin, was effective in retrograde destruction of sensory neurons. Many affected neurons were devoid of polysomes but packed with mitochondria; others had advanced to various stages in cytolysis. Despite its effectiveness, ricin cannot be recommended because of its extreme toxicity. The clinical use of retrograde transport in equine neurectomy will probably depend on future development of hybrid toxins with high neural specificity and low systemic toxicity.

Prognostic factors affecting survival of 507 horses with joint disease: (1983 to 1990).

Between July 1, 1983 and December 31, 1990, risk factors were determined for all horses with joint disease presented to a referral center, of being discharged, of ever becoming sound, or of being alive at 3 mo follow-up. Logistic multiple-regression models were done separately for foals (< or = 4 mo), yearlings (> 4-24 mo) and racing or nonracing adult horses (> 24 mo). The breakdown in this study was 53 foals, 87 yearlings, 141 nonracing adults, and 226 racing adults. Thirty-one foals (58%), 68 yearlings (78%), 119 non-racing adults (84%), and 213 racing adults (94%) were discharged. Foals with a less severe lameness, duration of illness of > 1 d, and infectious arthritis had increased odds of discharge. At follow-up, 12 of 18 (67%) were alive, 10 (56%) of which were sound. Yearlings with osteochondrosis had higher odds of discharge; at follow-up, 38 of 49 (78%) were alive, 32 (65%) of which were sound. For non-racing adults, horses with less severe lameness, without a miscellaneous diagnosis, or intended for pleasure use had increased odds of discharge. At follow-up, 55 of 78 (70%) were alive and 33 of 58 (57%) with soundness data became sound. Risk factors for higher odds of being alive at follow-up were carpal lameness, arthroscopic surgery, a prognosis other than poor, became sound, above-median hospitalization costs, and duration of follow-up. The 161 racing adults (76% of discharges), with follow-up, were more likely to have had osteoarthritis, higher hospital costs, hospitalization > 1 d, and arthroscopy. Sixty-four (60%) of these became sound; the odds increased if the horse was not severely lame at admission or was hospitalized for > 1 d. Risk factors and prognosis differed by age-use group among horses seen at our hospital.

Effect of intramuscularly administered polysulfated glycosaminoglycan on articular cartilage from equine joints injected with methylprednisolone acetate.

Intra-articularly administered, long-acting corticosteroids are a beneficial treatment for many equine joint disorders because they alleviate inflammation and signs of pain, but they also exert detrimental effects on the biochemical composition and morphologic features of articular cartilage. Chondroprotective drugs have been shown to mitigate some of the deleterious effects of intra-articularly administered corticosteroids on articular cartilage of laboratory animals. Twenty-one ponies were assigned at random to receive 1 of 3 treatments in the right middle carpal joint. Group-1 ponies (n = 8) had methylprednisolone acetate (MPA; 0.2 mg/kg of body weight) and saline solution administered intra-articularly and IM, respectively. Group-2 ponies (n = 9) received MPA (0.2 mg/kg) and polysulfated glycosaminoglycan (GAG; 2 mg/kg). Group-3 ponies (control; n = 4) had saline solution administered intra-articularly and IM. The corticosteroid or saline solution was injected into the right middle carpal joint on day 1. The IM administered polysulfated GAG or saline solution was administered at the same time, then was repeated every 3 days for 20 days. Ponies were euthanatized 21 days after initial injection by overdose of pentobarbital sodium. The cartilage of younger ponies was significantly (P < 0.05) more responsive to the proteoglycan-depleting effects of MPA. Ponies < 10 years old of groups 1 and 2 had significantly (P < 0.05) lower GAG content in the articular cartilage than did control ponies. Systemic treatment with polysulfated GAG did not result in a protective effect against proteoglycan loss from the articular cartilage.(ABSTRACT TRUNCATED AT 250 WORDS)

Structure of equine type I and type II collagens.

Collagen type I was purified from equine skin and flexor tendon, and type II collagen was purified from equine articular cartilage. The proteoglycans in these tissues were extracted, using guanidine HCl; the collagens were solubilized, using pepsin digestion, then were selectively precipitated with NaCl. Gel electrophoresis indicated that the precipitates contained only type I or type II collagen. Amino acid analysis indicated that collagen constituted > 97% of the total protein in the precipitates. Hydroxylation of proline was 42.0 +/- 0.6% (mean +/- SEM) in alpha 1(I) and alpha 2(I), and was 48.1 +/- 1.3% in alpha 1(II) chains. The hydroxylation of lysine was 23.2 +/- 0.7% in alpha 1(I) and 34.1 +/- 0.9% in alpha 2(I) chains from tendon, and 49.6 +/- 4.3% in alpha 1(II) chains from cartilage. The cyanogen bromide (CB)-peptide patterns of chromatographically purified equine alpha 2(I) and alpha 1(II) chains were similar to those published previously for rat, bovine, and human alpha 2 and alpha 1 chains. However, the CB-peptide pattern of the equine alpha 1(I) chain resembled the guinea pig alpha 1(I) chain, which has no methionine between CB7 and CB6. Purified equine alpha 1(I)CB7,6 contained no methionine, methionine sulfoxide, or homoserine lactone. Mass of 42.26 kd was determined by use of mass spectrometry, and N-terminal sequence analysis established that the first 12 amino acids of this CB7,6 were identical to the sequence of human alpha 1(I)CB7.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantitative ultrastructural anatomy of esophagus in different regions in the horse: effects of alternate methods of tissue processing.

A qualitative and quantitative study was made of 2 adult horses to determine the effect of specimen preparation and the influence of anatomically separate regions on the microscopic structure of the esophagus. The effects of 3 fixative solutions (formalin, Bouin's fixative and Karnovsky's fixative) and 4 histologic staining procedures (hematoxylin and eosin, Gomori's trichrome, Laidlow's reticulum, and orcein-elastin) were compared. Density-dependent image analysis was used to compare the relative contributions of elastic and reticular fibers in esophageal connective tissue for each region. Cross-sectional dimensions of the tissue layers which comprise the esophageal wall were obtained by light microscopic measurement and were compared for each region. The mucosal surface structure was examined by scanning electron microscopy. Our results indicate that specimen preparation has significant effects on the structural integrity of esophageal tissues. Bouin's fixative was inferior to the other fixatives in this regard. Furthermore, there are extensive regional differences in esophageal microstructure. Formalin-fixed tissues stained with either hematoxylin and eosin or Gomori's trichrome appear the most suitable for routine quantitative morphologic studies of the equine esophagus.

Keratan sulfate as a marker of articular cartilage catabolism and joint treatment in ponies.

Keratan sulfate (KS) is a glycosaminoglycan, distribution of which is confined mostly to hyaline cartilage. As such, it is a putative marker of hyaline cartilage catabolism. In experiment 1, a focal osteochondral defect was made arthroscopically in 1 radial carpal bone of 2 ponies, and in 2 other ponies, chymopapain was injected into the radiocarpal joint to induce cartilage catabolism. Sequential and concurrent plasma and synovial fluid concentrations of KS were measured, up to 13 months after induction of cartilage injury, to determine whether changes in KS concentrations reflected cartilage catabolism. In experiment 2, a large, bilateral osteochondral defect was made in the radial carpal bones of 18 ponies, which were subsequently given postoperative exercise and/or injected intra-articularly with 250 mg of polysulfated glycosaminoglycan (PSGAG). Medication was given at surgery, then weekly for 4 weeks. Blood samples were collected and synovial fluid was aspirated before surgery, when medication was given, and at postmortem examination (postoperative week 17). The KS concentration was measured in these fluids to determine whether changes in KS concentration indicated an effect of joint treatment. In experiment 1, the concentration of KS in synovial fluid was highest 1 day after joint injury, and the concentration in plasma peaked 2 days after joint injury. For ponies receiving chymopapain intra-articularly (generalized cartilage catabolism), a fivefold increase over baseline was observed in the concentration of KS in plasma (peak mean, 1.2 micrograms/ml), and a tenfold increase over baseline in synovial fluid (peak mean, 2.0 mg/ml) was observed. On average, these maxima were threefold higher than values in fluids of ponies with osteochondral defects (focal cartilage disease). In experiment 2, nonexercised ponies had lower KS concentration (as a percentage of the preoperative concentration) in synovial fluid than did exercised ponies at all postoperative times, and at postoperative week 17, this effect was significant (P < 0.05). This may be related to decreased turnover of KS in articular cartilage attributable to stall confinement and late increase in turnover related to exercise. Seventeen weeks after surgery, synovial fluid from exercised, medicated ponies had significantly (P < 0.05) higher KS content than did fluid from exercised, nomedicated ponies. This indicated that exercise, when combined with medication, may increase KS release from articular cartilage. Synovial fluid from medicated joints of nonexercised ponies had significantly (P < 0.05) lower KS concentration than did synovial fluid from nonmedicated joints of nonexercised ponies.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of scintimetry to assess effects of exercise and polysulfated glycosaminoglycan on equine carpal joints with osteochondral defects.

Combined blood pool and delayed images produced by use of 99mTc-methylene diphosphonate (99mTcMDP) were evaluated as an objective measurement of the response of equine joints with osteochondral defects to postoperative exercise and intra-articularly administered polysulfated glycosaminoglycan (PSGAG). Osteochondral defects (approx 2.4 x 0.9 cm) were induced arthroscopically in the dorsodistal radial carpal bones of 18 ponies. These ponies were randomized (while balancing for age [range 2 to 15; median, 5.0; mean, 5.1 years]) to 2 treatment groups. Nine ponies were assigned to be exercised, and 9 were stall-rested. Six ponies in each group were administered PSGAG (250 mg) in 1 joint (medicated) and lactated Ringer's solution (LRS) in the contralateral joint. The 3 remaining ponies in each group were administered LRS in both joints (nonmedicated). Medication was given at surgery, then weekly for 4 weeks. The exercise protocol (begun at postoperative day 6 and conducted twice daily) started with 30 minutes walking (approx 0.7 m/s), and, by postoperative month 3, the ponies were being walked for 15 minutes and trotted (approx 1.6 m/s) for 25 minutes. Simultaneous dorsal images of both carpi were made 2 to 3 minutes after IV administration of 99mTcMDP (blood pool image) and 90 to 120 minutes later (delayed image). Scintimetry, in counts per minute per pixel per millicurie, was done before, and at 1, 2, 4, 8, 10, 13, and 17 weeks after surgery, prior to euthanasia. Radionuclide uptake on blood pool images decreased faster than that on delayed images, in which uptake remained high for 17 weeks. This indicated that bone was metabolically active for at least 17 weeks after surgery. Exercise significantly (P < 0.05) decreased uptake on the blood pool images of medicated joints up to 1 month after surgery. Thus, exercise (in the presence of PSGAG) probably had a transient, beneficial effect on soft tissues of the joint. Exercise, without PSGAG, promoted increased bone remodeling, because the highest uptake on delayed images was observed in exercised, nonmedicated ponies up to 3 months after surgery. This was consistent with development of osteoarthritis in these ponies. Medication alone stimulated bone remodeling, and data indicated that an identical effect may take place in contralateral LRS-injected joints, because of systemic circulation of the drug. However, the combination of exercise and medication appeared to moderate the independent effects of each. The combination of exercise and medication in individual joints resulted in notably (P < 0.05) decreased bone remodeling.(ABSTRACT TRUNCATED AT 400 WORDS)

Comparison of the trotting gaits of Labrador Retrievers and Greyhounds.

OBJECTIVE: To compare the trotting gaits of Labrador Retrievers and Greyhounds to determine whether differences in locomotion are attributable to differences in their manner of moving or to body size and shape differences between these 2 breeds. ANIMALS: 8 healthy 5-month-old Greyhounds and 5 healthy Labrador Retrievers between 6 and 18 months old. PROCEDURE: A series of 4 force platforms was used to record independent ground reaction forces on the forelimbs and hind limbs during trotting. Values of stride parameters were compared between breeds before and after normalization for size differences. Standard values of absolute and normalized stride period and stride length were determined from linear regressions of these parameters on relative (normalized) velocity. Forces were normalized to body weight and compared at the same relative velocity. RESULTS: Greyhounds used fewer, longer strides than the Labrador Retrievers to travel at the same absolute speed. After normalization for body size differences, most measurable differences between breeds were eliminated. Subtle differences that did persist related to proportion of the stride that the forefoot was in contact with the ground, timing of initial hind foot contact relative to initial forefoot contact, and distribution of vertical force between the forelimbs and hind limbs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that apparent differences in the trotting gait between Labrador Retrievers and Greyhounds are mainly attributable to differences in size, and that dogs of these 2 breeds move in a dynamically similar manner at the trot.

Comparison of measurements of dorsolateral subluxation of the femoral head and maximal passive laxity for evaluation of the coxofemoral joint in dogs.

OBJECTIVE: To determine whether dorsolateral subluxation (DLS) of the femoral head reflects osseous conformation of the coxofemoral (hip) joint and represents a property distinct from maximum passive laxity of the hip joint in dogs. ANIMALS: 14 Labrador Retrievers, 16 Greyhounds, 58 Greyhound-Labrador Retriever mixed-breed dogs, and 1 Rottweiler. PROCEDURES: DLS of the femoral head (DLS score) and passive laxity of the hip joint (distraction index) were determined radiographically in 3 groups of dogs: not treated (167 joints of 84 dogs); before and after injecting 2 ml of hyaluronan into 25 hip joints of 13 dogs; and before and after unilateral triple pelvic osteotomy in 5 dogs. Results of the 2 methods were compared for each group. RESULTS: In untreated dogs, the correlation coefficient (r) of DLS score versus distraction index was -0.73 and -0.69 for 84 left and 83 right hip joints, respectively. Mean coefficient of determination (r2) for both hips was 0.5. Mean DLS score did not differ before and after intra-articular injection of hyaluronan into either hip joint, whereas mean distraction index increased significantly after intra-articular injection. Unilateral triple pelvic osteotomy resulted in a significant increase in DLS score, compared with values obtained before surgery. However, distraction index before and after surgery did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE: The DLS test assesses the congruity of the acetabulum and the femoral head in a canine hip joint and thus represents a characteristic distinct from maximum passive laxity. The DLS score and the distraction index evaluate different components of hip joint stability.