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  • Author or Editor: Riitta-Mari Tulamo x
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SUMMARY

Two recently developed direct methods, radioassay—125I-labeled hyaluronic acid binding protein (125I-habp)—and high-performance liquid chromatography (halc), were used to assess and compare the concentration of hyaluronate (ha) in synovial fluid of horses. Also determined were changes in the ha concentration in an experimental treatment model involving physiologic saline solution (pss)-irrigated or methylprednisolone acetate-injected tarsocrural joints of clinically normal horses. Serum ha concentration was determined simultaneously, using the 125I-habp assay.

Synovial fluid ha concentration values obtained by use of the hplc method were approximately double the values obtained by use of 125I-habp assay. Correlation (r = 0.819) between the 2 methods was highly significant (P < 0.001; linear regression analysis) for all samples studied and for various experimental subgroups. When pure ha standards were used, correlation between the 2 methods was close to 1 (r = 0.965; P < 0.001), with higher values obtained by use of the 125I-habp assay. It is suggested that the ha binding protein derived from endogenous cartilage proteoglycan interferes with the 125I-habp assay on synovial fluid, resulting in excessively low values, compared with those obtained using the hplc procedure.

Intra-articular injection of methylprednisolone acetate significantly (P < 0.01) increased synovial fluid ha concentration at 24 hours after injection. Increase was also detected after pss irrigation, but owing to wide intersubject variation, this increase was not significant. The hplc procedure, which provides simultaneous information about the concentration and degree of polymerization of ha, is recommended for the study of synovial fluid, whereas the 125I-habp assay is more suitable for serum ha analysis.

Free access
in American Journal of Veterinary Research

Abstract

Objective

To establish concentration of hyaluronate (HA) in tracheal lavage fluid from healthy horses and horses with chronic obstructive pulmonary disease (COPD).

Animals and Samples

Tracheal lavage fluid samples (n = 42) from 18 horses, 11 with COPD, and 7 control horses.

Procedure

Clinical examination of the respiratory tract, tracheal lavage, and blood sample collection were performed on horses without clinical signs of respiratory tract disease and horses with clinical signs of COPD. In some horses, 1 to 5 repeated examinations were performed at 1-week intervals. Tracheal lavage fluid samples were analyzed for cell numbers, and urea concentration (made in parallel with serum samples to evaluate sample dilution effect); HA was determined by radiometric assay.

Results

Mean (± SEM) HA concentration in tracheal lavage fluid samples was significantly (P = 0.005) higher in horses with COPD (1,880 [± 309] μg/L), compared with that in control horses (256 [± 72] μg/L). The increase in HA concentration in tracheal lavage fluid of COPD-affected horses was verified by repeated sample collection and analysis.

Conclusions

In horses with chronic respiratory tract inflammation such as COPD, tracheal lavage fluid HA concentration is about 7 times higher than reference values. High HA concentration in the tracheal or bronchoalveolar lavage fluid may reflect pathophysiologic changes in connective tissue around bronchi and bronchioli, leading to continuous increased production of HA in horses with advanced forms of COPD.

Clinical Relevance

Determination of tracheal lavage fluid HA concentration may be used as a marker of chronic inflammatory changes in the COPD-affected lung. (Am J Vet Res 1997;58:729–732)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine the validity, reliability, and sensitivity of a published chronic pain index by completion of a questionnaire in Finnish by owners of dogs with chronic signs of pain caused by osteoarthritis.

Animals—61 client-owned dogs with osteoarthritis.

Procedures—Validity, internal consistency, and repeatability testing of the questionnaire were evaluated by completion of questions in Finnish by owners of 61 dogs; the questionnaire was named the Helsinki chronic pain index (HCPI). Sensitivity testing of the questionnaire was determined from data of 2 smaller groups of dogs treated with carprofen (n = 17) or placebo (17). Owners completed the questionnaire 5 times during a 16-week period. Psychometric properties of the HCPI were evaluated.

Results—Principal component analysis yielded only a single stable component, indicating that the HCPI was best explained as an 11-item single construct index of chronic pain. Changes in the HCPI correlated well with change in quality of life and with change in the mobility visual analogue scale (r = 0.72 and r = 0.67, respectively), indicating a high predictive validity. Internal consistency (Cronbach A value = 0.82) and test-retest reliability (intraclass correlation coefficient = 0.84 for items and 0.92 for the HCPI) were high. Changes in scores (from baseline to treatment and back to no treatment) between the carprofen treatment group and placebo control group were significant, indicating high sensitivity.

Conclusions and Clinical Relevance—The Finnish version of the HCPI provided a valid, reliable, and responsive tool for assessment of response to treatment in dogs with osteoarthritis.

Full access
in American Journal of Veterinary Research

Summary

High molecular weight (mw) hyaluronate (ha) is an integral part of synovial fluid (sf), regulating many Important physiologic and pathophysiologic mechanisms. Many of its effects depend on, or are reflected in. the concentration and mw of ha. High-performance liquid chromatography was used to assess simultaneously the concentration and mw of ha in sf obtained from horses with various arthritides: acute traumatic arthritis; chronic traumatic arthritis, including degenerative joint disease (djd); and infectious arthritis. The size-exclusion column was calibrated, using appropriate ha concentration and mw standards, before the high-performance liquid chromatographic assays of the sf samples.

Calibration of the column disclosed that the maximal limit for mw estimation of ha was around 3 million. In control joints, mw of ha ranged from 2 to 3 × 106 (mean 2.5 × 106) and did not differ significantly from mw of ha in sf from horses with acute or chronic traumatic arthritis (mean 2 × 106; range 1.5 to 3 × 106). Interestingly, a small amount of ha of moderately high mw (approx 1 to 1.5 × 106) was detected in chromatograms of sf from infected joints. This degree of polymerization of sf ha was significantly (P < 0.01) lower, compared with that for control joints.

There was no difference in mean (± sd) concentration of ha between control joints and joints with acute or chronic traumatic arthritis (0.33 ± 0.12 g/L vs 0.18 ± 0.03 g/L. or 0.23 ± 0.12 g/L.), indicating that sf ha concentration probably should not be used as a diagnostic marker for the condition. However, the sf ha concentration was significantly (P < 0.01) lower in joints with infectious arthritis (0.07 ± 0.03 g/L) and in the joints with radiographic evidence of djd (0.12 ± 0.01 g/L), compared with control joints.

Free access
in American Journal of Veterinary Research

Abstract

Objective

To investigate the presence of large molecular weight (MW) proteoglycans (PG) and hyaluronate (HA) in synovial fluid (SF) from horses with various arthritides and from control joints.

Design

Horses with acute (< 2 weeks) or chronic (> 4 weeks) lameness were examined by clinical examination, intrasynovial anesthesia, radiography, arthroscopy, and SF analysis. Joints were grouped on the basis of diagnosis: acute traumatic arthritis, chronic traumatic arthritis (with a subgroup of degenerative joint disease), intra-articular fracture, and infectious arthritis.

Animals

31 horses with arthritis and 9 control horses; altogether 43 SF samples were analyzed.

Procedure

High-performance liquid chromatography was used to assess HA and large MW PG in SF samples.

Results.

A PG peak was identified in 8 of 23 SF samples of joints with chronic traumatic arthritis, 4 of which had no or minimal abnormal radiographic findings but mild articular cartilage fibrillation detected by arthroscopy, and in 3 joints with intra-articular fracture and 1 with resolving infectious arthritis, but not in joints with acute traumatic arthritis or in control joints. There was significant difference (P < 0.01) in mean (± SEM) HA concentration between control joints and joints with chronic traumatic arthritis (0.32 ± 0.04 g/L; n = 9 vs 0.18 ± 0.01 g/L; n = 23).

Conclusions

Large MW PG fragments are released into equine SF in the course of articular disease and can be detected simultaneously with HA by high-performance liquid chromatography.

Clinical Relevance

The SF HA concentration can be used as diagnostic marker for chronic traumatic arthritis. However, SF PG or other marker cannot be used for diagnosing or monitoring degenerative joint disease. (Am J Vet Res 1996;57:932–937)

Free access
in American Journal of Veterinary Research

Summary

Concentration of hyaluronate (ha) in equine serum was determined by a recently developed specific radioassay. The mean ± SD HA concentration in equine serum was 288 ± 145 μg/L, was age dependent, and varied widely between horses (range, 190 to 760 μg/L).

Light or moderate exercise increased serum ha concentration from baseline values by 1.5- to 3-fold. In all horses, serum ha concentration returned to or below the original resting values 1 and 2 hours after exercise.

Free access
in American Journal of Veterinary Research

Abstract

Objective

To evaluate effects of intra-articular (IA) injections of bufexamac on amphotericin B-induced aseptic arthritis in horses.

Animals

24 Standardbred horses.

Procedure

Aseptic arthritis was induced in the right intercarpal joint by IA injection of amphotericin B (20 mg). One week later (day 0), horses were randomly assigned to four 6-horse treatment groups and treated with IA injection of 10, 20, or 40 mg of bufexamac suspension (20 mg/ml) or 2.0 ml of sterile saline (0.9% NaCl) solution (control). The treatment was repeated once after 7 days. Clinical lameness examinations and synovial fluid (SF) analyses were done prior to induction and at weekly intervals for 5 weeks (days 0, 7, 14, 21 and 28).

Results

Intra-articular injection of amphotericin B consistently resulted in aseptic arthritis with a lameness index (mean ± SEM; scale 0 to 5) of 2.7 ± 0.17 on day 0. Intra-articular injections of 20 and 40 mg of bufexamac significantly reduced the day-28 lameness index, compared with control values. Amphotericin B administration also resulted in a significant increase in SF β-glucuronidase (BGLUC) activity, and IA injections of bufexamac significantly reduced day-28 activity of this enzyme, compared with control values.

Conclusions and Clinical Relevance

2 IA injections of 20 or 40 mg of bufexamac, at weekly intervals, were effective in reducing clinical signs of lameness and SF activity of BGLUC associated with amphotericin B-induced carpal joint arthritis. Bufexamac possesses anti-inflammatory properties useful for IA treatment of lameness associated with aseptic arthritis in horses. (Am J Vet Res 1999;60:1467–1473)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To evaluate the effects of intra-articular (IA) injections of bufexamac in horses, focusing particularly on the effects of bufexamac on articular cartilage.

Animals—20 Standardbreds.

Procedure—Horses were randomly allocated into 4 groups consisting of 5 horses each, and 20, 60, or 100 mg of bufexamac or 1 ml of sterile saline (0.9% NaCl) solution (control) was injected into 1 intercarpal joint at weekly intervals for 6 treatments (days 0, 7, 14, 21, 28, and 35). Clinical signs and results of hematologic, serum biochemical, and synovial fluid (SF) analyses and radiography were used to evaluate treatment effects. On day 49, all horses were euthanatized; gross necropsy and histologic examinations of internal organs and articular tissues were performed. Glycosaminoglycan concentration of the articular cartilage was evaluated in safranin O-stained sections by use of a semiquantitative microspectrophotometric method.

Results—No systemic signs were observed. Temporary mild to moderate heat and effusion were the only clinical signs observed in a number of joints after IA injections and more often only in the 100 mg group, compared with controls. The 100 mg dose resulted in significant increases in SF WBC counts, with relative neutrophilia and SF total protein concentration 24 hours after injection (day 1). No lesions suggestive of toxic effects were detected at necropsy or on histologic examination. No changes in articular cartilage glycosaminoglycan concentration were detected.

Conclusion and Clinical Relevance—Six injections of 20, 60, or 100 mg of bufexamac at weekly intervals did not cause any untoward systemic or local effects. These data suggest that bufexamac is a safe nonsteroidal anti-inflammatory drug for IA administration in horses. (Am J Vet Res 2001;62:1629–1635)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To identify variables and evaluate methods for assessing chronic pain in dogs.

Design—Prospective study.

Animals—41 dogs with canine hip dysplasia (CHD), and 24 apparently healthy dogs with no history of pain.

Procedure—2 veterinarians evaluated the dogs' locomotion and signs of pain. Owners of dogs with CHD and control dogs answered a questionnaire regarding their dogs' demeanor, behavior, and locomotion (descriptive scales) and assessed pain and locomotion (visual analog scales). Plasma concentrations of several stress-related hormones were determined, and 13 radiologic variables were assessed in affected hip joints.

Results—For many of the questions, answers provided by owners of dogs with CHD differed significantly from those of owners of control dogs. Stress hormone concentrations differed significantly between dogs with CHD and controls, but individual variation was too great for them to be of value in pain assessment. None of the radiologic variables examined correlated well with owner or veterinarian pain scores.

Conclusions and Clinical Relevance—Chronic pain could be assessed in dogs with CHD through completion of the study questionnaire by a person familiar with the pet (eg, owner) after receiving appropriate education in its use. Eleven variables were identified as being potentially useful in assessment of chronic pain in dogs. (J Am Vet Med Assoc 2003;222: 1552–1558)

Full access
in Journal of the American Veterinary Medical Association