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  • Author or Editor: Masahiro Okumura x
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Abstract

Objectives

To establish a sensitive and specific monoclonal antibody (MAB) against equine keratan sulfate (KS) and to develop an enzyme immunoassay for measurement of the concentration of KS in serum and synovial fluid from horses.

Sample Population

18 synovial fluid and 48 serum samples were obtained from clinically normal horses and horses with arthritis.

Procedure

BALB/c mice were immunized with chondroitinase-ABC-digested proteoglycan monomer from equine joint cartilage, and MAB were raised, using Sp2/O cells as a fusion partner. A competitive ELISA was optimized, using one of the established MAB, and KS concentration in synovial fluid and sera from horses was measured.

Results

The high-titer MAB1/14/16H9, which specifically recognizes the epitope on equine KS, was identified. This antibody had no reactivity with chondroitin sulfate and core protein of proteoglycan monomers, hyaluronan, heparin, dermatan sulfate, and heparan sulfate. A competitive ELISA for determination of KS concentration was optimized, using this antibody. Precision data were obtained for the range of 10 to 160 ng/ml. The within- and between-assay coefficients of variation were 10.0 and 12.7%, respectively.

Conclusions

MAB 1/14/16H9 that specifically recognized equine KS was established and was used to develop an enzyme immunoassay for measurement of the concentration of KS in synovial fluid and sera from horses. It is expected that the assay system using MAB 1/14/16H9 will contribute to evaluation of cartilage metabolic activity in horses. (Am J Vet Res 1998;59:1203-1208)

Free access
in American Journal of Veterinary Research

Abstract

OBJECTIVE

To describe outcomes for dogs that underwent balloon dilation for palliative treatment of urethral obstruction caused by urothelial carcinoma.

ANIMALS

12 client-owned dogs.

PROCEDURES

Medical records were searched to identify dogs with urothelial (bladder, urethra, or prostate) carcinoma that underwent balloon dilation for treatment of urethral obstruction between April 2010 and December 2015. Information regarding history, signalment, clinical signs, diagnostic imaging findings, balloon dilation technique, clinical outcomes, complications, and additional treatments was obtained by review of medical records.

RESULTS

Improvement in clinical signs of urethral obstruction was observed after the initial dilation procedure for 9 of 12 dogs. Urethral obstruction was known to recur in 5 dogs 48 to 296 days after the initial procedure. Three of these dogs underwent a second dilation procedure, with clinical improvement in all 3 dogs for 41 to 70 days. One of 2 dogs that had a third procedure after the second reobstruction had clinical improvement in urinary tract signs until subsequent death from metastatic disease 22 days later. Complications included hematuria, urinary incontinence, and dysuria; these resolved within a few days after treatment.

CONCLUSIONS AND CLINICAL RELEVANCE

Urethral balloon dilation was a minimally invasive procedure that provided relief of urethral obstruction from urothelial carcinoma in most dogs of the study population. Prospective studies are needed to identify optimal techniques for balloon dilation in dogs with neoplastic urethral obstructions and to identify patients that are likely to benefit most from the treatment.

Full access
in Journal of the American Veterinary Medical Association

SUMMARY

To measure the concentration of serum amyloid A (saa) protein in horses, a sensitive and highly reproducible sandwich (elisa) was established, using affinity purified saa antibody. Results of the elisa were found to have a high correlation (r = 0.95) with those of the single radial immunodiffusion test. Equine saa concentration was measured by use of this elisa. In clinically normal horses, the concentration of saa was high immediately after birth to 2 weeks of age. After that, saa concentration had periodic fluctuations in the range of approximately 10 to 30 μg/ml. Mean (± sd) concentrations of saa in foals (≤ 12 months old) and adult horses (≥ 18 months old) were 21.23 ± 12.20 and 14.93 ± 9.07 μg/ml, respectively. In mares during the perinatal period, saa concentration remained stable within the reference range before parturition. It increased quickly after delivery, and reached a peak value of 101.29 ± 98.82 μg/ml on postpartum day 3, then began to decrease, at postpartum week 2, to the reference range by the end of postpartum month 1. In horses with experimentally induced inflammation, saa concentration increased quickly and reached approximately four- to 40-fold increase over the pretreatment value on day 1 and remained high on days 2 to 6 after treatment. It then returned to the baseline value by 2 to 4 weeks in association with disappearance of local signs of inflammation. The saa concentration was high in most horses with clinical signs of inflammation. It was concluded from these data that this elisa is sensitive and reliable for measuring saa in horses.

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To determine keratan sulfate (KS) concentration in the serum of foals at the early stage of growing, and to evaluate the role of serum KS as a cartilage catabolic marker, comparing its values with the fluctuation of serum alkaline phosphatase (ALP) activity as a measurement of osteoblastic activity.

Animals

12 foals with normal growth and 3 foals with joint abnormalities within 18 months after birth.

Procedure

Measurement of KS concentration and ALP activity in serum and radiographic and physical examinations were done.

Results

In all foals, serum KS concentration was high from 1 week after birth to 3 months of age, while serum ALP decreased with aging. The value started to decrease rapidly from 3 to 5 months of age, then gradually reached adult values. During the first 3 months, KS concentration in male foals was significantly higher than that in female foals. In 3 foals which had joint problems, KS concentration was higher than that in normally growing foals at 1 week, and at 1, 2, and 3 months of age.

Conclusions

Cartilage catabolic activity is higher in developing foals up to 3 months of age, suggesting that the immature joint at this time could be easily affected by any factor of loading. Moreover, though only 3 diseased foals were examined, higher serum KS concentration in these foals suggest that this variable might be a useful measure of joint diseases, even at an early stage of life in foals. (Am J Vet Res 1997;58:925–929)

Free access
in American Journal of Veterinary Research

SUMMARY

Ceruloplasmin (Cp) was isolated from fresh equine plasma by precipitation, cellulose chromatography, and improved ion-exchange chromatography. Purified equine Cp is a glycoprotein having a molecular weight of approximately 115,000. In electrophoresis, equine Cp migrated to the α1-globulin region, its isoelectric point was about 4.15 and consisted of about 890 amino acid residues.

Serum Cp concentration was measured by use of the single radial immunodiffusion method. In clinically normal horses, the mean (± sd) serum Cp concentration of newborn foals was 2.87 ± 0.40 mg/ml and that of 3-month-old foals was 5.02 ± 0.92 mg/ml, which was similar to the adult value. It reached a peak of 6.06 ± 0.74 mg/ml in 2-year-old horses. The Cp concentration in mares was not statistically different for the perinatal period, but it decreased immediately before and after delivery.

Concentration of Cp increased at 6 days after im administration of turpentine oil, castration, or jejunojejunostomy in adult horses, and increased to peak values twice as high as baseline values at 7 to 14 days, returning to baseline values at 28 days after treatment.

We concluded that equine serum Cp is an acute-phase reactive protein increased in the intermediary or later phase of acute inflammation.

Free access
in American Journal of Veterinary Research

Summary

Equine α1-acid glycoprotein (α1 ag) was isolated from equine serum by successive ammonium precipitation, anion- and cation-exchange chromatographies, and gel filtration. Purified equine α1 ag had a molecular weight of 46,000 ± 1,000, and contained 31.4% carbohydrate. Gel isoelectric focusing revealed an isoelectric point range of 2.8 to 3.7. With immunoelectrophoresis, it was found that α1 ag migrated to the α1-globulin region.

Single radial immunodiffusion was used for quantitative measurement of α1 ag in equine serum. In clinically normal foals, serum α1 ag was undetectable (≤ 20 ng/ml) in ≤ 7-day-old foals, but was detected by 14 days. The α1 ag concentration (mean ± sd) increased to reach mean adult values of 99.23 ± 26.90 μg/ml by 1 year of age. The α1 ag concentration in pregnant mares decreased at 2 to 3 months before parturition, then gradually increased until 1 day after parturition, when a brief decrease was observed. The concentration increased again at 2 weeks after foaling, then a decrease was observed, after which the α1 ag concentration increased again by 2 to 4 months after parturition.

The concentration of serum α1 ag quickly rose to peak values 2 to 3 days after castration and jejunojejunostomy in adult horses, returning to baseline values by 14 to 28 days after surgery. The α1 ag was concluded to be an acute-phase reactive protein in horses.

Free access
in American Journal of Veterinary Research