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- Author or Editor: Philip James Johnson x
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Abstract
Objective—To evaluate expression of cyclooxygenase (COX)-1 and COX-2 in the cornea, eyelid, and third eyelid of healthy horses and those affected with squamous cell carcinoma (SCC) by use of immunohistochemical techniques.
Animals—15 horses with SCC involving ocular tissues and 5 unaffected control horses.
Procedures—SCC-affected tissues were obtained from the cornea (n = 5 horses), eyelid (5), and third eyelid (5). Site-matched control tissues were obtained from 5 horses unaffected with SCC. Tissue sections of affected and control cornea, eyelid, and third eyelid were stained immunohistochemically for COX-1 and COX-2 via standard techniques. Stain uptake was quantified by use of computer-assisted image analysis of digital photomicrographs.
Results—Immunoreactivity for both COX-1 and COX-2 was significantly greater in equine corneas with SCC than in control corneas. No significant differences in COX-1 or COX-2 immunoreactivity were detected in eyelid and third-eyelid SCC, compared with site-matched control tissues.
Conclusions and Clinical Relevance—Immunoreactivity for COX-1 and COX-2 is high in equine corneal SCC, possibly indicating that COX plays a role in oncogenesis or progression of this tumor type at this site. Pharmacologic inhibition of COX may represent a useful adjunctive treatment for corneal SCC in horses.
Summary
Seventy dogs were included in a randomized, controlled, multicenter trial to test the efficacy of carprofen (2.2 mg/kg of body weight, PO, q 12 h) for relief of clinical signs associated with osteoarthritis. Thirty-six dogs received carprofen, and 34 received a placebo. Response of the dogs was evaluated by comparing results of force plate examination and a graded lameness examination performed before and immediately after 2 weeks of treatment, and by obtaining a subjective assessment of the dog's posttreatment condition from owners and participating veterinarians. A physical examination, CBC, serum biochemical analyses, urinalysis, and fecal occult blood test were performed before and after treatment to monitor safety. For force plate evaluation, the odds ratio was 3.3, meaning that a dog treated with carprofen was 3.3 times more likely to have a positive response than was a dog treated with the placebo. For evaluation by a veterinarian, the odds ratio was 3.5, and for owner evaluation, the odds ratio was 4.2. Institution where dogs were treated did not have a significant effect on results. A variety of reactions that may have been related to the medication (placebo or carprofen) were recorded; however, none were considered serious. Serum alanine aminotransferase activity was high in 3 dogs (2 that received placebo and 1 that received carprofen) at the conclusion of treatment; none of the 3 dogs were clinically ill. Ten dogs © that received placebo and 5 that received carprofen) had negative pretreatment and positive posttreatment fecal occult blood test results.
Abstract
Objective—To determine the expression and distribution of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), its substrates aggrecan and versican, and their binding partner hyaluronan in laminae of healthy horses.
Sample—Laminae from the forelimb hooves of 8 healthy horses.
Procedures—Real-time quantitative PCR assay was used for gene expression analysis. Hyaluronidase, chondroitinase, and keratanase digestion of lamina extracts combined with SDS-PAGE and western blotting were used for protein and proteoglycan analysis. Immunofluorescent and immunohistochemical staining of tissue sections were used for protein and hyaluronan localization.
Results—Genes encoding ADAMTS-4, aggrecan, versican, and hyaluronan synthase II were expressed in laminae. The ADAMTS-4 was predominantly evident as a 51-kDa protein bearing a catalytic site neoepitope indicative of active enzyme and in situ activity, which was confirmed by the presence of aggrecan and versican fragments bearing ADAMTS-4 cleavage neoepitopes in laminar protein extracts. Aggrecan, versican, and hyaluronan were localized to basal epithelial cells within the secondary epidermal laminae. The ADAMTS-4 localized to these cells but was also present in some cells in the dermal laminae.
Conclusions and Clinical Relevance—Within digital laminae, versican exclusively and aggrecan primarily localized within basal epithelial cells and both were constitutively cleaved by ADAMTS-4, which therefore contributed to their turnover. On the basis of known properties of these proteoglycans, it is possible that they can protect the basal epithelial cells of horses from biomechanical and concussive stress.
Abstract
Objective—To determine whether increased gene expression of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) in laminae of horses with starch gruel–induced laminitis was accompanied by increased enzyme activity and substrate degradation.
Sample—Laminae from the forelimb hooves of 8 healthy horses and 17 horses with starch gruel–induced laminitis (6 at onset of fever, 6 at onset of Obel grade 1 lameness, and 5 at onset of Obel grade 3 lameness).
Procedures—Gene expression was determined by use of cDNA and real-time quantitative PCR assay. Protein expression and processing were determined via SDS-PAGE and quantitative western blotting. Protein distribution and abundance were determined via quantitative immunofluorescent staining.
Results—ADAMTS-4 gene expression was increased and that of versican decreased in laminitic laminae, compared with expression in healthy laminae. Catalytically active ADAMTS-4 also was increased in the tissue, as were ADAMTS-4–cleavage fragments of versican. Immunofluorescent analyses indicated that versican was depleted from the basal epithelia of laminae of horses at onset of Obel grade 3 lameness, compared with results for healthy laminae, and this was accompanied by regional separation of basal epithelial cells from the basement membrane. Aggrecan gene and protein expression were not significantly affected.
Conclusions and Clinical Relevance—Changes in gene and protein expression of ADAMTS-4 and versican in the basal epithelium of laminitic laminae indicated a fundamental change in the physiology of basal epithelial cells. This was accompanied by and may have caused detachment of these cells from the basement membrane.
