ABSTRACT
Platelet endothelial cell adhesion molecule (PECAM, CD31) and vascular cell adhesion molecule-1 (VCAM-1, CD106) are essential for leukocyte emigration and diapedesis. PECAM is an essential histologic marker of endothelial cells; VCAM-1 is a prototype marker for endothelial cell activation. In this study, equine PECAM and VCAM mRNA were cloned and sequenced. Both genes are highly conserved amongst several species. This study also revealed conserved structural and regulatory motifs, emphasizing the importance of these genes' physiological roles in immunological responses.
Subject(s)
Horses , Platelet Endothelial Cell Adhesion Molecule-1/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Vascular Cell Adhesion Molecule-1/genetics , Vascular Cell Adhesion Molecule-1/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Molecular Sequence Data , PhylogenyABSTRACT
OBJECTIVE: To evaluate the effects of interleukin (IL)-1beta on proteoglycan metabolism in equine cartilage explants when cultured in the presence of synoviocytes. SAMPLE POPULATION: Samples of cartilage and synovium collected from the femoropatellar joints of three 2- to 3-year-old horses. PROCEDURES: 3 experimental groups were established: cartilage explants only, synoviocytes only, and cartilage explants-synoviocytes in coculture. In each group, samples were cultured with or without IL-1beta (10 ng/mL) for 96 hours. Glycosaminoglycan (GAG) content of cartilage and medium samples was measured by use of a spectrophotometric assay; RNA was isolated from synoviocytes and cartilage and analyzed for expression of matrix metalloproteinases (MMP)-3 and -13 (cartilage and synoviocytes), aggrecan (cartilage), collagen type IIB (cartilage), and 18S as a control (cartilage and synoviocytes) by use of quantitative PCR assays. Cartilage matrix metachromasia was assessed histochemically. RESULTS: IL-1beta-induced GAG loss from cartilage was significantly less in cocultures than in cartilage-only cultures. Cartilage aggrecan gene expression was also significantly less downregulated and synoviocyte MMP-3 expression was less upregulated by IL-1beta in cocultures, compared with cartilage- and synoviocyte only cultures. Histochemical findings supported the molecular and biochemical results and revealed maintenance of matrix metachromasia in cocultured cartilage treated with IL-1beta. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that synoviocytes secrete 1 or more mediators that preferentially protect matrix GAG metabolism from the degradative effects of IL-1beta. Further studies involving proteomic and microarray approaches in similar coculture systems may elucidate novel therapeutic targets for the treatment of osteoarthritis.
Subject(s)
Cartilage, Articular/metabolism , Horses/metabolism , Interleukin-1beta/pharmacology , Proteoglycans/metabolism , Synovial Fluid/cytology , Aggrecans/genetics , Aggrecans/metabolism , Animals , Cartilage, Articular/drug effects , Collagen Type II/genetics , Collagen Type II/metabolism , Gene Expression Regulation/drug effects , Glycosaminoglycans/metabolism , Matrix Metalloproteinase 13/genetics , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 3/genetics , Matrix Metalloproteinase 3/metabolism , Synovial Fluid/drug effects , Tissue Culture TechniquesABSTRACT
OBJECTIVES: To determine the requirement for 2nd surgical interventions and mortality after 3 different surgical techniques (percutaneous tube cystostomy [10 goats], surgical tube cystostomy [25 goats], urinary bladder marsupialization [10 goats]) for caprine obstructive urolithiasis, and to determine whether pre- or 24-hour postoperative physical examination findings or serum chemistry results could be used as predictors of mortality. STUDY DESIGN: Retrospective study. ANIMALS: Male goats (45) with obstructive urolithiasis. METHODS: Medical records for all male goats admitted and operated for obstructive urolithiasis between 1993 and 2003 were reviewed. Data retrieved included signalment, pre- and 24-hour postoperative values for temperature, pulse, respiratory rate, packed cell volume, serum K(+), serum creatinine, and blood urea nitrogen [BUN]. The type of initial surgical procedure, time to 2nd surgical intervention, time to death, and duration and cost of hospitalization were also obtained. RESULTS: Percutaneous tube cystostomy was associated with a significantly increased requirement for (5.6-fold increased hazard) and decreased time to a 2nd intervention (P=.002). There were no differences between the 3 procedures for time to mortality. Increased BUN and respiratory rate at admission were associated with increased mortality (hazards ratio of 4.8 and 5.0, respectively). Urinary bladder marsupialization was associated with significantly decreased hospitalization time (P=.02) and cost (P=.04) compared with surgical tube cystostomy and percutaneous tube cystostomy. CONCLUSION: Surgical tube cystostomy and bladder marsupialization are both acceptable surgical methods for treatment of caprine obstructive urolithiasis. Each procedure has inherent complications that should be discussed with the owner before choosing the surgical treatment. CLINICAL RELEVANCE: There are acceptable options for treatment of caprine obstructive urolithiasis; however, percutaneous tube cystostomy should be avoided.
