Flow cytometric analysis of peripheral blood mononuclear cells induced by experimental endotoxemia in horse.

Cellular activation and functional cell surface markers were evaluated during experimentally-induced endotoxemia in healthy horses. Eight healthy adult horses were infused a low dose of endotoxin (lipopolysaccharide from Escherichia coli O26: B6, 30 ng/kg of body weight, IV) and five control horses were given an equivalent volume of sterile saline solution. Venous blood samples were collected for flow cytometric analysis of peripheral blood mononuclear cells (PBMCs) and to measure plasma endotoxin concentrations. Clinical signs of endotoxemia were recorded at 10, 20, 30, 40, 50 min, 1, 2, 3, 4, 8, 16, 24 and 48 hr after endotoxin or saline solution administration. Clinical findings characteristic of endotoxemia (tachycardia, tachypnea, increased rectal temperature, and leukopenia) occurred transiently in all horses administered endotoxin; however, plasma endotoxin concentrations were detectable in only 50% (4/8) of the endotoxin-infused horses. The percentage of CD4(+), CD5(+), and CD8(+) cells decreased while the percentage of CD14(+), IgM(+), and MHC class II(+) cells increased significantly after endotoxin infusion. Alterations in the immunophenotype of PBMCs from horses with experimentally-induced endotoxemia were associated with changes in vital signs, indicating that endotoxin altered the immuno balance.

Magnetic resonance imaging application to live horse for diagnosis of tendinitis.

Six live horses with various stages of acute to chronic superficial digital flexor (SDF) tendinitis were examined using magnetic resonance imaging (MRI). In each case, MRI findings were compared to the corresponding ultrasonographic (USD) and histologic findings, to establish the usefulness of MRI. In the acute cases, lesions characterized by hemorrhage were well defined as high signal intensity on MRI and hypoechoic regions on USD. Chronic tendon fibrosis was slightly hyperechoic and difficult to distinguish from the normal tendon tissue around the original injury by using USD. In contrast, MRI visualized the chronic lesion as a low intensity signal, which could be distinguished from the black background of the normal SDF tendon tissue. This study clearly demonstrated MRI was the better imaging modality for the objective detection of chronic scar tissue in live horses. These findings, from living horses, suggest an advantage of MRI in the clinical application to diagnose tendinitis in cases where there is chronic scar tissue that is difficult to discern on USD.

TGF-beta alters collagen XII and XIV mRNA levels in cultured equine tenocytes.

The effects of TGF-beta 1, beta 2 and beta 3 (TGF-beta) on levels of mRNA corresponding to the alpha chains of type XII and type XIV collagens in equine tenocyte cultures were assessed using the ribonuclease protection assay (RPA). The level of alpha1(XII) mRNA in untreated monolayer cultures was approximately 50- to 100-fold greater than alpha1(XIV) mRNA level. Addition of TGF-beta resulted in an increase in the amount of alpha1(XII) present and a decrease of alpha1(XIV) mRNA in a dose-dependent manner. Specifically, the level of alpha1(XII) mRNA was doubled, but alpha1(XIV) was decreased to 30% of control by the addition of 2 ng/ml of TGF-beta 1 to the cultures. These effects were completely abrogated by neutralizing antibody specific for TGF-beta. In addition, electron microscopy demonstrated that bundled collagen fibers were formed in the intercellular spaces of multilayered tenocytes which had been cultured for 2 weeks in the presence of exogenous TGF-beta 1 together with ascorbic acid phosphate. These results suggest that type XII and/or type XIV collagens modulate the structure of ECM formed by tenocytes in culture.